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<title>bioRxiv Channel: Oregon Health & Science University</title>
<link>https://biorxiv.org</link>
<description>
This feed contains articles for bioRxiv Channel "Oregon Health & Science University"
</description>

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<prism:publicationName>bioRxiv</prism:publicationName>
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<title>bioRxiv</title>
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<link>https://biorxiv.org</link>
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<item rdf:about="https://biorxiv.org/content/10.1101/2023.02.26.529963v1?rss=1">
<title>
<![CDATA[
The classic psychedelic DOI induces a persistent desynchronized state in medial prefrontal cortex 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.02.26.529963v1?rss=1</link>
<description><![CDATA[
Administration or consumption of classic psychedelics (CPs) leads to profound changes in experience which are often described as highly novel and meaningful. They have shown substantial promise in treating depressive symptoms and may be therapeutic in other situations. Although research suggests that the therapeutic response is correlated with the intensity of the experience, the neural circuit basis for the alterations in experience caused by CPs requires further study. The medial prefrontal cortex (mPFC), where CPs have been shown to induce rapid, 5-HT2A receptor-dependent structural and neurophysiological changes, is believed to be a key site of action. To investigate the acute neural circuit changes induced by CPs, we recorded single neurons and local field potentials in the mPFC of freely behaving mice after administration of the 5-HT2A/2C receptor-selective CP, 2,5-Dimethoxy-4-iodoamphetamine (DOI). We segregated recordings into active and rest periods in order to examine cortical activity during desynchronized (active) and synchronized (rest) states. We found that DOI induced a robust decrease in low frequency power and decoupled rhythmic activity from neural population dynamics when animals were at rest, attenuating the usual synchronization that occurs during less active behavioral states. DOI also increased broadband gamma power and suppressed activity in fast-spiking neurons in both active and rest periods. Together, these results show that the CP DOI induces persistent desynchronization in mPFC, including during rest when mPFC typically exhibits more synchronized activity. This shift in cortical dynamics may in part underlie the longer-lasting effects of CPs on plasticity, and may be critical to their therapeutic properties.
]]></description>
<dc:creator>Olson, R.</dc:creator>
<dc:creator>Bartlett, L.</dc:creator>
<dc:creator>Sonneborn, A.</dc:creator>
<dc:creator>Bretton-Granatoor, Z.</dc:creator>
<dc:creator>Firdous, A.</dc:creator>
<dc:creator>Harris, A.</dc:creator>
<dc:creator>Abbas, A.</dc:creator>
<dc:date>2023-02-27</dc:date>
<dc:identifier>doi:10.1101/2023.02.26.529963</dc:identifier>
<dc:title><![CDATA[The classic psychedelic DOI induces a persistent desynchronized state in medial prefrontal cortex]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-02-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.02.28.529432v1?rss=1">
<title>
<![CDATA[
Spontaneous allelic variant in Ush1g resulting in an expanded phenotype 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.02.28.529432v1?rss=1</link>
<description><![CDATA[
Strategies to reveal the discovery of the relationships between novel phenotypic behaviors and specific genetic alterations can be achieved via either target-specific, directed mutagenesis or phenotypic selection following random chemical mutagenesis. As an alternative approach, one can exploit deficiencies in DNA repair pathways that are responsible for the maintenance of genetic integrity in response to spontaneously-induced damage. In the genetic background of mice deficient in the DNA glycosylase NEIL1, elevated numbers of spontaneous mutations arise from translesion DNA synthesis past unrepaired, oxidatively-induced base damage. Several litters of Neil1 knockout mice included animals that were distinguished by their backwards-walking behavior in open-field environments, while maintaining frantic forward movements in their home cage environment. Other phenotypic manifestations included swim test failures, head tilting, and circling. Mapping of the mutation that conferred these behaviors revealed the introduction of a stop codon at amino acid 4 of the Ush1g gene; the allele was Ush1gbw, reflecting the backwards-walking phenotype. Ush1gbw/bw null mice displayed auditory and vestibular defects that are commonly seen with mutations affecting inner-ear hair-cell function, including a complete lack of auditory brainstem responses and vestibular-evoked potentials. As in other Usher syndrome type I mutant mouse lines, hair-cell phenotypes included disorganized and split hair bundles, as well as altered distribution of proteins for stereocilia that localize to the tips of row 1 or row 2. Disruption to the bundle and kinocilium displacement suggested that USH1G is essential for forming the hair cells kinocilial links. Due to the vestibular dysfunction, however, visual behavior as measured with optokinetic tracking could not be assessed in Ush1gbw/bw mice. Consistent with other Usher type 1 models, however, Ush1gbw/bw mice had no substantial retinal degeneration compared to Ush1gbw/+ controls out to six months. In contrast to previously-described Ush1g alleles, this new allele provides the first knockout model for this gene.
]]></description>
<dc:creator>Vartanian, V.</dc:creator>
<dc:creator>Krey, J. F.</dc:creator>
<dc:creator>Chatterjee, P.</dc:creator>
<dc:creator>Jones, S. M.</dc:creator>
<dc:creator>Curtis, A.</dc:creator>
<dc:creator>Ryals, R.</dc:creator>
<dc:creator>Lloyd, R. S.</dc:creator>
<dc:creator>Barr-Gillespie, P. G.</dc:creator>
<dc:date>2023-03-01</dc:date>
<dc:identifier>doi:10.1101/2023.02.28.529432</dc:identifier>
<dc:title><![CDATA[Spontaneous allelic variant in Ush1g resulting in an expanded phenotype]]></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/10.1101/2023.02.28.530466v1?rss=1">
<title>
<![CDATA[
GIPC3 couples to MYO6 and PDZ domain proteins and shapes the hair cell apical region 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.02.28.530466v1?rss=1</link>
<description><![CDATA[
GIPC3 has been implicated in auditory function. Initially localized to the cytoplasm of inner and outer hair cells of the cochlea, GIPC3 increasingly concentrated in cuticular plates and at cell junctions during postnatal development. Early postnatal Gipc3KO/KO mice had mostly normal mechanotransduction currents, but had no auditory brainstem response at one month of age. Cuticular plates of Gipc3KO/KO hair cells did not flatten during development as did those of controls; moreover, hair bundles were squeezed along the cochlear axis in mutant hair cells. Junctions between inner hair cells and adjacent inner phalangeal cells were also severely disrupted in Gipc3KO/KO cochleas. GIPC3 bound directly to MYO6, and the loss of MYO6 led to altered distribution of GIPC3. Immunoaffinity purification of GIPC3 from chicken inner ear extracts identified co-precipitating proteins associated with adherens junctions, intermediate filament networks, and the cuticular plate. Several of immunoprecipitated proteins contained GIPC-family consensus PDZ binding motifs (PBMs), including MYO18A, which binds directly to the PDZ domain of GIPC3. We propose that GIPC3 and MYO6 couple to PBMs of cytoskeletal and cell-junction proteins to shape the cuticular plate.

Summary statementThe PDZ-domain protein GIPC3 couples the molecular motors MYO6 and MYO18A to actin cytoskeleton structures in hair cells. GIPC3 is necessary for shaping the hair cells cuticular plate and hence the arrangement of the stereocilia in the hair bundle.
]]></description>
<dc:creator>Chatterjee, P.</dc:creator>
<dc:creator>Morgan, C. P.</dc:creator>
<dc:creator>Krey, J. F.</dc:creator>
<dc:creator>Benson, C.</dc:creator>
<dc:creator>Goldsmith, J.</dc:creator>
<dc:creator>Bateschell, M.</dc:creator>
<dc:creator>Ricci, A.</dc:creator>
<dc:creator>Barr-Gillespie, P. G.</dc:creator>
<dc:date>2023-03-01</dc:date>
<dc:identifier>doi:10.1101/2023.02.28.530466</dc:identifier>
<dc:title><![CDATA[GIPC3 couples to MYO6 and PDZ domain proteins and shapes the hair cell apical region]]></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/10.1101/2023.03.07.531415v1?rss=1">
<title>
<![CDATA[
Structurally divergent and recurrently mutated regions of primate genomes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.03.07.531415v1?rss=1</link>
<description><![CDATA[
To better understand the pattern of primate genome structural variation, we sequenced and assembled using multiple long-read sequencing technologies the genomes of eight nonhuman primate species, including New World monkeys (owl monkey and marmoset), Old World monkey (macaque), Asian apes (orangutan and gibbon), and African ape lineages (gorilla, bonobo, and chimpanzee). Compared to the human genome, we identified 1,338,997 lineage-specific fixed structural variants (SVs) disrupting 1,561 protein-coding genes and 136,932 regulatory elements, including the most complete set of human-specific fixed differences. Across 50 million years of primate evolution, we estimate that 819.47 Mbp or ~27% of the genome has been affected by SVs based on analysis of these primate lineages. We identify 1,607 structurally divergent regions (SDRs) wherein recurrent structural variation contributes to creating SV hotspots where genes are recurrently lost (CARDs, ABCD7, OLAH) and new lineage-specific genes are generated (e.g., CKAP2, NEK5) and have become targets of rapid chromosomal diversification and positive selection (e.g., RGPDs). High-fidelity long-read sequencing has made these dynamic regions of the genome accessible for sequence-level analyses within and between primate species for the first time.
]]></description>
<dc:creator>Mao, Y.</dc:creator>
<dc:creator>Harvey, W. T.</dc:creator>
<dc:creator>Porubsky, D.</dc:creator>
<dc:creator>Munson, K. M.</dc:creator>
<dc:creator>Hoekzema, K.</dc:creator>
<dc:creator>Lewis, A. P.</dc:creator>
<dc:creator>Audano, P. A.</dc:creator>
<dc:creator>Rozanski, A.</dc:creator>
<dc:creator>Yang, X.</dc:creator>
<dc:creator>Zhang, S.</dc:creator>
<dc:creator>Gordon, D. S.</dc:creator>
<dc:creator>Wei, X.</dc:creator>
<dc:creator>Logsdon, G. A.</dc:creator>
<dc:creator>Haukness, M.</dc:creator>
<dc:creator>Dishuck, P. C.</dc:creator>
<dc:creator>Jeong, H.</dc:creator>
<dc:creator>del Rosario, R.</dc:creator>
<dc:creator>Bauer, V. L.</dc:creator>
<dc:creator>Fattor, W. T.</dc:creator>
<dc:creator>Wilkerson, G. K.</dc:creator>
<dc:creator>Lu, Q.</dc:creator>
<dc:creator>Paten, B.</dc:creator>
<dc:creator>Feng, G.</dc:creator>
<dc:creator>Sawyer, S. L.</dc:creator>
<dc:creator>Warren, W. C.</dc:creator>
<dc:creator>Carbone, L.</dc:creator>
<dc:creator>Eichler, E. E.</dc:creator>
<dc:date>2023-03-07</dc:date>
<dc:identifier>doi:10.1101/2023.03.07.531415</dc:identifier>
<dc:title><![CDATA[Structurally divergent and recurrently mutated regions of primate genomes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-03-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.03.07.531534v1?rss=1">
<title>
<![CDATA[
TAD Evolutionary and functional characterization reveals diversity in mammalian TAD boundary properties and function 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.03.07.531534v1?rss=1</link>
<description><![CDATA[
Topological associating domains (TADs) are self-interacting genomic units crucial for shaping gene regulation patterns. Despite their importance, the extent of their evolutionary conservation and its functional implications remain largely unknown. In this study, we generate Hi-C and ChIP-seq data and compare TAD organization across four primate and four rodent species, and characterize the genetic and epigenetic properties of TAD boundaries in correspondence to their evolutionary conservation. We find that only 14% of all human TAD boundaries are shared among all eight species (ultraconserved), while 15% are human-specific. Ultraconserved TAD boundaries have stronger insulation strength, CTCF binding, and enrichment of older retrotransposons, compared to species-specific boundaries. CRISPR-Cas9 knockouts of two ultraconserved boundaries in mouse models leads to tissue-specific gene expression changes and morphological phenotypes. Deletion of a human-specific boundary near the autism-related AUTS2 gene results in upregulation of this gene in neurons. Overall, our study provides pertinent TAD boundary evolutionary conservation annotations, and showcase the functional importance of TAD evolution.
]]></description>
<dc:creator>Okhovat, M.</dc:creator>
<dc:creator>VanCampen, J.</dc:creator>
<dc:creator>Lima, A. C.</dc:creator>
<dc:creator>Nevonen, K.</dc:creator>
<dc:creator>Layman, C. E.</dc:creator>
<dc:creator>Ward, S.</dc:creator>
<dc:creator>Herrera, J.</dc:creator>
<dc:creator>Stendahl, A. M.</dc:creator>
<dc:creator>Yang, R.</dc:creator>
<dc:creator>Harshman, L.</dc:creator>
<dc:creator>Li, W.</dc:creator>
<dc:creator>Sheng, R. R.</dc:creator>
<dc:creator>Mao, Y.</dc:creator>
<dc:creator>Fedorov, L.</dc:creator>
<dc:creator>Ndjamen, B.</dc:creator>
<dc:creator>Vigh-Conrad, K. A.</dc:creator>
<dc:creator>Matthews, I. R.</dc:creator>
<dc:creator>Easow, S. A.</dc:creator>
<dc:creator>Chan, D. K.</dc:creator>
<dc:creator>Jan, T. A.</dc:creator>
<dc:creator>Eichler, E. E.</dc:creator>
<dc:creator>Rugonyi, S.</dc:creator>
<dc:creator>Conrad, D. F.</dc:creator>
<dc:creator>Ahituv, N.</dc:creator>
<dc:creator>Carbone, L.</dc:creator>
<dc:date>2023-03-07</dc:date>
<dc:identifier>doi:10.1101/2023.03.07.531534</dc:identifier>
<dc:title><![CDATA[TAD Evolutionary and functional characterization reveals diversity in mammalian TAD boundary properties and function]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-03-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.03.08.531794v1?rss=1">
<title>
<![CDATA[
CEP76 is a critical regulator of male germ cell transition zone function and tail composition 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.03.08.531794v1?rss=1</link>
<description><![CDATA[
The transition zone is a specialised gate at the base of cilia/flagella, which segregates the ciliary compartment from the cytoplasm and strictly regulates protein entry. In this study, we have identified CEP76 as an essential germ cell transition zone protein, involved in the selective entry and incorporation of key proteins required for sperm function and fertility into the ciliary compartment and ultimately the sperm tail. In its absence sperm tails are shorter and immotile as a consequence of deficits in essential sperm motility proteins including DNAH2 and AKAP4, which accumulate at the sperm neck in the mutant. We demonstrate CEP76 is required for sperm tail fibrous sheath formation, outer dense fibre loading and axoneme stability in the principal piece and ultimately sperm motility. Finally, we identify that CEP76 dictates annulus positioning and composition, adding further evidence that the spermatid transition zone and annulus are part of the same functional structure.
]]></description>
<dc:creator>Houston, B. J.</dc:creator>
<dc:creator>Merriner, D. J.</dc:creator>
<dc:creator>Stathatos, G. G.</dc:creator>
<dc:creator>O'Connor, A. E.</dc:creator>
<dc:creator>Lopes, A. M.</dc:creator>
<dc:creator>Conrad, D. F.</dc:creator>
<dc:creator>Baker, M.</dc:creator>
<dc:creator>Dunleavy, J. E.</dc:creator>
<dc:creator>O'Bryan, M. K.</dc:creator>
<dc:date>2023-03-09</dc:date>
<dc:identifier>doi:10.1101/2023.03.08.531794</dc:identifier>
<dc:title><![CDATA[CEP76 is a critical regulator of male germ cell transition zone function and tail composition]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-03-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.03.11.532242v1?rss=1">
<title>
<![CDATA[
Characterization of extracellular matrix deposited by segmental trabecular meshwork cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.03.11.532242v1?rss=1</link>
<description><![CDATA[
Biophysical and biochemical attributes of the extracellular matrix are major determinants of cell fate in homeostasis and disease. Ocular hypertension and glaucoma are diseases where the trabecular meshwork tissue responsible for aqueous humor egress becomes stiffer accompanied by changes in its matrisome in a segmental manner with regions of high or low flow. Prior studies demonstrate these alterations in the matrix are dynamic in response to age and pressure changes. The underlying reason for segmentation or differential response to pressure and stiffening are unknown. This is largely due to a lack of appropriate models (in vitro or ex vivo) to study this phenomena. In this study, we characterize the biomechanical attributes, matrisome, and incidence of crosslinks in the matrix deposited by primary cells isolated from segmental flow regions and when treated with glucocorticosteroid. Data demonstrate that matrix deposited by cells from low flow regions are stiffer and exhibit a greater number of immature and mature crosslinks, and that these are exacerbated in the presence of steroid. We also show a differential response of high or low flow cells to steroid via changes observed in the matrix composition. We conclude that although a mechanistic basis for matrix stiffness was undetermined in this study, it is a viable tool to study cell-matrix interactions and further our understanding of trabecular meshwork pathobiology.
]]></description>
<dc:creator>Raghunathan, V.</dc:creator>
<dc:creator>Nartey, A.</dc:creator>
<dc:creator>Dhamodaran, K.</dc:creator>
<dc:creator>Baidouri, H.</dc:creator>
<dc:creator>Staverosky, J. A.</dc:creator>
<dc:creator>Keller, K.</dc:creator>
<dc:creator>Zientek, K.</dc:creator>
<dc:creator>Reddu, A.</dc:creator>
<dc:creator>Acott, T.</dc:creator>
<dc:creator>Vranka, J. A.</dc:creator>
<dc:date>2023-03-12</dc:date>
<dc:identifier>doi:10.1101/2023.03.11.532242</dc:identifier>
<dc:title><![CDATA[Characterization of extracellular matrix deposited by segmental trabecular meshwork cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-03-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.03.13.532291v1?rss=1">
<title>
<![CDATA[
The landscape of cancer rewired GPCR signaling axes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.03.13.532291v1?rss=1</link>
<description><![CDATA[
We explored the dysregulation of GPCR ligand signaling systems in cancer transcriptomics datasets to uncover new therapeutics opportunities in oncology. We derived an interaction network of receptors with ligands and their biosynthetic enzymes, which revealed that multiple GPCRs are differentially regulated together with their upstream partners across cancer subtypes. We showed that biosynthetic pathway enrichment from enzyme expression recapitulated pathway activity signatures from metabolomics datasets, providing valuable surrogate information for GPCRs responding to organic ligands. We found that several GPCRs signaling components were significantly associated with patient survival in a cancer type-specific fashion. The expression of both receptor-ligand (or enzymes) partners improved patient stratification, suggesting a synergistic role for the activation of GPCR networks in modulating cancer phenotypes. Remarkably, we identified many such axes across several cancer molecular subtypes, including many pairs involving receptor- biosynthetic enzymes for neurotransmitters. We found that GPCRs from these actionable axes, including e.g., muscarinic, adenosine, 5-hydroxytryptamine and chemokine receptors, are the targets of multiple drugs displaying anti-growth effects in large-scale, cancer cell drug screens. We have made the results generated in this study freely available through a webapp (gpcrcanceraxes.bioinfolab.sns.it).

SignificanceComprehensive analysis of GPCR extracellular network in cancer transcriptomics datasets reveals signaling axes associated to patient survival, whose targeting is associated with growth inhibition in cancer cell lines drug sensitivity assays.
]]></description>
<dc:creator>Raimondi, F.</dc:creator>
<dc:creator>Arora, C.</dc:creator>
<dc:creator>Matic, M.</dc:creator>
<dc:creator>Rosa, N. D. O.</dc:creator>
<dc:creator>Nemati, L.</dc:creator>
<dc:creator>Clubb, L.</dc:creator>
<dc:creator>Kargas, G.</dc:creator>
<dc:creator>Vukotic, R.</dc:creator>
<dc:creator>Licata, L.</dc:creator>
<dc:creator>Wu, G.</dc:creator>
<dc:creator>Gutkind, J. S.</dc:creator>
<dc:date>2023-03-14</dc:date>
<dc:identifier>doi:10.1101/2023.03.13.532291</dc:identifier>
<dc:title><![CDATA[The landscape of cancer rewired GPCR signaling axes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-03-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.03.13.532348v1?rss=1">
<title>
<![CDATA[
Vertical transmission of African-lineage Zika virus through the fetal membranes in a rhesus Macaque (Macaca mulatta) model 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.03.13.532348v1?rss=1</link>
<description><![CDATA[
Zika virus (ZIKV) can be transmitted vertically from mother to fetus during pregnancy, resulting in a range of outcomes, including severe birth defects and fetal/infant death. Potential pathways of vertical transmission in utero have been proposed but remain undefined. Identifying the timing and routes of vertical transmission of ZIKV may help us identify when interventions would be most effective. Furthermore, understanding what barriers ZIKV overcomes to effect vertical transmission may help improve models for evaluating infection by other pathogens during pregnancy. To determine the pathways of vertical transmission, we inoculated 12 pregnant rhesus macaques with an African-lineage ZIKV at gestational day 30 (term is 165 days). Eight pregnancies were surgically terminated at either seven or 14 days post-maternal infection. Maternal-fetal interface and fetal tissues and fluids were collected and evaluated with RT-qPCR, in situ hybridization for ZIKV RNA, immunohistochemistry, and plaque assays. Four additional pregnant macaques were inoculated and terminally perfused with 4% paraformaldehyde at three, six, nine, or ten days post-maternal inoculation. For these four cases, the entire fixed pregnant uterus was evaluated with in situ hybridization for ZIKV RNA. We determined that ZIKV can reach the MFI by six days post-infection and infect the fetus by ten days. Infection of the chorionic membrane and the extraembryonic coelomic fluid preceded infection of the fetus and the mesenchymal tissue of the placental villi. We did not find evidence to support a transplacental route of ZIKV vertical transmission via infection of syncytiotrophoblasts or villous cytotrophoblasts. The pattern of infection observed in the maternal-fetal interface provides evidence of vertical ZIKV transmission through the fetal membranes.

Authors SummaryZika virus (ZIKV) can be vertically transmitted from mother to fetus during pregnancy resulting in adverse pregnancy outcomes. For vertical transmission to occur, ZIKV must overcome the barriers of the maternal-fetal interface, yet the exact pathway ZIKV takes remains undefined. The maternal-fetal interface consists of the maternal decidua, the placenta, and the fetal membranes. ZIKV could reach the fetus through the placenta if it can infect the layer of cells that are directly exposed to maternal blood. ZIKV could also reach the fetus by infecting the decidua and then the adjacent fetal membranes. To determine the pathways of ZIKV vertical transmission, we infected pregnant macaques and evaluated ZIKV burden in the maternal-fetal interface and fetus shortly after maternal infection. The pattern of infection observed suggests that ZIKV vertically transmits through the fetal membranes, not the placenta. This finding is significant because it challenges the assumption that vertical transmission occurs exclusively across the placenta. By including the fetal membranes in our models of vertical transmission, we can more accurately determine which pathogens can be vertically transmitted. Ultimately, this study demonstrates that fetal membranes are an essential barrier to pathogens that warrant further investigation.
]]></description>
<dc:creator>Koenig, M. R.</dc:creator>
<dc:creator>Mitzey, A. M.</dc:creator>
<dc:creator>Zeng, X.</dc:creator>
<dc:creator>Reyes, L.</dc:creator>
<dc:creator>Simmons, H. A.</dc:creator>
<dc:creator>Morgan, T. K.</dc:creator>
<dc:creator>Bohm, E. K.</dc:creator>
<dc:creator>Pritchard, J. C.</dc:creator>
<dc:creator>Schmidt, J. A.</dc:creator>
<dc:creator>Ren, E.</dc:creator>
<dc:creator>Jaimes, F. L.</dc:creator>
<dc:creator>Winston, E.</dc:creator>
<dc:creator>Basu, P.</dc:creator>
<dc:creator>Weiler, A. M.</dc:creator>
<dc:creator>Friedrich, T. C.</dc:creator>
<dc:creator>Aliota, M. T.</dc:creator>
<dc:creator>Mohr, E. L.</dc:creator>
<dc:creator>Golos, T. G.</dc:creator>
<dc:date>2023-03-14</dc:date>
<dc:identifier>doi:10.1101/2023.03.13.532348</dc:identifier>
<dc:title><![CDATA[Vertical transmission of African-lineage Zika virus through the fetal membranes in a rhesus Macaque (Macaca mulatta) model]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-03-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.03.22.533774v1?rss=1">
<title>
<![CDATA[
Thermogenetic control of Ca2+ levels in cells and tissues 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.03.22.533774v1?rss=1</link>
<description><![CDATA[
Virtually all major processes in cells and tissues are regulated by calcium ions (Ca2+). Understanding the influence of Ca2+ on cell function requires technologies that allow for non-invasive manipulation of intracellular calcium levels including the formation of calcium patterns, ideally in a way that is expandable to intact organisms. The currently existing tools for optical and optogenetic Ca2+ manipulation are limited with respect to response time, and tissue penetration depth. Here we present Genetically Encoded Calcium Controller (GECCO), a system for thermogenetic Ca2+ manipulation based on snake TRP channels optically controlled by infrared illumination. GECCO is functional in animal and plant cells and allows studying how cells decode different profiles of Ca2+ signals. GECCO enabled the shaping of insulin release from {beta}-cells, the identification of drugs that potentiate Ca2+-induced insulin release, and the generation of synthetic Ca2+ signatures in plants.
]]></description>
<dc:creator>Ermakova, Y. G.</dc:creator>
<dc:creator>Waadt, R.</dc:creator>
<dc:creator>Ozturk, M. S.</dc:creator>
<dc:creator>Roshchin, M.</dc:creator>
<dc:creator>Lanin, A. A.</dc:creator>
<dc:creator>Chebotarev, A.</dc:creator>
<dc:creator>Pochechuev, M.</dc:creator>
<dc:creator>Pak, V.</dc:creator>
<dc:creator>Kelmanson, I.</dc:creator>
<dc:creator>Smolyarovasmol, D.</dc:creator>
<dc:creator>Keutler, K.</dc:creator>
<dc:creator>Matyushenko, A. M.</dc:creator>
<dc:creator>Tischer, C. V.</dc:creator>
<dc:creator>Balaban, P. M.</dc:creator>
<dc:creator>Nikitin, E. S.</dc:creator>
<dc:creator>Schumacher, K.</dc:creator>
<dc:creator>Zheltikov, A.</dc:creator>
<dc:creator>Prevedel, R.</dc:creator>
<dc:creator>Schultz, C.</dc:creator>
<dc:creator>Belousov, V. V.</dc:creator>
<dc:date>2023-03-22</dc:date>
<dc:identifier>doi:10.1101/2023.03.22.533774</dc:identifier>
<dc:title><![CDATA[Thermogenetic control of Ca2+ levels in cells and tissues]]></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/10.1101/2023.03.23.533712v1?rss=1">
<title>
<![CDATA[
Supervised learning of high-confidence phenotypic subpopulations from single-cell data 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.03.23.533712v1?rss=1</link>
<description><![CDATA[
Accurately identifying phenotype-relevant cell subsets from heterogeneous cell populations is crucial for delineating the underlying mechanisms driving biological or clinical phenotypes. Here, by deploying a learning with rejection strategy, we developed a novel supervised learning framework called PENCIL to identify subpopulations associated with categorical or continuous phenotypes from single-cell data. By embedding a feature selection function into this flexible framework, for the first time, we were able to select informative features and identify cell subpopulations simultaneously, which enables the accurate identification of phenotypic subpopulations otherwise missed by methods incapable of concurrent gene selection. Furthermore, the regression mode of PENCIL presents a novel ability for supervised phenotypic trajectory learning of subpopulations from single-cell data. We conducted comprehensive simulations to evaluate PENCILs versatility in simultaneous gene selection, subpopulation identification and phenotypic trajectory prediction. PENCIL is fast and scalable to analyze 1 million cells within 1 hour. Using the classification mode, PENCIL detected T-cell subpopulations associated with melanoma immunotherapy outcomes. Moreover, when applied to scRNA-seq of a mantle cell lymphoma patient with drug treatment across multiple time points, the regression mode of PENCIL revealed a transcriptional treatment response trajectory. Collectively, our work introduces a scalable and flexible infrastructure to accurately identify phenotype-associated subpopulations from single-cell data.
]]></description>
<dc:creator>Ren, T.</dc:creator>
<dc:creator>Chen, C.</dc:creator>
<dc:creator>Danilov, A. V.</dc:creator>
<dc:creator>Liu, S.</dc:creator>
<dc:creator>Guan, X.</dc:creator>
<dc:creator>Du, S.</dc:creator>
<dc:creator>Wu, X.</dc:creator>
<dc:creator>Sherman, M. H.</dc:creator>
<dc:creator>Spellman, P. T.</dc:creator>
<dc:creator>Coussens, L. M.</dc:creator>
<dc:creator>Adey, A. C.</dc:creator>
<dc:creator>Mills, G. B.</dc:creator>
<dc:creator>Wu, L.-Y.</dc:creator>
<dc:creator>Xia, Z.</dc:creator>
<dc:date>2023-03-25</dc:date>
<dc:identifier>doi:10.1101/2023.03.23.533712</dc:identifier>
<dc:title><![CDATA[Supervised learning of high-confidence phenotypic subpopulations from single-cell data]]></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/10.1101/2023.03.27.534460v1?rss=1">
<title>
<![CDATA[
A naturally occurring variant of MBD4 causes maternal germline hypermutation in primates 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.03.27.534460v1?rss=1</link>
<description><![CDATA[
As part of an ongoing genome sequencing project at the Oregon National Primate Research Center, we identified a rhesus macaque with a rare homozygous frameshift mutation in the gene Methyl-CpG binding domain 4 (MBD4). MBD4 is responsible for the repair of C>T deamination mutations at CpG locations and has been linked to somatic hypermutation and cancer predisposition in humans. We show here that MBD4-associated hypermutation also affects the germline: the 6 offspring of the MBD4-null dam have a 4-6 fold increase in de novo mutation burden. This excess burden was predominantly C>T mutations at CpG locations consistent with MBD4 loss-of-function in the dam. There was also a significant excess of C>T at CpA sites, indicating an important, underappreciated role for MBD4 to repair deamination in CpA contexts. The MBD4-null dam developed sustained eosinophilia later in life, but we saw no other signs of neoplastic processes associated with MBD4 loss-of-function in humans, nor any obvious disease in the hypermutated offspring. This work provides what is likely the first evidence for a genetic factor causing hypermutation in the maternal germline of a mammal, and adds to the very small list of naturally occurring variants known to modulate germline mutation rates in mammals.
]]></description>
<dc:creator>Stendahl, A.</dc:creator>
<dc:creator>Sanghvi, R.</dc:creator>
<dc:creator>Peterson, S.</dc:creator>
<dc:creator>Ray, K.</dc:creator>
<dc:creator>Lima, A. C.</dc:creator>
<dc:creator>Rahbari, R.</dc:creator>
<dc:creator>Conrad, D.</dc:creator>
<dc:date>2023-03-29</dc:date>
<dc:identifier>doi:10.1101/2023.03.27.534460</dc:identifier>
<dc:title><![CDATA[A naturally occurring variant of MBD4 causes maternal germline hypermutation in primates]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-03-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.03.29.534715v1?rss=1">
<title>
<![CDATA[
Glycosylation differentially affects immune cell-specific tetraspanins CD37 and CD53 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.03.29.534715v1?rss=1</link>
<description><![CDATA[
Tetraspanin proteins play an important role in many cellular processes as they are key organizers of different receptors on the plasma membrane. Most tetraspanins are highly glycosylated at their large extracellular loop, but the function of this post-translational modification remains largely unstudied. In this study we investigated the effects of glycosylation of CD37 and CD53, two tetraspanins important for cellular and humoral immunity. Broad and cell-specific repertoires of N-glycosylated CD37 and CD53 were observed in human B cells. We generated different glycosylation mutants of CD37 and CD53 and analyzed their localization, nanoscale organization and partner protein interaction capacity. Abrogation of glycosylation in CD37 revealed the importance of this modification for CD37 surface expression, whereas neither surface expression nor nanoscale organization of CD53 was affected by its glycosylation. CD37 interaction with its known partner proteins, CD20 and IL-6R, was not affected by glycosylation, other than via its changed subcellular localization. Surprisingly, glycosylation was found to inhibit the interaction between CD53 and its partner proteins CD45 and CD20. Together, our data show that tetraspanin glycosylation affects their function in immune cells, which adds another layer of regulation to tetraspanin-mediated membrane organization.
]]></description>
<dc:creator>van Deventer, S. J.</dc:creator>
<dc:creator>Hoogvliet, I. A.</dc:creator>
<dc:creator>van de Voort, M.</dc:creator>
<dc:creator>Arnold, F.</dc:creator>
<dc:creator>van Spriel, A.</dc:creator>
<dc:date>2023-03-29</dc:date>
<dc:identifier>doi:10.1101/2023.03.29.534715</dc:identifier>
<dc:title><![CDATA[Glycosylation differentially affects immune cell-specific tetraspanins CD37 and CD53]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-03-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.03.29.534727v1?rss=1">
<title>
<![CDATA[
Integrated single cell analysis shows chronic alcohol drinking disrupts monocyte differentiation in the bone marrow niche 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.03.29.534727v1?rss=1</link>
<description><![CDATA[
Chronic alcohol drinking rewires circulating monocytes and tissue-resident macrophages towards heightened inflammatory states with compromised anti-microbial defenses. As these effects remain consistent in short-lived monocytes after a 1-month abstinence period it is unclear whether these changes are restricted to the periphery or mediated through alterations in the progenitor niche. To test this hypothesis, we profiled monocytes/macrophages and hematopoietic stem cell progenitors (HSCP) of the bone marrow compartment from rhesus macaques after 12 months of ethanol consumption using a combination of functional assays and single cell genomics. Bone marrow-resident monocytes/macrophages from ethanol-consuming animals exhibited heightened inflammation. Differentiation of HSCP in vitro revealed skewing towards monocytes expressing neutrophil-like markers with heightened inflammatory responses to bacterial agonists. Single cell transcriptional analysis of HSCPs showed reduced proliferation but increased inflammatory markers in mature myeloid progenitors. We observed transcriptional signatures associated with increased oxidative and cellular stress as well as oxidative phosphorylation in immature and mature myeloid progenitors. Single cell analysis of the chromatin landscape showed altered drivers of differentiation in monocytes and progenitors. Collectively, these data indicate that chronic ethanol drinking results in remodeling of the transcriptional and epigenetic landscapes of the bone marrow compartment leading to altered functions in the periphery.
]]></description>
<dc:creator>Lewis, S. A.</dc:creator>
<dc:creator>Doratt, B. M.</dc:creator>
<dc:creator>Qiao, Q.</dc:creator>
<dc:creator>Blanton, M. B.</dc:creator>
<dc:creator>Grant, K. A.</dc:creator>
<dc:creator>Messaoudi, I.</dc:creator>
<dc:date>2023-03-30</dc:date>
<dc:identifier>doi:10.1101/2023.03.29.534727</dc:identifier>
<dc:title><![CDATA[Integrated single cell analysis shows chronic alcohol drinking disrupts monocyte differentiation in the bone marrow niche]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-03-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.04.02.535293v1?rss=1">
<title>
<![CDATA[
Dopamine D1 and NMDA receptor co-regulation of protein translation in cultured nucleus accumbens neurons 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.04.02.535293v1?rss=1</link>
<description><![CDATA[
Protein translation is essential for some forms of synaptic plasticity. We used nucleus accumbens (NAc) medium spiny neurons (MSN), co-cultured with cortical neurons to restore excitatory synapses, to examine whether dopamine modulates protein translation in NAc MSN. FUNCAT was used to measure translation in MSNs under basal conditions and after disinhibiting excitatory transmission using the GABAA receptor antagonist bicuculline (2 hr). Under basal conditions, translation was not altered by the D1-class receptor (D1R) agonist SKF81297 or the D2-class receptor (D2R) agonist quinpirole. Bicuculline alone robustly increased translation. This was reversed by quinpirole but not SKF81297. It was also reversed by co-incubation with the D1R antagonist SCH23390, but not the D2R antagonist eticlopride, suggesting dopaminergic tone at D1Rs. This was surprising because no dopamine neurons are present. An alternative explanation is that bicuculline activates translation by increasing glutamate tone at NMDA receptors (NMDAR) within D1R/NMDAR heteromers, which have been described in other cell types. Supporting this, immunocytochemistry and proximity ligation assays revealed D1/NMDAR heteromers on NAc cells both in vitro and in vivo. Further, bicucullines effect was reversed to the same extent by SCH23390 alone, the NMDAR antagonist APV alone, or SCH23390+APV. These results suggest that: 1) excitatory synaptic transmission stimulates translation in NAc MSNs, 2) this is opposed when glutamate activates D1R/NMDAR heteromers, even in the absence of dopamine, and 3) antagonist occupation of D1Rs within the heteromers prevents their activation. Our study is the first to suggest a role for D2 receptors and D1R/NMDAR heteromers in regulating protein translation.
]]></description>
<dc:creator>Zimbelman, A. R.</dc:creator>
<dc:creator>Wong, B.</dc:creator>
<dc:creator>Murray, C. H.</dc:creator>
<dc:creator>Wolf, M. E.</dc:creator>
<dc:creator>Stefanik, M. T.</dc:creator>
<dc:date>2023-04-02</dc:date>
<dc:identifier>doi:10.1101/2023.04.02.535293</dc:identifier>
<dc:title><![CDATA[Dopamine D1 and NMDA receptor co-regulation of protein translation in cultured nucleus accumbens neurons]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-04-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.04.10.536057v1?rss=1">
<title>
<![CDATA[
Protective effect of pre-existing natural immunity in a nonhuman primate reinfection model of congenital cytomegalovirus infection 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.04.10.536057v1?rss=1</link>
<description><![CDATA[
Congenital cytomegalovirus (cCMV) is the leading infectious cause of neurologic defects in newborns with particularly severe sequelae in the setting of primary CMV infection in the first trimester of pregnancy. The majority of cCMV cases worldwide occur after non-primary infection in CMV-seropositive women; yet the extent to which pre-existing natural CMV-specific immunity protects against CMV reinfection or reactivation during pregnancy remains ill-defined. We previously reported on a novel nonhuman primate model of cCMV in rhesus macaques where 100% placental transmission and 83% fetal loss were seen in CD4+ T lymphocyte-depleted rhesus CMV (RhCMV)-seronegative dams after primary RhCMV infection. To investigate the protective effect of preconception maternal immunity, we performed reinfection studies in CD4+ T lymphocyte-depleted RhCMV-seropositive dams inoculated in late first / early second trimester gestation with RhCMV strains 180.92 (n=2), or RhCMV UCD52 and FL-RhCMV{Delta}Rh13.1/SIVgag, a wild-type-like RhCMV clone with SIVgag inserted as an immunological marker (n=3). An early transient increase in circulating monocytes followed by boosting of the pre-existing RhCMV-specific CD8+ T lymphocyte and antibody response was observed in the reinfected dams but not in control CD4+ T lymphocyte-depleted dams. Emergence of SIV Gag-specific CD8+ T lymphocyte responses in macaques inoculated with the FL-RhCMV{Delta}Rh13.1/SIVgag virus confirmed reinfection. Placental transmission was detected in only one of five reinfected dams and there were no adverse fetal sequelae. Viral whole genome, short-read, deep sequencing analysis confirmed transmission of both reinfection RhCMV strains across the placenta with [~]30% corresponding to FL-RhCMV{Delta}Rh13.1/SIVgag and [~]70% to RhCMV UCD52, consistent with the mixed human CMV infections reported in infants with cCMV. Our data showing reduced placental transmission and absence of fetal loss after non-primary as opposed to primary infection in CD4+ T lymphocyte-depleted dams indicates that preconception maternal CMV-specific CD8+ T lymphocyte and/or humoral immunity can protect against cCMV infection.

Author SummaryGlobally, pregnancies in CMV-seropositive women account for the majority of cases of congenital CMV infection but the immune responses needed for protection against placental transmission in mothers with non-primary infection remains unknown. Recently, we developed a nonhuman primate model of primary rhesus CMV (RhCMV) infection in which placental transmission and fetal loss occurred in RhCMV-seronegative CD4+ T lymphocyte-depleted macaques. By conducting similar studies in RhCMV-seropositive dams, we demonstrated the protective effect of pre-existing natural CMV-specific CD8+ T lymphocytes and humoral immunity against congenital CMV after reinfection. A 5-fold reduction in congenital transmission and complete protection against fetal loss was observed in dams with pre-existing immunity compared to primary CMV in this model. Our study is the first formal demonstration in a relevant model of human congenital CMV that natural pre-existing CMV-specific maternal immunity can limit congenital CMV transmission and its sequelae. The nonhuman primate model of non-primary congenital CMV will be especially relevant to studying immune requirements of a maternal vaccine for women in high CMV seroprevalence areas at risk of repeated CMV reinfections during pregnancy.
]]></description>
<dc:creator>Mostrom, M. J.</dc:creator>
<dc:creator>Yu, S.</dc:creator>
<dc:creator>Tran, D.</dc:creator>
<dc:creator>Saccoccio, F.</dc:creator>
<dc:creator>Versoza, C. J.</dc:creator>
<dc:creator>Malouli, D. J.</dc:creator>
<dc:creator>Mirza, A.</dc:creator>
<dc:creator>Valencia, S.</dc:creator>
<dc:creator>Gilbert, M.</dc:creator>
<dc:creator>Blair, R. V.</dc:creator>
<dc:creator>Hansen, S. G.</dc:creator>
<dc:creator>Barry, P. A.</dc:creator>
<dc:creator>Fruh, K.</dc:creator>
<dc:creator>Jensen, J. D.</dc:creator>
<dc:creator>Pfeifer, S. P.</dc:creator>
<dc:creator>Kowalik, T. F.</dc:creator>
<dc:creator>Permar, S. R.</dc:creator>
<dc:creator>Kaur, A.</dc:creator>
<dc:date>2023-04-10</dc:date>
<dc:identifier>doi:10.1101/2023.04.10.536057</dc:identifier>
<dc:title><![CDATA[Protective effect of pre-existing natural immunity in a nonhuman primate reinfection model of congenital cytomegalovirus infection]]></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/10.1101/2023.04.13.536793v1?rss=1">
<title>
<![CDATA[
Sequential chromogenic immunohistochemistry: spatial analysis of lymph nodes identifies contact interactions between plasmacytoid dendritic cells and plasmablasts 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.04.13.536793v1?rss=1</link>
<description><![CDATA[
Recent clinical observations highlight the importance of the spatial organization of immune cells into lymphoid structures for the success of cancer immunotherapy and patient survival. Sequential chromogenic immunohistochemistry (scIHC) supports the analysis of multiple biomarkers on a single tissue section thus providing unique information about relative location of cell types and assessment of disease states. Unfortunately, widespread implementation of scIHC is limited by lack of a standardized, rigorous guide to the development of customized biomarker panels and by the need for user-friendly analysis pipelines able to streamline the extraction of meaningful data. Here, we examine major steps from classical IHC protocols and highlight the impact they have on the scIHC procedure. We report practical examples and illustrations of the most common complications that can arise during the setup of a new biomarker panel and how to avoid them. We described in detail how to prevent and detect cross- reactivity between secondary reagents and carry over between detection antibodies. We developed a novel analysis pipeline based on non-rigid tissue deformation correction, Cellpose-inspired automated cell segmentation and computational network masking of low-quality data. The resulting biomarker panel and pipeline was used to study regional lymph nodes from head and neck cancer patients. We identified contact interactions between plasmablasts and plasmacytoid dendritic cells in vivo. Given that TLR receptors, which are highly expressed in plasmacytoid dendritic cells play a key role in vaccine efficacy, the significance of this cell-cell interaction decisively warrants further studies. In conclusion, this work streamlines the development of novel biomarker panels for scIHC, which will ultimately improve our understanding of immune responses in cancer.
]]></description>
<dc:creator>Claudio, N. M.</dc:creator>
<dc:creator>Nguyen, M.-T.</dc:creator>
<dc:creator>Wanner, A.</dc:creator>
<dc:creator>Pucci, F.</dc:creator>
<dc:date>2023-04-14</dc:date>
<dc:identifier>doi:10.1101/2023.04.13.536793</dc:identifier>
<dc:title><![CDATA[Sequential chromogenic immunohistochemistry: spatial analysis of lymph nodes identifies contact interactions between plasmacytoid dendritic cells and plasmablasts]]></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/10.1101/2023.04.17.537181v1?rss=1">
<title>
<![CDATA[
Structure-function Analyses Reveal Key Molecular Determinants of HIV-1 CRF01_AE Resistance to the Entry Inhibitor Temsavir 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.04.17.537181v1?rss=1</link>
<description><![CDATA[
The HIV-1 entry inhibitor temsavir prevents CD4 from interacting with the envelope glycoprotein (Env) and blocks its conformational changes. To do this temsavir relies on the presence of a residue with small side chain at position 375 in Env and is unable to neutralize viral strains like CRF01_AE carrying His375. Here we investigate the mechanism of temsavir-resistance and show that residue 375 is not the sole determinant of resistance. At least six additional residues within the gp120 inner domain layers, including five distant from the drug-binding pocket, contribute to resistance. A detailed structure-function analysis using engineered viruses and soluble trimer variants reveal that the molecular basis of resistance is mediated by crosstalk between His375 and the inner domain layers. Furthermore, our data confirm that temsavir can adjust its binding mode to accommodate changes in Env conformation, a property that likely contributes to its broad-antiviral activity.
]]></description>
<dc:creator>Pazgier, M.</dc:creator>
<dc:creator>Prevost, J.</dc:creator>
<dc:creator>Chen, Y.</dc:creator>
<dc:creator>Zhou, F.</dc:creator>
<dc:creator>Tolbert, W. D.</dc:creator>
<dc:creator>Gasser, R.</dc:creator>
<dc:creator>Medjahed, H.</dc:creator>
<dc:creator>Gottumukkala, S.</dc:creator>
<dc:creator>Hessell, A.</dc:creator>
<dc:creator>Rao, V. B.</dc:creator>
<dc:creator>Pozharski, E.</dc:creator>
<dc:creator>Huang, R.</dc:creator>
<dc:creator>Motthies, D.</dc:creator>
<dc:creator>Finzi, A.</dc:creator>
<dc:date>2023-04-17</dc:date>
<dc:identifier>doi:10.1101/2023.04.17.537181</dc:identifier>
<dc:title><![CDATA[Structure-function Analyses Reveal Key Molecular Determinants of HIV-1 CRF01_AE Resistance to the Entry Inhibitor Temsavir]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-04-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.04.15.536434v1?rss=1">
<title>
<![CDATA[
Gemcitabine plus nab-paclitaxel preserves skeletal and cardiac mass and function in a murine model of pancreatic cancer cachexia 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.04.15.536434v1?rss=1</link>
<description><![CDATA[
More than 85% of patients with pancreatic ductal adenocarcinoma (PDAC) suffer from cachexia, a debilitating syndrome characterized by the loss of muscle and fat and remains an unmet medical need. While chemotherapy remains an effective treatment option, it can also induce weight and muscle loss in patients with cancer. Gemcitabine combined with nab paclitaxel (GnP) is a first line treatment option for patients with PDAC but GnPs effect on cachexia has not been comprehensively investigated. We interrogated the effects of GnP in a murine model of pancreatic cancer cachexia. Mice were orthotopically implanted with the cachexia inducing pancreatic cell line (KPC) and were administered GnP or vehicle. The controls underwent sham surgery. We defined GnP effects on cachexia and tumor burden by evaluating muscle and cardiac mass and function, fat mass, bone morphometry, and hematology measurements. We completed RNA sequencing and deep proteome profiling in skeletal and cardiac muscle. KPC+GnP reduced tumor burden over 50% and increased survival compared to KPC. KPC vehicle group had more than 15% muscle mass loss and decreased left ventricular mass, this was not present in KPC+GnP when compared to controls. RNA Seq and deep proteomics analyses suggested that muscle and cardiac dysfunction pathways activated in KPC group were either reversed or decreased in KPC+GnP. In all, our data suggests that GnP protects against muscle and cardiac wasting in an experimental model of PDAC cachexia.



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]]></description>
<dc:creator>Narasimhan, A.</dc:creator>
<dc:creator>Jengelley, D.</dc:creator>
<dc:creator>Huot, J.</dc:creator>
<dc:creator>Umberger, T.</dc:creator>
<dc:creator>Doud, E.</dc:creator>
<dc:creator>Mosley, A. L.</dc:creator>
<dc:creator>Wang, M.</dc:creator>
<dc:creator>Zhong, X.</dc:creator>
<dc:creator>Counts, B.</dc:creator>
<dc:creator>Rupert, J.</dc:creator>
<dc:creator>Young, A.</dc:creator>
<dc:creator>Bonetto, A.</dc:creator>
<dc:creator>Horan, D.</dc:creator>
<dc:creator>Robling, A. G.</dc:creator>
<dc:creator>Fishel, M.</dc:creator>
<dc:creator>Kelley, M.</dc:creator>
<dc:creator>Koniaris, L.</dc:creator>
<dc:creator>Zimmers, T.</dc:creator>
<dc:date>2023-04-18</dc:date>
<dc:identifier>doi:10.1101/2023.04.15.536434</dc:identifier>
<dc:title><![CDATA[Gemcitabine plus nab-paclitaxel preserves skeletal and cardiac mass and function in a murine model of pancreatic cancer cachexia]]></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/10.1101/2023.04.19.537573v1?rss=1">
<title>
<![CDATA[
RhoA GEF Mcf2lb regulates rosette integrity during collective cell migration 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.04.19.537573v1?rss=1</link>
<description><![CDATA[
During development, multicellular rosettes serve as important cellular intermediates in the formation of diverse organ systems. Multicellular rosettes are transient epithelial structures that are defined by the apical constriction of cells towards the rosette center. Due to the important role these structures play during development, understanding the molecular mechanisms by which rosettes are formed and maintained is of high interest. Utilizing the zebrafish posterior lateral line primordium (pLLP) as a model system, we identify the RhoA GEF Mcf2lb as a regulator of rosette integrity. The pLLP is a group of [~]150 cells that migrates along the zebrafish trunk and is organized into epithelial rosettes; these are deposited along the trunk and will differentiate into sensory organs called neuromasts (NMs). Using single-cell RNA sequencing and whole-mount in situ hybridization, we showed that mcf2lb is expressed in the pLLP during migration. Given the known role of RhoA in rosette formation, we asked whether Mcf2lb plays a role in regulating apical constriction of cells within rosettes. Live imaging and subsequent 3D analysis of mcf2lb mutant pLLP cells showed disrupted apical constriction and subsequent rosette organization. This in turn resulted in a unique posterior Lateral Line phenotype: an excess number of deposited NMs along the trunk of the zebrafish. Cell polarity markers ZO-1 and Par-3 were apically localized, indicating that pLLP cells are normally polarized. In contrast, signaling components that mediate apical constriction downstream of RhoA, Rock-2a and non-muscle Myosin II were diminished apically. Altogether our results suggest a model whereby Mcf2lb activates RhoA, which in turn activates downstream signaling machinery to induce and maintain apical constriction in cells incorporated into rosettes.
]]></description>
<dc:creator>Olson, H. M.</dc:creator>
<dc:creator>Maxfield, A.</dc:creator>
<dc:creator>Calistri, N. L.</dc:creator>
<dc:creator>Heiser, L. M.</dc:creator>
<dc:creator>Nechiporuk, A. V.</dc:creator>
<dc:date>2023-04-20</dc:date>
<dc:identifier>doi:10.1101/2023.04.19.537573</dc:identifier>
<dc:title><![CDATA[RhoA GEF Mcf2lb regulates rosette integrity during collective cell migration]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-04-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.04.21.537769v1?rss=1">
<title>
<![CDATA[
Relationship of maternal cytomegalovirus-specific antibody responses and viral load to vertical transmission risk following primary maternal infection in a rhesus macaque model 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.04.21.537769v1?rss=1</link>
<description><![CDATA[
Cytomegalovirus (CMV) is the most common congenital infection and cause of birth defects worldwide. Primary CMV infection during pregnancy leads to a higher frequency of congenital CMV (cCMV) than maternal re-infection, suggesting that maternal immunity confers partial protection. However, poorly understood immune correlates of protection against placental transmission contributes to the current lack of an approved vaccine to prevent cCMV. In this study, we characterized the kinetics of maternal plasma rhesus CMV (RhCMV) viral load (VL) and RhCMV-specific antibody binding and functional responses in a group of 12 immunocompetent dams with acute, primary RhCMV infection. We defined cCMV transmission as RhCMV detection in amniotic fluid (AF) by qPCR. We then leveraged a large group of past and current primary RhCMV infection studies in late-first/early-second trimester RhCMV-seronegative rhesus macaque dams, including immunocompetent (n=15), CD4+ T cell-depleted with (n=6) and without (n=6) RhCMV-specific polyclonal IgG infusion before infection to evaluate differences between RhCMV AF-positive and AF-negative dams. During the first 3 weeks after infection, the magnitude of RhCMV VL in maternal plasma was higher in AF-positive dams in the combined cohort, while RhCMV glycoprotein B (gB)- and pentamer-specific binding IgG responses were lower magnitude compared to AF-negative dams. However, these observed differences were driven by the CD4+ T cell-depleted dams, as there were no differences in plasma VL or antibody responses between immunocompetent AF-positive vs AF-negative dams. Overall, these results suggest that levels of neither maternal plasma viremia nor humoral responses are associated with cCMV following primary maternal infection in healthy individuals. We speculate that other factors related to innate immunity are more important in this context as antibody responses to acute infection likely develop too late to influence vertical transmission. Yet, pre-existing CMV glycoprotein-specific and neutralizing IgG may provide protection against cCMV following primary maternal CMV infection even in high-risk, immunocompromised settings.

Author summaryCytomegalovirus (CMV) is the most common infectious cause of birth defects globally, but we still do not have licensed medical interventions to prevent vertical transmission of CMV. We utilized a non-human primate model of primary CMV infection during pregnancy to study virological and humoral factors that influence congenital infection. Unexpectedly, we found that the levels virus in maternal plasma were not predictive of virus transmission to the amniotic fluid (AF) in immunocompetent dams. In contrast, CD4+ T cell depleted pregnant rhesus macaques with virus detected in AF had higher plasma viral loads than dams not showing placental transmission. Virus-specific antibody binding, neutralizing, and Fc-mediated antibody effector antibody responses were not different in immunocompetent animals with and without virus detectable in AF, but passively infused neutralizing antibodies and antibodies binding to key glycoproteins were higher in CD4+ T cell-depleted dams who did not transmit the virus compared to those that did. Our data suggests that the natural development of virus-specific antibody responses is too slow to prevent congenital transmission following maternal infection, highlighting the need for the development of vaccines that confer levels of pre-existing immunity to CMV-naive mothers that can prevent congenital transmission to their infants during pregnancy.
]]></description>
<dc:creator>Otero, C. E.</dc:creator>
<dc:creator>Barfield, R.</dc:creator>
<dc:creator>Scheef, E.</dc:creator>
<dc:creator>Nelson, C. S.</dc:creator>
<dc:creator>Rodgers, N.</dc:creator>
<dc:creator>Wang, H.-Y.</dc:creator>
<dc:creator>Moström, M.</dc:creator>
<dc:creator>Manuel, T. D.</dc:creator>
<dc:creator>Sass, J.</dc:creator>
<dc:creator>Schmidt, K.</dc:creator>
<dc:creator>Taher, H.</dc:creator>
<dc:creator>Papen, C.</dc:creator>
<dc:creator>Sprehe, L.</dc:creator>
<dc:creator>Kendall, S.</dc:creator>
<dc:creator>Davalos, A.</dc:creator>
<dc:creator>Barry, P. A.</dc:creator>
<dc:creator>Früh, K.</dc:creator>
<dc:creator>Pollara, J.</dc:creator>
<dc:creator>Malouli, D.</dc:creator>
<dc:creator>Chan, C.</dc:creator>
<dc:creator>Kaur, A.</dc:creator>
<dc:creator>Permar, S. R.</dc:creator>
<dc:date>2023-04-21</dc:date>
<dc:identifier>doi:10.1101/2023.04.21.537769</dc:identifier>
<dc:title><![CDATA[Relationship of maternal cytomegalovirus-specific antibody responses and viral load to vertical transmission risk following primary maternal infection in a rhesus macaque model]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-04-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.04.24.538096v1?rss=1">
<title>
<![CDATA[
Layer 5a Corticostriatal Projection Neurons are Selectively Vulnerable in Huntington's Disease 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.04.24.538096v1?rss=1</link>
<description><![CDATA[
The properties of the cell types that are selectively vulnerable in Huntingtons disease (HD) cortex, the nature of somatic CAG expansions of mHTT in these cells, and their importance in CNS circuitry have not been delineated. Here we employed serial fluorescence activated nuclear sorting (sFANS), deep molecular profiling, and single nucleus RNA sequencing (snRNAseq) to demonstrate that layer 5a pyramidal neurons are vulnerable in primary motor cortex and other cortical areas of HD donors. Extensive mHTT-CAG expansions occur in vulnerable layer 5a pyramidal cells, and in Betz cells, layer 6a, layer 6b neurons that are resilient in HD. Retrograde tracing experiments in macaque brains identify the vulnerable layer 5a neurons as corticostriatal pyramidal cells. We propose that enhanced somatic mHTT-CAG expansion and altered synaptic function act together to cause corticostriatal disconnection and selective neuronal vulnerability in the HD cerebral cortex.
]]></description>
<dc:creator>Pressl, C.</dc:creator>
<dc:creator>Mätlik, K.</dc:creator>
<dc:creator>Kus, L.</dc:creator>
<dc:creator>Darnell, P.</dc:creator>
<dc:creator>Luo, J.-D.</dc:creator>
<dc:creator>Weiss, A. R.</dc:creator>
<dc:creator>Liguore, W.</dc:creator>
<dc:creator>Carroll, T.</dc:creator>
<dc:creator>Davis, D. A.</dc:creator>
<dc:creator>McBride, J.</dc:creator>
<dc:creator>Heintz, N.</dc:creator>
<dc:date>2023-04-24</dc:date>
<dc:identifier>doi:10.1101/2023.04.24.538096</dc:identifier>
<dc:title><![CDATA[Layer 5a Corticostriatal Projection Neurons are Selectively Vulnerable in Huntington's Disease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-04-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.05.05.539468v1?rss=1">
<title>
<![CDATA[
Dopamine D2 receptors in mossy cells reduce excitatory transmission and are essential for hippocampal function 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.05.05.539468v1?rss=1</link>
<description><![CDATA[
Hilar mossy cells (MCs) are principal excitatory neurons of the dentate gyrus (DG) that play critical roles in hippocampal function and have been implicated in brain disorders such as anxiety and epilepsy. However, the mechanisms by which MCs contribute to DG function and disease are poorly understood. Expression from the dopamine D2 receptor (D2R) gene (Drd2) promoter is a defining feature of MCs, and previous work indicates a key role for dopaminergic signaling in the DG. Additionally, the involvement of D2R signaling in cognition and neuropsychiatric conditions is well-known. Surprisingly, though, the function of MC D2Rs remain largely unexplored. In this study, we show that selective and conditional removal of Drd2 from MCs of adult mice impaired spatial memory, promoted anxiety-like behavior and was proconvulsant. To determine the subcellular expression of D2Rs in MCs, we used a D2R knockin mouse which revealed that D2Rs are enriched in the inner molecular layer of the DG, where MCs establish synaptic contacts with granule cells. D2R activation by exogenous and endogenous dopamine reduced MC to dentate granule cells (GC) synaptic transmission, most likely by a presynaptic mechanism. In contrast, removing Drd2 from MCs had no significant impact on MC excitatory inputs and passive and active properties. Our findings support that MC D2Rs are essential for proper DG function by reducing MC excitatory drive onto GCs. Lastly, impairment of MC D2R signaling could promote anxiety and epilepsy, therefore highlighting a potential therapeutic target.

SIGNIFICANCEGrowing evidence indicates that hilar mossy cells (MCs) of the dentate gyrus play critical but incompletely understood roles in memory and brain disorders, including anxiety and epilepsy. Dopamine D2 receptors (D2Rs), implicated in cognition and several psychiatric and neurological disorders, are considered to be characteristically expressed by MCs. Still, the subcellular localization and function of MC D2Rs are largely unknown. We report that removing the Drd2 gene specifically from MCs of adult mice impaired spatial memory and was anxiogenic and proconvulsant. We also found that D2Rs are enriched where MCs synaptically contact dentate granule cells (GC) and reduce MC-GC transmission. This work uncovered the functional significance of MC D2Rs, thus highlighting their therapeutic potential in D2R- and MC-associated pathologies.
]]></description>
<dc:creator>Gulfo, M. C.</dc:creator>
<dc:creator>Lebowitz, J. J.</dc:creator>
<dc:creator>Ramos, C.</dc:creator>
<dc:creator>Hwang, D.-W.</dc:creator>
<dc:creator>Nasrallah, K.</dc:creator>
<dc:creator>Castillo, P. E.</dc:creator>
<dc:date>2023-05-05</dc:date>
<dc:identifier>doi:10.1101/2023.05.05.539468</dc:identifier>
<dc:title><![CDATA[Dopamine D2 receptors in mossy cells reduce excitatory transmission and are essential for hippocampal function]]></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/10.1101/2023.05.10.540233v1?rss=1">
<title>
<![CDATA[
Mild/Asymptomatic Maternal SARS-CoV-2 Infection Leads to Immune Paralysis in Fetal Circulation and Immune Dysregulation in Fetal-Placental Tissues 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.05.10.540233v1?rss=1</link>
<description><![CDATA[
Few studies have addressed the impact of maternal mild/asymptomatic SARS-CoV-2 infection on the developing neonatal immune system. In this study, we analyzed umbilical cord blood and placental chorionic villi from newborns of unvaccinated mothers with mild/asymptomatic SARS-CoV-2 infection during pregnancy using flow cytometry, single-cell transcriptomics, and functional assays. Despite the lack of vertical transmission, levels of inflammatory mediators were altered in cord blood. Maternal infection was also associated with increased memory T, B cells, and non-classical monocytes as well as increased activation. However, ex vivo responses to stimulation were attenuated. Finally, within the placental villi, we report an expansion of fetal Hofbauer cells and infiltrating maternal macrophages and rewiring towards a heightened inflammatory state. In contrast to cord blood monocytes, placental myeloid cells were primed for heightened antiviral responses. Taken together, this study highlights dysregulated fetal immune cell responses in response to mild maternal SARS-CoV-2 infection during pregnancy.
]]></description>
<dc:creator>Doratt, B. M.</dc:creator>
<dc:creator>Sureshchandra, S.</dc:creator>
<dc:creator>True, H.</dc:creator>
<dc:creator>Rincon, M.</dc:creator>
<dc:creator>Marshall, N. E.</dc:creator>
<dc:creator>Messaoudi, I.</dc:creator>
<dc:date>2023-05-11</dc:date>
<dc:identifier>doi:10.1101/2023.05.10.540233</dc:identifier>
<dc:title><![CDATA[Mild/Asymptomatic Maternal SARS-CoV-2 Infection Leads to Immune Paralysis in Fetal Circulation and Immune Dysregulation in Fetal-Placental Tissues]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-05-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.05.10.540211v1?rss=1">
<title>
<![CDATA[
Mcam stabilizes a luminal progenitor-like breast cancer cell state via Ck2 control and Src/Akt/Stat3 attenuation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.05.10.540211v1?rss=1</link>
<description><![CDATA[
Breast cancers are categorized into subtypes with distinctive therapeutic vulnerabilities and prognoses based on their expression of clinically targetable receptors and gene expression patterns mimicking different cell types of the normal gland. Here, we tested the role of Mcam in breast cancer cell state control and tumorigenicity in a luminal progenitor-like murine tumor cell line (Py230) that exhibits lineage and tumor subtype plasticity. Mcam knockdown Py230 cells show augmented Stat3 and Pi3K/Akt activation associated with a lineage state switch away from a hormone-sensing/luminal progenitor state toward alveolar and basal cell related phenotypes that were refractory to growth inhibition by the anti-estrogen therapeutic, tamoxifen. Inhibition of Stat3, or the upstream activator Ck2, reversed these cell state changes. Mcam binds Ck2 and acts as a regulator of Ck2 substrate utilization across multiple mammary tumor cell lines. In Py230 cells this activity manifests as increased mesenchymal morphology, migration, and Src/Fak/Mapk/Paxillin adhesion complex signaling in vitro, in contrast to Mcams reported roles in promoting mesenchymal phenotypes. In vivo, Mcam knockdown reduced tumor growth and take rate and inhibited cell state transition to Sox10+/neural crest like cells previously been associated with tumor aggressiveness. This contrasts with human luminal breast cancers where MCAM copy number loss is highly coupled to Cyclin D amplification, increased proliferation, and the more aggressive Luminal B subtype. Together these data indicate a critical role for Mcam and its regulation of Ck2 in control of breast cancer cell state plasticity with implications for progression, evasion of targeted therapies and combination therapy design.

HIGHLIGHTSO_LIMcam is expressed by diverse breast cancer subtypes.
C_LIO_LIMcam regulates breast cancer cell state plasticity.
C_LIO_LIMcam maintains the mammary luminal progenitor state.
C_LIO_LIMcam attenuates Stat activity.
C_LIO_LIMcam dictates Ck2 substrate utilization.
C_LI
]]></description>
<dc:creator>Balcioglu, O.</dc:creator>
<dc:creator>Gates, B. L.</dc:creator>
<dc:creator>Freeman, D. W.</dc:creator>
<dc:creator>Hagos, B. M.</dc:creator>
<dc:creator>Mehrabad, E. M.</dc:creator>
<dc:creator>Ayala-Talavera, D.</dc:creator>
<dc:creator>Spike, B. T.</dc:creator>
<dc:date>2023-05-12</dc:date>
<dc:identifier>doi:10.1101/2023.05.10.540211</dc:identifier>
<dc:title><![CDATA[Mcam stabilizes a luminal progenitor-like breast cancer cell state via Ck2 control and Src/Akt/Stat3 attenuation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-05-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.05.10.540265v1?rss=1">
<title>
<![CDATA[
A Masked Image Modeling Approach to Cyclic Immunofluorescence (CyCIF) Panel Reduction and Marker Imputation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.05.10.540265v1?rss=1</link>
<description><![CDATA[
CyCIF quantifies multiple biomarkers, but panel capacity is compromised by technical challenges including tissue loss. We propose a computational panel reduction, inferring surrogate CyCIF data from a subset of biomarkers. Our model reconstructs the information content from 25 markers using only 9 markers, learning co-expression and morphological patterns. We demonstrate strong correlations in predictions and generalizability across breast and colorectal cancer tissue microarrays, illustrating broader applicability to diverse tissue types.
]]></description>
<dc:creator>Sims, Z.</dc:creator>
<dc:creator>Chang, Y. H.</dc:creator>
<dc:date>2023-05-13</dc:date>
<dc:identifier>doi:10.1101/2023.05.10.540265</dc:identifier>
<dc:title><![CDATA[A Masked Image Modeling Approach to Cyclic Immunofluorescence (CyCIF) Panel Reduction and Marker Imputation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-05-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.05.12.540615v1?rss=1">
<title>
<![CDATA[
Molecular Basis for Maternal Inheritance of Human Mitochondrial DNA 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.05.12.540615v1?rss=1</link>
<description><![CDATA[
Uniparental inheritance of mitochondrial DNA (mtDNA) is an evolutionary trait found in nearly all eukaryotes. In many species, including humans, the sperm mitochondria are introduced to the oocyte during fertilization1, 2. The mechanisms hypothesized to prevent paternal mtDNA transmission include ubiquitination of the sperm mitochondria and mitophagy3, 4. However, whether these mechanisms play a decisive role in paternal mtDNA elimination has been disputed5, 6. We found that mitochondria in human spermatozoa are devoid of mtDNA and lack mitochondrial transcription factor A (TFAM), the major nucleoid protein required to protect, maintain, and transcribe mtDNA. During spermatogenesis, sperm cells express an isoform of TFAM, which retains the mitochondrial pre-sequence, ordinarily removed upon mitochondrial import. Phosphorylation of this pre-sequence prevents mitochondrial import and directs TFAM to the spermatozoon nucleus. TFAM re-localization from the mitochondria of spermatogonia to the spermatozoa nucleus directly correlates with the elimination of mitochondrial DNA, thereby explaining maternal inheritance in this species.
]]></description>
<dc:creator>Lee, W.</dc:creator>
<dc:creator>Zamudio-Ochoa, A.</dc:creator>
<dc:creator>Buchel, G.</dc:creator>
<dc:creator>Podlesniy, P.</dc:creator>
<dc:creator>Marti Gutierrez, N.</dc:creator>
<dc:creator>Puigros, M.</dc:creator>
<dc:creator>Calderon, A.</dc:creator>
<dc:creator>Koski, A.</dc:creator>
<dc:creator>Tang, H.-Y.</dc:creator>
<dc:creator>Li, L.</dc:creator>
<dc:creator>Trullas, R.</dc:creator>
<dc:creator>Mitalipov, S.</dc:creator>
<dc:creator>Temiakov, D.</dc:creator>
<dc:date>2023-05-13</dc:date>
<dc:identifier>doi:10.1101/2023.05.12.540615</dc:identifier>
<dc:title><![CDATA[Molecular Basis for Maternal Inheritance of Human Mitochondrial DNA]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-05-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.05.11.540245v1?rss=1">
<title>
<![CDATA[
txci-ATAC-seq, a massive-scale single-cell technique to profile chromatin accessibility 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.05.11.540245v1?rss=1</link>
<description><![CDATA[
Measuring chromatin accessibility is a powerful method to identify cell types and states. Performed at single-cell resolution, this assay has generated catalogs of genome-wide DNA regulatory sites, whole-organism cell atlases, and dynamic chromatin reorganization through development. However, the limited throughput of current single-cell approaches poses a challenge for implementing proper study designs, population-scale profiling, and/or very deep profiling of complex samples. To this end, we developed a 10X-compatible combinatorial indexing ATAC sequencing ("txci-ATAC-seq"), which is a combinatorial indexing framework that initially indexes ("pre-indexes") chromatin within nuclei with barcoded transposases followed by encapsulation and further barcoding using a commercialized droplet-based microfluidics platform (10X Genomics). Leveraging this molecular hashing strategy, we demonstrate that txci-ATAC-seq enables the indexing of up to 200,000 nuclei across multiple samples in a single emulsion reaction, representing a [~]22-fold increase in throughput compared to the standard workflow at the same collision rate. To improve the efficiency of this new technique, we further developed a faster version of the protocol ("Fast-txci-ATAC-seq") that separates sample pre-processing from library generation and has the potential to profile up to 96 samples simultaneously. We initially benchmarked our assay by generating chromatin accessibility profiles for 230,018 cells from five native tissues across three experiments, including human cortex (28,513 cells), mouse brain (48,997 cells), human lung (15,799 cells), mouse lung (73,280 cells), and mouse liver (63,429 cells). We also applied our method to a club cell secretory protein knockout (CC16-/-) mouse model to examine the biological and technical limitations of the mouse line. By characterizing DNA regulatory landscapes in 76,498 wild-type and 77,638 CC16-/- murine lung nuclei, our investigations uncovered previously unappreciated residual genetic deviations from the reference strain that resulted from the method of gene targeting, which employed embryonic stem cells from the 129 strain. We found that these genetic remnants from the 129 strain led to profound cell-type-specific changes in chromatin accessibility in regulatory elements near a host of genes. Collectively, we defined single-cell chromatin signatures in 384,154 nuclei from 13 primary samples across different species, organs, biological replicates, and genetic backgrounds, establishing txci-ATAC-seq as a robust, high-quality, and highly multiplexable single-cell assay for large-scale chromatin studies.
]]></description>
<dc:creator>Zhang, H.</dc:creator>
<dc:creator>Mulqueen, R. M.</dc:creator>
<dc:creator>Iannuzo, N.</dc:creator>
<dc:creator>Farrera, D. O.</dc:creator>
<dc:creator>Polverino, F.</dc:creator>
<dc:creator>Galligan, J. J.</dc:creator>
<dc:creator>Ledford, J. G.</dc:creator>
<dc:creator>Adey, A. C.</dc:creator>
<dc:creator>Cusanovich, D. A.</dc:creator>
<dc:date>2023-05-14</dc:date>
<dc:identifier>doi:10.1101/2023.05.11.540245</dc:identifier>
<dc:title><![CDATA[txci-ATAC-seq, a massive-scale single-cell technique to profile chromatin accessibility]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-05-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.05.11.540443v1?rss=1">
<title>
<![CDATA[
NAIP-NLRC4 Inflammasome Activation in Tuft Cells Activates a PGD2-ILC3 Signaling Circuit that Protects Against Enteric Infection. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.05.11.540443v1?rss=1</link>
<description><![CDATA[
Intestinal epithelial cells (IEC) use innate sensing pathways to distinguish pathogens from commensals. One such pathway, the NAIP--NLRC4 inflammasome, initiates extrusion of infected IEC and mediator release upon cytosolic bacterial sensing. Tuft cells are primarily known for their function in anti-parasite immunity. We previously reported that activation of the inflammasome in tuft cells leads to release of prostaglandin D2 (PGD2). We test the hypothesis that tuft cell specific release of PGD2 after inflammasome activation initiates antibacterial responses. NAIP--NLRC4 inflammasome activation in tuft cells leads to a type 3 antimicrobial response with increased IL-22 and antimicrobial protein levels within the small intestine, which is dependent on PGD2 signaling. A subset of ILC3 express the PGD2 receptor CRTH2 and we show them as the source of the increased IL-22. Inflammasome activation in tuft cells also leads to better control of Salmonella Typhimurium. These data support that intestinal tuft cells can also induce antibacterial responses.

SummaryPGD2 release after NAIP--NLRC4 inflammasome activation in tuft cells signals onto ILC3s and mediates host defense mechanisms against Salmonella Typhimurium within the small intestine. Tuft cells therefore not only promote immune reactions against parasites, but also bacteria.
]]></description>
<dc:creator>Churchill, M. J.</dc:creator>
<dc:creator>Bauer, R.</dc:creator>
<dc:creator>Honodel, R.</dc:creator>
<dc:creator>Christopher, T.</dc:creator>
<dc:creator>Smita, S.</dc:creator>
<dc:creator>Warner, L.</dc:creator>
<dc:creator>Mooney, B. M.</dc:creator>
<dc:creator>Wojno, E. D. T.</dc:creator>
<dc:creator>Rauch, I.</dc:creator>
<dc:date>2023-05-14</dc:date>
<dc:identifier>doi:10.1101/2023.05.11.540443</dc:identifier>
<dc:title><![CDATA[NAIP-NLRC4 Inflammasome Activation in Tuft Cells Activates a PGD2-ILC3 Signaling Circuit that Protects Against Enteric Infection.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-05-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.05.14.540701v1?rss=1">
<title>
<![CDATA[
3D organoid modeling identified that targeting IGF1R signaling may overcome drug resistance in breast cancer 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.05.14.540701v1?rss=1</link>
<description><![CDATA[
Breast cancer is the most frequently diagnosed cancer and the second largest cause of cancer deaths in women. However, drug resistance and poor response to treatments are common. Thus, there is an unmet need to identify new treatments and effective lab-based drug testing methods. Here we established a novel 3-dimensional organoid method by co-culturing cancer cells with healthy endothelial cells for longer-term testing of new drug combinations that combat drug resistance. As a proof-of-concept we showed that paclitaxel efficacy can be improved by combining it with AKT inhibitors. In addition, we identified a new triple combination of paclitaxel, HER2 inhibitor, and IGF1R inhibitor, which is more effective in increasing cell death and reducing organoid growth. Interestingly, many IGF1R pathway members are upregulated in breast cancer patients, and high expression is associated with poor survival, indicating that IGF1R is an attractive therapeutic target. Overall, using this novel organoid method, we can mimic more accurate culture conditions and identify new targets to be tested in clinical trials. Our approach is applicable to various cancers to improve patients outcomes.
]]></description>
<dc:creator>Mittal, E.</dc:creator>
<dc:creator>Qian, D.</dc:creator>
<dc:date>2023-05-14</dc:date>
<dc:identifier>doi:10.1101/2023.05.14.540701</dc:identifier>
<dc:title><![CDATA[3D organoid modeling identified that targeting IGF1R signaling may overcome drug resistance in breast cancer]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-05-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/004242v1?rss=1">
<title>
<![CDATA[
Exact Reconstruction of Gene Regulatory Networks using Compressive Sensing 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/004242v1?rss=1</link>
<description><![CDATA[
BackgroundWe consider the problem of reconstructing a gene regulatory network structure from limited time series gene expression data, without any a priori knowledge of connectivity. We assume that the network is sparse, meaning the connectivity among genes is much less than full connectivity. We develop a method for network reconstruction based on compressive sensing, which takes advantage of the networks sparseness.nnResultsFor the case in which all genes are accessible for measurement, and there is no measurement noise, we show that our method can be used to exactly reconstruct the network. For the more general problem, in which hidden genes exist and all measurements are contaminated by noise, we show that our method leads to reliable reconstruction. In both cases, coherence of the model is used to assess the ability to reconstruct the network and to design new experiments. For each problem, a set of numerical examples is presented.nnConclusionsThe method provides a guarantee on how well the inferred graph structure represents the underlying system, reveals deficiencies in the data and model, and suggests experimental directions to remedy the deficiencies.
]]></description>
<dc:creator>Young Hwan Chang</dc:creator>
<dc:creator>Joe W. Gray</dc:creator>
<dc:creator>Claire J. Tomlin</dc:creator>
<dc:creator></dc:creator>
<dc:date>2014-04-20</dc:date>
<dc:identifier>doi:10.1101/004242</dc:identifier>
<dc:title><![CDATA[Exact Reconstruction of Gene Regulatory Networks using Compressive Sensing]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2014-04-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/004259v1?rss=1">
<title>
<![CDATA[
Disentangling Multidimensional Spatio-Temporal Data into their Common and Aberrant Responses 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/004259v1?rss=1</link>
<description><![CDATA[
With the advent of high-throughput measurement techniques, scientists and engineers are starting to grapple with massive data sets and encountering challenges with how to organize, process and extract information into meaningful structures. Multidimensional spatio-temporal biological data sets such as time series gene expression with various perturbations over different cell lines, or neural spike trains across many experimental trials, have the potential to acquire insight across multiple dimensions. For this potential to be realized, we need a suitable representation to understand the data. Since a wide range of experiments and the unknown complexity of the underlying system contribute to the heterogeneity of biological data, we propose a method based on Robust Principal Component Analysis (RPCA), which is well suited for extracting principal components when there are corrupted observations. The proposed method provides us a new representation of these data sets in terms of a common and aberrant response. This representation might help users to acquire a new insight from data.nnAuthor SummaryOne of the most exciting trends and important themes in science and engineering involves the use of high-throughput measurement data. With different dimensions, for example, various perturbations, different doses of drug or cell lines characteristics, such multidimensional data sets enable us to understand commonalities and differences across multiple dimensions. A general question is how to organize the observed data into meaningful structures and how to find an appropriate similarity measure. A natural way of viewing these complex high dimensional data sets is to examine and analyze the large-scale features and then to focus on the interesting details. With this notion, we propose an RPCA-based method which models common variations as approximately the low-rank component and anomalies as the sparse component. We show that the proposed method is able to find distinct subtypes and classify data sets in a robust way without any prior knowledge by separating these common responses and abnormal responses.
]]></description>
<dc:creator>Young Hwan Chang</dc:creator>
<dc:creator>Jim Korkola</dc:creator>
<dc:creator>Dhara N. Amin</dc:creator>
<dc:creator>Mark M. Moasser</dc:creator>
<dc:creator>Jose M. Carmena</dc:creator>
<dc:creator>Joe W. Gray</dc:creator>
<dc:creator>Claire J. Tomlin</dc:creator>
<dc:creator></dc:creator>
<dc:date>2014-04-23</dc:date>
<dc:identifier>doi:10.1101/004259</dc:identifier>
<dc:title><![CDATA[Disentangling Multidimensional Spatio-Temporal Data into their Common and Aberrant Responses]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2014-04-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/009498v1?rss=1">
<title>
<![CDATA[
An Approximate Bayesian Computation Approach to Examining the Phylogenetic Relationships among the Four Gibbon Genera using Whole Genome Sequence Data 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/009498v1?rss=1</link>
<description><![CDATA[
Gibbons are believed to have diverged from the larger great apes [~]16.8 Mya and today reside in the rainforests of Southeast Asia. Based on their diploid chromosome number, the family Hylobatidae is divided into four genera, Nomascus, Symphalangus, Hoolock and Hylobates. Genetic studies attempting to elucidate the phylogenetic relationships among gibbons using karyotypes, mtDNA, the Y chromosome, and short autosomal sequences have been inconclusive. To examine the relationships among gibbon genera in more depth, we performed 2nd generation whole genome sequencing to a mean of [~]15X coverage in two individuals from each genus. We developed a coalescent-based Approximate Bayesian Computation method incorporating a model of sequencing error generated by high coverage exome validation to infer the branching order, divergence times, and effective population sizes of gibbon taxa. Although Hoolock and Symphalangus are likely sister taxa, we could not confidently resolve a single bifurcating tree despite the large amount of data analyzed. Our combined results support the hypothesis that all four gibbon genera diverged at approximately the same time. Assuming an autosomal mutation rate of 1x10-9/site/year this speciation process occurred [~]5 Mya during a period in the Early Pliocene characterized by climatic shifts and fragmentation of the Sunda shelf forests. Whole genome sequencing of additional individuals will be vital for inferring the extent of gene flow among species after the separation of the gibbon genera.
]]></description>
<dc:creator>Krishna Veeramah</dc:creator>
<dc:creator>August E Woerner</dc:creator>
<dc:creator>Laurel Johnstone</dc:creator>
<dc:creator>Ivo Gut</dc:creator>
<dc:creator>Marta Gut</dc:creator>
<dc:creator>Tomas Marques-Bonet</dc:creator>
<dc:creator>Lucia Carbone</dc:creator>
<dc:creator>Jeff D Wall</dc:creator>
<dc:creator>Michael F Hammer</dc:creator>
<dc:creator></dc:creator>
<dc:date>2014-09-22</dc:date>
<dc:identifier>doi:10.1101/009498</dc:identifier>
<dc:title><![CDATA[An Approximate Bayesian Computation Approach to Examining the Phylogenetic Relationships among the Four Gibbon Genera using Whole Genome Sequence Data]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2014-09-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/012997v1?rss=1">
<title>
<![CDATA[
A Comprehensive Assessment of Somatic Mutation Calling in Cancer Genomes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/012997v1?rss=1</link>
<description><![CDATA[
The emergence of next generation DNA sequencing technology is enabling high-resolution cancer genome analysis. Large-scale projects like the International Cancer Genome Consortium (ICGC) are systematically scanning cancer genomes to identify recurrent somatic mutations. Second generation DNA sequencing, however, is still an evolving technology and procedures, both experimental and analytical, are constantly changing. Thus the research community is still defining a set of best practices for cancer genome data analysis, with no single protocol emerging to fulfil this role. Here we describe an extensive benchmark exercise to identify and resolve issues of somatic mutation calling. Whole genome sequence datasets comprising tumor-normal pairs from two different types of cancer, chronic lymphocytic leukaemia and medulloblastoma, were shared within the ICGC and submissions of somatic mutation calls were compared to verified mutations and to each other. Varying strategies to call mutations, incomplete awareness of sources of artefacts, and even lack of agreement on what constitutes an artefact or real mutation manifested in widely varying mutation call rates and somewhat low concordance among submissions. We conclude that somatic mutation calling remains an unsolved problem. However, we have identified many issues that are easy to remedy that are presented here. Our study highlights critical issues that need to be addressed before this valuable technology can be routinely used to inform clinical decision-making.nnAbbreviations and Definitionsaligner = mapper, these terms are used interchangeably
]]></description>
<dc:creator>Tyler S Alioto</dc:creator>
<dc:creator>Sophia Derdak</dc:creator>
<dc:creator>Timothy A Beck</dc:creator>
<dc:creator>Paul C Boutros</dc:creator>
<dc:creator>Lawrence Bower</dc:creator>
<dc:creator>Ivo Buchhalter</dc:creator>
<dc:creator>Matthew D Eldridge</dc:creator>
<dc:creator>Nicholas J Harding</dc:creator>
<dc:creator>Lawrence Edward Heisler</dc:creator>
<dc:creator>Eivind Hovig</dc:creator>
<dc:creator>David T W Jones</dc:creator>
<dc:creator>Andrew G Lynch</dc:creator>
<dc:creator>Sigve Nakken</dc:creator>
<dc:creator>Paolo Ribeca</dc:creator>
<dc:creator>Anne-Sophie Sertier</dc:creator>
<dc:creator>Jared T Simpson</dc:creator>
<dc:creator>Paul Spellman</dc:creator>
<dc:creator>Patrick Tarpey</dc:creator>
<dc:creator>Laurie Tonon</dc:creator>
<dc:creator>Daniel Vodák</dc:creator>
<dc:creator>Takafumi N Yamaguchi</dc:creator>
<dc:creator>Sergi Beltran Agullo</dc:creator>
<dc:creator>Marc Dabad</dc:creator>
<dc:creator>Robert E Denroche</dc:creator>
<dc:creator>Philip Ginsbach</dc:creator>
<dc:creator>Simon C Heath</dc:creator>
<dc:creator>Emanuele Raineri</dc:creator>
<dc:creator>Charlotte L Anderson</dc:creator>
<dc:creator>Benedikt Brors</dc:creator>
<dc:creator>Ruben Drews</dc:creator>
<dc:creator>Roland Eils</dc:creator>
<dc:creator>Akihiro Fujimoto</dc:creator>
<dc:creator>Francesc Castro Giner</dc:creator>
<dc:creator>Minghui He</dc:creator>
<dc:creator>Pablo Hennings-Yeomans</dc:creator>
<dc:creator>Barbara Hutter</dc:creator>
<dc:creator>Natalie Jäger</dc:creator>
<dc:creator>Rolf Kab</dc:creator>
<dc:date>2014-12-24</dc:date>
<dc:identifier>doi:10.1101/012997</dc:identifier>
<dc:title><![CDATA[A Comprehensive Assessment of Somatic Mutation Calling in Cancer Genomes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2014-12-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/039636v1?rss=1">
<title>
<![CDATA[
Context-specificity in causal signaling networks revealed by phosphoprotein profiling 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/039636v1?rss=1</link>
<description><![CDATA[
SummarySignaling networks downstream of receptor tyrosine kinases are among the most extensively studied biological networks. However, it remains unclear whether signaling networks depend on biological context. Signaling networks encode causal influences - and not just correlations - between network components. Here, using a causal framework and systematic time-course assays of signaling proteins, we investigate the context-specificity of signaling networks in a cell line system. We focus on a well-defined set of signaling proteins profiled in four breast cancer cell lines under eight stimulus conditions and inhibition of specific kinases. The data, spanning multiple pathways and comprising approximately 70,000 phosphoprotein and 260,000 protein measurements, provide a wealth of testable, context-specific hypotheses, several of which we validate in independent experiments. Furthermore, the data provide a resource for computational methods development, permitting empirical assessment of causal network learning in a complex, mammalian setting.
]]></description>
<dc:creator>Steven M. Hill</dc:creator>
<dc:creator>Nicole K. Nesser</dc:creator>
<dc:creator>Katie Johnson-Camacho</dc:creator>
<dc:creator>Mara Jeffress</dc:creator>
<dc:creator>Aimee Johnson</dc:creator>
<dc:creator>Chris Boniface</dc:creator>
<dc:creator>Simon E.F. Spencer</dc:creator>
<dc:creator>Yiling Lu</dc:creator>
<dc:creator>Laura M. Heiser</dc:creator>
<dc:creator>Yancey Lawrence</dc:creator>
<dc:creator>Nupur T. Pande</dc:creator>
<dc:creator>James E. Korkola</dc:creator>
<dc:creator>Joe W. Gray</dc:creator>
<dc:creator>Gordon B. Mills</dc:creator>
<dc:creator>Sach Mukherjee</dc:creator>
<dc:creator>Paul T. Spellman</dc:creator>
<dc:creator></dc:creator>
<dc:date>2016-02-15</dc:date>
<dc:identifier>doi:10.1101/039636</dc:identifier>
<dc:title><![CDATA[Context-specificity in causal signaling networks revealed by phosphoprotein profiling]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2016-02-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/043240v1?rss=1">
<title>
<![CDATA[
Whole-genome characterization in pedigreed non-human primates using Genotyping-By-Sequencing and imputation. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/043240v1?rss=1</link>
<description><![CDATA[
BackgroundRhesus macaques are widely used in biomedical research, but the application of genomic information in this species to better understand human disease is still undeveloped. Whole-genome sequence (WGS) data in pedigreed macaque colonies could provide substantial experimental power, but the collection of WGS data in large cohorts remains a formidable expense. Here, we describe a cost-effective approach that selects the most informative macaques in a pedigree for whole-genome sequencing, and imputes these dense marker data into all remaining individuals having sparse marker data, obtained using Genotyping-By-Sequencing (GBS).nnResultsWe developed GBS for the macaque genome using a single digest with PstI, followed by sequencing to 30X coverage. From GBS sequence data collected on all individuals in a 16-member pedigree, we characterized an optimal 22,455 sparse markers spaced ~125 kb apart. To characterize dense markers for imputation, we performed WGS at 30X coverage on 9 of the 16 individuals, yielding ~10.2 million high-confidence variants. Using the approach of "Genotype Imputation Given Inheritance" (GIGI), we imputed alleles at an optimized dense set of 4,920 variants on chromosome 19, using 490 sparse markers from GBS. We assessed changes in accuracy of imputed alleles, 1) across 3 different strategies for selecting individuals for WGS, i.e., a) using "GIGI-Pick" to select informative individuals, b) sequencing the most recent generation, or c) sequencing founders only; and 2) when using from 1-9 WGS individuals for imputation. We found that accuracy of imputed alleles was highest using the GIGI-Pick selection strategy (median 92%), and improved very little when using >4 individuals with WGS for imputation. We used this ratio of 4 WGS to 12 GBS individuals to impute an expanded set of ~14.4 million variants across all 20 macaque autosomes, achieving ~85-88% accuracy per chromosome.nnConclusionsWe conclude that an optimal tradeoff exists at the ratio of 1 individual selected for WGS using the GIGI-Pick algorithm, per 3-5 relatives selected for GBS, a cost savings of ~67-83% over WGS of all individuals. This approach makes feasible the collection of accurate, dense genome-wide sequence data in large pedigreed macaque cohorts without the need for expensive WGS data on all individuals.
]]></description>
<dc:creator>Ben N Bimber</dc:creator>
<dc:creator>Michael J Raboin</dc:creator>
<dc:creator>John Letaw</dc:creator>
<dc:creator>Kimberly Nevonen</dc:creator>
<dc:creator>Jennifer E Spindel</dc:creator>
<dc:creator>Susan McCouch</dc:creator>
<dc:creator>Rita Cervera-Juanes</dc:creator>
<dc:creator>Eliot Spindel</dc:creator>
<dc:creator>Lucia Carbone</dc:creator>
<dc:creator>Betsy Ferguson</dc:creator>
<dc:creator>Amanda Vinson</dc:creator>
<dc:creator></dc:creator>
<dc:date>2016-03-12</dc:date>
<dc:identifier>doi:10.1101/043240</dc:identifier>
<dc:title><![CDATA[Whole-genome characterization in pedigreed non-human primates using Genotyping-By-Sequencing and imputation.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2016-03-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/048843v1?rss=1">
<title>
<![CDATA[
k-BOOM: A Bayesian approach to ontology structure inference, with applications in disease ontology construction 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/048843v1?rss=1</link>
<description><![CDATA[
One strategy for building ontologies covering domains such as disease or anatomy is to weave together existing knowledge sources (databases, vocabularies and ontologies) into single cohesive whole. A first step in this process is to generate mappings between the elements of these different sources. There are a number of well-known techniques for generating mappings (also known as ontology alignemnt), both manual and automatic[7]. Sometimes mappings are seen as an end in themselves, with the sources remaining in a loosely connected state. However, if we want to take the next step and use the mappings to integrate the different sources into a cohesive reference ontology, then we need to translate the mappings into precise logical relationships. This will allow us to safely merge equivalent concepts, creating a unified ontology. This translation is a non-trivial step, as each mapping can be interpreted as multiple different logical relationships, with each interpretation affecting the likelihood of the others. There is a lack of automated methods to assist with this last step; this resolution is typically performed by expert ontologists.

Here we describe an ontology construction technique that takes two or more ontologies linked by hypothetical axioms, and estimates the most likely unified logical ontology. Hypothetical axioms can themselves be derived from semantically loose mappings. The method combines deductive reasoning and probabilistic inference and is called Bayesian OWL Ontology Merging (BOOM). We describe a special form k-BOOM that works by factorizing the probabilistic ontology into k submodules. We also briefly describe a supplemental lexical and knowledge-based technique for generating a set of hypothetical axioms from loose mappings.

We are currently using this technique to build a merged disease ontology (Monarch Disease Ontology; MonDO) that unifies a broad range of vocabularies into a consistent and coherent whole.
]]></description>
<dc:creator>Christopher J Mungall</dc:creator>
<dc:creator>Sebastian Koehler</dc:creator>
<dc:creator>Peter Robinson</dc:creator>
<dc:creator>Ian Holmes</dc:creator>
<dc:creator>Melissa Haendel</dc:creator>
<dc:creator></dc:creator>
<dc:date>2016-04-15</dc:date>
<dc:identifier>doi:10.1101/048843</dc:identifier>
<dc:title><![CDATA[k-BOOM: A Bayesian approach to ontology structure inference, with applications in disease ontology construction]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2016-04-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/060046v1?rss=1">
<title>
<![CDATA[
Architecture of fully occupied GluA2 AMPA receptor - TARP complex elucidated by single particle cryo-electron microscopy 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/060046v1?rss=1</link>
<description><![CDATA[
Fast excitatory neurotransmission in the mammalian central nervous system is largely carried out by AMPA-sensitive ionotropic glutamate receptors. Localized within the postsynaptic density of glutamatergic spines, AMPA receptors are composed of heterotetrameric receptor assemblies associated with auxiliary subunits, the most common of which are transmembrane AMPA-receptor regulatory proteins (TARPs). The association of TARPs with AMPA receptors modulates the kinetics of receptor gating and pharmacology, as well as trafficking. Here we report the cryo-EM structure of the homomeric GluA2 AMPA receptor saturated with TARP {gamma}2 subunits, showing how the TARPs are arranged with four-fold symmetry around the ion channel domain, making extensive interactions with the M1, M2 and M4 TM helices. Poised like partially opened  hands underneath the two-fold symmetric ligand binding domain (LBD)  clamshells, one pair of TARPs are juxtaposed near the LBD dimer interface, while the other pair are near the LBD dimer-dimer interface. The extracellular  domains of TARP are positioned to not only modulate LBD  clamshell closure, but also to affect conformational rearrangements of the LBD layer associated with receptor activation and desensitization, while the TARP transmembrane (TM) domains buttress the ion channel pore.
]]></description>
<dc:creator>Yan Zhao</dc:creator>
<dc:creator>Shanshuang Chen</dc:creator>
<dc:creator>Craig Yoshioka</dc:creator>
<dc:creator>Isabelle Baconguis</dc:creator>
<dc:creator>Eric Gouaux</dc:creator>
<dc:creator></dc:creator>
<dc:date>2016-06-21</dc:date>
<dc:identifier>doi:10.1101/060046</dc:identifier>
<dc:title><![CDATA[Architecture of fully occupied GluA2 AMPA receptor - TARP complex elucidated by single particle cryo-electron microscopy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2016-06-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/065482v1?rss=1">
<title>
<![CDATA[
Construction of thousands of single cell genome sequencing libraries using combinatorial indexing 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/065482v1?rss=1</link>
<description><![CDATA[
Single cell genome sequencing has proven to be a valuable tool for the detection of somatic variation, particularly in the context of tumor evolution and neuronal heterogeneity. Current technologies suffer from high per-cell library construction costs which restrict the number of cells that can be assessed, thus imposing limitations on the ability to quantitatively measure genomic heterogeneity within a tissue. Here, we present Single cell Combinatorial Indexed Sequencing (SCI-seq) as a means of simultaneously generating thousands of low-pass single cell libraries for the purpose of somatic copy number variant detection. In total, we constructed libraries for 16,698 single cells from a combination of cultured cell lines, frontal cortex tissue from Macaca mulatta, and two human adenocarcinomas. This novel technology provides the opportunity for low-cost, deep characterization of somatic copy number variation in single cells, providing a foundational knowledge across both healthy and diseased tissues.
]]></description>
<dc:creator>Sarah A Vitak</dc:creator>
<dc:creator>Kristof A Torkenczy</dc:creator>
<dc:creator>Jimi L Rosenkrantz</dc:creator>
<dc:creator>Andrew J Fields</dc:creator>
<dc:creator>Lena Christiansen</dc:creator>
<dc:creator>Melissa H Wong</dc:creator>
<dc:creator>Lucia Carbone</dc:creator>
<dc:creator>Frank J Steemers</dc:creator>
<dc:creator>Andrew Adey</dc:creator>
<dc:creator></dc:creator>
<dc:date>2016-07-23</dc:date>
<dc:identifier>doi:10.1101/065482</dc:identifier>
<dc:title><![CDATA[Construction of thousands of single cell genome sequencing libraries using combinatorial indexing]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2016-07-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/071217v1?rss=1">
<title>
<![CDATA[
Network Identification Methods 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/071217v1?rss=1</link>
<description><![CDATA[
Recently, network inference algorithms have grown tremendously in the field of systems biology because network identification is essential for understanding relationships between regulation mechanisms for genes, elucidating functional mechanisms underlying cellular processes, as well as identifying molecular targets for discoveries in medicines. This article provides a brief overview of different approaches used to identify biological networks and reviews recent advances in network identification.
]]></description>
<dc:creator>Young Hwan Chang</dc:creator>
<dc:creator>Claire Tomlin</dc:creator>
<dc:creator></dc:creator>
<dc:date>2016-08-23</dc:date>
<dc:identifier>doi:10.1101/071217</dc:identifier>
<dc:title><![CDATA[Network Identification Methods]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2016-08-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/079350v1?rss=1">
<title>
<![CDATA[
Reproducibility and replicability of rodent phenotyping in preclinical studies 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/079350v1?rss=1</link>
<description><![CDATA[
The scientific community is increasingly concerned with cases of published "discoveries" that are not replicated in further studies. The field of mouse behavioral phenotyping was one of the first to raise this concern, and to relate it to other complicated methodological issues: the complex interaction between genotype and environment; the definitions of behavioral constructs; and the use of the mouse as a model animal for human health and disease mechanisms. In January 2015, researchers from various disciplines including genetics, behavior genetics, neuroscience, ethology, statistics and bioinformatics gathered in Tel Aviv University to discuss these issues. The general consent presented here was that the issue is prevalent and of concern, and should be addressed at the statistical, methodological and policy levels, but is not so severe as to call into question the validity and the usefulness of model organisms as a whole. Well-organized community efforts, coupled with improved data and metadata sharing, were agreed by all to have a key role to play in identifying specific problems and promoting effective solutions. As replicability is related to validity and may also affect generalizability and translation of findings, the implications of the present discussion reach far beyond the issue of replicability of mouse phenotypes but may be highly relevant throughout biomedical research.
]]></description>
<dc:creator>Neri Kafkafi</dc:creator>
<dc:creator>Joseph Agassi</dc:creator>
<dc:creator>Elissa J Chesler</dc:creator>
<dc:creator>John C Crabbe</dc:creator>
<dc:creator>Wim E Crusio</dc:creator>
<dc:creator>David Eilam</dc:creator>
<dc:creator>Robert Gerlai</dc:creator>
<dc:creator>Ilan Golani</dc:creator>
<dc:creator>Alex Gomez-Marin</dc:creator>
<dc:creator>Ruth Heller</dc:creator>
<dc:creator>Fuad Iraqi</dc:creator>
<dc:creator>Iman Jaljuli</dc:creator>
<dc:creator>Natasha A Karp</dc:creator>
<dc:creator>Hugh Morgan</dc:creator>
<dc:creator>George Nicholson</dc:creator>
<dc:creator>Donald W Pfaff</dc:creator>
<dc:creator>Helene S Richter</dc:creator>
<dc:creator>Philip B Stark</dc:creator>
<dc:creator>Oliver Stiedl</dc:creator>
<dc:creator>Victoria Stodden</dc:creator>
<dc:creator>Lisa M Tarantino</dc:creator>
<dc:creator>Valter Tucci</dc:creator>
<dc:creator>William Valdar</dc:creator>
<dc:creator>Robert W Williams</dc:creator>
<dc:creator>Hanno Wurbel</dc:creator>
<dc:creator>Yoav Benjamini</dc:creator>
<dc:creator></dc:creator>
<dc:date>2016-10-05</dc:date>
<dc:identifier>doi:10.1101/079350</dc:identifier>
<dc:title><![CDATA[Reproducibility and replicability of rodent phenotyping in preclinical studies]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2016-10-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/093583v1?rss=1">
<title>
<![CDATA[
Analysis of copy number variants on chromosome 21 in Down syndrome-associated congenital heart defects 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/093583v1?rss=1</link>
<description><![CDATA[
One in five people with Down syndrome (DS) are born with an atrioventricular septal defect (AVSD), an incidence 2,000 times higher than in the euploid population. The genetic loci that contribute to this risk are poorly understood. In this study, we tested two hypotheses: 1) individuals with DS carrying chromosome 21 copy number variants (CNVs) that interrupt exons may be protected from AVSD, because these CNVs return AVSD susceptibility loci back to disomy, and 2) individuals with DS carrying chromosome 21 genes spanned by microduplications are at greater risk for AVSD because these microduplications boost the dosage of AVSD susceptibility loci beyond a tolerable threshold. We tested 198 case individuals with DS+AVSD and 211 control individuals with DS and a normal heart using a custom microarray with dense probes tiled on chromosome 21 for array CGH. We found that neither an individual chromosome 21 CNV nor any individual gene intersected by a CNV was associated with AVSD in DS. Burden analyses revealed that African American controls had more bases covered by rare deletions than did African American cases. Inversely, we found that Caucasian cases had more genes intersected by rare duplications than did Caucasian controls. Pathway analyses indicated copy number perturbations of genes involved in protein heterotrimerization and histone methylating proteins. Finally, we showed that previously DS+AVSD-associated common CNVs on chromosome 21 are likely false positives. This research adds to the swell of evidence indicating that DS-associated AVSD is similarly heterogeneous, as is AVSD in the euploid population.
]]></description>
<dc:creator>Rambo-Martin, B. L.</dc:creator>
<dc:creator>Mulle, J. G.</dc:creator>
<dc:creator>Cutler, D. J.</dc:creator>
<dc:creator>Bean, L. J. H.</dc:creator>
<dc:creator>Rosser, T. C.</dc:creator>
<dc:creator>Dooley, K. J.</dc:creator>
<dc:creator>Cua, C.</dc:creator>
<dc:creator>Capone, G.</dc:creator>
<dc:creator>Maslen, C. L.</dc:creator>
<dc:creator>Reeves, R. H.</dc:creator>
<dc:creator>Sherman, S. L.</dc:creator>
<dc:creator>Zwick, M. E.</dc:creator>
<dc:date>2016-12-28</dc:date>
<dc:identifier>doi:10.1101/093583</dc:identifier>
<dc:title><![CDATA[Analysis of copy number variants on chromosome 21 in Down syndrome-associated congenital heart defects]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2016-12-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/094649v1?rss=1">
<title>
<![CDATA[
Cryo-EM structure of the ATP-sensitive potassium channel illuminates mechanisms of assembly and gating 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/094649v1?rss=1</link>
<description><![CDATA[
ATP-sensitive potassium (KATP) channels are metabolic sensors that couple cell energetics to membrane excitability. In pancreatic {beta}-cells, channels formed by SUR1 and Kir6.2 regulate insulin secretion and are the targets of antidiabetic sulfonylureas. Here, we used cryo-EM to elucidate structural basis of channel assembly and gating. The structure, determined in the presence of ATP and the sulfonylurea glibenclamide, at ~6[A] resolution reveals a closed Kir6.2 tetrameric core with four peripheral SUR1s each anchored to a Kir6.2 by its N-terminal transmembrane domain (TMD0). Intricate interactions between TMD0, the loop following TMD0, and Kir6.2 near the proposed PIP2 binding site, and where ATP density is observed, suggest SUR1 may contribute to ATP and PIP2 binding to enhance Kir6.2 sensitivity to both. The SUR1-ABC core is found in an unusual inward-facing conformation whereby the two nucleotide binding domains are misaligned along a two-fold symmetry axis, revealing a possible mechanism by which glibenclamide inhibits channel activity.
]]></description>
<dc:creator>Martin, G. M.</dc:creator>
<dc:creator>Yoshioka, C.</dc:creator>
<dc:creator>Rex, E. A.</dc:creator>
<dc:creator>Fay, J. F.</dc:creator>
<dc:creator>Xie, Q.</dc:creator>
<dc:creator>Whorton, M. R.</dc:creator>
<dc:creator>Chen, J. Z.</dc:creator>
<dc:creator>Shyng, S.-L.</dc:creator>
<dc:date>2016-12-16</dc:date>
<dc:identifier>doi:10.1101/094649</dc:identifier>
<dc:title><![CDATA[Cryo-EM structure of the ATP-sensitive potassium channel illuminates mechanisms of assembly and gating]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2016-12-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/102905v1?rss=1">
<title>
<![CDATA[
Interactive analysis of Long-read RNA isoforms with Iso-Seq Browser 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/102905v1?rss=1</link>
<description><![CDATA[
BackgroundLong-read RNA sequencing, such as Pacific Biosciences Iso-Seq method, enables generation of sequencing reads that are 10 kilobases or even longer. These reads are ideal for discovering splice junctions and resolving full-length gene transcripts without time-consuming and error-prone techniques such as transcript assembly and junction inference.nnResultsIso-Seq Browser is a Web-based visual analytics tool for long-read RNA sequencing data produced by Pacific Biosciences isoform sequencing (Iso-Seq) techniques. Key features of the Iso-Seq Browser are: 1) an exon-only web-based interface with zooming and exon highlighting for exploring reference gene transcripts and novel gene isoforms, 2) automated grouping of transcripts and isoforms by similarity, 3) many customization features for data exploration and creating publication ready figures, and 4) exporting selected isoforms into fasta files for further analysis. Iso-Seq Browser is written in Python using several scientific libraries. The application and analyses described in this paper are freely available to both academic and commercial users at https://github.com/goeckslab/isoseq-browsernnConclusionsIso-Seq Browser enables interactive genome-wide visual analysis of long RNA sequence reads. Through visualization, highlighting, clustering, and filtering of gene isoforms, ISB makes it simple to identify novel isoforms and novel isoform features such as exons, introns and untranslated regions.
]]></description>
<dc:creator>Hu, J.</dc:creator>
<dc:creator>Uapinyoying, P.</dc:creator>
<dc:creator>Goecks, J.</dc:creator>
<dc:date>2017-01-25</dc:date>
<dc:identifier>doi:10.1101/102905</dc:identifier>
<dc:title><![CDATA[Interactive analysis of Long-read RNA isoforms with Iso-Seq Browser]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-01-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/109272v1?rss=1">
<title>
<![CDATA[
Poly(A)-ClickSeq: click-chemistry for next-generation 3´-end sequencing without RNA enrichment or fragmentation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/109272v1?rss=1</link>
<description><![CDATA[
The recent emergence of alternative polyadenylation (APA) as an engine driving transcriptomic diversity has stimulated the development of sequencing methodologies designed to assess genome-wide polyadenylation events. The goal of these approaches is to enrich, partition, capture, and ultimately sequence poly(A) site junctions. However, these methods often require poly(A) enrichment, 3{acute} linker ligation steps, and RNA fragmentation, which can necessitate higher levels of starting RNA, increase experimental error, and potentially introduce bias. We recently reported a click-chemistry based method for generating RNAseq libraries called "ClickSeq". Here, we adapt this method to direct the cDNA synthesis specifically toward the 3{acute} UTR/poly(A) tail junction of cellular RNA. With this novel approach, we demonstrate sensitive and specific enrichment for poly(A) site junctions without the need for complex sample preparation, fragmentation or purification. Poly(A)-ClickSeq (PAC-seq) is therefore a simple procedure that generates high-quality RNA-seq poly(A) libraries. As a proof-of-principle, we utilized PAC-seq to explore the poly(A) landscape of both human and Drosophila cells in culture and observed outstanding overlap with existing poly(A) databases and also identified previously unannotated poly(A) sites. Moreover, we utilize PAC-seq to quantify and analyze APA events regulated by CFIm25 illustrating how this technology can be harnessed to identify alternatively polyadenylated RNA.
]]></description>
<dc:creator>Routh, A.</dc:creator>
<dc:creator>Ji, P.</dc:creator>
<dc:creator>Jaworski, E.</dc:creator>
<dc:creator>Xia, Z.</dc:creator>
<dc:creator>Li, W.</dc:creator>
<dc:creator>Wagner, E. J.</dc:creator>
<dc:date>2017-02-24</dc:date>
<dc:identifier>doi:10.1101/109272</dc:identifier>
<dc:title><![CDATA[Poly(A)-ClickSeq: click-chemistry for next-generation 3´-end sequencing without RNA enrichment or fragmentation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-02-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/117812v1?rss=1">
<title>
<![CDATA[
Identifiers for the 21st century:How to design, provision, and reuse persistent identifiers to maximize utility and impact of life science data 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/117812v1?rss=1</link>
<description><![CDATA[
In many disciplines, data is highly decentralized across thousands of online databases (repositories, registries, and knowledgebases). Wringing value from such databases depends on the discipline of data science and on the humble bricks and mortar that make integration possible; identifiers are a core component of this integration infrastructure. Drawing on our experience and on work by other groups, we outline ten lessons we have learned about the identifier qualities and best practices that facilitate large-scale data integration. Specifically, we propose actions that identifier practitioners (database providers) should take in the design, provision and reuse of identifiers; we also outline important considerations for those referencing identifiers in various circumstances, including by authors and data generators. While the importance and relevance of each lesson will vary by context, there is a need for increased awareness about how to avoid and manage common identifier problems, especially those related to persistence and web-accessibility/resolvability. We focus strongly on web-based identifiers in the life sciences; however, the principles are broadly relevant to other disciplines.
]]></description>
<dc:creator>McMurry, J.</dc:creator>
<dc:creator>Juty, N.</dc:creator>
<dc:creator>Blomberg, N.</dc:creator>
<dc:creator>Burdett, A.</dc:creator>
<dc:creator>Conlin, T.</dc:creator>
<dc:creator>Conte, N.</dc:creator>
<dc:creator>Courtot, M.</dc:creator>
<dc:creator>Deck, J.</dc:creator>
<dc:creator>Dumontier, M.</dc:creator>
<dc:creator>Fellows, D.</dc:creator>
<dc:creator>Gonzalez-Beltran, A.</dc:creator>
<dc:creator>Gormanns, P.</dc:creator>
<dc:creator>Grethe, J.</dc:creator>
<dc:creator>Hastings, J.</dc:creator>
<dc:creator>Hermjakob, H.</dc:creator>
<dc:creator>Heriche, J.-K.</dc:creator>
<dc:creator>Ison, J.</dc:creator>
<dc:creator>Jimenez, R.</dc:creator>
<dc:creator>Jupp, S.</dc:creator>
<dc:creator>Kunze, J.</dc:creator>
<dc:creator>Laibe, C.</dc:creator>
<dc:creator>Le Novere, N.</dc:creator>
<dc:creator>Malone, J. R.</dc:creator>
<dc:creator>Martin, M.-J.</dc:creator>
<dc:creator>McEntyre, J.</dc:creator>
<dc:creator>Morris, C.</dc:creator>
<dc:creator>Muilu, J.</dc:creator>
<dc:creator>Mueller, W.</dc:creator>
<dc:creator>Rocca-Serra, P.</dc:creator>
<dc:creator>Sansone, S.-A.</dc:creator>
<dc:creator>Sariyar, M.</dc:creator>
<dc:creator>Snoep, J.</dc:creator>
<dc:creator>Stanford, N. J.</dc:creator>
<dc:creator>Soiland-Reyes, S.</dc:creator>
<dc:creator>Swainston, N.</dc:creator>
<dc:creator>Washington, N.</dc:creator>
<dc:creator>Williams, A.</dc:creator>
<dc:creator>Wimalaratne, S.</dc:creator>
<dc:creator>Winfree, L.</dc:creator>
<dc:creator>Wolstencroft, K.</dc:creator>
<dc:creator>Goble, C.</dc:creator>
<dc:creator>Mungall, C.</dc:creator>
<dc:creator>Haendel, M.</dc:creator>
<dc:creator>Parkinson,</dc:creator>
<dc:date>2017-03-20</dc:date>
<dc:identifier>doi:10.1101/117812</dc:identifier>
<dc:title><![CDATA[Identifiers for the 21st century:How to design, provision, and reuse persistent identifiers to maximize utility and impact of life science data]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-03-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/121756v1?rss=1">
<title>
<![CDATA[
Dystroglycan Maintains Inner Limiting Membrane Integrity To Coordinate Retinal Development 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/121756v1?rss=1</link>
<description><![CDATA[
Proper neural circuit formation requires the precise regulation of neuronal migration, axon guidance and dendritic arborization. Mutations affecting the function of the transmembrane glycoprotein dystroglycan cause a form of congenital muscular dystrophy that is frequently associated with neurodevelopmental abnormalities. Despite its importance in brain development, the role for dystroglycan in regulating retinal development remains poorly understood. Using a mouse model of dystroglycanopathy (ISPDL79*) and conditional dystroglycan mutants of both sexes, we show that dystroglycan is critical for the proper migration, axon guidance and dendritic stratification of neurons in the inner retina. Using genetic approaches, we show that dystroglycan functions in neuroepithelial cells as an extracellular scaffold to maintain the integrity of the retinal inner limiting membrane (ILM). Surprisingly, despite the profound disruptions in inner retinal circuit formation, spontaneous retinal activity is preserved. These results highlight the importance of dystroglycan in coordinating multiple aspects of retinal development.nnSignificance StatementThe extracellular environment plays a critical role in coordinating neuronal migration and neurite outgrowth during neural circuit development. The transmembrane glycoprotein dystroglycan functions as a receptor for multiple extracellular matrix proteins, and its dysfunction leads to a form of muscular dystrophy frequently associated with neurodevelopmental defects. Our results demonstrate that dystroglycan is required for maintaining the structural integrity of the inner limiting membrane (ILM) in the developing retina. In the absence of functional dystroglycan, ILM degeneration leads to defective migration, axon guidance and mosaic spacing of neurons, and a loss of multiple neuron types during retinal development. These results demonstrate that disorganization of retinal circuit development is a likely contributor to visual dysfunction in patients with dystroglycanopathy.
]]></description>
<dc:creator>Clements, R.</dc:creator>
<dc:creator>Turk, R.</dc:creator>
<dc:creator>Campbell, K.</dc:creator>
<dc:creator>Wright, K. M.</dc:creator>
<dc:date>2017-03-29</dc:date>
<dc:identifier>doi:10.1101/121756</dc:identifier>
<dc:title><![CDATA[Dystroglycan Maintains Inner Limiting Membrane Integrity To Coordinate Retinal Development]]></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/10.1101/125062v1?rss=1">
<title>
<![CDATA[
Burst Activation Of Dopamine Neurons Produces Prolonged Post-Burst Availability Of Actively Released Dopamine 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/125062v1?rss=1</link>
<description><![CDATA[
Both phasic and tonic modes of neurotransmission are implicated in critical functions assigned to dopamine. In learning, for example, sub-second phasic responses of ventral tegmental area (VTA) dopamine neurons to salient events serve as teaching signals, but learning is also interrupted by dopamine antagonists administered minutes after training. Our findings bridge the multiple timescales of dopamine neurotransmission by demonstrating that burst stimulation of VTA dopamine neurons produces a prolonged post-burst increase (> 20 min) of extracellular dopamine in nucleus accumbens and prefrontal cortex. This elevation is not due to spillover from the stimulation surge but depends on impulse flow-mediated dopamine release. We identified Rho-mediated internalization of dopamine transporter as a mechanism responsible for prolonged availability of actively released dopamine. These results demonstrate that phasic and tonic dopamine neurotransmission can be a continuum and may explain why both modes of signaling are critical for motivational and cognitive functions associated with dopamine.
]]></description>
<dc:creator>Lohani, S.</dc:creator>
<dc:creator>Martig, A.</dc:creator>
<dc:creator>Underhill, S.</dc:creator>
<dc:creator>DeFrancesco, A.</dc:creator>
<dc:creator>Roberts, M.</dc:creator>
<dc:creator>Rinaman, L.</dc:creator>
<dc:creator>Amara, S.</dc:creator>
<dc:creator>Moghaddam, B.</dc:creator>
<dc:date>2017-04-06</dc:date>
<dc:identifier>doi:10.1101/125062</dc:identifier>
<dc:title><![CDATA[Burst Activation Of Dopamine Neurons Produces Prolonged Post-Burst Availability Of Actively Released Dopamine]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-04-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/132258v1?rss=1">
<title>
<![CDATA[
MicroRNA perturbation of the MRN complex buffers DNA damage response from VEGF signaling 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/132258v1?rss=1</link>
<description><![CDATA[
MicroRNAs contribute to biological robustness by buffering cellular processes from external perturbations. Here we report an unexpected link between DNA damage response and angiogenic signaling that is buffered by two distinct microRNAs. We demonstrate that genotoxic stress-induced miR-494 and miR-99b inhibit the DNA repair machinery by targeting the MRE11a-RAD50-NBN (MRN) complex. Functionally, gain and loss of function experiments show that miR-494 and miR-99b affect telomerase activity, activate p21 and Rb pathways and diminish angiogenic sprouting in vitro and in vivo. Genetic and pharmacological disruption of VEGFR-2 signaling and the MRN complex reveal a surprising co-dependency of these pathways in regulating endothelial senescence and proliferation. Vascular-targeted delivery of miR-494 decreases both growth factor-induced and tumor angiogenesis in mouse models. Mechanistically, disruption of the MRN complex induced CD44, a known driver of senescence and regulator of VEGF signaling in addition to suppressing IL-13 a stimulator of VEGF signaling. Our work identifies a putative miR-facilitated mechanism by which endothelial cells can be insulated against VEGF signaling to facilitate the onset of senescence and highlight the potential of targeting DNA repair to disrupt pathological angiogenesis.
]]></description>
<dc:creator>Espinosa-Diez, C.</dc:creator>
<dc:creator>Wilson, R.</dc:creator>
<dc:creator>Chatterjee, N.</dc:creator>
<dc:creator>Hudson, C.</dc:creator>
<dc:creator>Ruhl, R.</dc:creator>
<dc:creator>Hipfinger, C.</dc:creator>
<dc:creator>Helms, E.</dc:creator>
<dc:creator>Khan, O. F.</dc:creator>
<dc:creator>Anderson, D. G.</dc:creator>
<dc:creator>Anand, S.</dc:creator>
<dc:date>2017-04-28</dc:date>
<dc:identifier>doi:10.1101/132258</dc:identifier>
<dc:title><![CDATA[MicroRNA perturbation of the MRN complex buffers DNA damage response from VEGF signaling]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-04-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/135541v1?rss=1">
<title>
<![CDATA[
Distinct Temporal Filters In Mitral Cells And External Tufted Cells Of The Olfactory Bulb 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/135541v1?rss=1</link>
<description><![CDATA[
Short-term synaptic plasticity is a critical regulator of neural circuits, and largely determines how information is temporally processed. In the olfactory bulb, afferent olfactory receptor neurons respond to increasing concentrations of odorants with barrages of action potentials, and their terminals have an extraordinarily high release probability (Sicard, 1986; Murphy et al., 2004). These features suggest that during naturalistic stimuli, afferent input to the olfactory bulb is subject to strong synaptic depression, presumably truncating the postsynaptic response to afferent stimuli. To examine this issue, we used single glomerular stimulation in mouse olfactory bulb slices to measure the synaptic dynamics of afferent-evoked input at physiological stimulus frequencies. In cell-attached recordings, mitral cells responded to high frequency stimulation with sustained responses, whereas external tufted cells responded transiently. Consistent with previous reports (Murphy et al., 2004), olfactory nerve terminals onto both cell types had a high release probability (0.7), from a single pool of slowly recycling vesicles, indicating that the distinct responses of mitral and external tufted cells to high frequency stimulation did not originate presyaptically. Rather, distinct temporal response profiles in mitral cells and external tufted cells could be attributed to slow dendrodendritic responses in mitral cells, as blocking this slow current in mitral cells converted mitral cell responses to a transient response profile, typical of external tufted cells. Our results suggest that despite strong axodendritic synaptic depression, the balance of axodendritic and dendrodendritic circuitry in external tufted cells and mitral cells, respectively, tunes the postsynaptic responses to high frequency, naturalistic stimulation.nnKey PointsO_LIThe release probability of the ORN is reportedly one of the highest in the brain (Murphy et al., 2004), which is predicted to impose a transient temporal filter on postsynaptic cells.nC_LIO_LIMitral cells responded to high frequency ORN stimulation with sustained transmission, whereas external tufted cells responded transiently.nC_LIO_LIThe release probability of ORNs (0.7) was equivalent across mitral and external tufted cells and could be explained by a single pool of slowly recycling vesicles.nC_LIO_LIThe sustained response in mitral cells resulted from dendrodendritic amplification in mitral cells, which was blocked by NMDA and mGluR1 receptor antagonists, converting mitral cell responses to transient response profiles.nC_LIO_LIOur results suggest that although the afferent ORN synapse shows strong synaptic depression, dendrodendritic circuitry in mitral cells produces robust amplification of brief afferent input, thus the relative strength of axodendritic and dendrodendritic input determines the postsynaptic response profile.nC_LI
]]></description>
<dc:creator>Vaaga, C. E.</dc:creator>
<dc:creator>Westbrook, G. L.</dc:creator>
<dc:date>2017-05-08</dc:date>
<dc:identifier>doi:10.1101/135541</dc:identifier>
<dc:title><![CDATA[Distinct Temporal Filters In Mitral Cells And External Tufted Cells Of The Olfactory Bulb]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-05-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/136234v1?rss=1">
<title>
<![CDATA[
MicroRNA-451a regulates colorectal cancer radiosensitivity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/136234v1?rss=1</link>
<description><![CDATA[
Colorectal cancer (CRC) is a leading cause of cancer-related death. The responses of CRC to standard of care adjuvant therapies such as radiation or chemotherapy are poorly understood. MicroRNAs (miRs) are small non-coding RNAs that affect gene expression programs in cells by downregulating specific mRNAs. In this study, we discovered a set of microRNAs upregulated rapidly in response to a single 2 Gy dose fraction of {gamma}-radiation in a mouse colorectal carcinoma xenograft model. The most upregulated candidate in our signature, miR-451a inhibits tumor cell proliferation and attenuated surviving fraction in longer-term cultures. Conversely, inhibition of miR-451a increased proliferation, tumorsphere formation and surviving fraction of tumor cells. Using a bioinformatics approach, we identified four genes-CAB39, EMSY, MEX3C and EREG as targets of miR-451a. Transfection of miR-451a decreased both mRNA and protein levels of these targets. Importantly, we found miR-451a expression was decreased with tumor stage in a small subset of CRC patients. Finally, analysis of a TCGA colorectal cancer dataset reveals that the CAB39 and EMSY are upregulated at the protein level in a significant number of CRC patients and correlates with poorer overall survival. Taken together, our data indicates miR-451a influences the radiation sensitivity of colorectal carcinomas.
]]></description>
<dc:creator>Ruhl, R.</dc:creator>
<dc:creator>Rana, S.</dc:creator>
<dc:creator>Kelley, K.</dc:creator>
<dc:creator>Espinosa-Diez, C.</dc:creator>
<dc:creator>Hudson, C.</dc:creator>
<dc:creator>Lanciault, C.</dc:creator>
<dc:creator>Thomas, C.</dc:creator>
<dc:creator>Tsikitis, L. V.</dc:creator>
<dc:creator>Anand, S.</dc:creator>
<dc:date>2017-05-09</dc:date>
<dc:identifier>doi:10.1101/136234</dc:identifier>
<dc:title><![CDATA[MicroRNA-451a regulates colorectal cancer radiosensitivity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-05-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/153767v1?rss=1">
<title>
<![CDATA[
Preferential targeting of lateral entorhinal inputs onto newly integrated granule cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/153767v1?rss=1</link>
<description><![CDATA[
Mature dentate granule cells in the hippocampus receive input from the entorhinal cortex via the perforant path in precisely arranged lamina, with medial entorhinal axons innervating the middle molecular layer and lateral entorhinal cortex axons innervating the outer molecular layer. Although vastly outnumbered by mature granule cells, adult-generated newborn granule cells play a unique role in hippocampal function, which has largely been attributed to their enhanced excitability and plasticity (Schmidt-Hieber et al., 2004; Ge et al., 2007). Inputs from the medial and lateral entorhinal cortex carry different informational content, thus the distribution of inputs onto newly integrated granules will affect their function in the circuit. Therefore we examined the functional innervation and synaptic maturation of newly-generated dentate granule cells using retroviral labeling in combination with selective optogenetic activation of medial or lateral entorhinal inputs. Our results indicate that lateral entorhinal inputs provide nearly all the functional innervation of newly integrated granule cells. Despite preferential functional targeting, the dendritic spine density of immature granule cells was not increased in the outer molecular layer compared to the middle molecular layer. However, chronic blockade of neurotransmitter release in medial entorhinal axons with tetanus toxin disrupted normal synapse development from both medial and lateral entorhinal inputs. Our results support a role for preferential lateral perforant path input onto newly generated neurons in mediating pattern separation, but also indicates that medial perforant path input is necessary for normal synaptic development.nnSignificance StatementThe formation of episodic memories involves the integration of contextual and spatial information. Newly integrated neurons in the dentate gyrus of the hippocampus play a critical role in this process, despite constituting only a minor fraction of total granule cells. Here we demonstrate that these neurons preferentially receive information thought to convey the context of an experience - a unique role that each newly integrated granule cell serves for about a month before reaching maturity.
]]></description>
<dc:creator>Woods, N. I.</dc:creator>
<dc:creator>Vaaga, C. E.</dc:creator>
<dc:creator>Chatzi, C.</dc:creator>
<dc:creator>Adelson, J. D.</dc:creator>
<dc:creator>Collie, M. F.</dc:creator>
<dc:creator>Perederiy, J. V.</dc:creator>
<dc:creator>Tovar, K. R.</dc:creator>
<dc:creator>Westbrook, G. L.</dc:creator>
<dc:date>2017-06-21</dc:date>
<dc:identifier>doi:10.1101/153767</dc:identifier>
<dc:title><![CDATA[Preferential targeting of lateral entorhinal inputs onto newly integrated granule cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-06-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/157032v1?rss=1">
<title>
<![CDATA[
Discrete and coordinated encoding of punishment contingent on rewarded actions by prefrontal cortex and VTA 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/157032v1?rss=1</link>
<description><![CDATA[
Actions motivated by a rewarding outcome are often associated with a risk of punishment. Little is known about the neural representation of punishment that is contingent on reward-guided behavior. We modeled this circumstance by using a task where actions were consistently rewarded but probabilistically punished. Spike activity and local field potentials were recorded during this task simultaneously from VTA and mPFC, two reciprocally connected regions implicated in both reward-seeking and aversive behavioral states. At the single unit level, we found that ensembles of VTA and mPFC neurons encode the contingency between action and punishment. At the network level, we found that coherent theta oscillations synchronize the VTA and mPFC in a bottom-up direction, effectively phase-modulating the neuronal spike activity in the two regions during punishment-free actions. This synchrony declined as a function of punishment contingency, suggesting that during reward-seeking actions, risk of punishment diminishes VTA-driven neural synchrony between the two regions.
]]></description>
<dc:creator>Park, J.</dc:creator>
<dc:creator>Moghaddam, B.</dc:creator>
<dc:date>2017-06-28</dc:date>
<dc:identifier>doi:10.1101/157032</dc:identifier>
<dc:title><![CDATA[Discrete and coordinated encoding of punishment contingent on rewarded actions by prefrontal cortex and VTA]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-06-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/157230v1?rss=1">
<title>
<![CDATA[
Scalable and efficient single-cell DNA methylation sequencing by combinatorial indexing. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/157230v1?rss=1</link>
<description><![CDATA[
Here we present a novel method: single-cell combinatorial indexing for methylation analysis (sci-MET), which is the first highly scalable assay for whole genome methylation profiling of single cells. We use sci-MET to produce 2,697 total single-cell bisulfite sequencing libraries and achieve read alignment rates of 69 {+/-} 7%, comparable to those of bulk cell methods. As a proof of concept, we applied sci-MET to successfully deconvolve the cellular identity of a mixture of three human cell lines.
]]></description>
<dc:creator>Mulqueen, R. M.</dc:creator>
<dc:creator>Pokholok, D.</dc:creator>
<dc:creator>Norberg, S.</dc:creator>
<dc:creator>Fields, A. J.</dc:creator>
<dc:creator>Sun, D.</dc:creator>
<dc:creator>Torkenczy, K. A.</dc:creator>
<dc:creator>Shendure, J.</dc:creator>
<dc:creator>Trapnell, C.</dc:creator>
<dc:creator>O'Roak, B. J.</dc:creator>
<dc:creator>Xia, Z.</dc:creator>
<dc:creator>Steemers, F. J.</dc:creator>
<dc:creator>Adey, A. C.</dc:creator>
<dc:date>2017-06-28</dc:date>
<dc:identifier>doi:10.1101/157230</dc:identifier>
<dc:title><![CDATA[Scalable and efficient single-cell DNA methylation sequencing by combinatorial indexing.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-06-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/158006v1?rss=1">
<title>
<![CDATA[
Neutral tumor evolution? 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/158006v1?rss=1</link>
<description><![CDATA[
Tumor growth is an evolutionary process governed by somatic mutation, clonal selection and random genetic drift, constrained by the co-evolution of the microenvironment1,2. Tumor subclones are subpopulations of tumor cells with a common set of mutations resulting from the expansion of a single cell during tumor development, and have been observed in a significant fraction of cancers and across multiple cancer types3. Peter Nowell proposed that tumors evolve through sequential genetic events4, whereby one cell acquires a selective advantage so that its lineage becomes predominant. According to this traditional model, the selective advantage is conferred by a small set of driver mutations, but, as the subclones that bear them expand successively, they accumulate passenger mutations as well, which can be detected in sequencing experiments1. Genomes of indiv ...
]]></description>
<dc:creator>Tarabichi, M.</dc:creator>
<dc:creator>Martincorena, I.</dc:creator>
<dc:creator>Gerstung, M.</dc:creator>
<dc:creator>Markowetz, F.</dc:creator>
<dc:creator>Spellman, P. T.</dc:creator>
<dc:creator>Morris, Q. D.</dc:creator>
<dc:creator>Lingjaerde, O. C.</dc:creator>
<dc:creator>Wedge, D. C.</dc:creator>
<dc:creator>Van Loo, P.</dc:creator>
<dc:creator>- PCAWG Evolution and Heterogeneity Working Group,</dc:creator>
<dc:date>2017-06-30</dc:date>
<dc:identifier>doi:10.1101/158006</dc:identifier>
<dc:title><![CDATA[Neutral tumor evolution?]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-06-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/158402v1?rss=1">
<title>
<![CDATA[
Activation and desensitization mechanism of AMPA receptor-TARP complex by cryo-EM 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/158402v1?rss=1</link>
<description><![CDATA[
Summary Summary Structure determination Agonists expand LBD gating... Activation by full and... Architecture of the ion... Ion-permeation profile of the... Asymmetric water/ion accessible... TARP modulation of receptor... TARP restricts LBD rearrangement... TARP modulation of AMPA... Materials and methods AUTHOR CONTRIBUTIONS AUTHOR INFORMATION References AMPA receptors mediate the majority of fast excitatory neurotransmission in the mammalian brain and transduce the binding of presynaptically released glutamate to the opening of a transmembrane cation channel. Within the postsynaptic density, however, AMPA receptors coassemble with transmembran ...
]]></description>
<dc:creator>Gouaux, E.</dc:creator>
<dc:creator>Chen, S.</dc:creator>
<dc:creator>Zhao, Y.</dc:creator>
<dc:creator>Wang, Y. S.</dc:creator>
<dc:creator>Shekhar, M.</dc:creator>
<dc:creator>Tajkhorshid, E.</dc:creator>
<dc:date>2017-07-01</dc:date>
<dc:identifier>doi:10.1101/158402</dc:identifier>
<dc:title><![CDATA[Activation and desensitization mechanism of AMPA receptor-TARP complex by cryo-EM]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-07-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/161752v1?rss=1">
<title>
<![CDATA[
Sexually divergent DNA methylation programs with hippocampal aging 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/161752v1?rss=1</link>
<description><![CDATA[
DNA methylation is a central regulator of genome function and altered methylation patterns are indicative of biological aging and mortality. Age-related cellular, biochemical, and molecular changes in the hippocampus lead to cognitive impairments and greater vulnerability to neurodegenerative disease that varies between the sexes. The role of hippocampal epigenomic changes with aging in these processes is unknown as no genome-wide analyses of age-related methylation changes have considered the factor of sex in a controlled animal model. High-depth, genome-wide bisulfite sequencing of young (3 month) and old (24 month) male and female mouse hippocampus revealed that while total genomic methylation amounts did not change with aging, specific sites in CG and non-CG (CH) contexts demonstrated age-related increases or decreases in methylation that were predominantly sexually divergent. Differential methylation with age for both CG and CH sites was enriched in intergenic, and intronic regions and under-represented in promoters, CG islands and specific enhancer regions in both sexes suggesting that certain genomic elements are especially labile with aging, even if the exact genomic loci altered are predominantly sex-specific. Life-long sex differences in autosomal methylation at CG and CH sites were also observed. The lack of genome-wide hypomethylation, sexually divergent aging response, and autosomal sex differences at CG sites were confirmed in human data. These data reveal sex as a previously unappreciated central factor of hippocampal epigenomic changes with aging. In total, these data demonstrate an intricate regulation of DNA methylation with aging by sex, cytosine context, genomic location, and methylation level.
]]></description>
<dc:creator>Masser, D.</dc:creator>
<dc:creator>Hadad, N.</dc:creator>
<dc:creator>Porter, H.</dc:creator>
<dc:creator>Mangold, C.</dc:creator>
<dc:creator>Unnikrishnan, A.</dc:creator>
<dc:creator>Ford, M.</dc:creator>
<dc:creator>Giles, C.</dc:creator>
<dc:creator>Georgescu, C.</dc:creator>
<dc:creator>Dozmorov, M.</dc:creator>
<dc:creator>Wren, J.</dc:creator>
<dc:creator>Richardson, A.</dc:creator>
<dc:creator>Stanford, D.</dc:creator>
<dc:creator>Freeman, W.</dc:creator>
<dc:date>2017-07-10</dc:date>
<dc:identifier>doi:10.1101/161752</dc:identifier>
<dc:title><![CDATA[Sexually divergent DNA methylation programs with hippocampal aging]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-07-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/167304v1?rss=1">
<title>
<![CDATA[
Correlated gene expression and anatomical communication support synchronized brain activity in the mouse functional connectome 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/167304v1?rss=1</link>
<description><![CDATA[
Cognition and behavior depend on synchronized intrinsic brain activity which is organized into functional networks across the brain. Research has investigated how anatomical connectivity both shapes and is shaped by these networks, but not how anatomical connectivity interacts with intra-areal molecular properties to drive functional connectivity. Here, we present a novel linear model to explain functional connectivity in the mouse brain by integrating systematically obtained measurements of axonal connectivity, gene expression, and resting state functional connectivity MRI. The model suggests that functional connectivity arises from synergies between anatomical links and inter-areal similarities in gene expression. By estimating these interactions, we identify anatomical modules in which correlated gene expression and anatomical connectivity cooperatively, versus distinctly, support functional connectivity. Along with providing evidence that not all genes equally contribute to functional connectivity, this research establishes new insights regarding the biological underpinnings of coordinated brain activity measured by BOLD fMRI.
]]></description>
<dc:creator>Mills, B. D.</dc:creator>
<dc:creator>Grayson, D.</dc:creator>
<dc:creator>Shunmugavel, A.</dc:creator>
<dc:creator>Miranda-Dominguez, O.</dc:creator>
<dc:creator>Feczko, E.</dc:creator>
<dc:creator>Earl, E.</dc:creator>
<dc:creator>Neve, K.</dc:creator>
<dc:creator>Fair, D.</dc:creator>
<dc:date>2017-07-22</dc:date>
<dc:identifier>doi:10.1101/167304</dc:identifier>
<dc:title><![CDATA[Correlated gene expression and anatomical communication support synchronized brain activity in the mouse functional connectome]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-07-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/169979v1?rss=1">
<title>
<![CDATA[
Image velocimetry and spectral analysis enable quantitative characterization of larval zebrafish gut motility 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/169979v1?rss=1</link>
<description><![CDATA[
Summary StatementWe present a new image analysis technique using image velocimetry and spectral analysis that returns quantitative measures of gut contraction strength, frequency, and wave speed that can be used to study gut motility and other cellular movements.nnAbstractNormal gut function requires rhythmic and coordinated movements that are affected by developmental processes, physical and chemical stimuli, and many debilitating diseases. The imaging and characterization of gut motility, especially regarding periodic, propagative contractions driving material transport, are therefore critical goals. Whereas previous image analysis approaches have successfully extracted properties related to temporal frequency of motility modes, robust measures of contraction magnitude remain elusive. We developed a new image analysis method based on image velocimetry and spectral analysis that reveals temporal characteristics such as frequency and wave propagation speed, while also providing quantitative measures of the amplitude of gut motions. We validate this approach using several challenges to larval zebrafish, imaged with differential interference contrast microscopy. Both acetylcholine exposure and feeding increase frequency and amplitude of motility. Larvae lacking enteric nervous system gut innervation show the same average motility frequency, but reduced and less variable amplitude compared to wild-types. Our image analysis approach enables insights into gut dynamics in a wide variety of developmental and physiological contexts and can also be extended to analyze other types of cell movements.
]]></description>
<dc:creator>Ganz, J.</dc:creator>
<dc:creator>Baker, R. P.</dc:creator>
<dc:creator>Hamilton, M. K.</dc:creator>
<dc:creator>Melancon, E.</dc:creator>
<dc:creator>Diba, P.</dc:creator>
<dc:creator>Eisen, J. S.</dc:creator>
<dc:creator>Parthasarathy, R.</dc:creator>
<dc:date>2017-07-28</dc:date>
<dc:identifier>doi:10.1101/169979</dc:identifier>
<dc:title><![CDATA[Image velocimetry and spectral analysis enable quantitative characterization of larval zebrafish gut motility]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-07-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/172908v1?rss=1">
<title>
<![CDATA[
Anti-diabetic drug binding site in KATP channels revealed by Cryo-EM 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/172908v1?rss=1</link>
<description><![CDATA[
Sulfonylureas are anti-diabetic medications that act by inhibiting pancreatic KATP channels composed of SUR1 and Kir6.2. The mechanism by which these drugs interact with and inhibit the channel has been extensively investigated, yet it remains unclear where the drug binding pocket resides. Here, we present a cryo-EM structure of the channel bound to a high-affinity sulfonylurea drug glibenclamide and ATP at 3.8[A] resolution, which reveals in unprecedented details of the ATP and glibenclamide binding sites. Importantly, the structure shows for the first time that glibenclamide is lodged in the transmembrane bundle of the SUR1-ABC core connected to the first nucleotide binding domain near the inner leaflet of the lipid bilayer. Mutation of residues predicted to interact with glibenclamide in our model led to reduced sensitivity to glibenclamide. Our structure provides novel mechanistic insights of how sulfonylureas and ATP interact with the KATP channel complex to inhibit channel activity.
]]></description>
<dc:creator>Martin, G. M.</dc:creator>
<dc:creator>Yoshioka, C.</dc:creator>
<dc:creator>Kandasamy, B.</dc:creator>
<dc:creator>DiMaio, F.</dc:creator>
<dc:creator>Shyng, S.-L.</dc:creator>
<dc:date>2017-08-05</dc:date>
<dc:identifier>doi:10.1101/172908</dc:identifier>
<dc:title><![CDATA[Anti-diabetic drug binding site in KATP channels revealed by Cryo-EM]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-08-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/185058v1?rss=1">
<title>
<![CDATA[
Ectopic Expression of DNA Repair Enzymes Modulates Survival following Ultraviolet Irradiation Challenge 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/185058v1?rss=1</link>
<description><![CDATA[
In Escherichia coli, the nucleotide excision repair (NER) pathway removes ultraviolet (UV) light-induced cyclobutane pyrimidine dimers (CPDs) and 6-4 dipyrimidine photoproducts (6-4 PPs). Activation of alternative repair pathways, such as base excision repair (BER) and nucleotide incision repair (NIR), is inoperative because this organism lacks both the necessary BER DNA glycosylase and NIR UV endonuclease to initiate repair of these lesions. To determine if initiation of either pathway would enhance survival to biologically-relevant UV irradiation, the BER and NIR pathways were activated by expression of Chlorella virus-1 pyrimidine dimer glycosylase (cv-pdg) and Schizosaccharomyces pombe UV endonuclease (UVDE), respectively. The substrate specificity of cv-pdg includes CPDs and ring-fragmented purines, 4,6-diamino-5-formamidopyrimidine and 2,6-diamino-4-hydroxy-5-formamidopyrimidine, but not 6-4 PPs. In contrast, while UVDE incises DNA containing CPDs and 6-4 PPs, it was not previously known if the substrate specificity of UVDE included DNA containing ring-fragmented purines. Mass spectrometry was used to establish that these oxidatively-induced lesions were not substrates for UVDE. Expression of either cv-pdg or UVDE in NER-deficient E. coli significantly enhanced survival following UVB irradiation, but not to the levels of wild type (WT) cells. Survival of NER-proficient, homologous recombination-deficient cells could also be significantly enhanced by expression of either enzyme, suggesting that in response to UVB exposure, interactions between NER and activated BER or NIR pathways could be additive. Further, expression of cv-pdg or UVDE in WT E. coli enhanced survival following solar-simulated light (SSL) exposures.
]]></description>
<dc:creator>Garg, S. P.</dc:creator>
<dc:creator>Minko, I. G.</dc:creator>
<dc:creator>Coskun, E.</dc:creator>
<dc:creator>Erdem, O.</dc:creator>
<dc:creator>Jaruga, P.</dc:creator>
<dc:creator>Dizdaroglu, M.</dc:creator>
<dc:creator>Lloyd, R. S.</dc:creator>
<dc:date>2017-09-06</dc:date>
<dc:identifier>doi:10.1101/185058</dc:identifier>
<dc:title><![CDATA[Ectopic Expression of DNA Repair Enzymes Modulates Survival following Ultraviolet Irradiation Challenge]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-09-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/189621v1?rss=1">
<title>
<![CDATA[
ELMOD1 stimulates ARF6-GTP hydrolysis to stabilize apical structures in developing vestibular hair cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/189621v1?rss=1</link>
<description><![CDATA[
Sensory hair cells require control of physical properties of their apical plasma membranes for normal development and function. Members of the ARF small GTPase family regulate membrane trafficking and cytoskeletal assembly in many cells. We identified ELMOD1, a guanine nucleoside triphosphatase activating protein (GAP) for ARF6, as the most highly enriched ARF regulator in hair cells. To characterize ELMOD1 control of trafficking, we used a mouse strain lacking functional ELMOD1 (roundabout; rda). In rda/rda mice, cuticular plates of utricle hair cells initially formed normally, then degenerated after postnatal day 5 (P5); large numbers of vesicles invaded the compromised cuticular plate. Hair bundles initially developed normally, but the cells apical membrane lifted away from the cuticular plate, and stereocilia elongated and fused. Membrane trafficking in type I hair cells, measured by FM1-43 dye labeling, was altered in rda/rda mice. Consistent with the proposed GAP role for ELMOD1, the ARF6 GTP/GDP ratio was significantly elevated in rda/rda utricles as compared to controls, and the level of ARF6-GTP was correlated with the severity of the rda/rda phenotype. These results suggest that conversion of ARF6 to its GDP-bound form is necessary for final stabilization of the hair bundle.nnSignificance StatementAssembly of the mechanically sensitive hair bundle of sensory hair cells requires growth and reorganization of apical actin and membrane structures. Hair bundles and apical membranes in mice with mutations in the Elmod1 gene degenerate after formation, suggesting that the ELMOD1 protein stabilizes these structures. We show that ELMOD1 is a GTPase-activating protein in hair cells for the small GTP-binding protein ARF6, known to participate in actin assembly and membrane trafficking. We propose that conversion of ARF6 into the GDP-bound form in the apical domain of hair cells is essential for stabilizing apical actin structures like the hair bundle and ensuring that the apical membrane forms appropriately around the stereocilia.
]]></description>
<dc:creator>Krey, J. F.</dc:creator>
<dc:creator>Dumont, R. A.</dc:creator>
<dc:creator>Wilmarth, P. A.</dc:creator>
<dc:creator>David, L. L.</dc:creator>
<dc:creator>Johnson, K. R.</dc:creator>
<dc:creator>Barr-Gillespie, P. G.</dc:creator>
<dc:date>2017-09-15</dc:date>
<dc:identifier>doi:10.1101/189621</dc:identifier>
<dc:title><![CDATA[ELMOD1 stimulates ARF6-GTP hydrolysis to stabilize apical structures in developing vestibular hair cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-09-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/191783v1?rss=1">
<title>
<![CDATA[
Enabling Precision Medicine via standard communication of NGS provenance, analysis, and results 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/191783v1?rss=1</link>
<description><![CDATA[
A personalized approach based on a patients or pathogens unique genomic sequence is the foundation of precision medicine. Genomic findings must be robust and reproducible, and experimental data capture should adhere to FAIR guiding principles. Moreover, effective precision medicine requires standardized reporting that extends beyond wet lab procedures to computational methods. The BioCompute framework (https://osf.io/zm97b/) enables standardized reporting of genomic sequence data provenance, including provenance domain, usability domain, execution domain, verification kit, and error domain. This framework facilitates communication and promotes interoperability. Bioinformatics computation instances that employ the BioCompute framework are easily relayed, repeated if needed and compared by scientists, regulators, test developers, and clinicians. Easing the burden of performing the aforementioned tasks greatly extends the range of practical application. Large clinical trials, precision medicine, and regulatory submissions require a set of agreed upon standards that ensures efficient communication and documentation of genomic analyses. The BioCompute paradigm and the resulting BioCompute Objects (BCO) offer that standard, and are freely accessible as a GitHub organization (https://github.com/biocompute-objects) following the "Open-Stand.org principles for collaborative open standards development". By communication of high-throughput sequencing studies using a BCO, regulatory agencies (e.g., FDA), diagnostic test developers, researchers, and clinicians can expand collaboration to drive innovation in precision medicine, potentially decreasing the time and cost associated with next generation sequencing workflow exchange, reporting, and regulatory reviews.
]]></description>
<dc:creator>Alterovitz, G.</dc:creator>
<dc:creator>Dean, D. A.</dc:creator>
<dc:creator>Goble, C.</dc:creator>
<dc:creator>Crusoe, M. R.</dc:creator>
<dc:creator>Soiland-Reyes, S.</dc:creator>
<dc:creator>Bell, A.</dc:creator>
<dc:creator>Hayes, A.</dc:creator>
<dc:creator>King, C. H. H.</dc:creator>
<dc:creator>Johanson, E.</dc:creator>
<dc:creator>Thompson, E. E.</dc:creator>
<dc:creator>Donaldson, E.</dc:creator>
<dc:creator>Tsang, H. S.</dc:creator>
<dc:creator>Goecks, J.</dc:creator>
<dc:creator>Almeida, J. S.</dc:creator>
<dc:creator>Guo, L.</dc:creator>
<dc:creator>Walderhaug, M.</dc:creator>
<dc:creator>Walsh, P.</dc:creator>
<dc:creator>Kahsay, R.</dc:creator>
<dc:creator>Bloom, T.</dc:creator>
<dc:creator>Lai, Y.</dc:creator>
<dc:creator>Simonyan, V.</dc:creator>
<dc:creator>Mazumder, R.</dc:creator>
<dc:date>2017-09-21</dc:date>
<dc:identifier>doi:10.1101/191783</dc:identifier>
<dc:title><![CDATA[Enabling Precision Medicine via standard communication of NGS provenance, analysis, and results]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-09-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/192716v1?rss=1">
<title>
<![CDATA[
Population-level distribution and putative immunogenicity of cancer neoepitopes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/192716v1?rss=1</link>
<description><![CDATA[
BackgroundTumor neoantigens are a driver of cancer immunotherapy response; however, current neoantigen prediction tools produce many candidates that require further prioritization for research/clinical applications. Additional filtration criteria and population-level understanding may help to produce refined lists of putative neoantigens. Herein, we show neoepitope immunogenicity is likely related to measures of peptide novelty and report population-level behavior of these and other metrics.nnMethodsWe propose four peptide novelty metrics to refine predicted neoantigenicity: tumor vs. paired normal peptide binding affinity difference, tumor vs. paired normal peptide sequence similarity, tumor vs. closest human peptide sequence similarity, and tumor vs. closest microbial peptide sequence similarity. We apply these metrics to tumor neoepitopes predicted from somatic missense mutations in The Cancer Genome Atlas (TCGA) and a cohort of melanoma patients, as well as to a group of peptides with neoepitope-specific immune response data using an extension of pVAC-Seq [1].nnResultsWe show neoepitope burden varies across TCGA disease sites and HLA alleles, with surprisingly low repetition of neoepitope sequences across patients or neoepitope preferences among sets of HLA alleles. Only 20.3% of predicted neoepitopes across TCGA patients displayed novel binding change based on our binding affinity difference criteria. Similarity of amino acid sequence was typically high between paired tumor-normal epitopes, but in 24.6% of cases, neoepitopes were more similar to other human peptides, or even to bacterial (56.8% of cases) or viral peptides (15.5% of cases), than their paired normal counterparts. Applied to peptides with neoepitope-specific immune response, a linear model incorporating neoepitope binding affinity, protein sequence similarity between neoepitopes and their closest viral peptides, and paired binding affinity difference was able to predict immunogenicity with an AUROC of 0.66.nnConclusionsOur proposed neoepitope prioritization criteria emphasize neoepitope novelty and refine patient neoepitope predictions for focus on biologically meaningful candidate neoantigens. We have demonstrated that neoepitopes should be considered not only with respect to their paired normal epitope, but with respect to the entire human proteome, as well as bacterial and viral peptides, with potential implications for neoepitope immunogenicity and personalized vaccines for cancer treatment. We conclude that putative neoantigens are highly variable across individuals as a function of both cancer genetics and personalized HLA repertoire, while the overall behavior of filtration criteria reflects predictable patterns.
]]></description>
<dc:creator>Wood, M. A.</dc:creator>
<dc:creator>Paralkar, M.</dc:creator>
<dc:creator>Paralkar, M. P.</dc:creator>
<dc:creator>Nguyen, A.</dc:creator>
<dc:creator>Struck, A. J.</dc:creator>
<dc:creator>Ellrott, K.</dc:creator>
<dc:creator>Margolin, A.</dc:creator>
<dc:creator>Nellore, A.</dc:creator>
<dc:creator>Thompson, R. F.</dc:creator>
<dc:date>2017-09-22</dc:date>
<dc:identifier>doi:10.1101/192716</dc:identifier>
<dc:title><![CDATA[Population-level distribution and putative immunogenicity of cancer neoepitopes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-09-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2019.12.16.877852v1?rss=1">
<title>
<![CDATA[
Re-evaluation of lysyl hydroxylation in the collagen triple helix: lysyl hydroxylase 1 and prolyl 3-hydroxylase 3 have site-differential and collagen type-dependent roles in lysine hydroxylation. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2019.12.16.877852v1?rss=1</link>
<description><![CDATA[
Collagen is the most abundant protein in humans and is heavily post-translationally modified. Its biosynthesis is very complex and requires three different types of hydroxylation (two for proline and one for lysine) that are generated in the rough endoplasmic reticulum (rER). These processes involve many enzymes and chaperones which were collectively termed the molecular ensemble for collagen biosynthesis. However, the function of some of the proteins in this molecular ensemble is controversial. While prolyl 3-hydroxylase 1 and 2 (P3H1, P3H2) are bona fide collagen prolyl 3-hydroxylases, the function of prolyl 3-hydroxylase 3 (P3H3) is less clear. A recent study of P3H3 null mice demonstrated that this enzyme had no activity as prolyl 3-hydroxylase but may instead act as a chaperone for lysyl hydroxylase 1 (LH1). LH1 is required to generate hydroxylysine for crosslinking within collagen triple helical sequences. If P3H3 is a LH1 chaperone that is critical for LH1 activity, P3H3 and LH1 null mice should have similar deficiency in lysyl hydroxylation. To test this hypothesis, we compared lysyl hydroxylation in type I and V collagen from P3H3 and LH1 null mice. Our results indicate LH1 plays a global role for lysyl hydroxylation in triple helical domain of type I collagen while P3H3 is indeed involved in lysyl hydroxylation particularly at crosslink formation sites but is not required for all lysyl hydroxylation sites in type I collagen triple helix. Furthermore, although type V collagen from LH1 null mice surprisingly contained as much hydroxylysine as type V collagen from wild type, the amount of hydroxylysine in type V collagen was clearly suppressed in P3H3 null mice. In summary, our study suggests that P3H3 and LH1 likely have two distinct mechanisms to distinguish crosslink formation sites from other sites in type I collagen and to recognize different collagen types in the rER.

Author summaryCollagen is one of the most heavily post-translationally modified proteins in the human body and its post-translational modifications provide biological functions to collagen molecules. In collagen post-translational modifications, crosslink formation on a collagen triple helix adds important biomechanical properties to the collagen fibrils and is mediated by hydroxylation of very specific lysine residues. LH1 and P3H3 show the similar role in lysine hydroxylation for specific residues at crosslink formation sites of type I collagen. Conversely, they have very distinct rules in lysine hydroxylation at other residues in type I collagen triple helix. Furthermore, they demonstrate preferential recognition and modification of different collagen types. Our findings provide a better understanding of the individual functions of LH1 and P3H3 in the rER and also offer new directions for the mechanism of lysyl hydroxylation followed by crosslink formation in different tissues and collagens.
]]></description>
<dc:creator>Ishikawa, Y.</dc:creator>
<dc:creator>Taga, Y.</dc:creator>
<dc:creator>Zientek, K.</dc:creator>
<dc:creator>Mizuno, N.</dc:creator>
<dc:creator>Salo, A. M.</dc:creator>
<dc:creator>Semenova, O.</dc:creator>
<dc:creator>Tufa, S.</dc:creator>
<dc:creator>Keene, D. R.</dc:creator>
<dc:creator>Holden, P.</dc:creator>
<dc:creator>Mizuno, K.</dc:creator>
<dc:creator>Myllyharju, J.</dc:creator>
<dc:creator>Bächinger, H. P.</dc:creator>
<dc:date>2019-12-16</dc:date>
<dc:identifier>doi:10.1101/2019.12.16.877852</dc:identifier>
<dc:title><![CDATA[Re-evaluation of lysyl hydroxylation in the collagen triple helix: lysyl hydroxylase 1 and prolyl 3-hydroxylase 3 have site-differential and collagen type-dependent roles in lysine hydroxylation.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-12-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2019.12.18.448985v1?rss=1">
<title>
<![CDATA[
Analysis of model organism viability through an interspecies pathway comparison pipeline using the dynamic impact approach 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2019.12.18.448985v1?rss=1</link>
<description><![CDATA[
BackgroundComputational biologists investigate gene expression time-series data using estimation, clustering, alignment, and enrichment methods to make biological sense of the data and provide compelling visualization. While there is an abundance of microarray and RNA-seq data available, interpreting the data while capturing the dynamism of a time-course experiment remains a difficult challenge. Advancements in RNA-seq technologies have allowed us to collect extensive profiles of diverse developmental processes but also requires additional methods for analysis and data integration to capture the increased dynamism. An approach that can both capture the dynamism and direction of change in a time-course experiment in a holistic manner and simultaneously identify which biological pathways are significantly altered is necessary for the interpretation of systems biology data. In addition, there is a need for a method to evaluate the viability of model organisms across different treatments and conditions. By comparing effects of a specific treatment (e.g., a drug) on the target pathway between multiple species and determining pathways with a similar response to biological cues between organisms, we can determine the best animal model for that treatment for future studies.

MethodsHere, we present Dynamic Impact Approach with Normalization (DIA-norm), a dynamic pathway analysis tool for the analysis of time-course data without unsupervised dimensionality reduction. We analyzed five datasets of mesenchymal stem cells retrieved from the Gene Expression Omnibus data repository (3 human, 1 mouse cell line, 1 pig) which were differentiated in vitro towards adipogenesis. In the first step, DIA-norm calculated an impact and flux score for each biological term using p-value and fold change. In the second step, these scores were normalized and interpolated using cubic spline. Cross-correlation was then performed between all the data sets with r[&ge;]0.6 as a benchmark for high correlation as r = 0.7 is the limit of experimental reproducibility.

ResultsDIA-norm predicted that the pig was a better model for humans than a mouse for the study of adipogenesis. The pig model had a higher number of correlating pathways with humans (64.5 to 30.5) and higher average correlation (r = 0.51 vs r = 0.46) as compared to mouse model vs human. While not a definitive conclusion, the results are in accordance with prior phylogenetic and disease studies in which pigs are a good model for studying humans, specifically regarding obesity. In addition, DIA-norm identified a larger number of biologically important pathways (approximately 2x number of pathways) versus a comparable enrichment analysis tool, DAVID. DIA-norm also identified some possible pathways of interests for adipogenesis, namely, nitrogen metabolism (r = 0.86), where there is little to no existing literature.

ConclusionDIA-norm captured 80+% of biological important pathways and achieved high pathway correlation between species for the vast majority of important adipogenesis pathways. DIA-norm can be used for both time-series pathway analysis and the determination of a model organism. Our findings indicate that DIA-norm can be used to study the effect of any treatment, including drugs, on specific pathways between multiple species to determine the best animal model for that treatment for future studies. The reliability of DIA-norm to provide biological insights compared to enrichment approach tools has been demonstrated in the selected transcriptomic studies by identifying a higher number of total and biologically relevant pathways. DIA-norms final advantage was its easily interpretable graphical outputs that aid in visualizing dynamic changes in expression.
]]></description>
<dc:creator>Nguyen, A. D.</dc:creator>
<dc:creator>Bionaz, M.</dc:creator>
<dc:date>2019-12-19</dc:date>
<dc:identifier>doi:10.1101/2019.12.18.448985</dc:identifier>
<dc:title><![CDATA[Analysis of model organism viability through an interspecies pathway comparison pipeline using the dynamic impact approach]]></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/10.1101/2019.12.19.881631v1?rss=1">
<title>
<![CDATA[
Investigating β-lactam drug targets in Mycobacterium tuberculosis using chemical probes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2019.12.19.881631v1?rss=1</link>
<description><![CDATA[
Tuberculosis is a deadly disease that requires a flexible arsenal of drugs to treat it. Although {beta}-lactam antibiotics are rarely used to treat Mycobacterium tuberculosis (Mtb) infections today, the targets of these drugs are present in the bacterium. Moreover, the cell wall peptidoglycan of Mtb contains an abundance of unusual (3[-&gt;]3) cross-links. These cross-links are formed by enzymes called L,D-transpeptidases, which are susceptible to inhibition by the carbapenem class of antibiotics. We developed new small molecule probes to investigate the L,D-transpeptidases and other {beta}-lactam drug targets in Mtb. We synthesized probes based on three classes of antibiotics, a monobactam, cephalosporin, and carbapenem. For the carbapenem, we synthesized a meropenem analogue conjugated to a far-red fluorophore. This probe was particularly useful in identifying active L,D-transpeptidases in protein gel-resolved lysates. Next we analyzed {beta}-lactam targets in lysates from both hypoxic and actively-replicating cultures of Mtb. We identified numerous targets, including transpeptidases, carboxypeptidases, and the {beta}-lactamase BlaC. Overall, we provide evidence that Mtb dynamically regulates the enzymes responsible for maintaining cell wall peptidoglycan and that meropenem is a good inhibitor of those enzymes.
]]></description>
<dc:creator>Beatty, K. E.</dc:creator>
<dc:creator>Levine, S. R.</dc:creator>
<dc:date>2019-12-19</dc:date>
<dc:identifier>doi:10.1101/2019.12.19.881631</dc:identifier>
<dc:title><![CDATA[Investigating β-lactam drug targets in Mycobacterium tuberculosis using chemical probes]]></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/10.1101/2019.12.19.883165v1?rss=1">
<title>
<![CDATA[
Short-term Effects of Vagus Nerve Stimulationon Learning and Evoked Activity in AuditoryCortex 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2019.12.19.883165v1?rss=1</link>
<description><![CDATA[
Chronic vagus nerve stimulation (VNS) can facilitate learning of sensory and motor behaviors. VNS is believed to trigger release of neuromodulators, including norepinephrine and acetylcholine, which can mediate cortical plasticity associated with learning. Most previous work has studied effects of VNS over many days, and less is known about how acute VNS influences neural coding and behavior over the shorter term. To explore this question, we measured effects of VNS on learning of an auditory discrimination over 1-2 days. Ferrets implanted with cuff electrodes on the vagus nerve were trained by classical conditioning on a tone frequency-reward association. One tone was associated with reward while another tone, was not. The frequencies and reward associations of the tones were changed every two days, requiring learning of a new relationship. When the tones (both rewarded and non-rewarded) were paired with VNS, rates of learning increased on the first day following a change in reward association. To examine VNS effects on auditory coding, we recorded single- and multi-unit neural activity in primary auditory cortex (A1) of passively listening animals following brief periods of VNS (20 trials/session) paired with tones. Because afferent VNS induces changes in pupil size associated with fluctuations in neuromodulation, we also measured pupil during recordings. After pairing VNS with a neurons best-frequency (BF) tone, responses in a subpopulation of neurons were reduced. Pairing with an off-BF tone or performing VNS during the inter-trial interval had no effect on responses. We separated the change in A1 activity into two components, one that could be predicted by fluctuations in pupil and one that persisted after VNS and was not accounted for by pupil. The BF-specific reduction in neural responses remained, even after regressing out changes that could be explained by pupil. In addition, the size of VNS-mediated changes in pupil predicted the magnitude of persistent changes in the neural response. This interaction suggests that changes in neuromodulation associated with arousal gate the long-term effects of VNS on neural activity. Taken together, these results support a role for VNS in auditory learning and help establish VNS as a tool to facilitate neural plasticity.
]]></description>
<dc:creator>Lai, J.</dc:creator>
<dc:creator>David, S. V.</dc:creator>
<dc:date>2019-12-20</dc:date>
<dc:identifier>doi:10.1101/2019.12.19.883165</dc:identifier>
<dc:title><![CDATA[Short-term Effects of Vagus Nerve Stimulationon Learning and Evoked Activity in AuditoryCortex]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-12-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2019.12.20.882746v1?rss=1">
<title>
<![CDATA[
Postnatal expansion, maturation and functionality of MR1 T cells in humans 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2019.12.20.882746v1?rss=1</link>
<description><![CDATA[
MR1-restricted T (MR1T) cells are defined by their recognition of metabolite antigens presented by the monomorphic MHC class 1-related molecule, MR1, the most highly conserved MHC class I related molecule in mammalian species. Mucosal-associated invariant T (MAIT) cells are the predominant subset of MR1T cells expressing an invariant TCR -chain, TRAV1-2. These cells comprise a T cell subset that recognizes and mediates host immune responses to a broad array of microbial pathogens, including Mycobacterium tuberculosis. Here, we sought to characterize development of circulating human MR1T cells as defined by MR1-5-OP-RU tetramer labelling and of the TRAV1-2+ MAIT cells defined by expression of TRAV1-2 and high expression of CD26 and CD161 (TRAV1-2+CD161++CD26++ cells). We analysed postnatal expansion, maturation and functionality of peripheral blood MR1-5-OP-RU tetramer+ MR1T cells in cohorts from three different geographic settings with different tuberculosis (TB) vaccination practices, levels of exposure to and infection with M. tuberculosis. Early after birth, frequencies of MR1-5-OP-RU tetramer+ MR1T cells increased rapidly by several fold. This coincided with the transition from a predominantly CD4+ and TRAV1-2- population in neonates, to a predominantly TRAV1-2+CD161++CD26++ CD8+ population. We also observed that tetramer+ MR1T cells that expressed TNF upon mycobacterial stimulation were very low in neonates, but increased [~]10-fold in the first year of life. These functional MR1T cells in all age groups were MR1-5-OP-RU tetramer+TRAV1-2+ and highly expressed CD161 and CD26, markers that appeared to signal phenotypic and functional maturation of this cell subset. This age-associated maturation was also marked by the loss of naive T cell markers on tetramer+ TRAV1-2+ MR1T cells more rapidly than tetramer+TRAV1-2- MR1T cells and non-MR1T cells. These data suggest that neonates have infrequent populations of MR1T cells with diverse phenotypic attributes; and that exposure to the environment rapidly and preferentially expands the MR1-5-OP-RU tetramer+TRAV1-2+ population of MR1T cells, which becomes the predominant population of functional MR1T cells early during childhood.
]]></description>
<dc:creator>Swarbrick, G. M.</dc:creator>
<dc:creator>Gela, A.</dc:creator>
<dc:creator>Cansler, M. E.</dc:creator>
<dc:creator>Null, M. D.</dc:creator>
<dc:creator>Duncan, R. B.</dc:creator>
<dc:creator>Nemes, E.</dc:creator>
<dc:creator>Shey, M.</dc:creator>
<dc:creator>Nsereko, M.</dc:creator>
<dc:creator>Mayanja-Kizza, H.</dc:creator>
<dc:creator>Kiguli, S.</dc:creator>
<dc:creator>Koh, J.</dc:creator>
<dc:creator>Hanekom, W.</dc:creator>
<dc:creator>Hatherill, M.</dc:creator>
<dc:creator>Lancioni, C.</dc:creator>
<dc:creator>Lewinsohn, D. M.</dc:creator>
<dc:creator>Scriba, T.</dc:creator>
<dc:creator>Lewinsohn, D. A.</dc:creator>
<dc:date>2019-12-21</dc:date>
<dc:identifier>doi:10.1101/2019.12.20.882746</dc:identifier>
<dc:title><![CDATA[Postnatal expansion, maturation and functionality of MR1 T cells in humans]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-12-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2019.12.26.888875v1?rss=1">
<title>
<![CDATA[
KCNQ channels enable reliable presynaptic spiking and synaptic transmission at high-frequency 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2019.12.26.888875v1?rss=1</link>
<description><![CDATA[
The presynaptic action potential (AP) results in calcium influx which triggers neurotransmitter release. For this reason, the AP waveform is crucial in determining the timing and strength of synaptic transmission. The calyx of Held nerve terminals of rat show minimum changes in AP waveform during high-frequency AP firing. We found that the stability of the calyceal AP waveform requires KCNQ K+ channel activated during high-frequency spiking activity. High-frequency presynaptic spikes gradually led to accumulation of KCNQ channels in open states which kept interspike membrane potential sufficiently negative to maintain Na+ channel availability. Accordingly, blocking KCNQ channels during stimulus trains led to inactivation of presynaptic Na+, and to a lesser extent KV1 channels, thereby reducing the AP height and broadening AP duration. Thus, while KCNQ channels are generally thought to prevent hyperactivity of neurons, we find that in axon terminals these channels function to facilitate high-frequency firing needed for sensory coding.

HIGHLIGHTSO_LIKCNQ channels are activated during high-frequency firing
C_LIO_LIThe activity of KCNQ channels helps the recovery of Na+ and KV1 channels from inactivation and maintains action potential waveform
C_LIO_LIReliable presynaptic action potential waveform preserves stable Ca2+ influx and reliable synaptic signaling
C_LI
]]></description>
<dc:creator>Zhang, Y.</dc:creator>
<dc:creator>Li, D.</dc:creator>
<dc:creator>Darwish, Y.</dc:creator>
<dc:creator>Trussell, L. O.</dc:creator>
<dc:creator>Huang, H.</dc:creator>
<dc:date>2019-12-27</dc:date>
<dc:identifier>doi:10.1101/2019.12.26.888875</dc:identifier>
<dc:title><![CDATA[KCNQ channels enable reliable presynaptic spiking and synaptic transmission at high-frequency]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-12-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.01.05.895201v1?rss=1">
<title>
<![CDATA[
A sample preparation protocol for high throughput immunofluorescence of suspension cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.01.05.895201v1?rss=1</link>
<description><![CDATA[
Imaging is a powerful approach for studying protein expression and has the advantage over other methodologies in providing spatial information in situ at single cell level. Using immunofluorescence and confocal microscopy, detailed information of subcellular distribution of proteins can be obtained. While adherent cells of different tissue origin are relatively easy to prepare for imaging applications, nonadherent cells from hematopoietic origin, present a challenge due to their poor attachment to surfaces and subsequent loss of a substantial fraction of the cells. Still, these cell types represent an important part of the human proteome and express genes that are not expressed in adherent cell types. In the era of cell mapping efforts, overcoming the challenge with suspension cells for imaging applications would enable systematic profiling of hematopoietic cells. In this work, we successfully established an immunofluorescence protocol for preparation of suspension cell lines and peripheral blood mononucleated cells (PBMC) and human platelets. The protocol is based on a multi-well plate format with automated sample preparation, allowing for robust high throughput imaging applications. In combination with confocal microscopy, the protocol enables systematic exploration of protein localization to all major subcellular structures.
]]></description>
<dc:creator>Backstrom, A.</dc:creator>
<dc:creator>Kugel, L.</dc:creator>
<dc:creator>Gnann, C.</dc:creator>
<dc:creator>Xu, H.</dc:creator>
<dc:creator>Aslan, J. E.</dc:creator>
<dc:creator>Lundberg, E.</dc:creator>
<dc:creator>Stadler, C.</dc:creator>
<dc:date>2020-01-06</dc:date>
<dc:identifier>doi:10.1101/2020.01.05.895201</dc:identifier>
<dc:title><![CDATA[A sample preparation protocol for high throughput immunofluorescence of suspension cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-01-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.01.06.891127v1?rss=1">
<title>
<![CDATA[
Cerebellar Purkinje cell activity modulates aggressive behavior 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.01.06.891127v1?rss=1</link>
<description><![CDATA[
Although the cerebellum is traditionally associated with balance and motor function, it also plays wider roles in affective and cognitive behaviors. Evidence suggests that the cerebellar vermis may regulate aggressive behavior, though the cerebellar circuits and patterns of activity that influence aggression remain unclear. We used optogenetic methods to bidirectionally modulate the activity of spatially-delineated cerebellar Purkinje cells to evaluate the impact on aggression in mice. Increasing Purkinje cell activity in the vermis significantly reduced the frequency of attacks in a resident-intruder assay. Reduced aggression was not a consequence of impaired motor function, because optogenetic stimulation did not alter motor performance. In complementary experiments, optogenetic inhibition of Purkinje cells in the vermis increased the frequency of attacks. These results establish Purkinje cell activity in the cerebellar vermis regulates aggression, and further support the importance of the cerebellum in driving affective behaviors that could contribute to neurological disorders.
]]></description>
<dc:creator>Jackman, S. L.</dc:creator>
<dc:creator>Chen, C. H.</dc:creator>
<dc:creator>Offermann, H. L.</dc:creator>
<dc:creator>Drew, I. R.</dc:creator>
<dc:creator>Harrison, B. M.</dc:creator>
<dc:creator>Bowman, A. M.</dc:creator>
<dc:creator>Flick, K. M.</dc:creator>
<dc:creator>Flaquer, I.</dc:creator>
<dc:creator>Regehr, W. G.</dc:creator>
<dc:date>2020-01-06</dc:date>
<dc:identifier>doi:10.1101/2020.01.06.891127</dc:identifier>
<dc:title><![CDATA[Cerebellar Purkinje cell activity modulates aggressive behavior]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-01-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.01.06.895474v1?rss=1">
<title>
<![CDATA[
Individual variability in functional organization of the human and monkey auditory cortex 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.01.06.895474v1?rss=1</link>
<description><![CDATA[
Accumulating evidence shows that auditory cortex (AC) of humans, and other primates, is involved in more complex cognitive processes than feature segregation only, which are shaped by experience-dependent plasticity and thus likely show substantial individual variability. However, thus far, individual variability of ACs has been considered a methodological impediment rather than a phenomenon of theoretical importance. Here, we examined the variability of ACs using intrinsic functional connectivity patterns in humans and macaques. Our results demonstrate that in humans, functional variability is 1) greater near the non-primary than primary ACs, 2) greater in ACs than comparable visual areas, and 3) greater in the left than right ACs. Remarkably similar modality differences and lateralization of variability were observed in macaques. These connectivity-based findings are consistent with a confirmatory task-based fMRI analysis. The quantitative proof of the exceptional variability of ACs has implications for understanding the evolution of advanced auditory functions in humans.
]]></description>
<dc:creator>Ren, J.</dc:creator>
<dc:creator>Liu, H.</dc:creator>
<dc:creator>Xu, T.</dc:creator>
<dc:creator>Schoeppe, F.</dc:creator>
<dc:creator>Wang, D.</dc:creator>
<dc:creator>Li, M.</dc:creator>
<dc:creator>Lin, Y.</dc:creator>
<dc:creator>Ramirez, J.</dc:creator>
<dc:creator>Lu, J.</dc:creator>
<dc:creator>Li, L.</dc:creator>
<dc:creator>Ahveninen, J.</dc:creator>
<dc:date>2020-01-07</dc:date>
<dc:identifier>doi:10.1101/2020.01.06.895474</dc:identifier>
<dc:title><![CDATA[Individual variability in functional organization of the human and monkey auditory cortex]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-01-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.01.06.896555v1?rss=1">
<title>
<![CDATA[
Regulation of HDAC2-PDX1 by RNF125 defines pancreatic cancer development 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.01.06.896555v1?rss=1</link>
<description><![CDATA[
There is an urgent need to define mechanisms underlying pancreatic adenocarcinoma (PDA) development. Our studies of ubiquitin ligases that may underlie PDA development led us to identify and characterize RNF125. We show that RNF125 exhibits nuclear expression in acinar cells, with reduced and largely cytosolic expression in ductal cells, PanIN and PDA specimens. We find that RNF125 interacts with histone deacetylase 2 (HDAC2) and promotes its non-canonical K63-linked ubiquitination. Inhibition of HDAC2 activity by RNF125 resulted in elevated expression of the pancreatic and duodenal homeobox 1 (PDX1). Correspondingly, inhibition of RNF125 expression enhanced organoid growth in culture and orthotopic tumor development. Conversely, restoration of PDX1 levels in human or mouse PDA cells and organoids depleted of RNF125, inhibited cell proliferation and growth, while expression of HDAC2 enhanced it. Notably, higher expression of RNF125 and PDX1 coincided with differentiated tumor phenotypes, and better outcome in PDA patients. In demonstrating the importance of RNF125 control of PDX1 expression via HDAC2 ubiquitination in PDA development, our findings highlight markers (RNF125, PDX1) and targets (HDAC2) for monitoring and possible treatment of PDA.
]]></description>
<dc:creator>Hasnis, E.</dc:creator>
<dc:creator>Kim, H.</dc:creator>
<dc:creator>Verma, S.</dc:creator>
<dc:creator>Feng, Y.</dc:creator>
<dc:creator>Almog, R.</dc:creator>
<dc:creator>Matsliah, S.</dc:creator>
<dc:creator>Vavinskaya, V.</dc:creator>
<dc:creator>Alelbaum, R.</dc:creator>
<dc:creator>Ben-Ishay, O.</dc:creator>
<dc:creator>Tuveson, D.</dc:creator>
<dc:creator>Sears, R.</dc:creator>
<dc:creator>Ronai, Z. A.</dc:creator>
<dc:date>2020-01-07</dc:date>
<dc:identifier>doi:10.1101/2020.01.06.896555</dc:identifier>
<dc:title><![CDATA[Regulation of HDAC2-PDX1 by RNF125 defines pancreatic cancer development]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-01-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.01.07.897132v1?rss=1">
<title>
<![CDATA[
Human cytomegalovirus evades ZAP detection by suppressing CpG dinucleotides in the major immediate early genes. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.01.07.897132v1?rss=1</link>
<description><![CDATA[
The genomes of RNA and small DNA viruses of vertebrates display significant suppression of CpG dinucleotide frequencies. Artificially increasing dinucleotide frequencies results in substantial attenuation of virus replication, suggesting that these compositional changes may facilitate recognition of non-self RNA sequences. Recently, the interferon inducible protein ZAP, was identified as the host factor responsible for sensing CpG in viral RNA, through direct binding and possibly downstream targeting for degradation. Using an arrayed interferon stimulated gene expression library screen, we identified ZAPS, and its associated factor TRIM25, as direct inhibitors of human cytomegalovirus (HCMV) replication. Exogenous expression of ZAPS and TRIM25 significantly reduced virus replication while knockdown resulted in increased virus replication. HCMV displays a strikingly heterogeneous pattern of CpG representation with a specific suppression of CpGs within the IE1 major immediate early transcript which is absent in subsequently expressed genes. We demonstrated that suppression of CpG dinucleotides in the IE1 gene allows evasion of inhibitory effects of ZAP. During HCMV infection, expression of ZAP and TRIM25 are rapidly reduced, removing pressure to suppress dinucleotide frequencies in viral genes expressed after the immediate early genes, while acute virus replication and high levels of ZAP are mutually exclusive. Finally, we show that TRIM25 regulates alternative splicing between the ZAP short and long isoforms during HCMV infection and interferon induction, with knockdown of TRIM25 resulting in decreased ZAPS and corresponding increased ZAPL expression. These results demonstrate for the first time that ZAP is a potent host restriction factor against large DNA viruses and that HCMV evades ZAP detection through suppression of CpG dinucleotides within the major immediate early transcripts. Furthermore, TRIM25 is required for efficient upregulation of the interferon inducible short isoform of ZAP through regulation of alternative splicing.
]]></description>
<dc:creator>Lee, C.-H.</dc:creator>
<dc:creator>Chiweshe, S.</dc:creator>
<dc:creator>McCormick, D.</dc:creator>
<dc:creator>Raper, A. R.</dc:creator>
<dc:creator>wickenhagen, A.</dc:creator>
<dc:creator>DeFilippis, V. R.</dc:creator>
<dc:creator>GAUNT, E.</dc:creator>
<dc:creator>Simmonds, P.</dc:creator>
<dc:creator>Wilson, S. J.</dc:creator>
<dc:creator>Grey, F.</dc:creator>
<dc:date>2020-01-08</dc:date>
<dc:identifier>doi:10.1101/2020.01.07.897132</dc:identifier>
<dc:title><![CDATA[Human cytomegalovirus evades ZAP detection by suppressing CpG dinucleotides in the major immediate early genes.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-01-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.01.07.897694v1?rss=1">
<title>
<![CDATA[
Local retinoic acid directs emergence of the extraocular muscle functional unit 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.01.07.897694v1?rss=1</link>
<description><![CDATA[
Coordinated development of muscles, tendons, and their attachment sites ensures emergence of functional musculoskeletal units that are adapted to diverse anatomical demands among different species. How these different tissues are patterned and functionally assembled during embryogenesis is poorly understood. Here, we investigated the morphogenesis of extraocular muscles (EOMs), an evolutionary conserved cranial muscle group that is crucial for the coordinated movement of the eyeballs and for visual acuity. By means of lineage analysis, we redefined the cellular origins of periocular connective tissues interacting with the EOMs, which do not arise exclusively from neural crest mesenchyme as previously thought. Using 3D imaging approaches, we established an integrative blueprint for the EOM functional unit. By doing so, we identified a developmental time window where individual EOMs emerge from a unique muscle anlage and establish insertions in the sclera, which sets these muscles apart from classical muscle-to-bone type of insertions. Further, we demonstrate that the eyeballs are a source of diffusible retinoic acid that allow their targeting by the EOMs in a temporal and dose dependent manner. Using genetically modified mice and inhibitor treatments, we find that endogenous local variations in the concentration of retinoids contribute to the establishment of tendon condensations and attachment sites that precede the initiation of muscle patterning. Collectively, our results highlight how global and site-specific programs are deployed for the assembly of muscle functional units with precise definition of muscle shapes and topographical wiring of their tendon attachments.
]]></description>
<dc:creator>Comai, G. E.</dc:creator>
<dc:creator>Tesarova, M.</dc:creator>
<dc:creator>Dupe, V.</dc:creator>
<dc:creator>Rhinn, M.</dc:creator>
<dc:creator>Vallecillo Garcia, P.</dc:creator>
<dc:creator>da Silva, F.</dc:creator>
<dc:creator>Feret, B.</dc:creator>
<dc:creator>Exelby, K.</dc:creator>
<dc:creator>Dolle, P.</dc:creator>
<dc:creator>Carlsson, L.</dc:creator>
<dc:creator>Pryce, B.</dc:creator>
<dc:creator>Spitz, F.</dc:creator>
<dc:creator>Stricker, S.</dc:creator>
<dc:creator>Zikmund, T.</dc:creator>
<dc:creator>Kaiser, J.</dc:creator>
<dc:creator>Briscoe, J.</dc:creator>
<dc:creator>Schedl, A.</dc:creator>
<dc:creator>Ghyselinck, N. B.</dc:creator>
<dc:creator>Schweitzer, R.</dc:creator>
<dc:creator>Tajbakhsh, S.</dc:creator>
<dc:date>2020-01-07</dc:date>
<dc:identifier>doi:10.1101/2020.01.07.897694</dc:identifier>
<dc:title><![CDATA[Local retinoic acid directs emergence of the extraocular muscle functional unit]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-01-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.01.08.898627v1?rss=1">
<title>
<![CDATA[
A new long-read RNA-seq analysis approach identifies and quantifies novel transcripts of very large genes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.01.08.898627v1?rss=1</link>
<description><![CDATA[
RNA-seq is widely used for studying gene expression, but commonly used sequencing platforms produce short reads that only span up to two exon-junctions per read. This makes it difficult to accurately determine the composition and phasing of exons within transcripts. Although long-read sequencing improves this issue, it is not amenable to precise quantitation, which limits its utility for differential expression studies. We used long-read isoform sequencing combined with a novel analysis approach to compare alternative splicing of large, repetitive structural genes in muscles. Analysis of muscle structural genes that produce medium (Nrap -5kb), large (nebulin - 22 kb) and very-large (titin - 106 kb) transcripts in cardiac muscle, and fast and slow skeletal muscles identified unannotated exons for each of these ubiquitous muscle genes. This also identified differential exon usage and phasing for these genes between the different muscle types. By mapping the in-phase transcript structures to known annotations, we also identified and quantified previously unannotated transcripts. Results were confirmed by endpoint PCR and Sanger sequencing, which revealed muscle-type specific differential expression of these novel transcripts. The improved transcript identification and quantification demonstrated by our approach removes previous impediments to studies aimed at quantitative differential expression of ultra-long transcripts.
]]></description>
<dc:creator>Uapinyoying, P. B.</dc:creator>
<dc:creator>Goecks, J.</dc:creator>
<dc:creator>Knoblach, S.</dc:creator>
<dc:creator>Panchapakesan, K.</dc:creator>
<dc:creator>Bonnemann, C.</dc:creator>
<dc:creator>Partridge, T.</dc:creator>
<dc:creator>Jaiswal, J.</dc:creator>
<dc:creator>Hoffman, E.</dc:creator>
<dc:date>2020-01-09</dc:date>
<dc:identifier>doi:10.1101/2020.01.08.898627</dc:identifier>
<dc:title><![CDATA[A new long-read RNA-seq analysis approach identifies and quantifies novel transcripts of very large genes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-01-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.01.08.899088v1?rss=1">
<title>
<![CDATA[
Portable Automated Rapid Testing ( PART ) for auditory research: Validation in a normal hearing population 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.01.08.899088v1?rss=1</link>
<description><![CDATA[
This study aims to determine the degree to which Portable Automated Rapid Testing (PART), a freely-available program running on a tablet computer, is capable of reproducing standard laboratory results. Undergraduate students were assigned to one of three within-subject conditions that examined repeatability of performance on a battery of psychoacoustical tests of temporal fine structure processing, spectro-temporal amplitude modulation, and targets in competition. The Repeatability condition examined test/retest with the same system, the Headphones condition examined the effects of varying headphones (passive and active noise-attenuating), and the Noise condition examined repeatability in the presence of recorded cafeteria noise. In general, performance on the test battery showed high repeatability, even across manipulated conditions, and was similar to that reported in the literature. These data serve as validation that suprathreshold psychoacoustical tests can be made accessible to run on consumer-grade hardware and performed in less controlled settings. This dataset also provides a distribution of thresholds that can be used as a normative baseline against which auditory dysfunction can be identified in future work.
]]></description>
<dc:creator>Lelo de Larrea-Mancera, E. S.</dc:creator>
<dc:creator>Stavropoulos, T.</dc:creator>
<dc:creator>Hoover, E. C.</dc:creator>
<dc:creator>Eddins, D. A.</dc:creator>
<dc:creator>Gallun, F. J.</dc:creator>
<dc:creator>Seitz, A. R.</dc:creator>
<dc:date>2020-01-09</dc:date>
<dc:identifier>doi:10.1101/2020.01.08.899088</dc:identifier>
<dc:title><![CDATA[Portable Automated Rapid Testing ( PART ) for auditory research: Validation in a normal hearing population]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-01-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.01.09.900092v1?rss=1">
<title>
<![CDATA[
Discovery of a novel inhibitory neuron class, the L-Stellate cells of the cochlear nucleus 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.01.09.900092v1?rss=1</link>
<description><![CDATA[
Auditory processing depends upon inhibitory signaling by interneurons, even at its earliest stages in the ventral cochlear nucleus (VCN). Remarkably, to date only a single subtype of inhibitory neuron has been documented in the VCN, a projection neuron termed the D-stellate cell. With the use of a transgenic mouse line, optical clearing and imaging techniques, combined with electrophysiological tools, we revealed a population of glycinergic cells in the VCN distinct from the D-stellate cell. These novel multipolar glycinergic cells were smaller in soma size and dendritic area, but over 10-fold more numerous than D-stellate cells. They were activated by auditory nerve fibers and T-stellate cells, and made local inhibitory synaptic contacts on principal cells of the VCN. Given their abundance, combined with their narrow dendritic fields and axonal projections, it is likely that these neurons, here termed L-stellate cells, play a significant role in frequency-specific processing of acoustic signals.
]]></description>
<dc:creator>Ngodup, T.</dc:creator>
<dc:creator>Romero, G. E.</dc:creator>
<dc:creator>Trussell, L.</dc:creator>
<dc:date>2020-01-09</dc:date>
<dc:identifier>doi:10.1101/2020.01.09.900092</dc:identifier>
<dc:title><![CDATA[Discovery of a novel inhibitory neuron class, the L-Stellate cells of the cochlear nucleus]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-01-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.01.10.902692v1?rss=1">
<title>
<![CDATA[
MATERNAL ZIKA VIRUS (ZIKV) INFECTION FOLLOWING VAGINAL INOCULATION WITH ZIKV-INFECTED SEMEN IN THE TIMED-PREGNANT OLIVE BABOON 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.01.10.902692v1?rss=1</link>
<description><![CDATA[
ZIKV infection is associated with pregnancy loss, fetal microcephaly and other malformations. While Aedes sp. of mosquito are the primary vector for ZIKV, sexual transmission of ZIKV is a significant route of infection. ZIKV has been documented in human, mouse and non-human primate (NHP) semen. It is critical to establish NHP models of vertical transfer of ZIKV that recapitulate human ZIKV pathogenesis. We hypothesized that vaginal deposition of ZIKV infected baboon semen would lead to maternal infection and vertical transfer in the olive baboon (Papio anubis). Timed pregnant baboons (n=6) were inoculated via vaginal deposition of baboon semen containing 106 ffu ZIKV (n=3, French Polynesian isolate:H/PF/2013, n=3 Puerto Rican isolate:PRVABC59) at mid-gestation (86-95 days gestation [dG]; term 183dG) on day (d) 0 (all dams), and then at 7 day intervals through three weeks. Maternal blood, saliva and cervico-vaginal washes were obtained at select days post-inoculation. Animals were euthanized at 28 days post initial inoculation (dpi; n=5) or 39 dpi (n=1) and maternal/fetal tissues collected. vRNA was quantified by qPCR. Viremia was achieved in 3/3 FP ZIKV infected dams and 2/3 PR ZIKV. ZIKV RNA was detected in cvw (5/6 dams;). ZIKV RNA was detected in lymph nodes, but not ovary, uterus, cervix or vagina in the FP ZIKV dams but was detected in uterus, vagina and lymph nodes. Placenta, amniotic fluid and all fetal tissues were ZIKV RNA negative in the FP infected dams whereas 2/3 PR infected dam placentas were ZIKV RNA positive. We conclude that ZIKV infected semen is a means of ZIKV transmission during pregnancy in primates. The PR isolate appeared more capable of wide spread dissemination to tissues, including placenta compared to the FP strain.

IMPORTANCEDue to its established link to pregnancy loss, microcephaly and other major congenital anomalies, Zika virus (ZIKV) remains a worldwide health threat. Although mosquitoes are the primary means of ZIVK transmission, sexual transmission in human populations is well documented and provides a means for widespread dissemination of the virus. Differences in viremia, tissue distribution, immune responses and pregnancy outcome from sexually transmitted ZIKV compared to the subcutaneous route of infection are needed to better clinically manage ZIKV in pregnancy. Through our previous work, we have developed the olive baboon as a non-human primate model of ZIKV infection that is permissible to ZIKV infection via the subcutaneous route of inoculation and transfer of ZIKV to the fetus in pregnancy. The current study evaluated the course of ZIKV infection after vaginal inoculation of ZIKV in pregnant baboons at mid-gestation using baboon semen as the carrier and comparing two isolates of ZIKV, the French Polynesian isolate first associated with microcephaly and the Puerto Rican isolate, associated with an increased risk of microcephaly observed in the Americas.
]]></description>
<dc:creator>Gurung, S.</dc:creator>
<dc:creator>Nadeau, H.</dc:creator>
<dc:creator>Maxted, M.</dc:creator>
<dc:creator>Peregrine, J.</dc:creator>
<dc:creator>Reuter, D. N.</dc:creator>
<dc:creator>Norris, A.</dc:creator>
<dc:creator>Edwards, R. K.</dc:creator>
<dc:creator>Hyatt, K.</dc:creator>
<dc:creator>Singleton, K.</dc:creator>
<dc:creator>Papin, J. F.</dc:creator>
<dc:creator>Myers, D. A.</dc:creator>
<dc:date>2020-01-11</dc:date>
<dc:identifier>doi:10.1101/2020.01.10.902692</dc:identifier>
<dc:title><![CDATA[MATERNAL ZIKA VIRUS (ZIKV) INFECTION FOLLOWING VAGINAL INOCULATION WITH ZIKV-INFECTED SEMEN IN THE TIMED-PREGNANT OLIVE BABOON]]></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/10.1101/2020.01.13.905471v1?rss=1">
<title>
<![CDATA[
Dietary Amino Acids Impact LRRK2-induced Neurodegeneration in Parkinson's Disease Models 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.01.13.905471v1?rss=1</link>
<description><![CDATA[
The G2019S mutation in leucine-rich repeat kinase 2 (LRRK2) is a common cause of Parkinsons disease (PD) and results in age-related dopamine neuron loss and locomotor dysfunction in Drosophila melanogaster through an aberrant increase in bulk neuronal protein synthesis. Under non-pathologic conditions, protein synthesis is tightly controlled by metabolic regulation. Whether nutritional and metabolic influences on protein synthesis can modulate the pathogenic effect of LRRK2 on protein synthesis and thereby impact neuronal loss is a key unresolved question. Here, we show that LRRK2 G2019S-induced neurodegeneration is critically dependent on dietary amino acid content. Low dietary amino acid concentration prevents aberrant protein synthesis and blocks LRRK2 G2019S-mediated neurodegeneration in Drosophila and rat primary neurons. Unexpectedly, a moderately high amino acid diet also blocks dopamine neuron loss and motor deficits in Drosophila through a separate mechanism involving stress-responsive activation of 5-AMP-activated protein kinase (AMPK) and neuroprotective induction of autophagy, implicating the importance of protein homeostasis to neuronal viability. At the highest amino acid diet of the range tested, PD-related neurodegeneration occurs in an age-related manner, but is also observed in control strains, suggesting that it is independent of mutant LRRK2 expression. We propose that dietary influences on protein synthesis and autophagy are critical determinants of LRRK2 neurodegeneration, opening up possibilities for future therapeutic intervention.
]]></description>
<dc:creator>Chittoor-Vinod, V. G.</dc:creator>
<dc:creator>Villalobos-Cantor, S.</dc:creator>
<dc:creator>Roshak, H.</dc:creator>
<dc:creator>Shea, K.</dc:creator>
<dc:creator>Martin, I.</dc:creator>
<dc:date>2020-01-16</dc:date>
<dc:identifier>doi:10.1101/2020.01.13.905471</dc:identifier>
<dc:title><![CDATA[Dietary Amino Acids Impact LRRK2-induced Neurodegeneration in Parkinson's Disease Models]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-01-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.01.14.905000v1?rss=1">
<title>
<![CDATA[
Complementary effects of adaptation and gain control on sound encoding in primary auditory cortex 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.01.14.905000v1?rss=1</link>
<description><![CDATA[
An important step toward understanding how the brain represents complex natural sounds is to develop accurate models of auditory coding by single neurons. A common model for auditory coding is the linear-nonlinear spectro-temporal receptive field (LN model). The LN model accounts for many features of auditory tuning, but it cannot account for long-lasting effects of sensory context on sound-evoked activity. Two mechanisms that may support these contextual effects are short-term plasticity (STP) and contrast-dependent gain control (GC), each of which has inspired an expanded version of the LN model. Both of these models improve performance over the LN model, but they have never been compared directly. Thus, it is unclear whether they account for distinct processes or describe the same phenomenon in different ways. To address this question, we recorded activity of neurons in primary auditory cortex of awake ferrets during presentation of natural sounds. We then fit models incorporating one nonlinear mechanism (GC or STP) or both (GC+STP) using this single dataset, and measured the correlation between the models predictions and the recorded neural activity. Both the STP and GC models performed significantly better than the LN model, but the GC+STP model performed better than either individual model. We also quantified the similarity between STP and GC model predictions and found only modest equivalence between them. Similar results were observed for a smaller dataset collected in clean and noisy acoustic contexts. These results suggest that the STP and GC models describe distinct, complementary processes in the auditory system.

Significance StatementComputational models are used widely to study neural sensory coding. However, models developed in separate studies are often difficult to compare because of differences in stimuli and experimental preparation. This study develops an approach for making systematic comparisons between models that measures the net benefit of incorporating additional nonlinear elements into models of auditory encoding. This approach was then used to compare two different hypotheses for how sensory context, that is, slow changes in the statistics of the acoustic environment, influences activity in auditory cortex. Both models accounted for complementary aspects of the neural response, indicating that a hybrid model incorporating elements of both models provides the most complete characterization of auditory processing.
]]></description>
<dc:creator>Pennington, J. R.</dc:creator>
<dc:creator>David, S. V.</dc:creator>
<dc:date>2020-01-15</dc:date>
<dc:identifier>doi:10.1101/2020.01.14.905000</dc:identifier>
<dc:title><![CDATA[Complementary effects of adaptation and gain control on sound encoding in primary auditory cortex]]></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/10.1101/2020.01.14.906198v1?rss=1">
<title>
<![CDATA[
Genetic ablation of the mitochondrial ribosome in Plasmodium falciparum sensitizes the human malaria parasite to antimalarial drugs targeting mitochondrial functions 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.01.14.906198v1?rss=1</link>
<description><![CDATA[
The mitochondrion of malaria parasites contains clinically validated drug targets. Within Plasmodium spp., the mitochondrial DNA (mtDNA) is only 6 kb long, being the smallest mitochondrial genome among all eukaryotes. The mtDNA encodes only three proteins of the mitochondrial electron transport chain and [~] 27 small, fragmented rRNA genes in length of 22-195 nucleotides. The rRNA fragments are thought to form a mitochondrial ribosome (mitoribosome), together with ribosomal proteins imported from the cytosol. The mitoribosome of Plasmodium falciparum has been shown to be essential for maintenance of the mitochondrial membrane potential and parasite viability. However, the role of mitoribosomes in sustaining the metabolic status of the parasite mitochondrion remains unknown. Here, among the 14 annotated mitoribosomal proteins of the small subunit of P. falciparum, we verified the localization and tested the essentiality of three candidates (PfmtRPS12, PfmtRPS17, PfmtRPS18), employing a CRISPR/Cas9 mediated conditional knockdown tool. Using immuno-electron microscopy, we provided evidence that the mitoribosome is closely associated with the mitochondrial inner membrane in the parasite. Upon knockdown of the mitoribosome, the parasites became hypersensitive to inhibitors targeting the bc1 complex, dihydroorotate dehydrogenase and F1Fo ATP synthase complex. Furthermore, knockdown of the mitoribosome blocked the pyrimidine biosynthesis pathway and reduced the pool of pyrimidine nucleotides. Together, our data suggest that disruption of the P. falciparum mitoribosome compromises the metabolic capability of the mitochondrion, rendering the parasite hypersensitive to a panel of inhibitors targeting mitochondrial functions.
]]></description>
<dc:creator>Ling, L.</dc:creator>
<dc:creator>Mulaka, M.</dc:creator>
<dc:creator>Munro, J.</dc:creator>
<dc:creator>Dass, S.</dc:creator>
<dc:creator>Mather, M. W.</dc:creator>
<dc:creator>Riscoe, M. K.</dc:creator>
<dc:creator>Llinas, M.</dc:creator>
<dc:creator>Zhou, J.</dc:creator>
<dc:creator>Ke, H.</dc:creator>
<dc:date>2020-01-14</dc:date>
<dc:identifier>doi:10.1101/2020.01.14.906198</dc:identifier>
<dc:title><![CDATA[Genetic ablation of the mitochondrial ribosome in Plasmodium falciparum sensitizes the human malaria parasite to antimalarial drugs targeting mitochondrial functions]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-01-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.01.16.907071v1?rss=1">
<title>
<![CDATA[
Transient Commensal Clonal Interactions Can Drive Tumor Metastasis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.01.16.907071v1?rss=1</link>
<description><![CDATA[
To interrogate functional heterogeneity and crosstalk between tumor cells, we generated clonal populations from a patient-derived ovarian clear cell carcinoma model which forms malignant ascites and solid peritoneal tumors upon intraperitoneal transplantation in mice. The clonal populations were engineered with secreted Gaussia luciferase to monitor tumor growth dynamics and tagged with a unique DNA barcode to track their fate in multiclonal mixtures during tumor progression. Only one clone, CL31, grew robustly, generating exclusively malignant ascites. However, multiclonal mixtures formed large solid peritoneal metastases, populated almost entirely by CL31, suggesting that transient cooperative interclonal interactions were sufficient to promote metastasis of CL31. CL31 uniquely harbored ERBB2 amplification, and its acquired metastatic trait was dependent on transient exposure to amphiregulin, which was exclusively secreted by non-tumorigenic clones. Amphiregulin enhanced CL31 mesothelial clearance, a prerequisite for metastasis. These findings demonstrate that transient, ostensibly innocuous tumor subpopulations can promote metastases via "hit- and-run" commensal interactions.
]]></description>
<dc:creator>Naffar-Abu Amara, S.</dc:creator>
<dc:creator>Kuiken, H. J.</dc:creator>
<dc:creator>Selfors, L. M.</dc:creator>
<dc:creator>Butler, T.</dc:creator>
<dc:creator>Leung, M. L.</dc:creator>
<dc:creator>Leung, C. T.</dc:creator>
<dc:creator>Kuhn, E. P.</dc:creator>
<dc:creator>Kolarova, T. R.</dc:creator>
<dc:creator>Hage, C.</dc:creator>
<dc:creator>Ganesh, K.</dc:creator>
<dc:creator>Foster, R.</dc:creator>
<dc:creator>Rueda, B. R.</dc:creator>
<dc:creator>Aktipis, A.</dc:creator>
<dc:creator>Spellman, P. T.</dc:creator>
<dc:creator>Ince, T.</dc:creator>
<dc:creator>Navin, N.</dc:creator>
<dc:creator>Mills, G. B.</dc:creator>
<dc:creator>Bronson, R. T.</dc:creator>
<dc:creator>Brugge, J. S. S.</dc:creator>
<dc:date>2020-01-16</dc:date>
<dc:identifier>doi:10.1101/2020.01.16.907071</dc:identifier>
<dc:title><![CDATA[Transient Commensal Clonal Interactions Can Drive Tumor Metastasis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-01-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.01.17.910893v1?rss=1">
<title>
<![CDATA[
Intra-striatal AAV2.retro administration leads to extensive retrograde transport in the rhesus macaque brain: implications for disease modeling and therapeutic development 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.01.17.910893v1?rss=1</link>
<description><![CDATA[
Recently, AAV2.retro, a new capsid variant capable of efficient retrograde transport in brain, was generated in mice using a directed evolution approach. However, it remains unclear to what degree transport will be recapitulated in the substantially larger and more complex nonhuman primate (NHP) brain. Here, we compared the biodistribution of AAV2.retro with its parent serotype, AAV2, in adult macaques following delivery into the caudate and putamen, brain regions which comprise the striatum. While AAV2 transduction was primarily limited to the injected brain regions, AAV2.retro transduced cells in the striatum and in dozens of cortical and subcortical regions with known striatal afferents. We then evaluated the capability of AAV2.retro to deliver disease-related gene cargo to biologically-relevant NHP brain circuits by packaging a fragment of human mutant HTT, the causative gene mutation in Huntingtons disease. Following intra-striatal delivery, pathological mHTT-positive protein aggregates were distributed widely among cognitive, motor, and limbic cortico-basal ganglia circuits. Together, these studies demonstrate strong retrograde transport of AAV2.retro in NHP brain, highlight its utility in developing novel NHP models of brain disease and suggest its potential for querying circuit function and delivering therapeutic genes in the brain, particularly where treating dysfunctional circuits, versus single brain regions, is warranted.
]]></description>
<dc:creator>Weiss, A. R.</dc:creator>
<dc:creator>Liguore, W. A.</dc:creator>
<dc:creator>Domire, J. S.</dc:creator>
<dc:creator>Button, D.</dc:creator>
<dc:creator>McBride, J. L.</dc:creator>
<dc:date>2020-01-18</dc:date>
<dc:identifier>doi:10.1101/2020.01.17.910893</dc:identifier>
<dc:title><![CDATA[Intra-striatal AAV2.retro administration leads to extensive retrograde transport in the rhesus macaque brain: implications for disease modeling and therapeutic development]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-01-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.01.26.920504v1?rss=1">
<title>
<![CDATA[
Purine-responsive expression of the Leishmania donovani NT3 purine nucleobase transporter is mediated by a conserved RNA stem-loop 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.01.26.920504v1?rss=1</link>
<description><![CDATA[
The ability to modulate gene expression in response to changes in the host environment is essential for survival of the kinetoplastid parasite Leishmania. Unlike most eukaryotes, gene expression in kinetoplastids is predominately regulated post-transcriptionally. Consequently, RNA-binding proteins (RBPs) and mRNA-encoded sequence elements serve as primary determinants of gene regulation in these organisms; however, few have been ascribed roles in specific stress-response pathways. Leishmania lack the capacity for de novo purine synthesis and must scavenge these essential nutrients from the host. Leishmania have evolved a robust stress response to withstand sustained periods of purine scarcity during their lifecycle. The purine nucleobase transporter, LdNT3, is among the most substantially upregulated proteins in purine-starved L. donovani. Here we report that the post-translational stability of the LdNT3 protein is unchanged in response to purine starvation. Instead, LdNT3 upregulation is primarily mediated by a 33 nucleotide (nt) sequence in the LdNT3 mRNA 3-untranslated region that is predicted to adopt a stem-loop structure. While this sequence is highly conserved within the mRNAs of orthologous transporters in multiple kinetoplastid species, putative stem-loops from L. donovani and Trypanosoma brucei nucleobase transporter mRNAs are not functionally interchangeable for purine-responsive regulation. Through mutational analysis of the element, we demonstrate that species specificity is attributable to just three variant bases within the predicted loop. Finally, we provide evidence that the abundance of the trans-acting factor that binds the LdNT3 stem-loop in vivo is substantially higher than required for regulation of LdNT3 alone, implying a potential role in regulating other purine-responsive genes.
]]></description>
<dc:creator>Licon, M. H.</dc:creator>
<dc:creator>Yates, P. A.</dc:creator>
<dc:date>2020-01-27</dc:date>
<dc:identifier>doi:10.1101/2020.01.26.920504</dc:identifier>
<dc:title><![CDATA[Purine-responsive expression of the Leishmania donovani NT3 purine nucleobase transporter is mediated by a conserved RNA stem-loop]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-01-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.01.27.910919v1?rss=1">
<title>
<![CDATA[
A global lipid map defines a network essential for Zika virus replication 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.01.27.910919v1?rss=1</link>
<description><![CDATA[
Zika virus (ZIKV), an arbovirus of global concern, remodels intracellular membranes to form replication sites. How ZIKV dysregulates lipid networks to allow this, and consequences for disease, is poorly understood. Here, we performed comprehensive lipidomics to create a lipid network map during ZIKV infection. We found that ZIKV significantly alters host lipid composition, with the most striking changes seen within subclasses of sphingolipids. Ectopic expression of ZIKV NS4B protein resulted in similar changes, demonstrating a role for NS4B in modulating sphingolipid pathways. Disruption of sphingolipid biosynthesis in various cell types, including human neural progenitor cells, blocked ZIKV infection. Additionally, the sphingolipid ceramide redistributes to ZIKV replication sites and increasing ceramide levels by multiple pathways sensitizes cells to ZIKV infection. Thus, we identify a sphingolipid metabolic network with a critical role in ZIKV replication and show that ceramide flux is a key mediator of ZIKV infection.
]]></description>
<dc:creator>Leier, H. C.</dc:creator>
<dc:creator>Weinstein, J. B.</dc:creator>
<dc:creator>Kyle, J. E.</dc:creator>
<dc:creator>Lee, J.-Y.</dc:creator>
<dc:creator>Bramer, L. M.</dc:creator>
<dc:creator>Stratton, K. G.</dc:creator>
<dc:creator>Kempthorne, D.</dc:creator>
<dc:creator>Navratil, A. R.</dc:creator>
<dc:creator>Tafesse, E. G.</dc:creator>
<dc:creator>Hornemann, T.</dc:creator>
<dc:creator>Messer, W. B.</dc:creator>
<dc:creator>Dennis, E. A.</dc:creator>
<dc:creator>Metz, T. O.</dc:creator>
<dc:creator>Barklis, E.</dc:creator>
<dc:creator>Tafesse, F. G.</dc:creator>
<dc:date>2020-01-27</dc:date>
<dc:identifier>doi:10.1101/2020.01.27.910919</dc:identifier>
<dc:title><![CDATA[A global lipid map defines a network essential for Zika virus replication]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-01-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.02.13.947226v1?rss=1">
<title>
<![CDATA[
Gut carriage of antimicrobial resistance genes in women exposed to small-scale poultry farms in rural Uganda: a feasibility study 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.02.13.947226v1?rss=1</link>
<description><![CDATA[
BackgroundAntibiotic use as growth promoters for livestock is presumed to be a major contributor to the acquisition of antimicrobial resistance (AMR) genes in humans, yet data evaluating AMR patterns in the setting of animal exposure are limited to observational studies that do not capture data from prior to livestock introduction.

MethodsWe performed a feasibility study by recruiting a subset of women in a delayed-start randomized controlled trial of small-scale chicken farming in order to examine the prevalence of clinically-relevant AMR genes. Stool samples were obtained at baseline and one year from five intervention women who received chickens at the start of the study, six control women who did not receive chickens until the end of the study, and from chickens provided to the control group at the end of the study. Stool was screened for 87 clinically significant AMR genes using a commercially available qPCR array (Qiagen).

ResultsChickens harbored 23 AMR genes from classes also found in humans as well as vancomycin and additional {beta}-lactamase resistance genes. After one year of exposure to chickens, six new AMR genes were detected in controls and seven new AMR genes were detected in the intervention group. Women who had direct contact with the chickens sampled in the study had greater similarities in AMR resistance gene patterns to chickens than those who did not have direct contact with chickens sampled (p = 0.006). There was a trend towards increased similarity in AMR gene patterns with chickens at one year (p = 0.12).

ConclusionsChickens and humans in this study harbored AMR genes from many antimicrobial classes at both baseline and follow up timepoints. Studies designed to evaluate human AMR genes in the setting of animal exposure should account for high baseline AMR rates, and consider collecting concomitant animal samples, human samples, and environmental samples over time to determine the directionality and source of AMR genes.

Trial registration: ClinicalTrials.gov Identifier: NCT02619227
]]></description>
<dc:creator>Weil, A. A.</dc:creator>
<dc:creator>Debela, M. D.</dc:creator>
<dc:creator>Muyanja, D.</dc:creator>
<dc:creator>Kakuhikire, B.</dc:creator>
<dc:creator>Baguma, C.</dc:creator>
<dc:creator>Bangsberg, D.</dc:creator>
<dc:creator>Tsai, A. C.</dc:creator>
<dc:creator>Lai, P. S.</dc:creator>
<dc:date>2020-02-13</dc:date>
<dc:identifier>doi:10.1101/2020.02.13.947226</dc:identifier>
<dc:title><![CDATA[Gut carriage of antimicrobial resistance genes in women exposed to small-scale poultry farms in rural Uganda: a feasibility study]]></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/10.1101/2020.02.13.948117v1?rss=1">
<title>
<![CDATA[
Composition of the Lipid Droplets of the Bovine Corpus Luteum 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.02.13.948117v1?rss=1</link>
<description><![CDATA[
2.Establishment and maintenance of pregnancy is dependent on progesterone synthesized by luteal tissue in the ovary. Our objective was to identify the characteristics of lipid droplets (LDs) in ovarian steroidogenic cells. We hypothesized that LDs are a major feature of steroidogenic luteal cells and store cholesteryl esters. Bovine luteal tissue was used for whole tissue analysis. Further analyses were performed on isolated ovarian steroidogenic cells: granulosa and theca cells of the follicle, and small/large luteal cells. Isolated luteal LDs were collected for lipid/protein analyses. Luteal tissue contained perilipins 2/3/5, hormone-sensitive lipase and abhydrolase domain containing 5. Luteal tissue was enriched in TGs compared to other tissues, except of adipose tissue. Large and small luteal cells were distinguished from follicular cells by the presence of LDs and LD-associated proteins. Furthermore, LDs from large luteal cells were numerous and small; whereas, LDs from small luteal cells were large and less numerous. Isolated LDs contained nearly all of the TGs and cholesteryl esters present in luteal tissue. Isolated luteal LDs were composed primarily of TG, with lesser amounts of cholesteryl esters, diglyceride and other phospholipids. Bovine luteal tissue LDs are distinct from LDs in other bovine tissues, including follicular steroidogenic cells.
]]></description>
<dc:creator>Talbott, H. A.</dc:creator>
<dc:creator>Plewes, M. R.</dc:creator>
<dc:creator>Krause, C.</dc:creator>
<dc:creator>Hou, X.</dc:creator>
<dc:creator>Zhang, P.</dc:creator>
<dc:creator>Rizzo, W. B.</dc:creator>
<dc:creator>Wood, J. R.</dc:creator>
<dc:creator>Cupp, A. S.</dc:creator>
<dc:creator>Davis, J. S.</dc:creator>
<dc:date>2020-02-13</dc:date>
<dc:identifier>doi:10.1101/2020.02.13.948117</dc:identifier>
<dc:title><![CDATA[Composition of the Lipid Droplets of the Bovine Corpus Luteum]]></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/10.1101/2020.02.13.948141v1?rss=1">
<title>
<![CDATA[
Inflammation promotes tumor aggression by stimulating stromal cell-dependent collagen crosslinking and stromal stiffening 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.02.13.948141v1?rss=1</link>
<description><![CDATA[
Collagen deposition and stromal stiffening accompany malignancy, compromise treatment, and promote tumor aggression. Clarifying the molecular nature of and the factors that regulate extracellular matrix stiffening in tumors should identify biomarkers to stratify patients for therapy and therapeutic interventions to improve outcome. We profiled lysyl hydroxylase- and lysyl oxidase-mediated collagen crosslinks and quantified the greatest abundance of total and complex collagen crosslinks in more aggressive human breast cancer subtypes with the stiffest stroma. These tissues also harbored the highest number of tumor-associated macrophages (TAM), whose therapeutic ablation not only reduced metastasis, but also concomitantly decreased accumulation of collagen crosslinks and stromal stiffening. Epithelial-targeted expression of the crosslinking enzyme lysyl oxidase had no impact on collagen crosslinking in PyMT mammary tumors, whereas stromal cell targeting did. Consistently, stromal cells in microdissected human tumors expressed the highest level of collagen crosslinking enzymes. Immunohistochemical analysis of a cohort of breast cancer patient biopsies revealed that stromal expression of lysyl hydroxylase two, an enzyme that induces hydroxylysine aldehyde-derived collagen crosslinks and stromal stiffening correlated significantly disease specific mortality. The findings link tissue inflammation, stromal cell-mediated collagen crosslinking and stiffening to tumor aggression and identify lysyl hydroxylase two as a novel stromal biomarker.

SignificanceWe show infiltrating macrophages induce stromal fibroblast, and not epithelial, expression of collagen crosslinking enzymes that drive tumor stiffening. Stromal enzyme LH2 is significantly upregulated in breast cancer patients with the stiffest stroma, the most trivalent HLCCs and the worst prognosis, underscoring its potential as a biomarker and therapeutic target.
]]></description>
<dc:creator>Maller, O.</dc:creator>
<dc:creator>Drain, A. P.</dc:creator>
<dc:creator>Barrett, A. S.</dc:creator>
<dc:creator>Borgquist, S.</dc:creator>
<dc:creator>Ruffell, B.</dc:creator>
<dc:creator>Thanh, P. T.</dc:creator>
<dc:creator>Gruosso, T.</dc:creator>
<dc:creator>Kuasne, H.</dc:creator>
<dc:creator>Lakins, J. N.</dc:creator>
<dc:creator>Acerbi, I.</dc:creator>
<dc:creator>Barnes, J. M.</dc:creator>
<dc:creator>Nemkov, T.</dc:creator>
<dc:creator>Chauhan, A.</dc:creator>
<dc:creator>Gruenberg, J.</dc:creator>
<dc:creator>Nasir, A.</dc:creator>
<dc:creator>Bjarnadottir, O.</dc:creator>
<dc:creator>Werb, Z.</dc:creator>
<dc:creator>Kabos, P.</dc:creator>
<dc:creator>Hwang, E. S.</dc:creator>
<dc:creator>Park, M.</dc:creator>
<dc:creator>Coussens, L. M.</dc:creator>
<dc:creator>Nelson, A. C.</dc:creator>
<dc:creator>Hansen, K. C.</dc:creator>
<dc:creator>Weaver, V. M.</dc:creator>
<dc:date>2020-02-13</dc:date>
<dc:identifier>doi:10.1101/2020.02.13.948141</dc:identifier>
<dc:title><![CDATA[Inflammation promotes tumor aggression by stimulating stromal cell-dependent collagen crosslinking and stromal stiffening]]></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/10.1101/2020.02.19.957118v1?rss=1">
<title>
<![CDATA[
Mucin 4 Protects Female Mice from Coronavirus Pathogenesis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.02.19.957118v1?rss=1</link>
<description><![CDATA[
Using incipient lines of the Collaborative Cross (CC), a murine genetic reference population, we previously identified a quantitative trait loci (QTL) associated with low SARS-CoV titer. In this study, we integrated sequence information and RNA expression of genes within the QTL to identify mucin 4 (Muc4) as a high priority candidate for controlling SARS-CoV titer in the lung. To test this hypothesis, we infected Muc4-/- mice and found that female, but not male, Muc4-/- mice developed more weight loss and disease following infection with SARS-CoV. Female Muc4-/- mice also had more difficulty breathing despite reduced lung pathology; however, no change in viral titers was observed. Comparing across viral families, studies with chikungunya virus, a mosquito-borne arthralgic virus, suggests that Muc4s impact on viral pathogenesis may be widespread. Although not confirming the original titer QTL, our data identifies a role for Muc4 in the SARS-CoV disease and viral pathogenesis.

ImportanceGiven the recent emergence of SARS-CoV-2, this work suggest that Muc4 expression plays a protective role in female mice not conserved in male mice following SARS-CoV infection. With the SARS-CoV-2 outbreak continuing, treatments that modulate or enhance Muc4 activity may provide an avenue for treatment and improved outcomes. In addition, the work highlights the importance of studying host factors including host genetics and biological sex as key parameters influencing infection and disease outcomes.
]]></description>
<dc:creator>Plante, J. A.</dc:creator>
<dc:creator>Plante, K.</dc:creator>
<dc:creator>Gralinski, L.</dc:creator>
<dc:creator>Beall, A.</dc:creator>
<dc:creator>Ferris, M. T.</dc:creator>
<dc:creator>Bottomly, D.</dc:creator>
<dc:creator>Green, R. R.</dc:creator>
<dc:creator>McWeeney, S.</dc:creator>
<dc:creator>Heise, M. T.</dc:creator>
<dc:creator>Baric, R. S.</dc:creator>
<dc:creator>Menachery, V. D.</dc:creator>
<dc:date>2020-02-20</dc:date>
<dc:identifier>doi:10.1101/2020.02.19.957118</dc:identifier>
<dc:title><![CDATA[Mucin 4 Protects Female Mice from Coronavirus Pathogenesis]]></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/10.1101/2020.02.21.960237v1?rss=1">
<title>
<![CDATA[
Accumulative deamidation of human lens protein γS-crystallin leads to partially unfolded intermediates with enhanced aggregation propensity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.02.21.960237v1?rss=1</link>
<description><![CDATA[
Age-related cataract is a major cause of blindness worldwide. Yet, the molecular mechanisms whereby large, light scattering aggregates form is poorly understood, because of the complexity of the aggregates isolated from human lenses. The predominant proteins in the lens are structural proteins called crystallins. The {gamma}S-crystallin is heavily modified in cataractous lenses by deamidation, which introduces a negative charge at labile asparagine residues. The effects of deamidation at asparagines, N14, N76, and N143, were mimicked by replacing the asparagine with aspartate using site-directed mutagenesis. The effects of these surface deamidations on the stability, unfolding, and aggregation properties of {gamma}S were determined using dynamic light scattering, chemical and thermal-denaturation, and hydrogen-deuterium exchange with mass spectrometry. We found that a small population of all the deamidation mimics aggregated directly into large light scattering bodies with a radius greater than 10 nm that contributed 14-60% of the total scattering intensity compared to 7% for WT under the same conditions. A possible mechanism was identified under partially denaturing conditions, where deamidation led to significantly more rapid unfolding and aggregation particularly for N76D compared to WT. The triple mutant was further destabilized, reflecting the enhanced aggregation properties of N14D and N143D. Thus, the effects of deamidation were both site-specific and cumulative. A-crystallin was ineffective at acting as a chaperone to prevent the aggregation of destabilized, deamidated {gamma}S. It is concluded that surface deamidations, while causing minimal structural disruption individually, progressively destabilize crystallin proteins, leading to their unfolding and precipitation in aged and cataractous lenses.
]]></description>
<dc:creator>Vetter, C. J.</dc:creator>
<dc:creator>Thorn, D.</dc:creator>
<dc:creator>Wheeler, S. G.</dc:creator>
<dc:creator>Mundorff, C.</dc:creator>
<dc:creator>Halverson-Kolkind, K.</dc:creator>
<dc:creator>Carver, J. A.</dc:creator>
<dc:creator>David, L. L.</dc:creator>
<dc:creator>Lampi, K. J.</dc:creator>
<dc:date>2020-02-22</dc:date>
<dc:identifier>doi:10.1101/2020.02.21.960237</dc:identifier>
<dc:title><![CDATA[Accumulative deamidation of human lens protein γS-crystallin leads to partially unfolded intermediates with enhanced aggregation propensity]]></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/10.1101/2020.02.24.963587v1?rss=1">
<title>
<![CDATA[
Mesoscopic-scale functional networks in the primate amygdala 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.02.24.963587v1?rss=1</link>
<description><![CDATA[
The primate amygdala performs multiple functions that may be related to the anatomical heterogeneity of its nuclei. At the level of single neurons, each function is reflected in stimulus- and task-specific responses. Given that neurons with a particular response profile are not clustered in any of the nuclei, single units may be too fine-grained to shed light on the mesoscale organization of the amygdala. We have extracted from local field potentials recorded simultaneously from multiple locations within the primate amygdala (Macaca mulatta) spatially defined and statistically separable responses to visual, tactile, and auditory stimuli. A generalized eigendecomposition-based method of source separation isolated coactivity patterns, or components, that in neurophysiological terms correspond to putative subnetworks. Some component spatial patterns mapped onto the anatomical organization of the amygdala, while other components reflected integration across nuclei. These components differentiated between visual, tactile, and auditory stimuli suggesting the presence of functionally distinct parallel subnetworks.
]]></description>
<dc:creator>Morrow, J. K.</dc:creator>
<dc:creator>Cohen, M. X.</dc:creator>
<dc:creator>Gothard, K. M.</dc:creator>
<dc:date>2020-02-26</dc:date>
<dc:identifier>doi:10.1101/2020.02.24.963587</dc:identifier>
<dc:title><![CDATA[Mesoscopic-scale functional networks in the primate amygdala]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-02-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.02.26.966747v1?rss=1">
<title>
<![CDATA[
Gut Microbiome Pattern Reflects Healthy Aging and Predicts Extended Survival in Humans 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.02.26.966747v1?rss=1</link>
<description><![CDATA[
The gut microbiome has important effects on human health, yet its importance in human aging remains unclear. Using two independent cohorts comprising 4582 individuals across the adult lifespan we demonstrate that, starting in mid-to-late adulthood, gut microbiomes become increasingly unique with age. This uniqueness pattern is strongly associated with gut microbial amino acid derivatives circulating within the bloodstream, many of which have been previously identified as longevity biomarkers. At the latest stages of human life, two distinct patterns emerge wherein individuals in good health show continued microbial drift toward a unique compositional state, while the same drift is absent in individuals who perform worse on a number of validated health measures. The identified healthy aging pattern is characterized by an overall depletion of core genera found across most humans - primarily a depletion in the nearly ubiquitous genus Bacteroides. Consistently, retaining a high Bacteroides dominance into extreme age, or, equivalently, having a low gut microbiome uniqueness score, predicts decreased survival in a four-year follow-up. Our comprehensive analysis identifies the gut microbiome as a novel component of healthy aging, with important implications for the worlds growing older population.
]]></description>
<dc:creator>Wilmanski, T.</dc:creator>
<dc:creator>Diener, C.</dc:creator>
<dc:creator>Rappaport, N.</dc:creator>
<dc:creator>Patwardhan, S.</dc:creator>
<dc:creator>Wiedrick, J.</dc:creator>
<dc:creator>Lapidus, J.</dc:creator>
<dc:creator>Earls, J. C.</dc:creator>
<dc:creator>Zimmer, A.</dc:creator>
<dc:creator>Glusman, G.</dc:creator>
<dc:creator>Robinson, M.</dc:creator>
<dc:creator>Yurkovich, J. T.</dc:creator>
<dc:creator>Kado, D. M.</dc:creator>
<dc:creator>Cauley, J. A.</dc:creator>
<dc:creator>Zmuda, J.</dc:creator>
<dc:creator>Lane, N. E.</dc:creator>
<dc:creator>Magis, A. T.</dc:creator>
<dc:creator>Lovejoy, J. C.</dc:creator>
<dc:creator>Gibbons, S. M.</dc:creator>
<dc:creator>Hood, L.</dc:creator>
<dc:creator>Orwoll, E. S.</dc:creator>
<dc:creator>Price, N. D.</dc:creator>
<dc:date>2020-02-28</dc:date>
<dc:identifier>doi:10.1101/2020.02.26.966747</dc:identifier>
<dc:title><![CDATA[Gut Microbiome Pattern Reflects Healthy Aging and Predicts Extended Survival in Humans]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-02-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.02.26.966762v1?rss=1">
<title>
<![CDATA[
Crosstalk between invadopodia and the extracellular matrix 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.02.26.966762v1?rss=1</link>
<description><![CDATA[
The scaffold protein Tks5 is required for invadopodia-mediated cancer invasion both in vitro and in vivo. We have previously also revealed a role for Tks5 in tumor cell growth using three-dimensional (3D) culture model systems and mouse transplantation experiments. Here we use both 3D and high-density fibrillar collagen (HDFC) culture to demonstrate that native type I collagen, but not a form lacking the telopeptides, stimulated Tks5-dependent growth, which was dependent on the DDR collagen receptors. We used microenvironmental microarray (MEMA) technology to determine that laminin, collagen I, fibronectin and tropoelastin also stimulated invadopodia formation. A Tks5-specific monoclonal antibody revealed its expression both on microtubules and at invadopodia. High- and super-resolution microscopy of cells in and on collagen was then used to place Tks5 at the base of invadopodia, separated from much of the actin and cortactin, but coincident with both matrix metalloprotease and cathepsin proteolytic activity. Inhibition of the Src family kinases, cathepsins or metalloproteases all reduced invadopodia length but each had distinct effects on Tks5 localization. These studies highlight the crosstalk between invadopodia and extracellular matrix components, and reveal the invadopodium to be a spatially complex structure.
]]></description>
<dc:creator>Iizuka, S.</dc:creator>
<dc:creator>Leon, R. P.</dc:creator>
<dc:creator>Gribbin, K. P.</dc:creator>
<dc:creator>Zhang, Y.</dc:creator>
<dc:creator>Navarro, J.</dc:creator>
<dc:creator>Smith, R.</dc:creator>
<dc:creator>Devlin, K.</dc:creator>
<dc:creator>Wang, L. G.</dc:creator>
<dc:creator>Gibbs, S. L.</dc:creator>
<dc:creator>Korkola, J.</dc:creator>
<dc:creator>Nan, X.</dc:creator>
<dc:creator>Courtneidge, S. A.</dc:creator>
<dc:date>2020-02-27</dc:date>
<dc:identifier>doi:10.1101/2020.02.26.966762</dc:identifier>
<dc:title><![CDATA[Crosstalk between invadopodia and the extracellular matrix]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-02-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.02.26.966820v1?rss=1">
<title>
<![CDATA[
In vivo repair of a protein underlying a neurological disorder by programmable RNA editing 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.02.26.966820v1?rss=1</link>
<description><![CDATA[
RNA base editing is gaining momentum as an approach to repair mutations, but its application to neurological disease has not been established. We have succeeded in directed transcript editing of a pathological mutation in a mouse model of the neurodevelopmental disease, Rett syndrome. Specifically, we directed editing of a guanosine to adenosine mutation in RNA encoding Methyl CpG Binding Protein 2 (MECP2). Repair was mediated by injecting the hippocampus of juvenile Rett mice with an adeno-associated virus expressing both an engineered enzyme containing the catalytic domain of Adenosine Deaminase Acting on RNA 2 and a Mecp2 targeting guide. After one month, 50% of Mecp2 RNA was recoded in three different hippocampal neuronal subtypes, and the ability of MeCP2 protein to associate with heterochromatin was similarly restored to 50% of wild-type levels. This study represents the first in vivo programmable RNA editing applied to a model of neurological disease.
]]></description>
<dc:creator>Sinnamon, J.</dc:creator>
<dc:creator>Kim, S. Y.</dc:creator>
<dc:creator>Fisk, J. R.</dc:creator>
<dc:creator>Song, Z.</dc:creator>
<dc:creator>Nakai, H.</dc:creator>
<dc:creator>Jeng, S.</dc:creator>
<dc:creator>McWeeney, S. K.</dc:creator>
<dc:creator>Mandel, G.</dc:creator>
<dc:date>2020-02-27</dc:date>
<dc:identifier>doi:10.1101/2020.02.26.966820</dc:identifier>
<dc:title><![CDATA[In vivo repair of a protein underlying a neurological disorder by programmable RNA editing]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-02-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.02.26.966895v1?rss=1">
<title>
<![CDATA[
Chemical and structural investigation of the paroxetine-human serotonin transporter complex 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.02.26.966895v1?rss=1</link>
<description><![CDATA[
Antidepressants target the serotonin transporter (SERT) by inhibiting serotonin reuptake. Structural and biochemical studies aiming to understand the binding of small-molecules to conformationally dynamic transporters like SERT often require thermostabilizing mutations and antibodies to stabilize a specific conformation. Such modifications to SERT have led to questions about the relationships of these structures to the bona fide conformation and inhibitor binding poses of the wild-type transporter. To address these concerns, we characterized wild-type SERT with truncated N- and C-termini and thermostabilized variants of SERT bound with paroxetine using x-ray crystallography, single particle cryo-EM and biochemical techniques. Moreover, using a C-H functionalization approach to synthesize enantiopure analogues, we replaced the halide of the fluorophenyl group in paroxetine with either bromine or iodine. We then exploited the anomalous scattering of Br and I to define the pose of the respective paroxetine analog. These structures provide mutually consistent insights into how paroxetine and its analogs bind to the central substrate-binding site of SERT, stabilize the outward-open conformation, and inhibit serotonin transport.
]]></description>
<dc:creator>Coleman, J. A.</dc:creator>
<dc:creator>Navratna, V.</dc:creator>
<dc:creator>Antermite, D.</dc:creator>
<dc:creator>Yang, D.</dc:creator>
<dc:creator>Bull, J. A.</dc:creator>
<dc:creator>Gouaux, E.</dc:creator>
<dc:date>2020-02-27</dc:date>
<dc:identifier>doi:10.1101/2020.02.26.966895</dc:identifier>
<dc:title><![CDATA[Chemical and structural investigation of the paroxetine-human serotonin transporter complex]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-02-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.02.27.968099v1?rss=1">
<title>
<![CDATA[
Subphase Material Stabilizes Films of Pulmonary Surfactant 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.02.27.968099v1?rss=1</link>
<description><![CDATA[
When compressed by the shrinking alveolar surface area during exhalation, films of pulmonary surfactant in situ reduce surface tension to levels, at which surfactant monolayers collapse from the surface in vitro. Vesicles of pulmonary surfactant added below these monolayers slow collapse. X-ray scattering here determined the structural changes that improve stability. Grazing incidence X-ray diffraction on monolayers of extracted calf surfactant detected an ordered phase. Mixtures of dipalmitoyl phosphatidylcholine and cholesterol, but not the phospholipid alone, mimic that structure. At concentrations that stabilize the monolayers, vesicles in the subphase had no effect on the unit cell, and the film remained monomolecular. The added vesicles, however, produced a concentration-dependent increase in the diffracted intensity. These results suggest that the enhanced resistance to collapse results from components of an ordered interfacial phase which partition from subphase to the surface, increasing the area of the ordered structure.

SIGNIFICANCELow alveolar surface tensions are essential for maintaining the integrity of the pulmonary air-sacks during normal breathing. Films of pulmonary surfactant cause the low tensions. The interfacial structures required for the low surface tensions remain uncertain. These studies used X-ray scattering to determine the initial structure of pulmonary surfactant monolayers, and to establish how vesicles of pulmonary surfactant enhance the ability of those initial monolayers to sustain low tensions. The initial monolayers contained ordered structures that differ from the crystalline forms widely speculated to occur in alveolar films. The added vesicles had no effect on the local structure of the initial monolayer, but substantially increased the area of the ordered regions. This structural change reasonably explains the functional improvement.
]]></description>
<dc:creator>Andreev, K.</dc:creator>
<dc:creator>Martynowycz, M.</dc:creator>
<dc:creator>Kuzmenko, I.</dc:creator>
<dc:creator>Bu, W.</dc:creator>
<dc:creator>Hall, S. B.</dc:creator>
<dc:creator>Gidalevitz, D.</dc:creator>
<dc:date>2020-02-28</dc:date>
<dc:identifier>doi:10.1101/2020.02.27.968099</dc:identifier>
<dc:title><![CDATA[Subphase Material Stabilizes Films of Pulmonary Surfactant]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-02-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.02.27.968305v1?rss=1">
<title>
<![CDATA[
The serine-threonine kinase TAO3 promotes cancer invasion and tumor growth by facilitating trafficking of endosomes containing the invadopodia scaffold TKS5α 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.02.27.968305v1?rss=1</link>
<description><![CDATA[
Invadopodia are actin-based proteolytic membrane protrusions required for invasive behavior and tumor growth. We used our high-content screening assay to identify kinases impacting invadopodia formation. Among the top hits we selected TAO3, a STE20-like kinase of the GCK subfamily, for further analysis. TAO3 was over-expressed in many human cancers, and regulated invadopodia formation in melanoma, breast and bladder cancers. Furthermore, TAO3 catalytic activity facilitated melanoma growth in 3-dimensional matrices and in vivo. We developed potent catalytic inhibitors of TAO3 that inhibited invadopodia formation and function, and tumor cell extravasation and growth. Using these inhibitors, we determined that TAO3 activity was required for endosomal trafficking of TKS5, an obligate invadopodia scaffold protein. A phosphoproteomics screen for TAO3 substrates revealed the dynein subunit protein LIC2 as a relevant substrate. Knockdown of LIC2 or expression of a phosphomimetic form promoted invadopodia formation. Thus, TAO3 is a new therapeutic target with a distinct mechanism of action.

SIGNIFICANCETargeting tumor invasive behavior represents an understudied opportunity. We used an unbiased screening approach to identify kinases required for invadopodia formation and function. We validated TAO3, both genetically and with a novel inhibitor, and determined TAO3 function. Our data support clinical development of this class of target.
]]></description>
<dc:creator>Iizuka, S.</dc:creator>
<dc:creator>Quintavalle, M.</dc:creator>
<dc:creator>Ceja Navarro, J.</dc:creator>
<dc:creator>Gribbin, K. P.</dc:creator>
<dc:creator>Ardecky, R. J.</dc:creator>
<dc:creator>Abelman, M.</dc:creator>
<dc:creator>Ma, C.-T.</dc:creator>
<dc:creator>Sergienko, E.</dc:creator>
<dc:creator>Zeng, F.-Y.</dc:creator>
<dc:creator>Pass, I.</dc:creator>
<dc:creator>Thomas, G.</dc:creator>
<dc:creator>McWeeney, S.</dc:creator>
<dc:creator>Hassig, C. A.</dc:creator>
<dc:creator>Pinkerton, A. B.</dc:creator>
<dc:creator>Courtneidge, S. A.</dc:creator>
<dc:date>2020-02-28</dc:date>
<dc:identifier>doi:10.1101/2020.02.27.968305</dc:identifier>
<dc:title><![CDATA[The serine-threonine kinase TAO3 promotes cancer invasion and tumor growth by facilitating trafficking of endosomes containing the invadopodia scaffold TKS5α]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-02-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.03.02.974063v1?rss=1">
<title>
<![CDATA[
BoneMA - Synthesis and Characterization of a Methacrylated Bone-derived Hydrogel for Bioprinting of Vascularized Tissues 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.03.02.974063v1?rss=1</link>
<description><![CDATA[
It has long been proposed that recapitulating the extracellular matrix (ECM) of native human tissues in the laboratory may enhance the regenerative capacity of engineered scaffolds in-vivo. Organ- and tissue-derived decellularized ECM biomaterials have been widely used for tissue repair, especially due to their intrinsic biochemical cues that can facilitate repair and regeneration. The main purpose of this study was to synthesize a new photocrosslinkable human bone-derived ECM hydrogel for bioprinting of vascularized scaffolds. To that end, we demineralized and decellularized human bone fragments to obtain a bone matrix, which was further processed and functionalized with methacrylate groups to form a photocrosslinkable methacrylate bone ECM hydrogel - BoneMA. The mechanical properties of BoneMA were tunable, with the elastic modulus increasing as a function of photocrosslinking time, while still retaining the nanoscale features of the polymer networks. The intrinsic cell-compatibility of the bone matrix ensured the synthesis of a highly cytocompatible hydrogel. The bioprinted BoneMA scaffolds supported vascularization of endothelial cells and within a day led to the formation of interconnected vascular networks. We propose that such a quick vascular network formation was due to the host of pro-angiogenic biomolecules present in the bone ECM matrix. Further, we also demonstrate the bioprintability of BoneMA in microdimensions as injectable ECM-based building blocks for microscale tissue engineering in a minimally invasive manner. We conclude that BoneMA may be a useful hydrogel system for tissue engineering and regenerative medicine.
]]></description>
<dc:creator>S., P. P.</dc:creator>
<dc:creator>Athirasala, A.</dc:creator>
<dc:creator>Tahayeri, A.</dc:creator>
<dc:creator>Abdelmoniem, R.</dc:creator>
<dc:creator>George, A.</dc:creator>
<dc:creator>Bertassoni, L. E.</dc:creator>
<dc:date>2020-03-04</dc:date>
<dc:identifier>doi:10.1101/2020.03.02.974063</dc:identifier>
<dc:title><![CDATA[BoneMA - Synthesis and Characterization of a Methacrylated Bone-derived Hydrogel for Bioprinting of Vascularized Tissues]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-03-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.03.02.974154v1?rss=1">
<title>
<![CDATA[
Conserved His-Gly motif of acid-sensing ion channels resides in a reentrant loop implicated in gating and ion selectivity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.03.02.974154v1?rss=1</link>
<description><![CDATA[
Acid-sensing ion channels (ASICs) are proton-gated members of the epithelial sodium channel/degenerin (ENaC/DEG) superfamily of ion channels and are expressed throughout central and peripheral nervous systems. The homotrimeric splice variant ASIC1a has been implicated in nociception, fear memory, mood disorders and ischemia. Here we extract full-length chicken ASIC1a (cASIC1a) from cell membranes using styrene maleic acid (SMA) copolymer, yielding structures of ASIC1a channels in both high pH resting and low pH desensitized conformations by single-particle cryo-electron microscopy (cryo-EM). The structures of resting and desensitized channels reveal a reentrant loop at the amino terminus of ASIC1a that includes the highly conserved  His-Gly (HG) motif. The reentrant loop lines the lower ion permeation pathway and buttresses the  Gly-Ala-Ser (GAS) constriction, thus providing a structural explanation for the role of the His-Gly dipeptide in the structure and function of ASICs.
]]></description>
<dc:creator>Yoder, N.</dc:creator>
<dc:creator>Gouaux, E.</dc:creator>
<dc:date>2020-03-03</dc:date>
<dc:identifier>doi:10.1101/2020.03.02.974154</dc:identifier>
<dc:title><![CDATA[Conserved His-Gly motif of acid-sensing ion channels resides in a reentrant loop implicated in gating and ion selectivity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-03-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.03.02.974204v1?rss=1">
<title>
<![CDATA[
3D printing of Microgel-loaded Modular LEGO-like Cages as Instructive Scaffolds for Tissue Engineering 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.03.02.974204v1?rss=1</link>
<description><![CDATA[
Biomaterial scaffolds have served as the foundation of tissue engineering and regenerative medicine. However, scaffold systems are often difficult to scale in size or shape in order to fit defect-specific dimensions, and thus provide only limited spatiotemporal control of therapeutic delivery and host tissue responses. Here, a lithography-based three-dimensional (3D) printing strategy is used to fabricate a novel miniaturized modular LEGO-like cage scaffold system, which can be assembled and scaled manually with ease. Scalability is based on an intuitive concept of stacking modules, like conventional LEGO blocks, allowing for literally thousands of potential geometric configurations, and without the need for specialized equipment. Moreover, the modular hollow-cage design allows each unit to be loaded with biologic cargo of different compositions, thus enabling controllable and easy patterning of therapeutics within the material in 3D. In summary, the concept of miniaturized cage designs with such straight-forward assembly and scalability, as well as controllable loading properties, is a flexible platform that can be extended to a wide range of materials for improved biological performance.

TOC3D printed LEGO-like hollow microcages can be easily assembled, adjoined, and stacked-up to suit the complexity of defect tissues; aid spatial loading of cells and biomolecules; instruct cells migration three-dimensionally; and facilitate cell invasion and neovascularization in-vivo, thus accelerating the process of tissue healing and new tissue formation.

O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=190 SRC="FIGDIR/small/974204v1_ufig1.gif" ALT="Figure 1">
View larger version (88K):
org.highwire.dtl.DTLVardef@1a97a72org.highwire.dtl.DTLVardef@1a60fbeorg.highwire.dtl.DTLVardef@1538f13org.highwire.dtl.DTLVardef@d6345b_HPS_FORMAT_FIGEXP  M_FIG C_FIG
]]></description>
<dc:creator>Hipfinger, C.</dc:creator>
<dc:creator>Subbiah, R.</dc:creator>
<dc:creator>Tahayeri, A.</dc:creator>
<dc:creator>Athirasala, A.</dc:creator>
<dc:creator>Horsophonphong, S.</dc:creator>
<dc:creator>Thrivikraman, G.</dc:creator>
<dc:creator>Franca, C. M.</dc:creator>
<dc:creator>Cunha, D. A.</dc:creator>
<dc:creator>Mansoorifar, A.</dc:creator>
<dc:creator>Zahariev, A.</dc:creator>
<dc:creator>Jones, J. M.</dc:creator>
<dc:creator>Coelho, P. G.</dc:creator>
<dc:creator>Witek, L.</dc:creator>
<dc:creator>Xie, H.</dc:creator>
<dc:creator>Guldberg, R. E.</dc:creator>
<dc:creator>Bertassoni, L. E.</dc:creator>
<dc:date>2020-03-04</dc:date>
<dc:identifier>doi:10.1101/2020.03.02.974204</dc:identifier>
<dc:title><![CDATA[3D printing of Microgel-loaded Modular LEGO-like Cages as Instructive Scaffolds for Tissue Engineering]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-03-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.03.03.973727v1?rss=1">
<title>
<![CDATA[
The physical form of microbial ligands bypasses the need for dendritic cell migration to stimulate adaptive immunity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.03.03.973727v1?rss=1</link>
<description><![CDATA[
A central paradigm of immunology is that the innate immune system first detects infectious agents in peripheral tissues, shortly after a pathogen has breached an epithelial barrier. This detection event is mediated by pattern recognition receptors in phagocytes, which then migrate to draining lymph nodes (dLNs), where information of a microbial encounter is conveyed to T and B lymphocytes to generate adaptive immunity. Through the study of fungal moieties, we present data that challenge this model. We found that soluble fungal polysaccharides are immunosilent in the periphery, but become potent immunogens in the dLN. These ligands completely bypass the need of phagocyte migration and, instead, directly activate an immune response that is most similar to those that typify viral infections. These data establish a class of microbial products that violate a central tenet of the immunological lexicon and illustrate that the physical form (not just the chemical structure) impacts innate and adaptive immunity.
]]></description>
<dc:creator>Borriello, F.</dc:creator>
<dc:creator>Spreafico, R.</dc:creator>
<dc:creator>Poli, V.</dc:creator>
<dc:creator>Chou, J.</dc:creator>
<dc:creator>Barrett, N.</dc:creator>
<dc:creator>Lacanfora, L.</dc:creator>
<dc:creator>Franco, M.</dc:creator>
<dc:creator>Marongiu, L.</dc:creator>
<dc:creator>Iwakura, Y.</dc:creator>
<dc:creator>Pucci, F.</dc:creator>
<dc:creator>Kruppa, M.</dc:creator>
<dc:creator>Ma, Z.</dc:creator>
<dc:creator>Williams, D.</dc:creator>
<dc:creator>Zanoni, I.</dc:creator>
<dc:date>2020-03-05</dc:date>
<dc:identifier>doi:10.1101/2020.03.03.973727</dc:identifier>
<dc:title><![CDATA[The physical form of microbial ligands bypasses the need for dendritic cell migration to stimulate adaptive immunity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-03-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.03.14.991042v1?rss=1">
<title>
<![CDATA[
Functional Identification of Specialized Basal Ganglia Circuits that Regulate Vocal Motor Sequences 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.03.14.991042v1?rss=1</link>
<description><![CDATA[
Disruption of the transcription factor FoxP2, which is enriched in the basal ganglia, impair vocal development in humans and songbirds. The basal ganglia are essential for the selection and sequencing of motor actions, but the circuit mechanisms governing accurate sequencing of learned vocalizations are unknown. Here, we show expression of FoxP2 in the basal ganglia is vital for the fluent initiation and termination of birdsong, and the maintenance of song syllable sequencing in adulthood. Knockdown of FoxP2 imbalances dopamine receptor expression across striatal direct-like and indirect-like pathways, suggesting a role of dopaminergic signaling in regulating vocal-motor sequencing. Confirming this prediction, we show that phasic dopamine activation, and not inhibition, during singing drives repetition of song syllables, thus also impairing fluent initiation and termination of birdsong. These findings demonstrate discrete circuit origins for the dysfluent repetition of vocal elements, a phenotype commonly observed in speech disorders.
]]></description>
<dc:creator>Xiao, L.</dc:creator>
<dc:creator>Merullo, D. P.</dc:creator>
<dc:creator>Cao, M.</dc:creator>
<dc:creator>Co, M.</dc:creator>
<dc:creator>Kulkarni, A.</dc:creator>
<dc:creator>Konopka, G.</dc:creator>
<dc:creator>Roberts, T. F.</dc:creator>
<dc:date>2020-03-16</dc:date>
<dc:identifier>doi:10.1101/2020.03.14.991042</dc:identifier>
<dc:title><![CDATA[Functional Identification of Specialized Basal Ganglia Circuits that Regulate Vocal Motor Sequences]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-03-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.03.14.992016v1?rss=1">
<title>
<![CDATA[
Autism-linked gene FoxP1 selectively regulates the cultural transmission of learned vocalizations 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.03.14.992016v1?rss=1</link>
<description><![CDATA[
Autism spectrum disorders (ASD) are characterized by impaired learning of culturally transmitted behaviors like social skills, speech, and language1-3. These behaviors are learned by copying parents and other social models during development, a two-stage process that involves forming memories of appropriate behaviors during social experiences and then using those memories to guide imitation. How ASD-linked genes impair these often-intertwined aspects of learning is not known, thereby limiting our understanding of the developmental progression of ASD and the targeting of therapeutic interventions. Here we show that these aspects of learning are dissociable and that the ASD-linked gene FoxP1 selectively impairs learning from social experience, but not behavioral imitation. Haploinsufficiency of FOXP1 in humans causes FOXP1 syndrome, a neurodevelopmental disorder typified by severe disruptions in speech and language development, and other ASD-associated symptoms4,5. We tested how knockdown of FoxP1 (FP1-KD) affects the cultural transmission of vocal behaviors in zebra finches, a songbird that learns by memorizing and vocally copying the song of an adult  song-tutor. We find that FP1-KD blocks song learning in juvenile birds by selectively impairing their ability to encode a memory during social experiences with a songtutor. These learning deficits are linked to disruptions in experience-driven structural and functional plasticity. However, if birds are exposed to tutor-song prior to FP1-KD, their ability to imitate that song during development is unaffected. Thus, FP1-KD impairs cultural transmission of vocalizations by disrupting the ability to form appropriate vocal memories, yet spares the ability to use previously acquired memories to guide vocal learning. This indicates that learning from social experience may be particularly vulnerable in FOXP1 syndrome.
]]></description>
<dc:creator>Garcia-Oscos, F.</dc:creator>
<dc:creator>Koch, T.</dc:creator>
<dc:creator>Pancholi, H.</dc:creator>
<dc:creator>Trusel, M.</dc:creator>
<dc:creator>Daliparthi, V.</dc:creator>
<dc:creator>Ayhan, F.</dc:creator>
<dc:creator>Co, M.</dc:creator>
<dc:creator>Alam, D. H.</dc:creator>
<dc:creator>Holdway, J. E.</dc:creator>
<dc:creator>Konopka, G.</dc:creator>
<dc:creator>Roberts, T. F.</dc:creator>
<dc:date>2020-03-16</dc:date>
<dc:identifier>doi:10.1101/2020.03.14.992016</dc:identifier>
<dc:title><![CDATA[Autism-linked gene FoxP1 selectively regulates the cultural transmission of learned vocalizations]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-03-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.03.19.999664v1?rss=1">
<title>
<![CDATA[
Orally Bioavailable Endochin-like Quinolone Carbonate Ester Prodrug Reduces Toxoplasma gondii Brain Cysts 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.03.19.999664v1?rss=1</link>
<description><![CDATA[
Toxoplasmosis is a potentially fatal infection for immunocompromised people and the developing fetus. Current medicines for toxoplasmosis have high rates of adverse effects that interfere with therapeutic and prophylactic regimens. Endochin-like quinolones (ELQs) are potent inhibitors of Toxoplasma gondii proliferation in vitro and in animal models of acute and latent infection. ELQ-316, in particular, was found to be effective orally against acute toxoplasmosis in mice and highly selective for the T. gondii cytochrome b over the human cytochrome b. Despite oral efficacy, the high crystallinity of ELQ-316 limits oral absorption, plasma concentrations and therapeutic potential. A carbonate ester prodrug of ELQ-316, ELQ-334, was created to decrease crystallinity and increase oral bioavailability, which resulted in a six-fold increase in both Cmax (maximum plasma concentration) and AUC (area under the curve) of ELQ-316. The increased bioavailability of ELQ-316, when administered as ELQ-334, resulted in greater efficacy than the equivalent dose of ELQ-316 against acute toxoplasmosis and had similar efficacy against latent toxoplasmosis compared to intraperitoneal administration of ELQ-316. Carbonate ester prodrugs are a successful strategy to overcome the limited oral bioavailability of ELQs for the treatment of toxoplasmosis.
]]></description>
<dc:creator>Doggett, J. S.</dc:creator>
<dc:creator>Schultz, T. L.</dc:creator>
<dc:creator>Miller, A.</dc:creator>
<dc:creator>Bruzual, I.</dc:creator>
<dc:creator>Pou, S.</dc:creator>
<dc:creator>Winter, R. W.</dc:creator>
<dc:creator>Dodean, R.</dc:creator>
<dc:creator>Zakharov, L. N.</dc:creator>
<dc:creator>Nilsen, A.</dc:creator>
<dc:creator>Riscoe, M. K.</dc:creator>
<dc:creator>Carruthers, V. B.</dc:creator>
<dc:date>2020-03-20</dc:date>
<dc:identifier>doi:10.1101/2020.03.19.999664</dc:identifier>
<dc:title><![CDATA[Orally Bioavailable Endochin-like Quinolone Carbonate Ester Prodrug Reduces Toxoplasma gondii Brain Cysts]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-03-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.03.19.999672v1?rss=1">
<title>
<![CDATA[
Efficacy of Guanabenz Combination Therapy Against Chronic Toxoplasmosis Across Multiple Mouse Strains 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.03.19.999672v1?rss=1</link>
<description><![CDATA[
Toxoplasma gondii, an obligate intracellular parasite that can cause life-threatening acute disease, differentiates into a quiescent cyst stage to establish lifelong chronic infections in animal hosts, including humans. This tissue cyst reservoir, which can reactivate into an acute infection, is currently refractory to clinically available therapeutics. Recently, we and others have discovered drugs capable of significantly reducing brain cyst burden in latently infected mice, but not to undetectable levels. In this study, we examined the use of novel combination therapies possessing multiple mechanisms of action in mouse models of latent toxoplasmosis. Our drug regimens included combinations of pyrimethamine, clindamycin, guanabenz, and endochin-like quinolones (ELQs), and were administered to two different mouse strains in an attempt to eradicate brain tissue cysts. We observed mouse strain-dependent effects with these drug treatments: pyrimethamine + guanabenz showed synergistic efficacy in C57BL/6 mice, yet did not improve upon guanabenz monotherapy in BALB/c mice. Contrary to promising in vitro results demonstrating toxicity to bradyzoites, we observed an antagonistic effect between guanabenz + ELQ-334 in vivo. While we were unable to completely eliminate brain cyst burden, we found that a combination treatment of ELQ-334 + pyrimethamine impressively reduced brain cysts to 95% in C57BL/6 mice, which approaches the limit of detection. These analyses highlight the importance of evaluating anti-infective drugs in multiple mouse strains and will help inform further preclinical cocktail therapy studies designed to treat chronic toxoplasmosis.
]]></description>
<dc:creator>Martynowicz, J.</dc:creator>
<dc:creator>Doggett, J. S.</dc:creator>
<dc:creator>Sullivan, W. J.</dc:creator>
<dc:date>2020-03-20</dc:date>
<dc:identifier>doi:10.1101/2020.03.19.999672</dc:identifier>
<dc:title><![CDATA[Efficacy of Guanabenz Combination Therapy Against Chronic Toxoplasmosis Across Multiple Mouse Strains]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-03-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.03.20.000604v1?rss=1">
<title>
<![CDATA[
Selective interferon responses of intestinal epithelial cells minimize TNFα cytotoxicity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.03.20.000604v1?rss=1</link>
<description><![CDATA[
Interferon (IFN) family cytokines stimulate genes (ISGs) that are integral to antiviral host defense. Type I IFNs act systemically whereas type III IFNs act preferentially at epithelial barriers. Among barrier cells, intestinal epithelial cells (IECs) are particularly dependent on type III IFN for control and clearance of virus infection, but the physiological basis of this selective IFN response is not well understood. Here, we confirm that type III IFN treatment elicits robust and uniform ISG expression in neonatal mouse IECs and inhibits replication of IEC-tropic rotavirus. In contrast, type I IFN elicits a marginal ISG response in neonatal mouse IECs and does not inhibit rotavirus replication. In vitro treatment of IEC organoids with type III IFN results in ISG expression that mirrors the in vivo type III IFN response. However, the response of IEC organoids to type I IFN is strikingly increased relative to type III IFN in magnitude and scope. The expanded type I IFN-specific response includes pro-apoptotic genes and potentiates toxicity triggered by tumor necrosis factor alpha (TNF). The ISGs stimulated in common by types I and III IFN have strong interferon-stimulated response element (ISRE) promoter motifs, whereas the expanded set of type I IFN-specific ISGs, including pro-apoptotic genes, have weak ISRE motifs. Thus, preferential responsiveness of IECs to type III IFN in vivo enables selective ISG expression during infection that confers antiviral protection but minimizes disruption of intestinal homeostasis.
]]></description>
<dc:creator>Van Winkle, J. A.</dc:creator>
<dc:creator>Constant, D. A.</dc:creator>
<dc:creator>Li, L.</dc:creator>
<dc:creator>Nice, T. J.</dc:creator>
<dc:date>2020-03-22</dc:date>
<dc:identifier>doi:10.1101/2020.03.20.000604</dc:identifier>
<dc:title><![CDATA[Selective interferon responses of intestinal epithelial cells minimize TNFα cytotoxicity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-03-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.03.20.998443v1?rss=1">
<title>
<![CDATA[
Sulfopin, a selective covalent inhibitor of Pin1, blocks Myc-driven tumor initiation and growth in vivo 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.03.20.998443v1?rss=1</link>
<description><![CDATA[
The peptidyl-prolyl cis-trans isomerase, Pin1, acts as a unified signaling hub that is exploited in cancer to activate oncogenes and inactivate tumor suppressors, in particular through up-regulation of c-Myc target genes. However, despite considerable efforts, Pin1 has remained an elusive drug target. Here, we screened an electrophilic fragment library to discover covalent inhibitors targeting Pin1s active site nucleophile - Cys113, leading to the development of Sulfopin, a double-digit nanomolar Pin1 inhibitor. Sulfopin is highly selective for Pin1, as validated by two independent chemoproteomics methods, achieves potent cellular and in vivo target engagement, and phenocopies genetic knockout of Pin1. Although Pin1 inhibition had a modest effect on viability in cancer cell cultures, Sulfopin induced downregulation of c-Myc target genes and reduced tumor initiation and tumor progression in murine and zebrafish models of MYCN-driven neuroblastoma. Our results suggest that Sulfopin is a suitable chemical probe for assessing Pin1-dependent pharmacology in cells and in vivo. Moreover, these studies indicate that Pin1 should be further investigated as a potential cancer target.
]]></description>
<dc:creator>Dubiella, C.</dc:creator>
<dc:creator>Pinch, B. J.</dc:creator>
<dc:creator>Zaidman, D.</dc:creator>
<dc:creator>Manz, T. D.</dc:creator>
<dc:creator>Poon, E.</dc:creator>
<dc:creator>He, S.</dc:creator>
<dc:creator>Resnick, E.</dc:creator>
<dc:creator>Langer, E. M.</dc:creator>
<dc:creator>Daniel, C. J.</dc:creator>
<dc:creator>Seo, H.-S.</dc:creator>
<dc:creator>Chen, Y.</dc:creator>
<dc:creator>Ficarro, S. B.</dc:creator>
<dc:creator>Jamin, Y.</dc:creator>
<dc:creator>Lian, X.</dc:creator>
<dc:creator>Kibe, S.</dc:creator>
<dc:creator>Kozono, S.</dc:creator>
<dc:creator>Koikawa, K.</dc:creator>
<dc:creator>Doctor, Z. M.</dc:creator>
<dc:creator>Nabet, B.</dc:creator>
<dc:creator>Browne, C. M.</dc:creator>
<dc:creator>Yang, A.</dc:creator>
<dc:creator>Stoler-Barak, L.</dc:creator>
<dc:creator>Shah, R. B.</dc:creator>
<dc:creator>Vangos, N. E.</dc:creator>
<dc:creator>Geffken, E. A.</dc:creator>
<dc:creator>Oren, R.</dc:creator>
<dc:creator>Sidi, S.</dc:creator>
<dc:creator>Shulman, Z.</dc:creator>
<dc:creator>Wang, C.</dc:creator>
<dc:creator>Marto, J. A.</dc:creator>
<dc:creator>Dhe-Paganon, S.</dc:creator>
<dc:creator>Look, T.</dc:creator>
<dc:creator>Zhou, X. Z.</dc:creator>
<dc:creator>Lu, K. P.</dc:creator>
<dc:creator>Sears, R. C.</dc:creator>
<dc:creator>Chesler, L.</dc:creator>
<dc:creator>Gray, N. S.</dc:creator>
<dc:creator>London, N.</dc:creator>
<dc:date>2020-03-21</dc:date>
<dc:identifier>doi:10.1101/2020.03.20.998443</dc:identifier>
<dc:title><![CDATA[Sulfopin, a selective covalent inhibitor of Pin1, blocks Myc-driven tumor initiation and growth in vivo]]></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/10.1101/2020.03.25.008813v1?rss=1">
<title>
<![CDATA[
Effects of Controlled Dual Growth Factor Delivery on Bone Regeneration Following Composite Bone-Muscle Injury 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.03.25.008813v1?rss=1</link>
<description><![CDATA[
The objective of this study was to investigate the controlled release of two growth factors (BMP-2 and VEGF) as a treatment strategy for clinically challenging composite injuries, consisting of a segmental bone defect and volumetric muscle loss. This is the first investigation of dual growth factor delivery in a composite injury model using an injectable smart delivery system consisting of heparin microparticles and alginate gel. The loading efficiency of growth factors into these biomaterials was found to be >90%, revealing a strong affinity of VEGF and BMP-2 to heparin and alginate. The system could achieve simultaneous or sequential release of VEGF and BMP-2 by varying the loading strategy. Single growth factor delivery (VEGF or BMP-2 alone) significantly enhanced vascular growth in vitro. However, no synergistic effect was observed for dual growth factor (BMP-2 + VEGF) delivery. Effective bone healing was achieved in all treatment groups (BMP-2, simultaneous or sequential delivery of BMP-2 and VEGF) in the composite injury model. The mechanics of the regenerated bone reached a maximum strength of [~]52% of intact bone with sequential delivery of VEGF and BMP-2. Overall, simultaneous or sequential co-delivery of low-dose BMP-2 and VEGF failed to fully restore the mechanics of bone in this injury model. Given the severity of the composite injury, VEGF alone may not be sufficient to establish mature and stable blood vessels when compared with previous studies co-delivering BMP-2+VEGF enhanced bone tissue regeneration. Hence, future studies are warranted to develop an alternative treatment strategy focusing on better control over growth factor dose, spatiotemporal delivery, and additional growth factors to regenerate fully functional bone tissue.

HighlightsO_LIWe developed a smart growth factor delivery system using heparin microparticles and alginate that facilitates tunable delivery of VEGF and BMP-2 in a simultaneous or sequential manner by merely varying the loading strategy.
C_LIO_LIIn vitro, both VEGF and BMP-2 alone promoted vascular growth; however, VEGF was significantly more potent, and there was no detectable benefit of co-delivery.
C_LIO_LIIn vivo, both BMP-2 alone and co-delivery of VEGF and BMP-2 promoted bone formation in the challenging bone/muscle polytrauma model; however, none of the treatment groups restored biomechanical properties to that of uninjured bone.
C_LI
]]></description>
<dc:creator>Subbiah, R.</dc:creator>
<dc:creator>Cheng, A.</dc:creator>
<dc:creator>Ruehle, M. A.</dc:creator>
<dc:creator>Hettiaratchi, M. H.</dc:creator>
<dc:creator>Bertassoni, L. E.</dc:creator>
<dc:creator>Guldberg, R. E.</dc:creator>
<dc:date>2020-03-26</dc:date>
<dc:identifier>doi:10.1101/2020.03.25.008813</dc:identifier>
<dc:title><![CDATA[Effects of Controlled Dual Growth Factor Delivery on Bone Regeneration Following Composite Bone-Muscle Injury]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-03-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.03.28.012567v1?rss=1">
<title>
<![CDATA[
In vivo photopharmacology enabled by multifunctional fibers 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.03.28.012567v1?rss=1</link>
<description><![CDATA[
To reversibly manipulate neural circuits with increased spatial and temporal control, photoswitchable ligands can add an optical switch to a target receptor or signaling cascade. This approach, termed photopharmacology, has been enabling to molecular neuroscience, however, its application to behavioral experiments has been impeded by a lack of integrated hardware capable of delivering both light and compounds to deep brain regions in moving subjects. Here, we devise a hybrid photochemical genetic approach to target neurons using a photoswitchable agonist of capsaicin receptor (TRPV1), red-AzCA-4. Using the thermal drawing process we created multifunctional fibers that can deliver viruses, photoswitchable ligands, and light to deep brain regions in awake, freely moving mice. We implanted our fibers into the ventral tegmental area (VTA), a midbrain hub of the mesolimbic pathway, and used them to deliver a transgene coding for TRPV1. This sensitized excitatory VTA neurons to red-AzCA-4, and allowed us to optically control conditioned place preference using a mammalian ion-channel, thus extending applications of photopharmacology to behavioral experiments. Applied to endogenous receptors, our approach may accelerate studies of molecular mechanisms underlying animal behavior.
]]></description>
<dc:creator>Frank, J. A.</dc:creator>
<dc:creator>Antonini, M.-J.</dc:creator>
<dc:creator>Chiang, P.-H.</dc:creator>
<dc:creator>Canales, A.</dc:creator>
<dc:creator>Konrad, D.</dc:creator>
<dc:creator>Garwood, I.</dc:creator>
<dc:creator>Rajic, G.</dc:creator>
<dc:creator>Koehler, F.</dc:creator>
<dc:creator>Fink, Y.</dc:creator>
<dc:creator>Anikeeva, P.</dc:creator>
<dc:date>2020-03-30</dc:date>
<dc:identifier>doi:10.1101/2020.03.28.012567</dc:identifier>
<dc:title><![CDATA[In vivo photopharmacology enabled by multifunctional fibers]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-03-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.03.28.013474v1?rss=1">
<title>
<![CDATA[
Identification and characterization of diverse OTU deubiquitinases in bacteria 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.03.28.013474v1?rss=1</link>
<description><![CDATA[
Manipulation of host ubiquitin signaling is becoming an increasingly apparent evolutionary strategy among bacterial and viral pathogens. By removing host ubiquitin signals, for example, invading pathogens can inactivate immune response pathways and evade detection. The Ovarian Tumor (OTU) family of deubiquitinases regulates diverse ubiquitin signals in humans. Viral pathogens have also extensively co-opted the OTU fold to subvert host signaling, but the extent to which bacteria utilize the OTU fold was unknown. We have predicted and validated a set of OTU deubiquitinases encoded by several classes of pathogenic bacteria. Biochemical assays highlight the ubiquitin and polyubiquitin linkage specificities of these bacterial deubiquitinases. By determining the ubiquitin-bound structures of two examples, we demonstrate the novel strategies that have evolved to both thread an OTU fold and to recognize a ubiquitin substrate. With these new examples, we perform the first cross-kingdom structural analysis of the OTU fold that highlights commonalities among distantly-related OTU deubiquitinases.
]]></description>
<dc:creator>Schubert, A. F.</dc:creator>
<dc:creator>Nguyen, J. V.</dc:creator>
<dc:creator>Franklin, T. G.</dc:creator>
<dc:creator>Geurink, P. P.</dc:creator>
<dc:creator>Roberts, C. G.</dc:creator>
<dc:creator>Sanderson, D. J.</dc:creator>
<dc:creator>Miller, L. N.</dc:creator>
<dc:creator>Ovaa, H.</dc:creator>
<dc:creator>Hofmann, K.</dc:creator>
<dc:creator>Pruneda, J. N.</dc:creator>
<dc:creator>Komander, D.</dc:creator>
<dc:date>2020-03-29</dc:date>
<dc:identifier>doi:10.1101/2020.03.28.013474</dc:identifier>
<dc:title><![CDATA[Identification and characterization of diverse OTU deubiquitinases in bacteria]]></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/10.1101/2020.04.01.020842v1?rss=1">
<title>
<![CDATA[
VISTA: Virtual ImmunoSTAining for pancreatic disease quantification in murine cohorts 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.04.01.020842v1?rss=1</link>
<description><![CDATA[
Mechanistic studies of pancreatic disease progression using animal models require objective and quantifiable assessment of tissue changes among animal cohorts. Disease state quantification, however, relies heavily on tissue immunostaining, which can be expensive, labor- and time-intensive, and all too often produces uneven staining that is prone to variable interpretation between experts and inaccurate quantification. Here we develop a fully automated semantic segmentation tool using deep learning for the rapid and objective quantification of histologic features using hematoxylin and eosin (H&E) stained pancreatic tissue sections acquired from murine pancreatic cancer models. The tool was successfully trained to segment and quantify multiple histopathologic features of pancreatic pre-cancer evolution, including normal acinar structures, the ductal phenotype of acinar-to ductal metaplasia (ADM), dysplasia, and the expanding stromal compartment. Disease quantifications produced by our computational tool were highly correlated to the results obtained by immunostaining markers of normal and diseased tissue (DAPI, amylase, and cytokeratins; correlation score= 0.9, 0.95, and 0.91, respectively) and were able to accurately reproduce immunostain patterns. Moreover, our tool was able to distinguish ADM from dysplasia, which are not reliably distinguished by immunostaining, and avoid the pitfalls of uneven or poor-quality staining. Using this tool, we quantified the changes in histologic feature abundance for murine cohorts with oncogenic Kras-driven disease at 2 months and 5 months of age (n=12, n=13). The calculated changes in histologic feature abundance were consistent with biological expectations, showing an expansion of the stromal compartment, a reduction of normal acinar tissue, and an increase in both ADM and dysplasia as disease progresses (p= 2e-6, 6e-7, 4e-4, and 3e-5, respectively). These results demonstrate the tools efficacy for accurate and rapid quantification of multiple histologic features using an objective and automated platform. Our tool promises to rapidly accelerate and improve the quantification of altered pancreatic disease progression in animal studies.
]]></description>
<dc:creator>Ternes, L.</dc:creator>
<dc:creator>Huang, G.</dc:creator>
<dc:creator>Lanciault, C.</dc:creator>
<dc:creator>Thibault, G.</dc:creator>
<dc:creator>Riggers, R.</dc:creator>
<dc:creator>Gray, J. W.</dc:creator>
<dc:creator>Muschler, J.</dc:creator>
<dc:creator>Chang, Y. H.</dc:creator>
<dc:date>2020-04-02</dc:date>
<dc:identifier>doi:10.1101/2020.04.01.020842</dc:identifier>
<dc:title><![CDATA[VISTA: Virtual ImmunoSTAining for pancreatic disease quantification in murine cohorts]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-04-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.04.14.036384v1?rss=1">
<title>
<![CDATA[
Connexin-46/50 in a dynamic lipid environment resolved by CryoEM at 1.9 A 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.04.14.036384v1?rss=1</link>
<description><![CDATA[
Gap junctions establish direct pathways for connected cells and tissues to transfer metabolic and electrical messages1. The local lipid environment is known to affect the structure, stability and intercellular channel activity of gap junctions2-5; however, the molecular basis for these effects remains unknown. To gain insight toward how gap junctions interact with their local membrane environment, we used lipid nanodisc technology to incorporate native connexin-46/50 (Cx46/50) intercellular channels into a dual lipid membrane system, closely mimicking a native cell-to-cell junction. Structural characterization of Cx46/50 lipid-embedded channels by single particle CryoEM revealed a lipid-induced stabilization to the channel, resulting in a 3D reconstruction at 1.9 [A] resolution. Together with all-atom molecular dynamics (MD) simulations and 3D heterogeneity analysis of the ensemble CryoEM data, it is shown that Cx46/50 in turn imparts long-range stabilization to the dynamic local lipid environment that is specific to the extracellular lipid leaflet of the two opposed membranes. In addition, nearly 400 water molecules are resolved in the CryoEM map, localized throughout the intercellular permeation pathway and contributing to the channel architecture. These results illustrate how the aqueous-lipid environment is integrated with the architectural stability, structure and function of gap junction communication channels, and demonstrates the ability of CryoEM to effectively characterize dynamical protein-lipid interactions.
]]></description>
<dc:creator>Flores, J. A.</dc:creator>
<dc:creator>Haddad, B. G.</dc:creator>
<dc:creator>Dolan, K. A.</dc:creator>
<dc:creator>Myers, J. B.</dc:creator>
<dc:creator>Yoshioka, C. C.</dc:creator>
<dc:creator>Copperman, J.</dc:creator>
<dc:creator>Zuckerman, D. M.</dc:creator>
<dc:creator>Reichow, S. L.</dc:creator>
<dc:date>2020-04-15</dc:date>
<dc:identifier>doi:10.1101/2020.04.14.036384</dc:identifier>
<dc:title><![CDATA[Connexin-46/50 in a dynamic lipid environment resolved by CryoEM at 1.9 A]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-04-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.04.14.041608v1?rss=1">
<title>
<![CDATA[
PD-1 Specific 'Blocking' Antibodies That Deplete PD-1+ T Cells Present An Inconvenient Variable In Pre-clinical Immunotherapy Experiments 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.04.14.041608v1?rss=1</link>
<description><![CDATA[
Therapeutic antibodies blocking PD-1-/PD-L1 interaction have achieved remarkable clinical success in cancer. In addition to blocking a target molecule, some isotypes of antibodies can activate complement, NK cells or phagocytes, resulting in death of the cell expressing the antibodys target. Human anti-PD-1 therapeutics use antibody isotypes designed to minimize such antibody-dependent lysis. In contrast, anti-PD-1 reagents used in mice are derived from multiple species, with different isotypes, and are not engineered to reduce target cell death: few studies analyze or discuss how antibody species and isotype may impact data interpretation. We demonstrate here that anti-PD-1 therapy to promote activation and proliferation of PD-1-expressing CD8 T cells sometimes led instead to a loss of antigen specific cells. This phenomenon was seen in two tumor models and a model of virus infection, and was varied with the clone of anti-PD-1 antibody. Additionally, we compared competition among anti-PD-1 clones to find a combination that allows detection of PD-1-expressing cells despite the presence of blocking anti-PD1 antibodies in vivo. These data bring attention to the possibility of unintended target cell depletion with some commonly used anti-mouse PD-1 clones, and should provide a valuable resource for the design and interpretation of anti-PD-1 studies in mice. (200)
]]></description>
<dc:creator>Polesso, F.</dc:creator>
<dc:creator>Munks, M. W.</dc:creator>
<dc:creator>Rott, K. H.</dc:creator>
<dc:creator>Smart, S.</dc:creator>
<dc:creator>Hill, A. B.</dc:creator>
<dc:creator>Moran, A. E.</dc:creator>
<dc:date>2020-04-16</dc:date>
<dc:identifier>doi:10.1101/2020.04.14.041608</dc:identifier>
<dc:title><![CDATA[PD-1 Specific 'Blocking' Antibodies That Deplete PD-1+ T Cells Present An Inconvenient Variable In Pre-clinical Immunotherapy Experiments]]></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/10.1101/2020.04.15.041566v1?rss=1">
<title>
<![CDATA[
Invasion of homogeneous and polyploid populations in nutrient-limiting environments 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.04.15.041566v1?rss=1</link>
<description><![CDATA[
Breast cancer progresses in a multistep process from primary tumor growth and stroma invasion to metastasis. Progression is accompanied by a switch to an invasive cell phenotype. Nutrient-limiting environments promote chemotaxis with aggressive morphologies characteristic of invasion. It is unknown how co-existing cells differ in their response to nutrient limitations and how this impacts invasion of the metapopulation as a whole. We integrate mathematical modeling with microenvironmental perturbation-data to investigate invasion in nutrient-limiting environments inhabited by one or two cancer cell subpopulations. Hereby, subpopulations are defined by their energy efficiency and chemotactic ability. We estimate the invasion-distance traveled by a homogeneous population. For heterogeneous populations, our results suggest that an imbalance between nutrient efficacy and chemotactic superiority accelerates invasion. Such imbalance will spatially segregate the two populations and only one type will dominate at the invasion front. Only if these two phenotypes are balanced do the two subpopulations compete for the same space, which decelerates invasion. We investigate ploidy as a candidate biomarker of this phenotypic heterogeneity to discern circumstances when inhibiting chemotaxis amplifies internal competition and decelerates tumor progression, from circumstances that render clinical consequences of chemotactic inhibition unfavorable.Significance A better understanding of the nature of the double-edged sword of high ploidy is a prerequisite to personalize combination-therapies with cytotoxic drugs and inhibitors of signal transduction pathways such as MTOR-Is.Competing Interest StatementThe authors have declared no competing interest.View Full Text
]]></description>
<dc:creator>Kimmel, G. J.</dc:creator>
<dc:creator>Dane, M.</dc:creator>
<dc:creator>Heiser, L.</dc:creator>
<dc:creator>Altrock, P. M.</dc:creator>
<dc:creator>Andor, N.</dc:creator>
<dc:date>2020-04-17</dc:date>
<dc:identifier>doi:10.1101/2020.04.15.041566</dc:identifier>
<dc:title><![CDATA[Invasion of homogeneous and polyploid populations in nutrient-limiting environments]]></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/10.1101/2020.04.15.043364v1?rss=1">
<title>
<![CDATA[
Analysis of SARS-CoV-2 Antibodies in COVID-19 Convalescent Plasma using a Coronavirus Antigen Microarray 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.04.15.043364v1?rss=1</link>
<description><![CDATA[
The current practice for diagnosis of COVID-19, based on SARS-CoV-2 PCR testing of pharyngeal or respiratory specimens in a symptomatic patient at high epidemiologic risk, likely underestimates the true prevalence of infection. Serologic methods can more accurately estimate the disease burden by detecting infections missed by the limited testing performed to date. Here, we describe the validation of a coronavirus antigen microarray containing immunologically significant antigens from SARS-CoV-2, in addition to SARS-CoV, MERS-CoV, common human coronavirus strains, and other common respiratory viruses. A comparison of antibody profiles detected on the array from control sera collected prior to the SARS-CoV-2 pandemic versus convalescent blood specimens from virologically confirmed COVID-19 cases demonstrates near complete discrimination of these two groups, with improved performance from use of antigen combinations that include both spike protein and nucleoprotein. This array can be used as a diagnostic tool, as an epidemiologic tool to more accurately estimate the disease burden of COVID-19, and as a research tool to correlate antibody responses with clinical outcomes.
]]></description>
<dc:creator>de Assis, R. R.</dc:creator>
<dc:creator>Jain, A.</dc:creator>
<dc:creator>Nakajima, R.</dc:creator>
<dc:creator>Jasinskas, A.</dc:creator>
<dc:creator>Felgner, J.</dc:creator>
<dc:creator>Obiero, J. M.</dc:creator>
<dc:creator>Adenaiye, O. O.</dc:creator>
<dc:creator>Tai, S.</dc:creator>
<dc:creator>Hong, F. H.</dc:creator>
<dc:creator>Norris, P.</dc:creator>
<dc:creator>Stone, M.</dc:creator>
<dc:creator>Simmons, G.</dc:creator>
<dc:creator>Bagri, A.</dc:creator>
<dc:creator>Schreiber, M.</dc:creator>
<dc:creator>Buser, A.</dc:creator>
<dc:creator>Holbro, A.</dc:creator>
<dc:creator>Battegay, M.</dc:creator>
<dc:creator>Milton, D. K.</dc:creator>
<dc:creator>Prometheus Study Group,</dc:creator>
<dc:creator>Davies, H.</dc:creator>
<dc:creator>Corash, L. M.</dc:creator>
<dc:creator>Busch, M. P.</dc:creator>
<dc:creator>Felgner, P. L.</dc:creator>
<dc:creator>Khan, S.</dc:creator>
<dc:date>2020-04-17</dc:date>
<dc:identifier>doi:10.1101/2020.04.15.043364</dc:identifier>
<dc:title><![CDATA[Analysis of SARS-CoV-2 Antibodies in COVID-19 Convalescent Plasma using a Coronavirus Antigen Microarray]]></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/10.1101/2020.04.17.047480v1?rss=1">
<title>
<![CDATA[
Coronavirus Infection and PARP Expression Dysregulate the NAD Metabolome: A Potentially Actionable Component of Innate Immunity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.04.17.047480v1?rss=1</link>
<description><![CDATA[
Poly-ADP-ribose polymerase (PARP) superfamily members covalently link either a single ADP-ribose (ADPR) or a chain of ADPR units to proteins using nicotinamide adenine dinucleotide (NAD) as the source of ADPR. While the well-known poly-ADP-ribosylating (PARylating) PARPs primarily function in the DNA damage response, many non-canonical mono-ADP-ribosylating (MARylating) PARPs are associated with cellular antiviral responses. We recently demonstrated robust upregulation of several PARPs following infection with Murine Hepatitis Virus (MHV), a model coronavirus. Here we show that SARS-CoV-2 infection strikingly upregulates MARylating PARPs and induces the expression of genes encoding enzymes for salvage NAD synthesis from nicotinamide (NAM) and nicotinamide riboside (NR), while downregulating other NAD biosynthetic pathways. We show that overexpression of PARP10 is sufficient to depress cellular NAD and that the activities of the transcriptionally induced enzymes PARP7, PARP10, PARP12 and PARP14 are limited by cellular NAD and can be enhanced by pharmacological activation of NAD synthesis. We further demonstrate that infection with MHV induces a severe attack on host cell NAD+ and NADP+. Finally, we show that NAMPT activation, NAM and NR dramatically decrease the replication of an MHV virus that is sensitive to PARP activity. These data suggest that the antiviral activities of noncanonical PARP isozyme activities are limited by the availability of NAD, and that nutritional and pharmacological interventions to enhance NAD levels may boost innate immunity to coronaviruses.
]]></description>
<dc:creator>Heer, C. D.</dc:creator>
<dc:creator>Sanderson, D. J.</dc:creator>
<dc:creator>Alhammad, Y. M. O.</dc:creator>
<dc:creator>Schmidt, M. S.</dc:creator>
<dc:creator>Trammell, S. A. J.</dc:creator>
<dc:creator>Perlman, S.</dc:creator>
<dc:creator>Cohen, M. S.</dc:creator>
<dc:creator>Fehr, A. R.</dc:creator>
<dc:creator>Brenner, C.</dc:creator>
<dc:date>2020-04-18</dc:date>
<dc:identifier>doi:10.1101/2020.04.17.047480</dc:identifier>
<dc:title><![CDATA[Coronavirus Infection and PARP Expression Dysregulate the NAD Metabolome: A Potentially Actionable Component of Innate Immunity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-04-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.04.22.055418v1?rss=1">
<title>
<![CDATA[
Multiscale cardiac imaging to capture the whole heart and its internal cellular architecture, with applications to congenital heart disease 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.04.22.055418v1?rss=1</link>
<description><![CDATA[
Efficient cardiac pumping depends on the morphological structure of the heart, but also on its sub-cellular (ultrastructural) architecture, which enables cardiac contraction. In cases of congenital heart defects, localized sub-cellular disruptions in architecture that increase the risk of heart failure are only starting to be discovered. This is in part due to a lack of technologies that can image the three dimensional (3D) heart structure, assessing malformations; and its ultrastructure, assessing disruptions. We present here a multiscale, correlative imaging procedure that achieves high-resolution images of the whole heart, using 3D micro-computed tomography (micro-CT); and its ultrastructure, using 3D scanning electron microscopy (SEM). This combination of technologies has not been possible before in imaging the same cardiac sample due to the heart large size, even when studying small fetal and neonatal animal models (~5x5x5mm3). Here, we achieved uniform fixation and staining of the whole heart, without losing ultrastructural preservation (at the nm resolution range). Our approach enables multiscale studies of cardiac architecture in models of congenital heart disease and beyond.
]]></description>
<dc:creator>Rykiel, G.</dc:creator>
<dc:creator>Lopez, C. S.</dc:creator>
<dc:creator>Riesterer, J. L.</dc:creator>
<dc:creator>Fries, I.</dc:creator>
<dc:creator>Deosthali, S.</dc:creator>
<dc:creator>Courchaine, K.</dc:creator>
<dc:creator>Maloyan, A.</dc:creator>
<dc:creator>Thornburg, K.</dc:creator>
<dc:creator>Rugonyi, S.</dc:creator>
<dc:date>2020-04-23</dc:date>
<dc:identifier>doi:10.1101/2020.04.22.055418</dc:identifier>
<dc:title><![CDATA[Multiscale cardiac imaging to capture the whole heart and its internal cellular architecture, with applications to congenital heart disease]]></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/10.1101/2020.04.29.068445v1?rss=1">
<title>
<![CDATA[
A Cytoskeletal Protein Complex is Essential for Division of Intracellular Amastigotes of Leishmania mexicana 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.04.29.068445v1?rss=1</link>
<description><![CDATA[
Previous studies in Leishmania mexicana have identified the cytoskeletal protein KHARON as being important for both flagellar trafficking of the glucose transporter GT1 and for successful cytokinesis and survival of infectious amastigote forms inside mammalian macrophages. KHARON is located in three distinct regions of the cytoskeleton: the base of the flagellum, the subpellicular microtubules, and the mitotic spindle. To deconvolve the different functions for KHARON, we have identified two partner proteins, KHAP1 and KHAP2, that associate with KHARON. KHAP1 is located only in the subpellicular microtubules, while KHAP2 is located at the subpellicular microtubules and the base of the flagellum. Both the KHAP1 and KHAP2 null mutants are unable to execute cytokinesis but are able to traffic GT1 to the flagellum. These results confirm that KHARON assembles into distinct functional complexes and that the subpellicular complex is essential for cytokinesis and viability of disease-causing amastigotes but not for flagellar membrane trafficking.
]]></description>
<dc:creator>Kelly, F. D.</dc:creator>
<dc:creator>Tran, K. D.</dc:creator>
<dc:creator>Hatfield, J.</dc:creator>
<dc:creator>Schmidt, K.</dc:creator>
<dc:creator>Sanchez, M. A.</dc:creator>
<dc:creator>Landfear, S. M.</dc:creator>
<dc:date>2020-04-30</dc:date>
<dc:identifier>doi:10.1101/2020.04.29.068445</dc:identifier>
<dc:title><![CDATA[A Cytoskeletal Protein Complex is Essential for Division of Intracellular Amastigotes of Leishmania mexicana]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-04-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.05.01.072124v1?rss=1">
<title>
<![CDATA[
Structural and molecular determinants of CCS-mediated copper activation of MEK1/2 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.05.01.072124v1?rss=1</link>
<description><![CDATA[
SummaryNormal physiology relies on the precise coordination of intracellular signal transduction pathways that respond to nutrient availability to balance cell growth and cell death. We recently established a critical mechanistic function for the redox-active micronutrient copper (Cu) in the canonical mitogen activated protein kinase (MAPK) pathway at the level of MEK1 and MEK2. Here we report the X-ray crystal structure of Cu-MEK1 and reveal active site chemical ligands and oxidation state specificity for MEK1 Cu coordination. Mechanistically, the Cu chaperone CCS selectively bound to and facilitated Cu transfer to MEK1. Mutations in MEK1 that disrupt Cu(I) affinity or a CCS small molecule inhibitor reduced Cu-stimulated MEK1 kinase activity. These atomic and molecular level data provide the first mechanistic insights of Cu kinase signaling and could be exploited for the development of novel MEK1/2 inhibitors that either target the Cu structural interface or blunt dedicated Cu delivery mechanisms via CCS.Competing Interest StatementD.C.B holds ownership in Merlon Inc. D.C.B. is an inventor on the patent application 20150017261 entitled Methods of treating and preventing cancer by disrupting the binding of copper in the MAP kinase pathway. No potential conflicts of interest were disclosed by the other authors.View Full Text
]]></description>
<dc:creator>Grasso, M.</dc:creator>
<dc:creator>Bond, G. J.</dc:creator>
<dc:creator>Kim, Y.-J.</dc:creator>
<dc:creator>Alwan, K. B.</dc:creator>
<dc:creator>Boyd, S.</dc:creator>
<dc:creator>Dzebo, M. M.</dc:creator>
<dc:creator>Valenzuela, S.</dc:creator>
<dc:creator>Tsang, T.</dc:creator>
<dc:creator>Schibrowsky, N. A.</dc:creator>
<dc:creator>Matthews, M. L.</dc:creator>
<dc:creator>Burslem, G. M.</dc:creator>
<dc:creator>Wittung-Stafshede, P.</dc:creator>
<dc:creator>Winkler, D. D.</dc:creator>
<dc:creator>Blackburn, N. J.</dc:creator>
<dc:creator>Marmorstein, R.</dc:creator>
<dc:creator>Brady, D. C.</dc:creator>
<dc:date>2020-05-02</dc:date>
<dc:identifier>doi:10.1101/2020.05.01.072124</dc:identifier>
<dc:title><![CDATA[Structural and molecular determinants of CCS-mediated copper activation of MEK1/2]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-05-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.05.12.088856v1?rss=1">
<title>
<![CDATA[
A miR-494 dependent feedback loop regulates ER stress 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.05.12.088856v1?rss=1</link>
<description><![CDATA[
Defects in stress responses are important contributors in many chronic conditions including cancer, cardiovascular disease, diabetes, and obesity-driven pathologies like non-alcoholic steatohepatitis (NASH). Specifically, endoplasmic reticulum (ER) stress is linked with these pathologies and control of ER stress can ameliorate tissue damage. MicroRNAs have a critical role in regulating diverse stress responses including ER stress. Here we show that miR-494 plays a functional role during ER stress. ER stress inducers (tunicamycin & thapsigargin) robustly increase the expression of miR-494 in vitro in an ATF6 dependent manner. Surprisingly, miR-494 pretreatment dampens the induction and magnitude of ER stress in response to tunicamycin in endothelial cells. Conversely, inhibition of miR-494 increases ER stress de novo and amplifies the effects of ER stress inducers. Using Mass Spectrometry (TMT-MS) we identified 23 proteins that are downregulated by both tunicamycin and miR-494. Among these, we found 6 transcripts which harbor a putative miR-494 binding site. We validated the anti-apoptotic gene BIRC5 (survivin) as one of the targets of miR-494 during ER stress. Finally, induction of ER stress in vivo increases miR-494 expression in the liver. Pretreatment of mice with a miR-494 plasmid via hydrodynamic injection decreased ER stress in response to tunicamycin in part by decreasing inflammatory chemokines and cytokines. In summary, our data indicates that ER stress driven miR-494 may act in a feedback inhibitory loop to dampen downstream ER stress signaling. We propose that RNA-based approaches targeting miR-494 or its targets may be attractive candidates for inhibiting ER stress dependent pathologies in human disease.
]]></description>
<dc:creator>Anand, S.</dc:creator>
<dc:creator>Chatterjee, N.</dc:creator>
<dc:creator>Espinosa-Diez, C.</dc:creator>
<dc:date>2020-05-13</dc:date>
<dc:identifier>doi:10.1101/2020.05.12.088856</dc:identifier>
<dc:title><![CDATA[A miR-494 dependent feedback loop regulates ER stress]]></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/10.1101/2020.05.12.088971v1?rss=1">
<title>
<![CDATA[
Exploiting KRAS-driven Ferroaddiction in Cancer Through Ferrous Iron-Activatable Drug Conjugates (FeADC) 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.05.12.088971v1?rss=1</link>
<description><![CDATA[
KRAS mutations cause a quarter of cancer mortality and most are undruggable. Several inhibitors of the MAPK pathway are FDA approved but poorly tolerated at dosages required to adequately extinguish RAS/RAF/MAPK signaling. We found that oncogenic KRAS signaling induces ferrous iron (Fe2+) accumulation early in and throughout KRAS-mediated transformation. We used an FDA-approved MEK inhibitor to produce a prototypical Ferrous Iron-Activatable Drug Conjugate (FeADC) which achieved potent MAPK blockade in tumor cells while sparing normal tissues. This innovation allowed sustainable, effective treatment of tumor bearing animals, with tumor-selective drug activation producing superior systemic tolerability. Ferrous iron accumulation is an exploitable feature of KRAS transformation and FeADCs hold promise for improving treatment of KRAS-driven solid tumors.
]]></description>
<dc:creator>Jiang, H.</dc:creator>
<dc:creator>Muir, R. K.</dc:creator>
<dc:creator>Gonciarz, R. L.</dc:creator>
<dc:creator>Olshen, A. B.</dc:creator>
<dc:creator>Yeh, I.</dc:creator>
<dc:creator>Hann, B. C.</dc:creator>
<dc:creator>Zhao, N.</dc:creator>
<dc:creator>Wang, Y.-h.</dc:creator>
<dc:creator>Korkola, J.</dc:creator>
<dc:creator>Behr, S. C.</dc:creator>
<dc:creator>Evans, M. J.</dc:creator>
<dc:creator>collisson, e.</dc:creator>
<dc:creator>Renslo, A.</dc:creator>
<dc:date>2020-05-14</dc:date>
<dc:identifier>doi:10.1101/2020.05.12.088971</dc:identifier>
<dc:title><![CDATA[Exploiting KRAS-driven Ferroaddiction in Cancer Through Ferrous Iron-Activatable Drug Conjugates (FeADC)]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-05-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.05.15.098608v1?rss=1">
<title>
<![CDATA[
Astrocytes close a critical period of motor circuit plasticity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.05.15.098608v1?rss=1</link>
<description><![CDATA[
Critical periods - brief intervals where neural circuits can be modified by sensory input - are necessary for proper neural circuit assembly. Extended critical periods are associated with neurodevelopmental disorders, including schizophrenia and autism; however, the mechanisms that ensure timely critical period closure remain unknown. Here, we define the extent of a critical period in the developing Drosophila motor circuit, and identify astrocytes as essential for proper critical period termination. During the critical period, decreased activity produces larger motor dendrites with fewer inhibitory inputs; conversely, increased motor neuron activity produces smaller motor dendrites with fewer excitatory inputs. Importantly, activity has little effect on dendrite morphology after critical period closure. Astrocytes invade the neuropil just prior to critical period closure, and astrocyte ablation prolongs the critical period. Finally, we use a genetic screen to identify astrocyte-motor neuron signaling pathways that close the critical period, including Neuroligin-Neurexin signaling. Reduced signaling destabilizes dendritic microtubules, increases dendrite dynamicity, and impairs locomotor behavior, underscoring the importance of critical period closure. Previous work defines astroglia as regulators of plasticity at individual synapses; here, we show that astrocytes also regulate large-scale structural plasticity to motor dendrite, and thus, circuit architecture to ensure proper locomotor behavior.
]]></description>
<dc:creator>Ackerman, S. D.</dc:creator>
<dc:creator>Perez-Catalan, N. A.</dc:creator>
<dc:creator>Freeman, M. R.</dc:creator>
<dc:creator>Doe, C. Q.</dc:creator>
<dc:date>2020-05-16</dc:date>
<dc:identifier>doi:10.1101/2020.05.15.098608</dc:identifier>
<dc:title><![CDATA[Astrocytes close a critical period of motor circuit plasticity]]></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/10.1101/2020.05.16.099366v1?rss=1">
<title>
<![CDATA[
A novel fragmented mitochondrial genome in the protist pathogen Toxoplasma gondii and related tissue coccidia 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.05.16.099366v1?rss=1</link>
<description><![CDATA[
Mitochondrial genome content and structure vary widely across the eukaryotic tree of life with protists displaying extreme examples. Apicomplexan and dinoflagellate protists have evolved highly-reduced mitochondrial genome sequences, mtDNA, consisting of only 3 cytochrome genes and fragmented rRNA genes. Here we report the independent evolution of fragmented cytochrome genes in Toxoplasma and related tissue coccidia and evolution of a novel genome architecture consisting minimally of 21 sequence blocks (SBs) that exist as non-random concatemers. Single-molecule Nanopore reads consisting entirely of SB concatemers ranging from 1-23 kb reveal both whole and fragmented cytochrome genes. Full-length cytochrome transcripts including a divergent coxIII are detected. The topology of the mitochondrial genome remains an enigma. Analysis of a cob point mutation reveals that homoplasmy of SBs is maintained. Tissue coccidia are important pathogens of man and animals and the mitochondrion represents an important therapeutic target. Their mtDNA sequence has remained elusive until now.
]]></description>
<dc:creator>Namasivayam, S.</dc:creator>
<dc:creator>Baptista, R. d. P.</dc:creator>
<dc:creator>Xiao, W.</dc:creator>
<dc:creator>Hall, E. M.</dc:creator>
<dc:creator>Doggett, J. S.</dc:creator>
<dc:creator>Troell, K.</dc:creator>
<dc:creator>Kissinger, J. C.</dc:creator>
<dc:date>2020-05-18</dc:date>
<dc:identifier>doi:10.1101/2020.05.16.099366</dc:identifier>
<dc:title><![CDATA[A novel fragmented mitochondrial genome in the protist pathogen Toxoplasma gondii and related tissue coccidia]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-05-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.05.16.099929v1?rss=1">
<title>
<![CDATA[
NAIP-NLRC4-deficient mice are susceptible to shigellosis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.05.16.099929v1?rss=1</link>
<description><![CDATA[
Bacteria of the genus Shigella cause shigellosis, a severe gastrointestinal disease that is a major cause of diarrhea-associated mortality in humans. Shigellosis develops upon oral ingestion of as few as 100 bacteria, but million-fold higher doses fail to cause disease in mice. The lack of a physiologically relevant mouse model of shigellosis has impeded our understanding of this important human disease, but why mice are resistant is unknown. Here we show that in human cells, but not in mice, Shigella evades detection by the NAIP-NLRC4 inflammasome, an immune sensor present in intestinal epithelial cells (IECs). We find that NAIP-NLRC4-deficient mice are highly susceptible to oral Shigella infection and recapitulate the clinical features of human shigellosis, including bacterial replication in IECs and neutrophilic inflammation of the colon. Confirming a role for bacterial replication in IECs in our new model, a Shigella mutant lacking IcsA, a factor required for cell-to-cell spread among IECs, is attenuated in otherwise susceptible NAIP-NLRC4-deficient mice. Although inflammasome-mediated cell death is widely held to promote Shigella infection and pathogenesis, we instead demonstrate that IEC-specific NAIP-NLRC4-induced cell death is sufficient to protect the host from shigellosis. Thus, NAIP-NLRC4-deficient mice are a physiologically relevant and experimentally tractable model for shigellosis. More broadly, our results suggest that the lack of an inflammasome response in IECs may help explain the extreme susceptibility of humans to shigellosis.
]]></description>
<dc:creator>Mitchell, P. S.</dc:creator>
<dc:creator>Roncaioli, J. L.</dc:creator>
<dc:creator>Turcotte, E. A.</dc:creator>
<dc:creator>Goers, L.</dc:creator>
<dc:creator>Chavez, R. A.</dc:creator>
<dc:creator>Lee, A. Y.</dc:creator>
<dc:creator>Lesser, C. F.</dc:creator>
<dc:creator>Rauch, I.</dc:creator>
<dc:creator>Vance, R. E.</dc:creator>
<dc:date>2020-05-17</dc:date>
<dc:identifier>doi:10.1101/2020.05.16.099929</dc:identifier>
<dc:title><![CDATA[NAIP-NLRC4-deficient mice are susceptible to shigellosis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-05-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.05.19.104307v1?rss=1">
<title>
<![CDATA[
Gbx2 identifies two amacrine cell subtypes with distinct molecular, morphological, and physiological properties 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.05.19.104307v1?rss=1</link>
<description><![CDATA[
Our understanding of how the nervous sytem works is limited by our ability to identify the neuronal subtypes that comprise functional circuits. Using a genetic approach, we show that the transcription factor Gbx2 labels two amacrine cell (AC) subtypes in the mouse retina that have distinct morphological, physiological, and molecular properties. One subtype of Gbx2+ ACs are likely the previously characterized On-type GABAergic CRH-1 AC. The other Gbx2+ AC population is a previously uncharacterized non-GABAergic, non-Glycinergic (nGnG) AC subtype. Gbx2+ nGnG ACs are On-Off type cells with asymmetric dendritic arbors. Gbx2+ nGnG ACs also exhibit tracer coupling to bipolar cells (BCs) through gap junctions that are modulated by dopamine signaling. This study genetically identifies a previously uncharacterized AC subtype and reveals an unusual AC-BC connectivity through gap junctions that may provide a novel model of synaptic communication and visual circuit function.
]]></description>
<dc:creator>Kerstein, P. C.</dc:creator>
<dc:creator>Leffler, J.</dc:creator>
<dc:creator>Sivyer, B.</dc:creator>
<dc:creator>Taylor, W. R.</dc:creator>
<dc:creator>Wright, K. M.</dc:creator>
<dc:date>2020-05-20</dc:date>
<dc:identifier>doi:10.1101/2020.05.19.104307</dc:identifier>
<dc:title><![CDATA[Gbx2 identifies two amacrine cell subtypes with distinct molecular, morphological, and physiological properties]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-05-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.05.20.104612v1?rss=1">
<title>
<![CDATA[
S-nitrosylated and non-nitrosylated COX2 has differential expression and distinct subcellular localization in normal and breast cancer tissue 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.05.20.104612v1?rss=1</link>
<description><![CDATA[
Immunohistochemical staining in breast cancer shows both gain and loss of COX2 expression with disease risk and progression. We investigated four common COX2 antibody clones and found high specificity for purified human COX2 for three clones; however, recognition of COX2 in cell lysates was clone dependent. Biochemical characterization revealed two distinct forms of COX2, with SP21 recognizing an S-nitrosylated form and CX229 and CX294 appearing to recognize the same non-nitrosylated COX2 antigen. We found S-nitrosylated and non-nitrosylated COX2 occupy different subcellular locations in normal and breast cancer tissue, implicating distinct synthetic/trafficking pathways and function. Dual stains of ~2000 breast cancer cases show early onset breast cancer has increased expression of both forms of COX2 compared to postmenopausal cases. Our results highlight the strengths of using multiple, highly characterized antibody clones for COX2 immunohistochemical studies and raise the prospect that S-nitrosylation of COX2 may play a role in breast cancer biology.
]]></description>
<dc:creator>Jindal, S.</dc:creator>
<dc:creator>Pennock, N. D.</dc:creator>
<dc:creator>Klug, A.</dc:creator>
<dc:creator>Narasimhan, J.</dc:creator>
<dc:creator>Calhoun, A.</dc:creator>
<dc:creator>Roberts, M. R.</dc:creator>
<dc:creator>Tamimi, R. M.</dc:creator>
<dc:creator>Eliassen, A. H.</dc:creator>
<dc:creator>Weinmann, S.</dc:creator>
<dc:creator>Borges, V. F.</dc:creator>
<dc:creator>Schedin, P. J.</dc:creator>
<dc:date>2020-05-21</dc:date>
<dc:identifier>doi:10.1101/2020.05.20.104612</dc:identifier>
<dc:title><![CDATA[S-nitrosylated and non-nitrosylated COX2 has differential expression and distinct subcellular localization in normal and breast cancer tissue]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-05-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.05.22.110833v1?rss=1">
<title>
<![CDATA[
Towards complete and error-free genome assemblies of all vertebrate species 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.05.22.110833v1?rss=1</link>
<description><![CDATA[
High-quality and complete reference genome assemblies are fundamental for the application of genomics to biology, disease, and biodiversity conservation. However, such assemblies are only available for a few non-microbial species1-4. To address this issue, the international Genome 10K (G10K) consortium5,6 has worked over a five-year period to evaluate and develop cost-effective methods for assembling the most accurate and complete reference genomes to date. Here we summarize these developments, introduce a set of quality standards, and present lessons learned from sequencing and assembling 16 species representing major vertebrate lineages (mammals, birds, reptiles, amphibians, teleost fishes and cartilaginous fishes). We confirm that long-read sequencing technologies are essential for maximizing genome quality and that unresolved complex repeats and haplotype heterozygosity are major sources of error in assemblies. Our new assemblies identify and correct substantial errors in some of the best historical reference genomes. Adopting these lessons, we have embarked on the Vertebrate Genomes Project (VGP), an effort to generate high-quality, complete reference genomes for all ~70,000 extant vertebrate species and help enable a new era of discovery across the life sciences.
]]></description>
<dc:creator>Rhie, A.</dc:creator>
<dc:creator>McCarthy, S. A.</dc:creator>
<dc:creator>Fedrigo, O.</dc:creator>
<dc:creator>Damas, J.</dc:creator>
<dc:creator>Formenti, G.</dc:creator>
<dc:creator>Koren, S.</dc:creator>
<dc:creator>Uliano-Silva, M.</dc:creator>
<dc:creator>Chow, W.</dc:creator>
<dc:creator>Fungtammasan, A.</dc:creator>
<dc:creator>Gedman, G. L.</dc:creator>
<dc:creator>Cantin, L. J.</dc:creator>
<dc:creator>Thibaud-Nissen, F.</dc:creator>
<dc:creator>Haggerty, L.</dc:creator>
<dc:creator>Lee, C.</dc:creator>
<dc:creator>Ko, B. J.</dc:creator>
<dc:creator>Kim, J.</dc:creator>
<dc:creator>Bista, I.</dc:creator>
<dc:creator>Smith, M.</dc:creator>
<dc:creator>Haase, B.</dc:creator>
<dc:creator>Mountcastle, J.</dc:creator>
<dc:creator>Winkler, S.</dc:creator>
<dc:creator>Paez, S.</dc:creator>
<dc:creator>Howard, J.</dc:creator>
<dc:creator>Vernes, S. C.</dc:creator>
<dc:creator>Lama, T. M.</dc:creator>
<dc:creator>Grutzner, F. C.</dc:creator>
<dc:creator>Warren, W. C.</dc:creator>
<dc:creator>Balakrishnan, C.</dc:creator>
<dc:creator>Burt, D.</dc:creator>
<dc:creator>George, J. M.</dc:creator>
<dc:creator>Biegler, M.</dc:creator>
<dc:creator>Iorns, D.</dc:creator>
<dc:creator>Digby, A.</dc:creator>
<dc:creator>Eason, D.</dc:creator>
<dc:creator>Edwards, T.</dc:creator>
<dc:creator>Wilkinson, M.</dc:creator>
<dc:creator>Turner, G. F.</dc:creator>
<dc:creator>Meyer, A.</dc:creator>
<dc:creator>Kautt, A. F.</dc:creator>
<dc:creator>Franchini, P.</dc:creator>
<dc:creator>Detrich, H. W.</dc:creator>
<dc:creator>Svardal, H.</dc:creator>
<dc:creator>Wagner, M.</dc:creator>
<dc:creator>Naylor, G. J. P.</dc:creator>
<dc:creator>Pippel, M</dc:creator>
<dc:date>2020-05-23</dc:date>
<dc:identifier>doi:10.1101/2020.05.22.110833</dc:identifier>
<dc:title><![CDATA[Towards complete and error-free genome assemblies of all vertebrate species]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-05-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.05.28.121905v1?rss=1">
<title>
<![CDATA[
Ventral tegmental area GABA neurons mediate stress-induced anhedonia. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.05.28.121905v1?rss=1</link>
<description><![CDATA[
Stressful experiences frequently precede depressive episodes1. Depression results in anhedonia, or disrupted reward-seeking, in most patients2. In humans3,4 and rodents5,6, stress can disrupt reward-seeking, providing a potential mechanism by which stress can precipitate depression7-9. Yet despite decades investigating how stress modulates dopamine neuron transmission between the ventral tegmental area (VTA) and nucleus accumbens (NAc), the underpinnings of the stress-anhedonia transition remain elusive10-13. Here we show that during restraint stress, VTA GABA neurons drive low frequency NAc LFP oscillations, rhythmically modulating NAc firing rates. The strength of these stress-induced NAc oscillations predict the degree of impaired reward-seeking upon release from restraint. Inhibiting VTA GABA neurons disrupts stress-induced NAc oscillations and reverses the effect of stress on reward-seeking. By contrast, mimicking these oscillations with rhythmic VTA GABA stimulation in the absence of stress blunts subsequent reward-seeking. These experiments demonstrate that VTA GABA inputs to the NAc are both necessary and sufficient for stress-induced decreases in reward seeking behavior, elucidating a key circuit-level mechanism underlying stress-induced anhedonia.
]]></description>
<dc:creator>Lowes, D. C.</dc:creator>
<dc:creator>Chamberlin, L. A.</dc:creator>
<dc:creator>Kretsge, L. N.</dc:creator>
<dc:creator>Holt, E. S.</dc:creator>
<dc:creator>Abbas, A. I.</dc:creator>
<dc:creator>Park, A. J.</dc:creator>
<dc:creator>Yusufova, L.</dc:creator>
<dc:creator>Bretton, Z. H.</dc:creator>
<dc:creator>Firdous, A.</dc:creator>
<dc:creator>Gordon, J. A.</dc:creator>
<dc:creator>Harris, A. Z.</dc:creator>
<dc:date>2020-05-31</dc:date>
<dc:identifier>doi:10.1101/2020.05.28.121905</dc:identifier>
<dc:title><![CDATA[Ventral tegmental area GABA neurons mediate stress-induced anhedonia.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-05-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.05.28.122333v1?rss=1">
<title>
<![CDATA[
A deep intronic variant activates a pseudoexon in the MTM1 gene in a family with X-linked myotubular myopathy 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.05.28.122333v1?rss=1</link>
<description><![CDATA[
We report a novel intronic variant in the MTM1 gene in four males in a family with severe X-linked myotubular myopathy. The A>G variant in deep intronic space activates a cryptic 5 donor splice site resulting in the inclusion of a 48bp pseudoexon into the mature MTM1 mRNA. The variant is present in all affected males, absent in unaffected males and heterozygous in the mother of the affected males. The included intronic sequence contains a premature stop codon and experiments using a translational inhibitor indicate that the mutant mRNAs undergo nonsense-mediate decay. We conclude that affected males produce no, or low, levels of myotubularin-1 protein leading to a severe neonatal myopathy. The study highlights the need to consider non-coding variants in genomic screening in families with X-linked myotubular myopathy.
]]></description>
<dc:creator>Fitzgerald, J.</dc:creator>
<dc:creator>Feist, C.</dc:creator>
<dc:creator>Dietz, P.</dc:creator>
<dc:creator>Moore, S.</dc:creator>
<dc:creator>Basel, D.</dc:creator>
<dc:date>2020-05-30</dc:date>
<dc:identifier>doi:10.1101/2020.05.28.122333</dc:identifier>
<dc:title><![CDATA[A deep intronic variant activates a pseudoexon in the MTM1 gene in a family with X-linked myotubular myopathy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-05-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.05.28.122341v1?rss=1">
<title>
<![CDATA[
CRISPR Knockdown of Kcnq3 Attenuates the M-current in NPY/AgRP Neurons 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.05.28.122341v1?rss=1</link>
<description><![CDATA[
Arcuate nucleus Neuropeptide Y/Agouti-related peptide (NPY/AgRP) neurons drive ingestive behavior in response to the internal and external environment of an organism. NPY/AgRP neurons are adjacent to the median eminence, a circumventricular organ, and circulating metabolic factors and hormones communicate the energy state of the animal via these neurons by altering the excitability of NPY/AgRP neurons, which produces an appropriate change in behavior to maintain homeostasis. One example of this plasticity is seen in the M-current, a subthreshold, non-inactivating K+ current that acts to modulate excitability. Fasting decreases while estradiol increases the M-current through regulation of subunit mRNA expression of Kcnq 2, 3, & 5. KCNQ2/3 heteromers are thought to mediate the majority of the M-current. Here we used a recently developed single adeno-associated viral (AAV) vector containing a recombinase-dependent Staphylococcus aureus Cas9 (SaCas9) and a single guide RNA against Kcnq3 to selectively delete Kcnq3 in NPY/AgRP neurons to produce a loss of function in the M-current. We found that this virus was effective at knocking down Kcnq3 but not Kcnq2 expression. With the reduced KCNQ3 channel expression NPY/AgRP neurons were more depolarized, exhibited a higher input resistance, and the rheobase current needed to induce firing was significantly reduced, indicative of increased excitability. Although the resulting decrease in the M-current did not overtly alter ingestive behavior, it did significantly reduce the locomotor activity as measured in open field testing. Therefore, the SaCas9-sgKcnq3 is efficient to knock down Kcnq3 expression thereby reducing the M-current and increasing the excitability of NPY/AgRP neurons.
]]></description>
<dc:creator>Stincic, T. L.</dc:creator>
<dc:creator>Bosch, M. A.</dc:creator>
<dc:creator>Hunker, A. C.</dc:creator>
<dc:creator>Juarez, B.</dc:creator>
<dc:creator>Connors, A. M.</dc:creator>
<dc:creator>Zweifel, L.</dc:creator>
<dc:creator>Rønnekleiv, O. K.</dc:creator>
<dc:creator>Kelly, M. J.</dc:creator>
<dc:date>2020-05-29</dc:date>
<dc:identifier>doi:10.1101/2020.05.28.122341</dc:identifier>
<dc:title><![CDATA[CRISPR Knockdown of Kcnq3 Attenuates the M-current in NPY/AgRP Neurons]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-05-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.05.28.122697v1?rss=1">
<title>
<![CDATA[
Competition and Caries on Enamel of a Dual-species Biofilm Model of Streptococcus mutans and Streptococcus sanguinis. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.05.28.122697v1?rss=1</link>
<description><![CDATA[
Imbalances within the dental biofilm trigger dental caries, currently considered a dysbiosis and the most prevalent non-communicable disease. There is still a gap in knowledge about the dynamics of enamel colonization by bacteria from the dental biofilm in caries. The aim, therefore, was to test whether the sequence of enamel colonization by a typically commensal and a cariogenic species modifies biofilms cariogenicity. Dual-species biofilms of Streptococcus mutans (Sm) and Streptococcus sanguinis (Ss) on saliva-coated enamel slabs were inoculated in different sequences: Sm followed by Ss (Sm-Ss), Ss followed by Sm (Ss-Sm), Sm and Ss inoculated at the same time (Sm=Ss) and the single-species controls Sm followed by Sm (Sm-Sm) and Ss followed by Ss (Ss-Ss). Biofilms were exposed to 10% sucrose, 3x/day for 5 days and the slabs/biofilms were retrieved to assess demineralization, viable cells, biomass, proteins, polysaccharides and H2O2 production. When compared with Sm-Sm, primary inoculation with Ss reduced demineralization (p<0.05). Both Ss-Sm and Sm=Ss sequences showed reduction in biomass, protein and polysaccharide content (p<0.05). The highest S. sanguinis viable cells and H2O2 production and the lowest acidogenicity were observed when Ss colonized enamel before Sm (p<0.05). Initial enamel adherence with commensal biofilms seems to induce more intense competition against more typically cariogenic species, reducing cariogenicity.

ImportanceThe concept of caries as an ecological disease implies the understanding of the intricate relationships among the populating microorganisms. Under frequent sugars exposure, some the bacteria from the oral biofilm develop pathogenic traits that lead to oral imbalances, known as dysbiosis. Depending on which microorganism colonizes the dental surface first, different competition strategies may be developed. Since the study of the interactions in the entire dental biofilm is not an easy task, in this article we model the interplay among these microorganisms using a caries-inducing (S. mutans) and a health-associated species (S. sanguinis). Initial enamel adherence with S. sanguinis seems to induce more intense competition against more typically caries-inducing species. Besides continuous exposure with sugars, early colonization of the enamel by highly cariogenic species, like S. mutans, appears to be needed to develop caries lesions, as well. Promoting early colonization by health-associated bacteria, such as S. sanguinis, could help maintaining oral health, delaying dysbiosis.
]]></description>
<dc:creator>Diaz-Garrido, N.</dc:creator>
<dc:creator>Lozano, C.</dc:creator>
<dc:creator>Kreth, J.</dc:creator>
<dc:creator>Giacaman, R. A.</dc:creator>
<dc:date>2020-05-31</dc:date>
<dc:identifier>doi:10.1101/2020.05.28.122697</dc:identifier>
<dc:title><![CDATA[Competition and Caries on Enamel of a Dual-species Biofilm Model of Streptococcus mutans and Streptococcus sanguinis.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-05-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.05.30.125468v1?rss=1">
<title>
<![CDATA[
Lmo7a Coordinates Neural Crest Migration and Lineage Specification by Regulating Cell Adhesion Dynamics 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.05.30.125468v1?rss=1</link>
<description><![CDATA[
Cell migration requires dynamic regulation of cell-cell signaling and cell adhesion. Neural crest (NC) cells are highly migratory cells, which undergo an epithelial-mesenchymal transition (EMT) to leave the neural epithelium and migrate throughout the body to give rise to many different derivatives. We have identified a Lim-domain only (Lmo) protein, Lmo7a, expressed in early NC cells that controls both actin cytoskeletal dynamics and Wnt signaling during NC migration. In embryos deficient in Lmo7a, many NC cells fail to migrate away from the dorsal midline, and form aggregates. Unlike the majority of NC cells that appear to migrate normally, cells that aggregate in Lmo7a-deficient embryos mislocalize paxillin (Pxn) and have reduced levels of phosphorylated focal adhesion kinase (pFAK). Lmo7a loss-of-function also disrupts canonical Wnt signaling such that after the onset of NC cell migration, Wnt responses and nuclear {beta}-catenin levels increase in the cells that aggregate. However, this increase in Wnt signaling appears secondary to the defect in migration. Similar to mutants for other Wnt regulators in NC cells, the NC cells in Lmo7a-deficient aggregates exhibit gene expression signatures of pigment cell progenitors, but also express markers of Schwann cell progenitors, suggesting a role for Lmo7a in pigment-glial specification. We propose that Lmo7a modulates cell adhesion to facilitate both robust NC cell migration and a subset of lineage decisions.
]]></description>
<dc:creator>Tatarakis, D. M.</dc:creator>
<dc:creator>Tuttle, A.</dc:creator>
<dc:creator>Schilling, T. F.</dc:creator>
<dc:date>2020-05-31</dc:date>
<dc:identifier>doi:10.1101/2020.05.30.125468</dc:identifier>
<dc:title><![CDATA[Lmo7a Coordinates Neural Crest Migration and Lineage Specification by Regulating Cell Adhesion Dynamics]]></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/10.1101/2020.06.02.128850v1?rss=1">
<title>
<![CDATA[
Systematic interrogation of mutation groupings reveals divergent downstream expression programs within key cancer genes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.06.02.128850v1?rss=1</link>
<description><![CDATA[
Genes implicated in tumorigenesis often exhibit diverse sets of genomic variants in the tumor cohorts within which they are frequently mutated. We sought to identify the downstream expression effects of these perturbations and to find whether or not this heterogeneity at the genomic level is reflected in a corresponding heterogeneity at the transcriptomic level. Applying a novel hierarchical framework for organizing the mutations present in a cohort along with machine learning pipelines trained on sample expression profiles we systematically interrogated the signatures associated with combinations of perturbations recurrent in cancer. This allowed us to catalogue the mutations with discernible downstream expression effects across a number of tumor cohorts as well as to uncover and characterize a multitude of cases where subsets of a genes mutations are clearly divergent in their function from the remaining mutations of the gene.
]]></description>
<dc:creator>Grzadkowski, M. R.</dc:creator>
<dc:creator>Manning, H.</dc:creator>
<dc:creator>Somers, J.</dc:creator>
<dc:creator>Demir, E.</dc:creator>
<dc:date>2020-06-03</dc:date>
<dc:identifier>doi:10.1101/2020.06.02.128850</dc:identifier>
<dc:title><![CDATA[Systematic interrogation of mutation groupings reveals divergent downstream expression programs within key cancer genes]]></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/10.1101/2020.06.02.129486v1?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/10.1101/2020.06.05.137240v1?rss=1">
<title>
<![CDATA[
Phenotype-guided subpopulation identification from single-cell sequencing data 
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</title>
<link>https://biorxiv.org/content/10.1101/2020.06.05.137240v1?rss=1</link>
<description><![CDATA[
Single-cell sequencing yields novel discoveries by distinguishing cell types, states and lineages within the context of heterogeneous tissues. However, interpreting complex single-cell data from highly heterogeneous cell populations remains challenging. Currently, most existing single-cell data analyses focus on cell type clusters defined by unsupervised clustering methods, which cannot directly link cell clusters with specific biological and clinical phenotypes. Here we present Scissor, a novel approach that utilizes disease phenotypes to identify cell subpopulations from single-cell data that most highly correlate with a given phenotype. This "phenotype-to-cell within a single step" strategy enables the utilization of a large amount of clinical information that has been collected for bulk assays to identify the most highly phenotype-associated cell subpopulations. When applied to a lung cancer single-cell RNA-seq (scRNA-seq) dataset, Scissor identified a subset of cells exhibiting high hypoxia activities, which predicted worse survival outcomes in lung cancer patients. Furthermore, in a melanoma scRNA-seq dataset, Scissor discerned a T cell subpopulation with low PDCD1/CTLA4 and high TCF7 expressions, which is associated with a favorable immunotherapy response. Thus, Scissor provides a novel framework to identify the biologically and clinically relevant cell subpopulations from single-cell assays by leveraging the wealth of phenotypes and bulk-omics datasets.
]]></description>
<dc:creator>Sun, D.</dc:creator>
<dc:creator>Guan, X.</dc:creator>
<dc:creator>Moran, A. E.</dc:creator>
<dc:creator>Qian, D. Z.</dc:creator>
<dc:creator>Schedin, P. J.</dc:creator>
<dc:creator>Adey, A.</dc:creator>
<dc:creator>Spellman, P. T.</dc:creator>
<dc:creator>Xia, Z.</dc:creator>
<dc:date>2020-06-07</dc:date>
<dc:identifier>doi:10.1101/2020.06.05.137240</dc:identifier>
<dc:title><![CDATA[Phenotype-guided subpopulation identification from single-cell sequencing data]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-06-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.06.07.138693v1?rss=1">
<title>
<![CDATA[
Kinome Profiling of Gastrointestinal Stromal Tumors Using Multiplexed Inhibitor Beads and Mass Spectrometry Identifies Wee1 as Candidate Therapeutic Target 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.06.07.138693v1?rss=1</link>
<description><![CDATA[
Management of gastrointestinal stromal tumor (GIST) has been revolutionized by the identification of activating mutations in KIT and PDGFRA, and the clinical application of receptor tyrosine kinase (RTK) inhibitors in the advanced disease setting. Stratification of GIST into molecularly defined subsets provides insight into clinical behavior and response to approved targeted therapies. Although these RTK inhibitors are effective in the majority of GIST, resistance to these agents remains a significant clinical problem. Development of effective treatment strategies for refractory GIST subtypes requires identification of novel targets to provide additional therapeutic options. Global kinome profiling has the potential to identify critical signaling networks and reveal protein kinases that are essential in GIST. Using Multiplexed Inhibitor Beads and Mass Spectrometry, we explored the majority of the kinome in GIST specimens from the three most common molecular subtypes to identify novel kinase targets. Kinome profiling revealed distinct signatures in GIST subtypes and identified kinases that are universally activated in all GIST, as well as kinases that are unique to each subtype. Kinome profiling in combination with loss-of-function assays identified a significant role for the G2-M tyrosine kinase, Wee1, in GIST cell survival. In vitro and in vivo studies revealed significant efficacy of MK-1775 (Wee1 inhibitor) in combination with avapritinib in KIT and PDGFRA-mutant GIST cell lines, and notable efficacy of MK-1775 as a single agent in the PDGFRA-mutant line. These studies provide strong preclinical justification for the use of MK-1775 in GIST.
]]></description>
<dc:creator>Ye, S.</dc:creator>
<dc:creator>Sharipova, D.</dc:creator>
<dc:creator>Kozinova, M.</dc:creator>
<dc:creator>Klug, L.</dc:creator>
<dc:creator>D'Souza, J.</dc:creator>
<dc:creator>Belinsky, M. G.</dc:creator>
<dc:creator>Johnson, K. J.</dc:creator>
<dc:creator>Einarson, M. B.</dc:creator>
<dc:creator>Cai, K.</dc:creator>
<dc:creator>Devarajan, K.</dc:creator>
<dc:creator>Zhou, Y.</dc:creator>
<dc:creator>Litwin, S.</dc:creator>
<dc:creator>Heinrich, M. C.</dc:creator>
<dc:creator>DeMatteo, R.</dc:creator>
<dc:creator>von Mehren, M.</dc:creator>
<dc:creator>Duncan, J.</dc:creator>
<dc:creator>Rink, L.</dc:creator>
<dc:date>2020-06-08</dc:date>
<dc:identifier>doi:10.1101/2020.06.07.138693</dc:identifier>
<dc:title><![CDATA[Kinome Profiling of Gastrointestinal Stromal Tumors Using Multiplexed Inhibitor Beads and Mass Spectrometry Identifies Wee1 as Candidate Therapeutic Target]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-06-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.06.16.155432v1?rss=1">
<title>
<![CDATA[
Dissociation of task engagement and arousal effects in auditory cortex and midbrain 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.06.16.155432v1?rss=1</link>
<description><![CDATA[
Both generalized arousal and engagement in a specific task influence sensory neural processing. To isolate effects of these state variables in the auditory system, we recorded single-unit activity from primary auditory cortex (A1) and inferior colliculus (IC) of ferrets during a tone detection task, while monitoring arousal via changes in pupil size. We used a generalized linear model to assess the influence of task engagement and pupil size on sound-evoked activity. In both areas, these two variables affected independent neural populations. Pupil size effects were more prominent in IC, while pupil and task engagement effects were equally likely in A1. Task engagement was correlated with larger pupil; thus, some apparent effects of task engagement should in fact be attributed to fluctuations in pupil size. These results indicate a hierarchy of auditory processing, where generalized arousal enhances activity in midbrain, and effects specific to task engagement become more prominent in cortex.
]]></description>
<dc:creator>Saderi, D.</dc:creator>
<dc:creator>Schwartz, Z. P.</dc:creator>
<dc:creator>Heller, C. R.</dc:creator>
<dc:creator>Pennington, J. R.</dc:creator>
<dc:creator>David, S. V.</dc:creator>
<dc:date>2020-06-17</dc:date>
<dc:identifier>doi:10.1101/2020.06.16.155432</dc:identifier>
<dc:title><![CDATA[Dissociation of task engagement and arousal effects in auditory cortex and midbrain]]></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/10.1101/2020.06.22.164418v1?rss=1">
<title>
<![CDATA[
Modeling heterogeneity of Triple-Negative Breast Cancer highlights potency of WEE1 and BCL-XL targeting 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.06.22.164418v1?rss=1</link>
<description><![CDATA[
Triple-negative breast cancer (TNBC) is a highly aggressive breast cancer subtype characterized by a remarkable molecular heterogeneity. Currently, there are no effective druggable targets and advanced preclinical models of the human disease. Here, we generated a unique mouse model (MMTV-R26Met mice) of mammary tumors driven by a subtle increase in the expression of the wild-type MET receptor. MMTV-R26Met mice develop spontaneous, exclusive TNBC tumors, recapitulating primary resistance to treatment of patients. Proteomic profiling of MMTV-R26Met tumors and machine learning approach showed that the model faithfully recapitulates inter-tumoral heterogeneity of human TNBC. Further signaling network analysis highlighted potential druggable targets, of which co-targeting of WEE1 and BCL-XL synergistically killed TNBC cells and efficiently induced tumor regression. Mechanistically, BCL-XL inhibition exacerbates the dependency of TNBC cells on WEE1 function, leading to Histone H3 and phosphoS33RPA32 upregulation, RRM2 downregulation, cell cycle perturbation, mitotic catastrophe and apoptosis. Our study introduces a unique, powerful mouse model for studying TNBC formation and evolution, its heterogeneity, and for identifying efficient therapeutic targets.
]]></description>
<dc:creator>Lamballe, F.</dc:creator>
<dc:creator>Ahmad, F.</dc:creator>
<dc:creator>Vinik, Y.</dc:creator>
<dc:creator>Castellanet, O.</dc:creator>
<dc:creator>Daian, F.</dc:creator>
<dc:creator>Mueller, A.</dc:creator>
<dc:creator>Koehler, U.</dc:creator>
<dc:creator>Bailly, A.-L.</dc:creator>
<dc:creator>Josselin, E.</dc:creator>
<dc:creator>Castellano, R.</dc:creator>
<dc:creator>Cayrou, C.</dc:creator>
<dc:creator>Charafe-Jauffret, E.</dc:creator>
<dc:creator>Mills, G. B.</dc:creator>
<dc:creator>Geli, V.</dc:creator>
<dc:creator>Borg, J.-P.</dc:creator>
<dc:creator>Lev, S.</dc:creator>
<dc:creator>Maina, F.</dc:creator>
<dc:date>2020-06-22</dc:date>
<dc:identifier>doi:10.1101/2020.06.22.164418</dc:identifier>
<dc:title><![CDATA[Modeling heterogeneity of Triple-Negative Breast Cancer highlights potency of WEE1 and BCL-XL targeting]]></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/10.1101/2020.06.25.172445v1?rss=1">
<title>
<![CDATA[
Accessible, Reproducible, and Scalable Machine Learning for Biomedicine 
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</title>
<link>https://biorxiv.org/content/10.1101/2020.06.25.172445v1?rss=1</link>
<description><![CDATA[
Supervised machine learning, where the goal is to predict labels of new instances by training on labeled data, has become an essential tool in biomedical data analysis. To make supervised machine learning more accessible to biomedical scientists, we have developed Galaxy-ML, a platform that enables scientists to perform end-to-end reproducible machine learning analyses at large scale using only a web browser. Galaxy-ML extends Galaxy, a biomedical computational workbench used by tens of thousands of scientists across the world, with a machine learning tool suite that supports end-to-end analysis.
]]></description>
<dc:creator>Gu, Q.</dc:creator>
<dc:creator>Kumar, A.</dc:creator>
<dc:creator>Bray, S. A.</dc:creator>
<dc:creator>Creason, A.</dc:creator>
<dc:creator>Khanteymoori, A.</dc:creator>
<dc:creator>Jalili, V.</dc:creator>
<dc:creator>Gruning, B.</dc:creator>
<dc:creator>Goecks, J.</dc:creator>
<dc:date>2020-06-28</dc:date>
<dc:identifier>doi:10.1101/2020.06.25.172445</dc:identifier>
<dc:title><![CDATA[Accessible, Reproducible, and Scalable Machine Learning for Biomedicine]]></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/10.1101/2020.06.25.172676v1?rss=1">
<title>
<![CDATA[
Myristoylation alone is sufficient for PKA catalytic subunits to fractionally associate with the plasma membrane to regulate neuronal functions 
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</title>
<link>https://biorxiv.org/content/10.1101/2020.06.25.172676v1?rss=1</link>
<description><![CDATA[
Myristoylation is a post-translational modification that plays diverse functional roles in many protein species. The myristate moiety is considered insufficient for protein-membrane associations unless additional membrane-affinity motifs, such as a stretch of positively charged residues, are present. Here, we report that the electrically neutral N-terminal fragment of the protein kinase A catalytic subunit (PKA-C), in which myristoylation is the only functional motif, is sufficient for membrane association. This myristoylation can associate a fraction of PKA-C molecules or fluorescent proteins (FPs) to the plasma membrane in neuronal dendrites. The net neutral charge of PKA-C is evolutionally conserved, even though its membrane affinity can be readily tuned by changing charges near the myristoylation site. The observed membrane association, while moderate, is sufficient to concentrate PKA activity at the membrane by nearly 20-fold, and is required for PKA regulation of AMPA receptors at neuronal synapses. Our results indicate that myristoylation alone may be sufficient to drive functionally significant membrane association in the absence of assisting motifs. This provides a revised foundation for the understanding of how myristoylation regulates protein functions.
]]></description>
<dc:creator>Xiong, W.-H.</dc:creator>
<dc:creator>Qin, M.</dc:creator>
<dc:creator>Zhong, H.</dc:creator>
<dc:date>2020-06-27</dc:date>
<dc:identifier>doi:10.1101/2020.06.25.172676</dc:identifier>
<dc:title><![CDATA[Myristoylation alone is sufficient for PKA catalytic subunits to fractionally associate with the plasma membrane to regulate neuronal functions]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-06-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.06.26.172965v1?rss=1">
<title>
<![CDATA[
Multi- and transgenerational disruption of maternal behavior and female puberty by Endocrine Disrupting Chemical (EDC) mixture exposure 
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</title>
<link>https://biorxiv.org/content/10.1101/2020.06.26.172965v1?rss=1</link>
<description><![CDATA[
Female reproductive development and maternal behavior are two intertwined phenotypes centrally controlled by the hypothalamus. Endocrine disrupting chemicals (EDC) can alter these processes especially when animals are exposed during development. We propose the concept that developmental exposure to a low environmentally relevant dose of EDC mixture induces a transgenerational alteration of female rat pubertal timing and ovarian physiology throughout epigenetic reprograming of hypothalamic Kiss1, Esr1 and Oxt1 loci. Such exposure also caused a multigenerational reduction of maternal behavior induced by the loss in hypothalamic dopaminergic signaling. Our results identify the hypothalamic Polycomb Group of epigenetic repressors as actors of this mechanism of transgenerational reproductive disruption. Using a cross-fostering approach, we identified that while the reduction in maternal phenotype was normalized in EDC exposed pups raised by unexposed dams, no reversal of the pubertal phenotype was achieved, suggesting a germline transmission of the reproductive phenotype.
]]></description>
<dc:creator>Lopez Rodriguez, D.</dc:creator>
<dc:creator>Aylwin, C. F.</dc:creator>
<dc:creator>Delli, V.</dc:creator>
<dc:creator>Sevrin, E.</dc:creator>
<dc:creator>Campanile, M.</dc:creator>
<dc:creator>Martin, M.</dc:creator>
<dc:creator>Franssen, D.</dc:creator>
<dc:creator>Gerard, A.</dc:creator>
<dc:creator>Blacher, S.</dc:creator>
<dc:creator>Tirelli, E.</dc:creator>
<dc:creator>Noel, A.</dc:creator>
<dc:creator>Lomniczi, A.</dc:creator>
<dc:creator>Parent, A.-S.</dc:creator>
<dc:date>2020-06-29</dc:date>
<dc:identifier>doi:10.1101/2020.06.26.172965</dc:identifier>
<dc:title><![CDATA[Multi- and transgenerational disruption of maternal behavior and female puberty by Endocrine Disrupting Chemical (EDC) mixture exposure]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-06-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.06.30.180356v1?rss=1">
<title>
<![CDATA[
Inhibition of CMP-sialic acid transport by endogenous 5-methyl CMP 
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</title>
<link>https://biorxiv.org/content/10.1101/2020.06.30.180356v1?rss=1</link>
<description><![CDATA[
Nucleotide-sugar transporters (NSTs) transport nucleotide-sugar conjugates into the Golgi lumen where they are then used in the synthesis of glycans. We previously reported crystal structures of a mammalian NST, the CMP-sialic acid transporter (CST) (Ahuja and Whorton 2019). These structures elucidated many aspects of substrate recognition, selectivity, and transport; however, one fundamental unaddressed question is how the transport activity of NSTs might be physiologically regulated as a means to produce the vast diversity of observed glycan structures. Here, we describe the discovery that an endogenous methylated form of cytidine monophosphate (m5CMP) binds and inhibits CST. The presence of m5CMP in cells results from the degradation of RNA that has had its cytosine bases post-transcriptionally methylated through epigenetic processes. Therefore, this work not only demonstrates that m5CMP represents a novel physiological regulator of CST, but it also establishes a link between epigenetic control of gene expression and regulation of glycosylation.View Full Text
]]></description>
<dc:creator>Ahjua, S.</dc:creator>
<dc:creator>Cahill, J.</dc:creator>
<dc:creator>Hartfield, K. A.</dc:creator>
<dc:creator>Whorton, M. R.</dc:creator>
<dc:date>2020-07-01</dc:date>
<dc:identifier>doi:10.1101/2020.06.30.180356</dc:identifier>
<dc:title><![CDATA[Inhibition of CMP-sialic acid transport by endogenous 5-methyl CMP]]></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/10.1101/2020.07.01.182584v1?rss=1">
<title>
<![CDATA[
Connexin 46 and connexin 50 gap junction channel open stability and unitary conductance are shaped by structural and dynamic features of their N-terminal domains 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.07.01.182584v1?rss=1</link>
<description><![CDATA[
The connexins form intercellular communication channels, known as gap junctions (GJs), that facilitate diverse physiological roles in vertebrate species, ranging from electrical coupling and long-range chemical signaling, to coordinating development and nutrient exchange. GJs formed by different connexins are expressed throughout the body and harbor unique channel properties that have not been fully defined mechanistically. Recent structural studies have implicated the amino-terminal (NT) domain as contributing to isoform-specific functional differences that exist between the lens connexins, Cx50 and Cx46. To better understand the structural and functional differences in the two closely related, yet functionally distinct GJs, we constructed models corresponding to CryoEM-based structures of the wildtype Cx50 and Cx46 GJs, NT domain swapped chimeras (Cx46-50NT and Cx50-46NT), and point variants at the 9th residue (Cx46-R9N and Cx50-N9R) for comparative MD simulation and electrophysiology studies. All of these constructs formed functional GJ channels, except Cx46-50NT, which correlated with increased dynamical behavior (instability) of the NT domain observed by MD simulation. Single channel conductance (γj) also correlated well with free-energy landscapes predicted by MD, where γj of Cx46-R9N was increased from Cx46 and the γjs of Cx50-46NT and Cx50-N9R was decreased from Cx50, but to a surprisingly greater degree. Additionally, we observed significant effects on transjunctional voltage-dependent gating (Vj-gating) and open-state dwell times induced by the designed NT domain variants. Together, these studies indicate that the NT domains of Cx46 and Cx50 play an important role in defining channel properties related to open-state stability and single channel conductance.Competing Interest StatementThe authors have declared no competing interest.AbbreviationsGJgap junctionGjgap junctional coupling conductanceγjsingle gap junction channel conductanceIjmacroscopic transjunctional currentijsingle gap junction channel currentCx46sheep connexin 46Cx50sheep connexin 50Vjtransjunctional voltageMDmolecular dynamicsr.m.s.d.root mean square deviationr.m.s.f.root mean square fluctuationPMFpotential of mean forceView Full Text
]]></description>
<dc:creator>Yue, B.</dc:creator>
<dc:creator>Haddad, B. G.</dc:creator>
<dc:creator>Khan, U.</dc:creator>
<dc:creator>Chen, H.</dc:creator>
<dc:creator>Atalla, M.</dc:creator>
<dc:creator>Zhang, Z.</dc:creator>
<dc:creator>Zuckerman, D. M.</dc:creator>
<dc:creator>Reichow, S. L.</dc:creator>
<dc:creator>Bai, D.</dc:creator>
<dc:date>2020-07-01</dc:date>
<dc:identifier>doi:10.1101/2020.07.01.182584</dc:identifier>
<dc:title><![CDATA[Connexin 46 and connexin 50 gap junction channel open stability and unitary conductance are shaped by structural and dynamic features of their N-terminal domains]]></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/10.1101/2020.07.06.185066v1?rss=1">
<title>
<![CDATA[
Genome sequencing analysis identifies new loci associated with Lewy body dementia and provides insights into the complex genetic architecture 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.07.06.185066v1?rss=1</link>
<description><![CDATA[
The genetic basis of Lewy body dementia (LBD) is not well understood. Here, we performed whole-genome sequencing in large cohorts of LBD cases and neurologically healthy controls to study the genetic architecture of this understudied form of dementia and to generate a resource for the scientific community. Genome-wide association analysis identified five independent risk loci, whereas genome-wide gene-aggregation tests implicated mutations in the gene GBA. Genetic risk scores demonstrate that LBD shares risk profiles and pathways with Alzheimer’s and Parkinson’s disease, providing a deeper molecular understanding of the complex genetic architecture of this age-related neurodegenerative condition.Competing Interest StatementThomas G. Beach is a consultant for Prothena, Vivid Genomics and Avid Radiopharmaceuticals. He is a scientific advisory board member for Vivid Genomics. John A. Hardy, Huw R. Morris, Stuart Pickering-Brown, Andrew B. Singleton, and Bryan J. Traynor hold US, EU and Canadian patents on the clinical testing and therapeutic intervention for the hexanucleotide repeat expansion of C9orf72. Michael A. Nalls is supported by a consulting contract between Data Tecnica International and the National Institute on Aging, NIH, Bethesda, MD, USA; as a possible conflict of interest Dr. Nalls also consults for Neuron23 Inc., Lysosomal Therapeutics Inc., Illumina Inc., the Michael J. Fox Foundation and Vivid Genomics among others. Jose A. Palma is an editorial board member of Movement Disorders, Parkinsonism &amp; Related Disorders, BMC Neurology, and Clinical Autonomic Research. Bradley F. Boeve, James Leverenz, and Sonja W. Scholz serve on the Scientific Advisory Council of the Lewy Body Dementia Association. Sonja W. Scholz is an editorial board member for the Journal of Parkinson's Disease. Bryan J. Traynor is an editorial board member for JAMA Neurology; Journal of Neurology, Neurosurgery, and Psychiatry; Brain; and Neurobiology of Aging. Zbigniew K. Wszolek serves as a principal investigator or co-principal investigator on Abbvie, Inc. (M15-562 and M15-563), Biogen, Inc. (228PD201) grant, and Biohaven Pharmaceuticals, Inc. (BHV4157-206 and BHV3241-301). Zbigniew K. Wszolek serves as the principal investigator of the Mayo Clinic American Parkinson Disease Association (APDA) Information and Referral Center, and as co-principal investigator of the Mayo Clinic APDA Center for Advanced Research. All other authors report no competing interests.View Full Text
]]></description>
<dc:creator>Chia, R.</dc:creator>
<dc:creator>Sabir, M. S.</dc:creator>
<dc:creator>Bandres-Ciga, S.</dc:creator>
<dc:creator>Saez-Atienzar, S.</dc:creator>
<dc:creator>Reynolds, R. H.</dc:creator>
<dc:creator>Gustavsson, E.</dc:creator>
<dc:creator>Walton, R. L.</dc:creator>
<dc:creator>Ahmed, S.</dc:creator>
<dc:creator>Viollet, C.</dc:creator>
<dc:creator>Ding, J.</dc:creator>
<dc:creator>Makarious, M. B.</dc:creator>
<dc:creator>Diez-Fairen, M.</dc:creator>
<dc:creator>Portley, M. K.</dc:creator>
<dc:creator>Shah, Z.</dc:creator>
<dc:creator>Abramzon, Y.</dc:creator>
<dc:creator>Hernandez, D. G.</dc:creator>
<dc:creator>Blauwendraat, C.</dc:creator>
<dc:creator>Stone, D. J.</dc:creator>
<dc:creator>Eicher, J.</dc:creator>
<dc:creator>Parkkinen, L.</dc:creator>
<dc:creator>Ansorge, O.</dc:creator>
<dc:creator>Clark, L.</dc:creator>
<dc:creator>Honig, L. S.</dc:creator>
<dc:creator>Marder, K.</dc:creator>
<dc:creator>Lemstra, A.</dc:creator>
<dc:creator>St. George-Hyslop, P.</dc:creator>
<dc:creator>Londos, E.</dc:creator>
<dc:creator>Morgan, K.</dc:creator>
<dc:creator>Lashley, T.</dc:creator>
<dc:creator>Warner, T. T.</dc:creator>
<dc:creator>Jaunmuktane, Z.</dc:creator>
<dc:creator>Galasko, D.</dc:creator>
<dc:creator>Santana, I.</dc:creator>
<dc:creator>Tienari, P.</dc:creator>
<dc:creator>Myllykangas, L.</dc:creator>
<dc:creator>Oinas, M.</dc:creator>
<dc:creator>Cairns, N. J.</dc:creator>
<dc:creator>Morris, J. C.</dc:creator>
<dc:creator>Halliday, G. M.</dc:creator>
<dc:creator>Van Deerlin, V. M.</dc:creator>
<dc:creator>Trojanowski, J.</dc:creator>
<dc:date>2020-07-06</dc:date>
<dc:identifier>doi:10.1101/2020.07.06.185066</dc:identifier>
<dc:title><![CDATA[Genome sequencing analysis identifies new loci associated with Lewy body dementia and provides insights into the complex genetic architecture]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-07-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.07.07.192229v1?rss=1">
<title>
<![CDATA[
Genome-wide assessment of REST binding profiles reveals distinctions between human and mouse hippocampus 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.07.07.192229v1?rss=1</link>
<description><![CDATA[
Background The transcriptional repressor, RE1 Silencing Transcription Factor (REST), recognized historically as a master regulator of neuronal gene expression during mouse development, has recently been ascribed roles in human aging and neurodegenerative disorders. However, REST’s role in healthy adult human brain, and how faithfully mouse models reproduce REST function in human brain, is not known.Results Here, we present the first genome-wide binding profile for REST in both mouse and human postnatal hippocampus. We find the majority of REST-bound sites in human hippocampus are unique compared to both mouse hippocampus and to all other reported human ENCODE cell types. Genes associated with unique REST-bound sites include previously unidentified categories related to innate immunity and inflammation signaling, suggesting species specific roles for REST in protecting human brain health.Conclusions Our results suggest newly evolved functions for REST in maintaining human brain health.Competing Interest StatementThe authors have declared no competing interest.View Full Text
]]></description>
<dc:creator>McGann, J. C.</dc:creator>
<dc:creator>Spinner, M. A.</dc:creator>
<dc:creator>Garg, S. K.</dc:creator>
<dc:creator>Mullendorf, K.</dc:creator>
<dc:creator>Woltjer, R. L.</dc:creator>
<dc:creator>Mandel, G.</dc:creator>
<dc:date>2020-07-07</dc:date>
<dc:identifier>doi:10.1101/2020.07.07.192229</dc:identifier>
<dc:title><![CDATA[Genome-wide assessment of REST binding profiles reveals distinctions between human and mouse hippocampus]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-07-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.07.09.194993v1?rss=1">
<title>
<![CDATA[
A targeted computational screen of the SWEETLEAD database reveals FDA-approved compounds with anti-dengue viral activity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.07.09.194993v1?rss=1</link>
<description><![CDATA[
Affordable and effective antiviral therapies are needed worldwide, especially against agents such as dengue virus that are endemic in underserved regions. Many antiviral compounds have been studied in cultured cells but are unsuitable for clinical applications due to pharmacokinetic profiles, side effects, or inconsistent efficacy across dengue serotypes. Such tool compounds can, however, aid in identifying clinically useful treatments. Here, computational screening (Rapid Overlay of Chemical Structures) was used to identify entries in an in silico database of safe-in-human compounds (SWEETLEAD) that display high chemical similarities to known inhibitors of dengue virus. Molecules known to inhibit dengue proteinase NS2B/3, dengue capsid, and the host autophagy pathway were used as query compounds. Following computational and initial virological screening, three FDA-approved compounds that resemble the tool molecules structurally, cause little toxicity and display strong antiviral activity in cultured cells were selected for further analysis. Pyrimethamine (IC50 = 1.2 µM), like the dengue proteinase inhibitor ARDP0006 to which it shows structural similarity, inhibited intramolecular NS2B/3 cleavage. Lack of toxicity allowed testing in mice, in which pyrimethamine also reduced viral loads. Niclosamide (IC50 = 0.28 µM), like dengue core inhibitor ST-148, affected structural components of the virion and inhibited early processes during infection. Vandetanib (IC50 = 1.6 µM), like cellular autophagy inhibitor spautin-1, blocked viral exit from cells and could further be shown to extend survival in vivo. Our approach confirmed previous studies which used extensive high-throughput screening to identify niclosamide and pyrimethamine as antivirals, and it also revealed their likely molecular targets. Thus, three FDA-approved compounds with promising utility for repurposing to treat dengue virus infections and their potential mechanisms were identified using computational tools and minimal phenotypic screening.Author Summary No antiviral therapeutics are currently available for dengue virus infections. By computationally overlaying the 3D chemical structures of compounds known to inhibit dengue virus with those of compounds known to be safe in humans, we identified three FDA-approved compounds that are attractive candidates for drug repurposing towards treatment of dengue virus infections. We identified potential targets of two previously identified antiviral compounds and revealed a previously unknown potential anti-dengue drug, vandetanib. This computational approach to analyze a highly curated library of structures has the benefits of speed and cost efficiency. It also leverages mechanistic work with query compounds used in biomedical research to provide strong hypotheses for the antiviral mechanisms of the safer hit compounds. This workflow to identify compounds with known safety profiles in humans can be expanded to any biological activity for which a small-molecule query compound has been identified, potentially expediting the translation of basic research to clinical interventions.View Full Text
]]></description>
<dc:creator>Jasmine Moshiri</dc:creator>
<dc:creator>David A. Constant</dc:creator>
<dc:creator>Bowen Liu</dc:creator>
<dc:creator>Roberto Mateo</dc:creator>
<dc:creator>Steven Kearnes</dc:creator>
<dc:creator>Paul Novick</dc:creator>
<dc:creator>Ritika Prasad</dc:creator>
<dc:creator>Claude Nagamine</dc:creator>
<dc:creator>Vijay Pande</dc:creator>
<dc:creator>Karla Kirkegaard</dc:creator>
<dc:date>2020-07-09</dc:date>
<dc:identifier>doi:10.1101/2020.07.09.194993</dc:identifier>
<dc:title><![CDATA[A targeted computational screen of the SWEETLEAD database reveals FDA-approved compounds with anti-dengue viral activity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-07-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.07.14.202853v1?rss=1">
<title>
<![CDATA[
Primary and secondary motoneurons use different calcium channel types to control escape and swimming behaviors in zebrafish 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.07.14.202853v1?rss=1</link>
<description><![CDATA[
The escape response and rhythmic swimming in zebrafish are distinct behaviors mediated by two functionally distinct motoneuron (Mn) types. The primary (1{degrees}Mn) type depresses, has a large quantal content (Qc), and a high release probability (Pr). Conversely, the secondary (2{degrees}Mn) type facilitates and has low and variable Qc and Pr. This functional duality matches well the distinct associated behaviors, with the 1{degrees}Mn providing the strong, singular C-bend initiating escape and the 2{degrees}Mn confers weaker, rhythmic contractions. Contributing to these functional distinctions is our identification of P/Q type calcium channels mediating transmitter release in 1{degrees}Mns and N type channels in 2{degrees}Mns. Remarkably, despite these functional and behavioral distinctions, all ~15 individual synapses on each muscle cell are shared by a 1{degrees}Mn bouton and at least one 2{degrees}Mn bouton. This novel blueprint of synaptic sharing provides an efficient way of controlling two different behaviors at the level of a single postsynaptic cell.
]]></description>
<dc:creator>Wen, H.</dc:creator>
<dc:creator>Eckenstein, K.</dc:creator>
<dc:creator>Weihrauch, V.</dc:creator>
<dc:creator>Stigloher, C.</dc:creator>
<dc:creator>Brehm, P.</dc:creator>
<dc:date>2020-07-14</dc:date>
<dc:identifier>doi:10.1101/2020.07.14.202853</dc:identifier>
<dc:title><![CDATA[Primary and secondary motoneurons use different calcium channel types to control escape and swimming behaviors in zebrafish]]></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/10.1101/2020.07.15.205229v1?rss=1">
<title>
<![CDATA[
The S1 protein of SARS-CoV-2 crosses the blood-brain barrier: Kinetics, distribution, mechanisms, and influence of ApoE genotype, sex, and inflammation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.07.15.205229v1?rss=1</link>
<description><![CDATA[
Evidence strongly suggests that SARS-CoV-2, the cause of COVID-19, can enter the brain. SARS-CoV-2 enters cells via the S1 subunit of its spike protein, and S1 can be used as a proxy for the uptake patterns and mechanisms used by the whole virus; unlike studies based on productive infection, viral proteins can be used to precisely determine pharmacokinetics and biodistribution. Here, we found that radioiodinated S1 (I-S1) readily crossed the murine blood-brain barrier (BBB). I-S1 from two commercial sources crossed the BBB with unidirectional influx constants of 0.287 {+/-} 0.024 L/g-min and 0.294 {+/-} 0.032 L/g-min and was also taken up by lung, spleen, kidney, and liver. I-S1 was uniformly taken up by all regions of the brain and inflammation induced by lipopolysaccharide reduced uptake in the hippocampus and olfactory bulb. I-S1 crossed the BBB completely to enter the parenchymal brain space, with smaller amounts retained by brain endothelial cells and the luminal surface. Studies on the mechanisms of transport indicated that I-S1 crosses the BBB by the mechanism of adsorptive transcytosis and that the murine ACE2 receptor is involved in brain and lung uptake, but not that by kidney, liver, or spleen. I-S1 entered brain after intranasal administration at about 1/10th the amount found after intravenous administration and about 0.66% of the intranasal dose entered blood. ApoE isoform or sex did not affect whole brain uptake, but had variable effects on olfactory bulb, liver, spleen, and kidney uptakes. In summary, I-S1 readily crosses the murine BBB, entering all brain regions and the peripheral tissues studied, likely by the mechanism of adsorptive transcytosis.

Graphical Abstract O_FIG_DISPLAY_L [Figure 1] M_FIG_DISPLAY C_FIG_DISPLAY
]]></description>
<dc:creator>Rhea, E. M.</dc:creator>
<dc:creator>Logsdon, A. F.</dc:creator>
<dc:creator>Hansen, K. M.</dc:creator>
<dc:creator>Williams, L.</dc:creator>
<dc:creator>Reed, M.</dc:creator>
<dc:creator>Baumann, K.</dc:creator>
<dc:creator>Holden, S.</dc:creator>
<dc:creator>Raber, J.</dc:creator>
<dc:creator>Banks, W. A.</dc:creator>
<dc:creator>Erickson, M. A.</dc:creator>
<dc:date>2020-07-15</dc:date>
<dc:identifier>doi:10.1101/2020.07.15.205229</dc:identifier>
<dc:title><![CDATA[The S1 protein of SARS-CoV-2 crosses the blood-brain barrier: Kinetics, distribution, mechanisms, and influence of ApoE genotype, sex, and inflammation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-07-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.07.20.211342v1?rss=1">
<title>
<![CDATA[
BMP signalling directs a fibroblast-to-myoblast conversion at the connective tissue/muscle interface to pattern limb muscles 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.07.20.211342v1?rss=1</link>
<description><![CDATA[
Positional information driving limb muscle patterning is contained in lateral plate mesoderm-derived tissues, such as tendon or muscle connective tissue but not in myogenic cells themselves. The long-standing consensus is that myogenic cells originate from the somitic mesoderm, while connective tissue fibroblasts originate from the lateral plate mesoderm. We challenged this model using cell and genetic lineage tracing experiments in birds and mice, respectively, and identified a subpopulation of myogenic cells at the muscle tips close to tendons originating from the lateral plate mesoderm and derived from connective tissue gene lineages. Analysis of single-cell RNA-sequencing data obtained from limb cells at successive developmental stages revealed a subpopulation of cells displaying a dual muscle and connective tissue signature, in addition to independent muscle and connective tissue populations. Active BMP signalling was detected in this junctional cell sub-population and at the tendon/muscle interface in developing limbs. BMP gain- and loss-of-function experiments performed in vivo and in vitro showed that this signalling pathway regulated a fibroblast-to-myoblast conversion. We propose that localised BMP signalling converts a subset of lateral plate mesoderm-derived fibroblasts to a myogenic fate and establishes a boundary of fibroblast-derived myonuclei at the muscle/tendon interface to control the muscle pattern during limb development.
]]></description>
<dc:creator>Esteves de Lima, J.</dc:creator>
<dc:creator>Blavet, C.</dc:creator>
<dc:creator>Bonnin, M.-A.</dc:creator>
<dc:creator>Hirsinger, E.</dc:creator>
<dc:creator>Comai, G.</dc:creator>
<dc:creator>Yvernogeau, L.</dc:creator>
<dc:creator>Bellenger, l.</dc:creator>
<dc:creator>Mella, S.</dc:creator>
<dc:creator>Nassari, S.</dc:creator>
<dc:creator>Robin, C.</dc:creator>
<dc:creator>Schweitzer, R.</dc:creator>
<dc:creator>Fournier-Thibault, C.</dc:creator>
<dc:creator>Tajbakhsh, S.</dc:creator>
<dc:creator>Relaix, F.</dc:creator>
<dc:creator>DUPREZ, D.</dc:creator>
<dc:date>2020-07-21</dc:date>
<dc:identifier>doi:10.1101/2020.07.20.211342</dc:identifier>
<dc:title><![CDATA[BMP signalling directs a fibroblast-to-myoblast conversion at the connective tissue/muscle interface to pattern limb muscles]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-07-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.07.24.205583v1?rss=1">
<title>
<![CDATA[
Rapid generation of circulating and mucosal decoy ACE2 using mRNA nanotherapeutics for the potential treatment of SARS-CoV-2 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.07.24.205583v1?rss=1</link>
<description><![CDATA[
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) enters through the airways and infects the lungs, causing lethal pulmonary damage in vulnerable patients. This virus contains spike proteins on its envelope that binds to human angiotensin-converting enzyme 2 (hACE2) expressed on the surface of airway cells, enabling entry of the virus for causing infection1,2. In severe cases, the virus enters the circulatory system, contributing to multiorgan failure. Soluble form of hACE2 binds to SARS-CoV-2 spike protein and prevents viral entry into target cells3. Moreover, soluble recombinant ACE2 ameliorates lung injury4 but its short half-life limits its therapeutic utility5. Here, we engineered synthetic mRNA to encode a soluble form of hACE2 (hsACE2) to prevent viral infection. Novel lipid nanoparticles (LNPs) were used to package mRNA and transfect mammalian cells for enhanced production of secreted proteins. Intravenously administered LNP led to hepatic delivery of the mRNA. This elicited secretion of hsACE2 into the blood circulation within 2 h, and levels of circulating hsACE2 peaked at 6 h and gradually decreased over several days. Since the primary site of entry and pathogenesis for SARS-CoV-2 is the lungs, we instilled LNPs into the lungs and were able to detect hsACE2 in the bronchoalveolar lavage fluid within 24 h and lasted for 48 h. Through co-immunoprecipitation, we found that mRNA-generated hsACE2 was able to bind with the receptor binding domain of the SARS-CoV-2 spike protein. Furthermore, hsACE2 was able to strongly inhibit (over 90%) SARS-CoV-2 pseudovirus infection. Our proof of principle study shows that mRNA-based nanotherapeutics can be potentially deployed for pulmonary and extrapulmonary neutralization of SARS-CoV-2 and open new treatment opportunities for COVID-19.
]]></description>
<dc:creator>Kim, J.</dc:creator>
<dc:creator>Mukherjee, A.</dc:creator>
<dc:creator>Nelson, D.</dc:creator>
<dc:creator>Jozic, A.</dc:creator>
<dc:creator>Sahay, G.</dc:creator>
<dc:date>2020-07-25</dc:date>
<dc:identifier>doi:10.1101/2020.07.24.205583</dc:identifier>
<dc:title><![CDATA[Rapid generation of circulating and mucosal decoy ACE2 using mRNA nanotherapeutics for the potential treatment of SARS-CoV-2]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-07-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.07.24.219907v1?rss=1">
<title>
<![CDATA[
Single cell tracing reveals heterogeneous drug-, dose-, and time-dependent effects on cancer cell fates 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.07.24.219907v1?rss=1</link>
<description><![CDATA[
Identifying effective therapeutic strategies that can prevent tumor cell proliferation is a major challenge to improving outcomes for patients with breast cancer. Here we sought to deepen our understanding of how clinically relevant anti-cancer agents modulate cell cycle progression. We genetically engineered breast cancer cell lines to express a cell cycle reporter and then tracked drug-induced changes in cell number and cell cycle phase, which revealed drug-specific cell cycle effects that varied across time. This suggested that a computational model that could account for cell cycle phase durations would provide a framework to explore drug-induced changes in cell cycle changes. Toward that goal, we developed a linear chain trick (LCT) computational model, in which the cell cycle was partitioned into subphases that faithfully captured drug-induced dynamic responses. The model inferred drug effects and localized them to specific cell cycle phases, which we confirmed experimentally. We then used our LCT model to predict the effect of unseen drug combinations that target cells in different cell cycle phases. Experimental testing confirmed several model predictions and identified combination treatment strategies that may improve therapeutic response in breast cancer patients. Overall, this integrated experimental and modeling approach opens new avenues for assessing drug responses, predicting effective drug combinations, and identifying optimal drug sequencing strategies.
]]></description>
<dc:creator>Gross, S. M.</dc:creator>
<dc:creator>Sanchez-Aguila, C.</dc:creator>
<dc:creator>Zhan, P. J.</dc:creator>
<dc:creator>Heiser, L. M.</dc:creator>
<dc:date>2020-07-25</dc:date>
<dc:identifier>doi:10.1101/2020.07.24.219907</dc:identifier>
<dc:title><![CDATA[Single cell tracing reveals heterogeneous drug-, dose-, and time-dependent effects on cancer cell fates]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-07-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.07.24.220475v1?rss=1">
<title>
<![CDATA[
Genotypic Complexity in Initial Cleavage Divisions of Mammalian Embryos is Contributed by Defective BUB1B/BUBR1 Signaling 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.07.24.220475v1?rss=1</link>
<description><![CDATA[
Embryonic aneuploidy is highly complex, often leading to developmental arrest, implantation failure, or spontaneous miscarriage in both natural and assisted reproduction. Despite our knowledge of mitotic mis-segregation in somatic cells, the molecular pathways regulating chromosome fidelity during the error-prone cleavage-stage of mammalian embryogenesis remain largely undefined. Using bovine embryos and live-cell fluorescent imaging, we observed frequent micro-/multi-nucleation of anaphase lagging or mis-segregated chromosomes in initial mitotic divisions that underwent unilateral inheritance, re-fused with the primary nucleus, or formed a chromatin bridge with neighboring cells. A correlation between a lack of maternal and paternal pronuclei fusion (syngamy), multipolar cytokinesis, and uniparental genome segregation was also revealed and single-cell DNA-seq showed propagation of primarily non-reciprocal mitotic errors in embryonic blastomeres. Depletion of the mitotic checkpoint protein, BUB1B/BUBR1, resulted in micro-/multi-nuclei formation, atypical cytokinesis, chaotic aneuploidy, and disruption of the kinase-substrate network regulating mitotic progression and exit, culminating in embryo arrest prior to genome activation. This demonstrates that embryonic micronuclei sustain multiple fates, provides a mechanism for blastomeres with uniparental origins, and substantiates the contribution of defective checkpoint signaling and/or the inheritance of other maternally-derived factors to the high genotypic complexity afflicting preimplantation development in higher-order mammals.
]]></description>
<dc:creator>Brooks, K. E.</dc:creator>
<dc:creator>Daughtry, B. L.</dc:creator>
<dc:creator>Davis, B.</dc:creator>
<dc:creator>Yan, M. Y.</dc:creator>
<dc:creator>Fei, S. S.</dc:creator>
<dc:creator>Carbone, L.</dc:creator>
<dc:creator>Chavez, S. L.</dc:creator>
<dc:date>2020-07-24</dc:date>
<dc:identifier>doi:10.1101/2020.07.24.220475</dc:identifier>
<dc:title><![CDATA[Genotypic Complexity in Initial Cleavage Divisions of Mammalian Embryos is Contributed by Defective BUB1B/BUBR1 Signaling]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-07-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.07.24.220483v1?rss=1">
<title>
<![CDATA[
Cross-Tissue Transcriptomic Analysis Leveraging Machine Learning Approaches Identifies New Biomarkers for Rheumatoid Arthritis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.07.24.220483v1?rss=1</link>
<description><![CDATA[
Background/PurposeThere is an urgent need to identify effective biomarkers for early diagnosis of rheumatoid arthritis (RA) and to accurately monitor disease activity. Here we define an RA meta-profile using publicly available cross-tissue gene expression data and apply machine learning to identify putative biomarkers, which we further validate on independent datasets.

MethodsWe carried out a comprehensive search for publicly available microarray gene expression data in the NCBI Gene Expression Omnibus database for whole blood and synovial tissues from RA patients and healthy controls. The raw data from 13 synovium datasets with 284 samples and 14 blood datasets with 1,885 samples were downloaded and processed. The datasets for each tissue were merged, batch corrected and split into training and test sets. We then developed and applied a robust feature selection pipeline to identify genes dysregulated in both tissues and highly associated with RA. From the training data, we identified a set of overlapping differentially expressed genes following the condition of co-directionality. The classification performance of each gene in the resulting set was evaluated on the testing sets using the area under a receiver operating characteristic curve. Five independent datasets were used to validate and threshold the feature selected (FS) genes. Finally, we defined the RA Score, composed of the geometric mean of the selected RA Score Panel genes, and demonstrated its clinical utility.

ResultsThis feature selection pipeline resulted in a set of 25 upregulated and 28 downregulated genes. To assess the robustness of these FS genes, we trained a Random Forest machine learning model with this set of 53 genes and then with the set of 33 overlapping genes differentially expressed in both tissues and tested on the validation cohorts. The model with FS genes outperformed the model with common DE genes with AUC 0.89 {+/-} 0.04 vs 0.87 {+/-} 0.04. The FS genes were further validated on the 5 independent datasets resulting in 10 upregulated genes, TNFAIP6, S100A8, TNFSF10, DRAM1, LY96, QPCT, KYNU, ENTPD1, CLIC1, and ATP6V0E1, which are involved in innate immune system pathways, including neutrophil degranulation and apoptosis. There were also three downregulated genes, HSP90AB1, NCL, and CIRBP, that are involved in metabolic processes and T-cell receptor regulation of apoptosis.

To investigate the clinical utility of the 13 validated genes, the RA Score was developed and found to be highly correlated with the disease activity score based on the 28 examined joints (DAS28) (r = 0.33 {+/-} 0.03, p = 7e-9) and able to distinguish osteoarthritis (OA) from RA samples (OR 0.57, 95% CI [0.34, 0.80], p = 8e-10). Moreover, the RA Score was not significantly different for rheumatoid factor (RF) positive and RF-negative RA sub-phenotypes (p = 0.9) and also distinguished polyarticular juvenile idiopathic arthritis (polyJIA) from healthy individuals in 10 independent pediatric cohorts (OR 1.15, 95% CI [1.01, 1.3], p = 2e-4) suggesting the generalizability of this score in clinical applications. The RA Score was also able to monitor the treatment effect among RA patients (t-test of treated vs untreated, p = 2e-4). Finally, we performed immunoblotting analysis of 6 proteins in unstimulated PBMC lysates from an independent cohort of 8 newly diagnosed RA patients and 7 healthy controls, where two proteins, TNFAIP6/TSG6 and HSP90AB1/HSP90, were validated and the S100A8 protein showed near significant up-regulation.

ConclusionThe RA Score, consisting of 13 putative biomarkers identified through a robust feature selection procedure on public data and validated using multiple independent data sets, could be useful in the diagnosis and treatment monitoring of RA.
]]></description>
<dc:creator>Rychkov, D.</dc:creator>
<dc:creator>Neely, J.</dc:creator>
<dc:creator>Oskotsky, T.</dc:creator>
<dc:creator>Sirota, M.</dc:creator>
<dc:date>2020-07-26</dc:date>
<dc:identifier>doi:10.1101/2020.07.24.220483</dc:identifier>
<dc:title><![CDATA[Cross-Tissue Transcriptomic Analysis Leveraging Machine Learning Approaches Identifies New Biomarkers for Rheumatoid Arthritis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-07-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.07.25.221507v1?rss=1">
<title>
<![CDATA[
Identification of clinical combination therapies to induce durable responses in kidney cancers 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.07.25.221507v1?rss=1</link>
<description><![CDATA[
The lack of effective treatment options for advanced non-clear cell renal cell carcinoma (NCCRCC) is a critical unmet clinical need. Applying a high throughput drug screen to multiple human kidney cancer cells, we identified the combination of the VEGFR-MET inhibitor cabozantinib and the SRC inhibitor dasatinib acted synergistically in cells to markedly reduce cell viability. Importantly, the combination was well tolerated and caused tumor regression in vivo. Transcriptional and phosphoproteomic profiling revealed that the combination converged to downregulate the MAPK-ERK signaling pathway, a result not predicted by single agent analysis alone. Correspondingly, the addition of a MEK inhibitor synergized with either dasatinib or cabozantinib to increase its efficacy. This study, by employing approved, clinically relevant drugs provides the rationale for the design of effective combination treatments in NCCRCC that can be rapidly translated to the clinic.
]]></description>
<dc:creator>Thomas, G. V.</dc:creator>
<dc:date>2020-07-26</dc:date>
<dc:identifier>doi:10.1101/2020.07.25.221507</dc:identifier>
<dc:title><![CDATA[Identification of clinical combination therapies to induce durable responses in kidney cancers]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-07-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.07.28.220509v1?rss=1">
<title>
<![CDATA[
Natural food intake patterns do not synchronize peripheral clocks 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.07.28.220509v1?rss=1</link>
<description><![CDATA[
Food is thought to synchronize circadian clocks in the body, but this is based on time-restricted feeding (TRF) protocols. To test whether naturalistic feeding patterns are sufficient to phase-shift and entrain peripheral tissues, we measured circadian rhythms of the liver, kidney, and submandibular gland in mPer2Luc mice under different feeding schedules. In ad lib feeding as well as in a schedule designed to mimic the ad lib pattern, PER2::LUC bioluminescence peaked during the night as expected. Surprisingly, shifting the scheduled feeding by 12h caused only small advances (<3h). To isolate the effects of feeding from the light-dark cycle, clock phase was then measured in mice acclimated to scheduled feeding and housed in constant darkness. In these conditions, peripheral clock phases were better predicted by the rest-activity cycle than the food schedule. Under natural feeding patterns, the master pacemaker in the brain sets the phase of peripheral organs independent of feeding behavior.
]]></description>
<dc:creator>Xie, X.</dc:creator>
<dc:creator>Kukino, A.</dc:creator>
<dc:creator>Calcagno, H. E.</dc:creator>
<dc:creator>Berman, A.</dc:creator>
<dc:creator>Garner, J. P.</dc:creator>
<dc:creator>Butler, M. P.</dc:creator>
<dc:date>2020-07-29</dc:date>
<dc:identifier>doi:10.1101/2020.07.28.220509</dc:identifier>
<dc:title><![CDATA[Natural food intake patterns do not synchronize peripheral clocks]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-07-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.07.30.229377v1?rss=1">
<title>
<![CDATA[
Cross-reactivity of SARS-CoV structural protein antibodies against SARS-CoV-2 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.07.30.229377v1?rss=1</link>
<description><![CDATA[
There is currently a lack of biological tools to study the replication cycle and pathogenesis of SARS-CoV-2, the etiological agent of COVID-19. Repurposing the existing tools, including antibodies of SARS-CoV, is an effective way to accelerate the development of therapeutics for COVID-19. Here, we extensively characterized antibodies of the SARS-CoV structural proteins for their cross-reactivity, experimental utility, and neutralization of SARS-CoV-2. We assessed a total of 10 antibodies (six for Spike, two for Membrane, and one for Nucleocapsid and Envelope viral protein). We evaluated the utility of these antibodies against SARS-CoV-2 in a variety of assays, including immunofluorescence, ELISA, biolayer interferometry, western blots, and micro-neutralization. Remarkably, a high proportion of the antibodies we tested showed cross-reactivity, indicating a potentially generalizable theme of cross-reactivity between SARS-CoV and SARS-CoV-2 antibodies. These antibodies should help facilitate further research into SARS-CoV-2 basic biology. Moreover, our study provides critical information about the propensity of SARS-CoV antibodies to cross-react with SARS-CoV-2 and highlights its relevance in defining the clinical significance of such antibodies to improve testing and guide the development of novel vaccines and therapeutics.
]]></description>
<dc:creator>Bates, T. A.</dc:creator>
<dc:creator>Weinstein, J. B.</dc:creator>
<dc:creator>Farley, S. E.</dc:creator>
<dc:creator>Leier, H. C.</dc:creator>
<dc:creator>Messer, W. B.</dc:creator>
<dc:creator>Tafesse, F. G.</dc:creator>
<dc:date>2020-07-30</dc:date>
<dc:identifier>doi:10.1101/2020.07.30.229377</dc:identifier>
<dc:title><![CDATA[Cross-reactivity of SARS-CoV structural protein antibodies against SARS-CoV-2]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-07-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.08.11.236372v1?rss=1">
<title>
<![CDATA[
The identity of human tissue-emigrant CD8+ T cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.08.11.236372v1?rss=1</link>
<description><![CDATA[
Lymphocyte migration is essential for human adaptive immune surveillance. However, our current understanding of this process is rudimentary, because most human studies to date have been restricted to immunological analyses of blood and various tissues. To address this issue, we used an integrated approach to characterize tissue-emigrant immune cells in thoracic duct lymph (TDL). In humans and non-human primates, lymphocytes were by far the most abundant immune lineage population in efferent lymph, and a vast majority of these lymphocytes were T cells. Cytolytic CD8+ T cell subsets were clonotypically discrete and selectively confined to the intravascular circulation, persisting for months after inhibition of S1P-dependent tissue egress by FTY-720. In contrast, non-cytolytic CD8+ T cell subsets with stem-like epigenetic and transcriptional signatures predominated in tissues and TDL. Collectively, these data provide an atlas of the migratory immune system and define the nature of tissue-emigrant CD8+ T cells that recirculate via TDL.
]]></description>
<dc:creator>Buggert, M.</dc:creator>
<dc:creator>Vella, L. A.</dc:creator>
<dc:creator>Nguyen, S.</dc:creator>
<dc:creator>Wu, V.</dc:creator>
<dc:creator>Sekine, T.</dc:creator>
<dc:creator>Perez-Potti, A.</dc:creator>
<dc:creator>Maldini, C. R.</dc:creator>
<dc:creator>Manne, S.</dc:creator>
<dc:creator>Darko, S.</dc:creator>
<dc:creator>Ransier, A.</dc:creator>
<dc:creator>Kuri-Cervantes, L.</dc:creator>
<dc:creator>Sada Japp, A.</dc:creator>
<dc:creator>Bukh Brody, I.</dc:creator>
<dc:creator>Ivarsson, M. A.</dc:creator>
<dc:creator>Hertwig, L.</dc:creator>
<dc:creator>Antel, J. P.</dc:creator>
<dc:creator>Johnson, M. E.</dc:creator>
<dc:creator>Okoye, A.</dc:creator>
<dc:creator>Picker, L. J.</dc:creator>
<dc:creator>Vahedi, G.</dc:creator>
<dc:creator>Sparrelid, E.</dc:creator>
<dc:creator>Llewellyn-Lacey, S.</dc:creator>
<dc:creator>Gostick, E.</dc:creator>
<dc:creator>Bjorkstrom, N. K.</dc:creator>
<dc:creator>Bar-Or, A.</dc:creator>
<dc:creator>Dori, Y.</dc:creator>
<dc:creator>Naji, A.</dc:creator>
<dc:creator>Canaday, D.</dc:creator>
<dc:creator>Laufer, T. M.</dc:creator>
<dc:creator>Wells, A. D.</dc:creator>
<dc:creator>Price, D.</dc:creator>
<dc:creator>Frank, I.</dc:creator>
<dc:creator>Douek, D.</dc:creator>
<dc:creator>Wherry, E. J.</dc:creator>
<dc:creator>Itkin, M. G.</dc:creator>
<dc:creator>Betts, M. R.</dc:creator>
<dc:date>2020-08-11</dc:date>
<dc:identifier>doi:10.1101/2020.08.11.236372</dc:identifier>
<dc:title><![CDATA[The identity of human tissue-emigrant CD8+ T cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-08-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.08.12.248716v1?rss=1">
<title>
<![CDATA[
Postembryonic screen for mutations affecting spine development in zebrafish 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.08.12.248716v1?rss=1</link>
<description><![CDATA[
The spinal vertebral column gives structural support for the adult body plan, protects the spinal cord, and provides muscle attachment and stability, which allows the animal to move within its environment. The development and maturation of the spine and its physiology involve the integration of multiple musculoskeletal tissues including bone, cartilage, and fibrocartilaginous joints, as well as innervation and control by the nervous system. One of the most common disorders of the spine in human is adolescent idiopathic scoliosis (AIS), which is characterized by the onset of an abnormal lateral curvature of the spine of <10{degrees} around adolescence, in otherwise healthy children. The genetic basis of AIS is largely unknown. Systematic genome-wide mutagenesis screens for embryonic phenotypes in zebrafish have been instrumental in the understanding of early patterning of embryonic tissues necessary to build and pattern the embryonic spine. However, the mechanisms required for postembryonic maturation and homeostasis of the spine remain poorly understood. Here we report the results from a small-scale forward genetic screen for adult-viable recessive and dominant mutant zebrafish, displaying overt morphological abnormalities of the adult spine. Germline mutations induced with N-ethyl N-nitrosourea (ENU) were transmitted and screened for dominant phenotypes in 1,229 F1 animals, and subsequently bred to homozygosity in F3 families, from these, 314 haploid genomes were screened for recessive phenotypes. We cumulatively found 39 adult-viable (3 dominant and 36 recessive) mutations each leading to a defect in the morphogenesis of the spine. The largest phenotypic group displayed larval onset axial curvatures, leading to whole-body scoliosis without vertebral dysplasia in adult fish. Pairwise complementation testing within this phenotypic group revealed at least 16 independent mutant loci. Using massively-parallel whole genome or whole exome sequencing and meiotic mapping we defined the molecular identity of several loci for larval onset whole-body scoliosis in zebrafish. We identified a new mutation in the skolios/kinesin family member 6 (kif6) gene, causing neurodevelopmental and ependymal cilia defects in mouse and zebrafish. We also report several recessive alleles of the scospondin and a disintegrin and metalloproteinase with thrombospondin motifs 9 (adamts9) genes, which all display defects in spine morphogenesis. Many of the alleles characterized thus far are non-synonymous mutations in known essential scospondin and adamts9 genes. Our results provide evidence of monogenic traits that are critical for normal spine development in zebrafish, that may help to establish new candidate risk loci for spine disorders in humans.
]]></description>
<dc:creator>GRAY, R.</dc:creator>
<dc:creator>Gonzalez, R.</dc:creator>
<dc:creator>Ackerman, S. D.</dc:creator>
<dc:creator>Minowa, R. C.</dc:creator>
<dc:creator>Griest, J. F.</dc:creator>
<dc:creator>Bayrak, M. N.</dc:creator>
<dc:creator>Troutwine, B. R.</dc:creator>
<dc:creator>Canter, S.</dc:creator>
<dc:creator>Monk, K. R.</dc:creator>
<dc:creator>Sepich, D. S.</dc:creator>
<dc:creator>Solnica-Krezel, L.</dc:creator>
<dc:date>2020-08-12</dc:date>
<dc:identifier>doi:10.1101/2020.08.12.248716</dc:identifier>
<dc:title><![CDATA[Postembryonic screen for mutations affecting spine development in zebrafish]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-08-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.08.19.258467v1?rss=1">
<title>
<![CDATA[
CD300lf conditional knockout mouse reveals strain-specific cellular tropism for murine norovirus 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.08.19.258467v1?rss=1</link>
<description><![CDATA[
Noroviruses are a leading cause of gastrointestinal infection in humans and mice. Understanding human norovirus (HuNoV) cell tropism has important implications for our understanding of viral pathogenesis. Murine norovirus (MNoV) is extensively used as a surrogate model for HuNoV. We previously identified CD300lf as the receptor for MNoV. Here, we generated a Cd300lf conditional knockout (CD300lfF/F) mouse to elucidate the cell tropism of persistent and non-persistent strains of murine norovirus. Using this mouse model, we demonstrate that CD300lf expression on intestinal epithelial cells (IECs), and on tuft cells in particular, is essential for transmission of the persistent MNoV strain CR6 (MNoVCR6) in vivo. In contrast, the nonpersistent MNoV strain CW3 (MNoVCW3) does not require CD300lf expression on IECs for infection. However, deletion of CD300lf in myelomonocytic cells (LysM Cre+) partially reduces CW3 viral load in lymphoid and intestinal tissues. Disruption of CD300lf expression on B cells (CD19 Cre), neutrophils (Mrp8 Cre), and dendritic cells (CD11c Cre) did not affect CW3 viral RNA levels. Finally, we show that the transcription factor STAT1, which is critical for the innate immune response, partially restricts the cell tropism of MNoVCW3 to LysM+ cells. Taken together, these data demonstrate that CD300lf expression on tuft cells is essential for MNoVCR6, that myelomonocytic cells are a major, but not exclusive, target cell of MNoVCW3, and that STAT1 signaling restricts the cellular tropism of MNoVCW3. This provides the first genetic system to study the cell type-specific role of CD300lf in norovirus pathogenesis.

IMPORTANCEHuman noroviruses (HuNoVs) are a leading cause of gastroenteritis resulting in up to 200,000 deaths each year. The receptor and cell tropism of HuNoV in immunocompetent humans are unclear. We use murine norovirus (MNoV) as a model for HuNoV. We recently identified CD300lf as the sole physiologic receptor for MNoV. Here, we leverage this finding to generate a Cd300lf conditional knockout mouse to decipher the contributions of specific cell types to MNoV infection. We demonstrate that persistent MNoVCR6 requires CD300lf expression on tuft cells. In contrast, multiple CD300lf+ cell types, dominated by myelomonocytic cells, are sufficient for non-persistent MNoVCW3 infection. CD300lf expression on epithelial cells, B cells, neutrophils, and dendritic cells is not critical for MNoVCW3 infection. Mortality associated with MNoVCW3 strain in Stat1-/- mice does not require CD300lf expression on LysM+ cells, highlighting that both CD300lf receptor expression and innate immunity regulate MNoV cell tropism in vivo.
]]></description>
<dc:creator>Graziano, V. R.</dc:creator>
<dc:creator>Alfajaro, M. M.</dc:creator>
<dc:creator>Schmitz, C.</dc:creator>
<dc:creator>Filler, R. B.</dc:creator>
<dc:creator>Strine, M. S.</dc:creator>
<dc:creator>Wei, J.</dc:creator>
<dc:creator>Hsieh, L. L.</dc:creator>
<dc:creator>Baldridge, M. T.</dc:creator>
<dc:creator>Nice, T. J.</dc:creator>
<dc:creator>Lee, S.</dc:creator>
<dc:creator>Orchard, R. C.</dc:creator>
<dc:creator>Wilen, C. B.</dc:creator>
<dc:date>2020-08-21</dc:date>
<dc:identifier>doi:10.1101/2020.08.19.258467</dc:identifier>
<dc:title><![CDATA[CD300lf conditional knockout mouse reveals strain-specific cellular tropism for murine norovirus]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-08-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.08.20.259408v1?rss=1">
<title>
<![CDATA[
Single cell tracking based on Voronoi partition via stable matching 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.08.20.259408v1?rss=1</link>
<description><![CDATA[
Live-cell imaging is an important technique to study cell migration and proliferation as well as image-based profiling of drug perturbations over time. To gain biological insights from live-cell imaging data, it is necessary to identify individual cells, follow them over time and extract quantitative information. However, since often biological experiment does not allow the high temporal resolution to reduce excessive levels of illumination or minimize unnecessary oversampling to monitor long-term dynamics, it is still a challenging task to obtain good tracking results with coarsely sampled imaging data. To address this problem, we consider cell tracking problem as "stable matching problem" and propose a robust tracking method based on Voronoi partition which adapts parameters that need to be set according to the spatio-temporal characteristics of live cell imaging data such as cell population and migration. We demonstrate the performance improvement provided by the proposed method using numerical simulations and compare its performance with proximity-based tracking and nearest neighbor-based tracking.
]]></description>
<dc:creator>Chang, Y. H.</dc:creator>
<dc:creator>Linsley, J. W.</dc:creator>
<dc:creator>Lamstein, J.</dc:creator>
<dc:creator>Kalra, J.</dc:creator>
<dc:creator>Epstein, I.</dc:creator>
<dc:creator>Barch, M.</dc:creator>
<dc:creator>Daily, K.</dc:creator>
<dc:creator>Snyder, P.</dc:creator>
<dc:creator>Omberg, L.</dc:creator>
<dc:creator>Heiser, L.</dc:creator>
<dc:creator>Finkbeiner, S.</dc:creator>
<dc:date>2020-08-20</dc:date>
<dc:identifier>doi:10.1101/2020.08.20.259408</dc:identifier>
<dc:title><![CDATA[Single cell tracking based on Voronoi partition via stable matching]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-08-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.08.21.257758v1?rss=1">
<title>
<![CDATA[
Towards Reproducible Brain-Wide Association Studies 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.08.21.257758v1?rss=1</link>
<description><![CDATA[
Magnetic resonance imaging (MRI) continues to drive many important neuroscientific advances. However, progress in uncovering reproducible associations between individual differences in brain structure/function and behavioral phenotypes (e.g., cognition, mental health) may have been undermined by typical neuroimaging sample sizes (median N=25)1,2. Leveraging the Adolescent Brain Cognitive Development (ABCD) Study3 (N=11,878), we estimated the effect sizes and reproducibility of these brain-wide associations studies (BWAS) as a function of sample size. The very largest, replicable brain-wide associations for univariate and multivariate methods were r=0.14 and r=0.34, respectively. In smaller samples, typical for brain-wide association studies (BWAS), irreproducible, inflated effect sizes were ubiquitous, no matter the method (univariate, multivariate). Until sample sizes started to approach consortium-levels, BWAS were underpowered and statistical errors assured. Multiple factors contribute to replication failures4-6; here, we show that the pairing of small brain-behavioral phenotype effect sizes with sampling variability is a key element in wide-spread BWAS replication failure. Brain-behavioral phenotype associations stabilize and become more reproducible with sample sizes of N{gap}2,000. While investigator-initiated brain-behavior research continues to generate hypotheses and propel innovation, large consortia are needed to usher in a new era of reproducible human brain-wide association studies.
]]></description>
<dc:creator>Marek, S.</dc:creator>
<dc:creator>Tervo-Clemmens, B.</dc:creator>
<dc:creator>Calabro, F. J.</dc:creator>
<dc:creator>Montez, D. F.</dc:creator>
<dc:creator>Kay, B. P.</dc:creator>
<dc:creator>Hatoum, A. S.</dc:creator>
<dc:creator>Donohue, M. R.</dc:creator>
<dc:creator>Foran, W.</dc:creator>
<dc:creator>Miller, R. L.</dc:creator>
<dc:creator>Feczko, E.</dc:creator>
<dc:creator>Miranda Dominguez, O.</dc:creator>
<dc:creator>Graham, A.</dc:creator>
<dc:creator>Earl, E. A.</dc:creator>
<dc:creator>Perrone, A.</dc:creator>
<dc:creator>Cordova, M.</dc:creator>
<dc:creator>Doyle, O.</dc:creator>
<dc:creator>Moore, L. A.</dc:creator>
<dc:creator>Conan, G.</dc:creator>
<dc:creator>Uriarte, J.</dc:creator>
<dc:creator>Snider, K.</dc:creator>
<dc:creator>Tam, A.</dc:creator>
<dc:creator>Chen, J.</dc:creator>
<dc:creator>Newbold, D. J.</dc:creator>
<dc:creator>Zheng, A.</dc:creator>
<dc:creator>Seider, N. A.</dc:creator>
<dc:creator>Van, A. N.</dc:creator>
<dc:creator>Laumann, T. O.</dc:creator>
<dc:creator>Thompson, W. K.</dc:creator>
<dc:creator>Greene, D. J.</dc:creator>
<dc:creator>Petersen, S. E.</dc:creator>
<dc:creator>Nichols, T.</dc:creator>
<dc:creator>Yeo, B. T. T.</dc:creator>
<dc:creator>Barch, D. M.</dc:creator>
<dc:creator>Garavan, H.</dc:creator>
<dc:creator>Luna, B.</dc:creator>
<dc:creator>Fair, D. A.</dc:creator>
<dc:creator>Dosenbach, N. U. F.</dc:creator>
<dc:date>2020-08-22</dc:date>
<dc:identifier>doi:10.1101/2020.08.21.257758</dc:identifier>
<dc:title><![CDATA[Towards Reproducible Brain-Wide Association Studies]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-08-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.08.21.261578v1?rss=1">
<title>
<![CDATA[
A sort-seq approach to the development of single fluorescent protein biosensors 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.08.21.261578v1?rss=1</link>
<description><![CDATA[
The utility of single fluorescent protein biosensors (SFPBs) in biological research is offset by the difficulty in engineering these tools. SFPBs generally consist of three basic components: a circularly permuted fluorescent protein, a ligand-binding domain, and a pair of linkers connecting the two domains. In the absence of predictive methods for biosensor engineering, most designs combining these three components will fail to produce allosteric coupling between ligand binding and fluorescence emission. Methods to construct libraries of biosensor designs with variations in the site of GFP insertion and linker sequences have been developed, however, our ability to construct new variants has exceeded our ability to test them for function. Here, we address this challenge by applying a massively parallel assay termed "sort-seq" to the characterization of biosensor libraries. Sort-seq combines binned fluorescence-activated cell sorting, next-generation sequencing, and maximum likelihood estimation to quantify the dynamic range of many biosensor variants in parallel. We applied this method to two common biosensor optimization tasks: choice of insertion site and optimization of linker sequences. The sort-seq assay applied to a maltose-binding protein domain-insertion library not only identified previously described high-dynamic-range variants but also discovered new functional insertion-sites with diverse properties. A sort-seq assay performed on a pyruvate biosensor linker library expressed in mammalian cell culture identified linker variants with substantially improved dynamic range. Machine learning models trained on the resulting data can predict dynamic range from linker sequence. This high-throughput approach will accelerate the design and optimization of SFPBs, expanding the biosensor toolbox.
]]></description>
<dc:creator>Koberstein, J. N.</dc:creator>
<dc:creator>Stewart, M. L.</dc:creator>
<dc:creator>Mighell, T. L.</dc:creator>
<dc:creator>Smith, C. B.</dc:creator>
<dc:creator>Cohen, M. S.</dc:creator>
<dc:date>2020-08-22</dc:date>
<dc:identifier>doi:10.1101/2020.08.21.261578</dc:identifier>
<dc:title><![CDATA[A sort-seq approach to the development of single fluorescent protein biosensors]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-08-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.08.21.262030v1?rss=1">
<title>
<![CDATA[
Transcriptomic analysis of primate placentas and novel rhesus trophoblast cell lines informs investigations of human placentation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.08.21.262030v1?rss=1</link>
<description><![CDATA[
Proper placentation, including trophoblast differentiation and function, is essential for the health and well-being of both the mother and baby throughout pregnancy. Placental abnormalities that occur during the early stages of development are thought to contribute to pre-eclampsia and other placenta-related pregnancy complications. However, relatively little is known about these stages in humans due to obvious ethical and technical limitations. Rhesus macaques are considered an ideal surrogate for studying human placentation, but the unclear translatability of known human placental markers and lack of accessible rhesus trophoblast cell lines can impede the use of this animal model. Here, we performed a cross-species transcriptomic comparison of human and rhesus placenta and determined that while the majority of known placental markers were similarly expressed, 952 differentially expressed genes (DEGs) were identified between the two species. Pathway enrichment analysis of the 447 human-upregulated DEGs, including ADAM12, ERVW-1, KISS1, LGALS13, PAPPA2, PGF, and SIGLEC6, revealed over-representation of functional terms associated with pre-eclampsia and other pregnancy disorders. Additionally, to enable in vitro functional studies of early placentation, we generated and thoroughly characterized two highly-pure first-trimester telomerase (TERT) immortalized rhesus trophoblast cell lines (iRP-D26 and iRP-D28A) that retained crucial features of isolated primary trophoblasts. Overall, our findings help elucidate the molecular translatability between human and rhesus placenta and reveal notable expression differences in human placental markers and genes associated with pregnancy complications that should be considered when using the rhesus animal model to study normal and pathological human placentation.
]]></description>
<dc:creator>Rosenkrantz, J. L.</dc:creator>
<dc:creator>Gaffney, J. E.</dc:creator>
<dc:creator>Roberts, V. H.</dc:creator>
<dc:creator>Carbone, L.</dc:creator>
<dc:creator>Chavez, S. L.</dc:creator>
<dc:date>2020-08-22</dc:date>
<dc:identifier>doi:10.1101/2020.08.21.262030</dc:identifier>
<dc:title><![CDATA[Transcriptomic analysis of primate placentas and novel rhesus trophoblast cell lines informs investigations of human placentation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-08-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.08.25.267120v1?rss=1">
<title>
<![CDATA[
BAX is necessary for neuronal death following exposure to isoflurane during the neonatal period 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.08.25.267120v1?rss=1</link>
<description><![CDATA[
Exposure to volatile anesthetics during the neonatal period results in acute neuronal death in rodent and non-human primate models, potentially leading to lasting cognitive deficits. We used Bax-/- mice to show that neuronal death following neonatal exposure to isoflurane is mediated by the apoptotic pathway, and that GABAergic interneurons are selectively vulnerable. Neonatal Bax-/- mice also showed attenuated microglial activation after exposure to isoflurane, indicating that neuroinflammatory response is secondary to neuronal apoptosis. Isoflurane-induced neuronal apoptosis in neonates appeared to have little effect on seizure threshold or cognitive function later in life. Collectively, these findings define the acute injury mechanism of volatile anesthetics during the neonatal period.
]]></description>
<dc:creator>Slupe, A. M.</dc:creator>
<dc:creator>Villasana, L.</dc:creator>
<dc:creator>Wright, K. M.</dc:creator>
<dc:date>2020-08-26</dc:date>
<dc:identifier>doi:10.1101/2020.08.25.267120</dc:identifier>
<dc:title><![CDATA[BAX is necessary for neuronal death following exposure to isoflurane during the neonatal period]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-08-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.08.27.270322v1?rss=1">
<title>
<![CDATA[
Base Pairing and Functional Insights into N3-methylcytidine (m3C) in RNA 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.08.27.270322v1?rss=1</link>
<description><![CDATA[
N3-methylcytidine (m3C) is present in both eukaryotic tRNA and mRNA and plays critical roles in many biological processes. We report the synthesis of the m3C phosphoramidite building block and its containing RNA oligonucleotides. The base-pairing stability and specificity studies show that the m3C modification significantly disrupts the stability of the Watson-Crick C:G pair. Further m3C decreases the base pairing discrimination between C:G and the other mismatched C:A, C:U, and C:C pairs. Our molecular dynamic simulation study further reveals the detailed structural insights into the m3C:G base pairing pattern in an RNA duplex. More importantly, the biochemical investigation of m3C using reverse transcription shows that N3-methylation specifies the C:A pair and induces a G to A mutation using HIV-1-RT, MMLV-RT and MutiScribe-RT enzymes, all with relatively low replication fidelity. For other reverse transcriptases with higher fidelity like AMV-RT, the methylation could completely shut down DNA synthesis.
]]></description>
<dc:creator>Mao, S.</dc:creator>
<dc:creator>Haruehanroengra, P.</dc:creator>
<dc:creator>Ranganathan, S. V.</dc:creator>
<dc:creator>Shen, F.</dc:creator>
<dc:creator>Begley, T.</dc:creator>
<dc:creator>Sheng, J.</dc:creator>
<dc:date>2020-08-28</dc:date>
<dc:identifier>doi:10.1101/2020.08.27.270322</dc:identifier>
<dc:title><![CDATA[Base Pairing and Functional Insights into N3-methylcytidine (m3C) in RNA]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-08-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.08.28.272641v1?rss=1">
<title>
<![CDATA[
Targeting uncoupling of copy number and gene expression in cancers 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.08.28.272641v1?rss=1</link>
<description><![CDATA[
The high degree of aneuploidy in cancer is likely tolerated via extensive uncoupling of copy number (CN) and mRNA expression (UCNE) of deleterious genes located in copy number aberrations (CNAs). To test the extent and role of UCNE in cancer, we performed integrative analysis of multiomics data across The Cancer Genome Atlas (TCGA), encompassing [~] 5000 individual tumors. We found many genes having UCNE, the degree of which are associated with increased oncogenic signaling, proliferation and immune-suppression. The occurrence of UCNE appears to be orchestrated by complex epigenetic and regulatory changes, with transcription factors (TFs) playing a prominent role. To further dissect the regulatory mechanisms, we developed a systems-biological approach to identify candidate TFs, which upon perturbation can offset UCNE and reduce tumor fitness. Applying our approach on TCGA data, we identified 20 putative targets, 45% of which were validated by independent sources. Among them are IRF1, which plays a prominent role in anti-tumor immunity and response to immune checkpoint therapy, ETS1, TRIM21 and GATA3, which are associated with anti-tumor immunity, tumor proliferation and metastasis. Together, our study indicates that UCNE is likely an important mechanism in cancer development that can be exploited therapeutically.
]]></description>
<dc:creator>Mohanty, V.</dc:creator>
<dc:creator>Wang, F.</dc:creator>
<dc:creator>Mills, G. B.</dc:creator>
<dc:creator>Chen, K.</dc:creator>
<dc:date>2020-08-30</dc:date>
<dc:identifier>doi:10.1101/2020.08.28.272641</dc:identifier>
<dc:title><![CDATA[Targeting uncoupling of copy number and gene expression in cancers]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-08-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.08.31.276584v1?rss=1">
<title>
<![CDATA[
Selective effects of arousal on population coding of natural sounds in primary auditory cortex 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.08.31.276584v1?rss=1</link>
<description><![CDATA[
The ability to discriminate between complex natural sounds is critical for survival. Changes in arousal and other aspects of behavioral state can impact the accuracy of sensory coding, affecting both the reliability of single neuron responses and the degree of correlated noise between neurons. However, it is unclear how these effects interact to influence coding of diverse natural stimuli. We recorded the spiking activity of neural populations in primary auditory cortex (A1) evoked by a large library of natural sounds while monitoring changes in pupil size as an index of arousal. Heightened arousal increased response magnitude and reduced noise correlations between neurons, improving coding accuracy on average. Rather than suppressing shared noise along all dimensions of neural activity, the change in noise correlations occurred via coherent, low-dimensional modulation of response variability in A1. The modulation targeted a different group of neurons from those undergoing changes in response magnitude. Thus, changes in response magnitude and correlation are mediated by distinct mechanisms. The degree to which these low-dimensional changes were aligned with the high-dimensional natural sound-evoked activity was variable, resulting in stimulus-dependent improvements in coding accuracy.
]]></description>
<dc:creator>Heller, C. R.</dc:creator>
<dc:creator>Schwartz, Z. P.</dc:creator>
<dc:creator>Saderi, D.</dc:creator>
<dc:creator>David, S. V.</dc:creator>
<dc:date>2020-09-01</dc:date>
<dc:identifier>doi:10.1101/2020.08.31.276584</dc:identifier>
<dc:title><![CDATA[Selective effects of arousal on population coding of natural sounds in primary auditory cortex]]></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/10.1101/2020.09.04.282269v1?rss=1">
<title>
<![CDATA[
QSIPrep: An integrative platform for preprocessing and reconstructing diffusion MRI 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.09.04.282269v1?rss=1</link>
<description><![CDATA[
Diffusion-weighted magnetic resonance imaging (dMRI) has become the primary method for non-invasively studying the organization of white matter in the human brain. While many dMRI acquisition sequences have been developed, they all sample q-space in order to characterize water diffusion. Numerous software platforms have been developed for processing dMRI data, but most work on only a subset of sampling schemes or implement only parts of the processing workflow. Reproducible research and comparisons across dMRI methods are hindered by incompatible software, diverse file formats, and inconsistent naming conventions. Here we introduce QSIPrep, an integrative software platform for the processing of diffusion images that is compatible with nearly all dMRI sampling schemes. Drawing upon a diverse set of software suites to capitalize upon their complementary strengths, QSIPrep automatically applies best practices for dMRI preprocessing, including denoising, distortion correction, head motion correction, coregistration, and spatial normalization. Throughout, QSIPrep provides both visual and quantitative measures of data quality as well as "glass-box" methods reporting. Taken together, these features facilitate easy implementation of best practices for processing of diffusion images while simultaneously ensuring reproducibility.
]]></description>
<dc:creator>Cieslak, M.</dc:creator>
<dc:creator>Cook, P. A.</dc:creator>
<dc:creator>He, X.</dc:creator>
<dc:creator>Yeh, F.-C.</dc:creator>
<dc:creator>Dhollander, T.</dc:creator>
<dc:creator>Adebimpe, A.</dc:creator>
<dc:creator>Aguirre, G. K.</dc:creator>
<dc:creator>Bassett, D. S.</dc:creator>
<dc:creator>Betzel, R. F.</dc:creator>
<dc:creator>Bourque, J.</dc:creator>
<dc:creator>Cabral, L.</dc:creator>
<dc:creator>Davatzikos, C.</dc:creator>
<dc:creator>Detre, J.</dc:creator>
<dc:creator>Earl, E.</dc:creator>
<dc:creator>Elliott, M. A.</dc:creator>
<dc:creator>Fadnavis, S.</dc:creator>
<dc:creator>Fair, D. A.</dc:creator>
<dc:creator>Foran, W.</dc:creator>
<dc:creator>Fotiadis, P.</dc:creator>
<dc:creator>Garyfallidis, E.</dc:creator>
<dc:creator>Giesbrecht, B.</dc:creator>
<dc:creator>Gur, R. C.</dc:creator>
<dc:creator>Gur, R. E.</dc:creator>
<dc:creator>Kelz, M.</dc:creator>
<dc:creator>Keshavan, A.</dc:creator>
<dc:creator>Larsen, B. S.</dc:creator>
<dc:creator>Luna, B.</dc:creator>
<dc:creator>Mackey, A. P.</dc:creator>
<dc:creator>Milham, M.</dc:creator>
<dc:creator>Oathes, D. J.</dc:creator>
<dc:creator>Perrone, A.</dc:creator>
<dc:creator>Pines, A. R.</dc:creator>
<dc:creator>Roalf, D. R.</dc:creator>
<dc:creator>Richie-Halford, A.</dc:creator>
<dc:creator>Rokem, A.</dc:creator>
<dc:creator>Sydnor, V.</dc:creator>
<dc:creator>Tapera, T. M.</dc:creator>
<dc:creator>Tooley, U. A.</dc:creator>
<dc:creator>Vettel, J. M.</dc:creator>
<dc:creator>Yeatman, J.</dc:creator>
<dc:creator>Grafton, S. T.</dc:creator>
<dc:creator>Satterthwaite, T. D.</dc:creator>
<dc:date>2020-09-04</dc:date>
<dc:identifier>doi:10.1101/2020.09.04.282269</dc:identifier>
<dc:title><![CDATA[QSIPrep: An integrative platform for preprocessing and reconstructing diffusion MRI]]></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/10.1101/2020.09.04.283234v1?rss=1">
<title>
<![CDATA[
DISSECT: DISentangle SharablE ConTent for Multimodal Integration and Crosswise-mapping 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.09.04.283234v1?rss=1</link>
<description><![CDATA[
Deep learning systems have emerged as powerful mechanisms for learning domain translation models. However, in many cases, complete information in one domain is assumed to be necessary for sufficient cross-domain prediction. In this work, we motivate a formal justification for domain-specific information separation in a simple linear case and illustrate that a self-supervised approach enables domain translation between data domains while filtering out domain-specific data features. We introduce a novel approach to identify domainspecific information from sets of unpaired measurements in complementary data domains by considering a deep learning cross-domain autoencoder architecture designed to learn shared latent representations of data while enabling domain translation. We introduce an orthogonal gate block designed to enforce orthogonality of input feature sets by explicitly removing non-sharable information specific to each domain and illustrate separability of domain-specific information on a toy dataset.
]]></description>
<dc:creator>Schau, G.</dc:creator>
<dc:creator>Burlingame, E. A.</dc:creator>
<dc:creator>Chang, Y. H.</dc:creator>
<dc:date>2020-09-06</dc:date>
<dc:identifier>doi:10.1101/2020.09.04.283234</dc:identifier>
<dc:title><![CDATA[DISSECT: DISentangle SharablE ConTent for Multimodal Integration and Crosswise-mapping]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-09-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.09.09.289017v1?rss=1">
<title>
<![CDATA[
Insulin potentiates the synchronous firing of arcuate nucleus Kiss1 neurons that protects against diet-induced obesity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.09.09.289017v1?rss=1</link>
<description><![CDATA[
Kisspeptin (Kiss1) neurons are essential for reproduction, but their role in the control of energy balance and other homeostatic functions remains unclear. High frequency firing of hypothalamic arcuate Kiss1 (Kiss1ARH) neurons releases kisspeptin into the median eminence, and neurokinin B (NKB) and dynorphin onto neighboring Kiss1ARH neurons to generate a slow excitatory postsynaptic potential (EPSP) mediated by TRPC5 channels that entrains intermittent, synchronous firing of Kiss1ARH neurons. High frequency optogenetic stimulation of Kiss1ARH neurons releases glutamate to excite the anorexigenic proopiomelanocortin (POMC) neurons and inhibit the orexigenic neuropeptide Y/agouti-related peptide (AgRP) neurons via metabotropic glutamate receptors. At the molecular level, the endoplasmic reticulum calcium-sensing protein stromal interaction molecule 1 (STIM1) is critically involved in the regulation of neuronal Ca2+ signaling and neuronal excitability through its interaction with plasma membrane calcium (e.g., TRPC) channels. 17{beta}-estradiol (E2) downregulates Stim1 mRNA expression in female arcuate neurons. Therefore, we hypothesized that deletion of Stim1 in Kiss1ARH neurons would increase neuronal excitability and their synchronous firing, which ultimately would affect energy homeostasis. Using optogenetics in combination with whole-cell recording and GCaMP6 imaging in slices, we discovered that the deletion of Stim1 in Kiss1 neurons significantly increased the amplitude of the slow EPSP and augmented synchronous [Ca2+]i oscillations in Kiss1ARH neurons. Deletion of Stim1 in Kiss1ARH neurons amplified the actions of NKB and protected ovariectomized female mice from developing obesity and glucose intolerance with high-fat dieting. Therefore, STIM1 appears to play a critical role in regulating synchronous firing of Kiss1ARH neurons, which ultimately affects energy homeostasis.

Significance StatementHypothalamic arcuate kisspeptin (Kiss1ARH) neurons are essential for stimulating the pulsatile release of gonadotropin releasing hormone (GnRH) and maintaining fertility. However, Kiss1ARH neurons appear to be a key player in coordinating energy balance with reproduction. The regulation of calcium channels and hence calcium signaling is critically dependent on the endoplasmic reticulum calcium-sensing protein stromal interaction molecule 1 (STIM1), which interacts with the plasma membrane calcium channels. We have conditionally deleted Stim1 in Kiss1ARH neurons and found that it significantly increased the excitability of Kiss1ARH neurons and protected ovariectomized female mice from developing obesity and glucose intolerance with high-fat dieting.
]]></description>
<dc:creator>Qiu, J.</dc:creator>
<dc:creator>Stincic, T. L.</dc:creator>
<dc:creator>Bosch, M. A.</dc:creator>
<dc:creator>Connors, A. M.</dc:creator>
<dc:creator>Petrie, S. K.</dc:creator>
<dc:creator>Ronnekleiv, O. K.</dc:creator>
<dc:creator>Kelly, M. J.</dc:creator>
<dc:date>2020-09-11</dc:date>
<dc:identifier>doi:10.1101/2020.09.09.289017</dc:identifier>
<dc:title><![CDATA[Insulin potentiates the synchronous firing of arcuate nucleus Kiss1 neurons that protects against diet-induced obesity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-09-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.09.10.286252v1?rss=1">
<title>
<![CDATA[
Supraphysiological testosterone induces ferroptosis and activates NF-kappaB mediated immune pathways in prostate cancer through nucleophagy 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.09.10.286252v1?rss=1</link>
<description><![CDATA[
The discovery that androgens play an important role in the progression of prostate cancer (PCa) has led to the development of androgen deprivation therapy as a first line of treatment against PCa. However, paradoxical growth inhibition has been observed, both experimentally and clinically, in a subset of PCa upon administration of supraphysiological levels of testosterone (SupraT). Here we report that SupraT activates cytoplasmic nucleic acid sensors and induces growth inhibition of SupraT-sensitive PCa cells. This is initiated by induction of two parallel autophagy-mediated processes, namely, ferritinophagy and nucleophagy. Consequently, autophagosomal DNA activates nucleic acid sensors that converge on NF-kappaB to drive immune signaling pathways. Chemokines and cytokines secreted by the tumor cells in response to SupraT results in increased migration of cytotoxic immune cells to tumor beds of animal xenografts and patient tumors. Collectively, our findings indicate that SupraT may inhibit a subset of PCa by activating nucleic acid sensors and downstream immune signaling.
]]></description>
<dc:creator>Mendonca, J.</dc:creator>
<dc:creator>Kumar, R.</dc:creator>
<dc:creator>Owoyemi, O.</dc:creator>
<dc:creator>Boyapati, K.</dc:creator>
<dc:creator>Thomas, N.</dc:creator>
<dc:creator>Kanacharoen, S.</dc:creator>
<dc:creator>Coffey, M.</dc:creator>
<dc:creator>Topiwala, D.</dc:creator>
<dc:creator>Gomes, C.</dc:creator>
<dc:creator>Ozbek, B.</dc:creator>
<dc:creator>Jones, T.</dc:creator>
<dc:creator>Rosen, M.</dc:creator>
<dc:creator>Dong, L.</dc:creator>
<dc:creator>Wiens, S.</dc:creator>
<dc:creator>Brennen, W. N.</dc:creator>
<dc:creator>Isaacs, J.</dc:creator>
<dc:creator>De Marzo, A. M.</dc:creator>
<dc:creator>Markowski, M.</dc:creator>
<dc:creator>Antonarakis, E. S.</dc:creator>
<dc:creator>Qian, D.</dc:creator>
<dc:creator>Pienta, K.</dc:creator>
<dc:creator>Pardoll, D.</dc:creator>
<dc:creator>Carducci, M.</dc:creator>
<dc:creator>Denmeade, S.</dc:creator>
<dc:creator>Kachhap, S.</dc:creator>
<dc:date>2020-09-10</dc:date>
<dc:identifier>doi:10.1101/2020.09.10.286252</dc:identifier>
<dc:title><![CDATA[Supraphysiological testosterone induces ferroptosis and activates NF-kappaB mediated immune pathways in prostate cancer through nucleophagy]]></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/10.1101/2020.09.10.291823v1?rss=1">
<title>
<![CDATA[
Microbial colonization induces histone acetylation critical for inherited gut-germline-neural signaling 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.09.10.291823v1?rss=1</link>
<description><![CDATA[
The gut-neural axis plays a critical role in the control of several physiological processes, including the communication of signals from the microbiome to the nervous system, which affects learning, memory, and behavior. However, the pathways involved in gut-neural signaling of gut-governed behaviors remain unclear. We found that the intestinal distension caused by the bacterium Pseudomonas aeruginosa induces histone H4 Lys8 acetylation (H4K8ac) in the germline of Caenorhabditis elegans, which is required for both a bacterial aversion behavior and its transmission to the next generation. We show that induction of H4K8ac in the germline is essential for bacterial aversion and that a 14-3-3 chaperone protein family member, PAR-5, is required for H4K8ac. Our findings highlight a role for H4K8ac in the germline not only in the intergenerational transmission of pathogen avoidance but also in the transmission of pathogenic cues that travel through the gut-neural axis to control the aversive behavior.
]]></description>
<dc:creator>Aballay, A.</dc:creator>
<dc:creator>Hong, C.</dc:creator>
<dc:date>2020-09-11</dc:date>
<dc:identifier>doi:10.1101/2020.09.10.291823</dc:identifier>
<dc:title><![CDATA[Microbial colonization induces histone acetylation critical for inherited gut-germline-neural signaling]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-09-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.09.14.295295v1?rss=1">
<title>
<![CDATA[
ASXL1 Directs Neutrophilic Differentiation via Modulation of MYC and RNA Polymerase II. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.09.14.295295v1?rss=1</link>
<description><![CDATA[
Mutations in the gene Additional Sex-Combs Like 1 (ASXL1) are recurrent in myeloid malignancies as well as the pre-malignant condition clonal hematopoiesis, where they are universally associated with poor prognosis. An epigenetic regulator, ASXL1 canonically directs the deposition of H3K27me3 via the polycomb repressive complex 2. However, its precise role in myeloid lineage maturation is incompletely described. We utilized single cell RNA sequencing (scRNA-seq) on a murine model of hematopoietic-specific ASXL1 deletion and identified a specific role for ASXL1 in terminal granulocyte maturation. Terminal maturation is accompanied by down regulation of Myc expression and cell cycle exit. ASXL1 deletion leads to hyperactivation of Myc in granulocyte precursors and a quantitative decrease in neutrophil production. This failure of normal developmentallyassociated Myc suppression is not accompanied by significant changes in the landscape of covalent histone modifications including H3K27me3. Examining the genome-wide localization of ASXL1 in myeloid progenitors revealed strong co-localization with RNA Polymerase II (RNAPII) at the promoters and spread across the gene bodies of transcriptionally active genes. ASXL1 deletion results in a decrease in RNAPII promoter-proximal pausing in granulocyte progenitors, indicative of a global increase in productive transcription, consistent with the known role of ASXL1 as a mediator of RNAPII pause release. These results suggest that ASXL1 inhibits productive transcription in granulocyte progenitors, identifying a new role for this epigenetic regulator and highlighting a novel potential oncogenic mechanism for ASXL1 mutations in myeloid malignancies.
]]></description>
<dc:creator>Braun, T. P.</dc:creator>
<dc:creator>Estabrook, J.</dc:creator>
<dc:creator>Coleman, D. J.</dc:creator>
<dc:creator>Schonrock, Z.</dc:creator>
<dc:creator>Smith, B. M.</dc:creator>
<dc:creator>Enright, T.</dc:creator>
<dc:creator>Coblentz, C.</dc:creator>
<dc:creator>Callahan, R.</dc:creator>
<dc:creator>Mohammed, H.</dc:creator>
<dc:creator>Druker, B. J.</dc:creator>
<dc:creator>Lusardi, T.</dc:creator>
<dc:creator>Maxson, J. E.</dc:creator>
<dc:date>2020-09-14</dc:date>
<dc:identifier>doi:10.1101/2020.09.14.295295</dc:identifier>
<dc:title><![CDATA[ASXL1 Directs Neutrophilic Differentiation via Modulation of MYC and RNA Polymerase II.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-09-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.09.14.297135v1?rss=1">
<title>
<![CDATA[
Polycomb represses a gene network controlling puberty via modulation of histone demethylase Kdm6b expression 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.09.14.297135v1?rss=1</link>
<description><![CDATA[
Female puberty is subject to Polycomb Group (PcG)-dependent transcriptional repression. Kiss1, a puberty-activating gene, is a key target of this silencing mechanism. Using a gain-of-function approach and a systems biology strategy we now show that EED, an essential PcG component, acts in the arcuate nucleus of the hypothalamus to alter the functional organization of a gene network involved in the stimulatory control of puberty. A central node of this network is Kdm6b, which encodes an enzyme that erases the PcG-dependent histone modification H3K27me3. Kiss1 is a first neighbor in the network; genes encoding glutamatergic receptors and potassium channels are second neighbors. By repressing Kdm6b expression, EED increases H3K27me3 abundance at these gene promoters, reducing gene expression throughout a gene network controlling puberty activation. These results indicate that Kdm6b repression is a basic mechanism used by PcG to modulate the biological output of puberty-activating gene networks.
]]></description>
<dc:creator>Wright, H.</dc:creator>
<dc:creator>Aylwin, C. F.</dc:creator>
<dc:creator>Toro, C. A.</dc:creator>
<dc:creator>Ojeda, S. R.</dc:creator>
<dc:creator>Lomniczi, A.</dc:creator>
<dc:date>2020-09-14</dc:date>
<dc:identifier>doi:10.1101/2020.09.14.297135</dc:identifier>
<dc:title><![CDATA[Polycomb represses a gene network controlling puberty via modulation of histone demethylase Kdm6b expression]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-09-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.09.16.300277v1?rss=1">
<title>
<![CDATA[
Baricitinib treatment resolves lower airway inflammation and neutrophil recruitment in SARS-CoV-2-infected rhesus macaques 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.09.16.300277v1?rss=1</link>
<description><![CDATA[
Effective therapeutics aimed at mitigating COVID-19 symptoms are urgently needed. SARS-CoV-2 induced hypercytokinemia and systemic inflammation are associated with disease severity. Baricitinib, a clinically approved JAK1/2 inhibitor with potent anti-inflammatory properties is currently being investigated in COVID-19 human clinical trials. Recent reports suggest that baricitinib may also have antiviral activity in limiting viral endocytosis. Here, we investigated the immunologic and virologic efficacy of baricitinib in a rhesus macaque model of SARS-CoV-2 infection. Viral shedding measured from nasal and throat swabs, bronchoalveolar lavages and tissues was not reduced with baricitinib. Type I IFN antiviral responses and SARS-CoV-2 specific T cell responses remained similar between the two groups. Importantly, however, animals treated with baricitinib showed reduced immune activation, decreased infiltration of neutrophils into the lung, reduced NETosis activity, and more limited lung pathology. Moreover, baricitinib treated animals had a rapid and remarkably potent suppression of alveolar macrophage derived production of cytokines and chemokines responsible for inflammation and neutrophil recruitment. These data support a beneficial role for, and elucidate the immunological mechanisms underlying, the use of baricitinib as a frontline treatment for severe inflammation induced by SARS-CoV-2 infection.
]]></description>
<dc:creator>Hoang, T. N.</dc:creator>
<dc:creator>Pino, M.</dc:creator>
<dc:creator>Boddapati, A. K.</dc:creator>
<dc:creator>Viox, E. G.</dc:creator>
<dc:creator>Starke, C. E.</dc:creator>
<dc:creator>Upadhyay, A. A.</dc:creator>
<dc:creator>Gumber, S.</dc:creator>
<dc:creator>Busman-Sahay, K.</dc:creator>
<dc:creator>Strongin, Z.</dc:creator>
<dc:creator>Harper, J. L.</dc:creator>
<dc:creator>Tharp, G. K.</dc:creator>
<dc:creator>Pellegrini, K. L.</dc:creator>
<dc:creator>Kirejczyk, S.</dc:creator>
<dc:creator>Zandi, K.</dc:creator>
<dc:creator>Tao, S.</dc:creator>
<dc:creator>Horton, T. R.</dc:creator>
<dc:creator>Beagle, E. N.</dc:creator>
<dc:creator>Mahar, E. A.</dc:creator>
<dc:creator>Lee, M. Y.</dc:creator>
<dc:creator>Cohen, J.</dc:creator>
<dc:creator>Jean, S.</dc:creator>
<dc:creator>Wood, J. S.</dc:creator>
<dc:creator>Connor-Stroud, F.</dc:creator>
<dc:creator>Stammen, R. L.</dc:creator>
<dc:creator>Delmas, O. M.</dc:creator>
<dc:creator>Wang, S.</dc:creator>
<dc:creator>Cooney, K. A.</dc:creator>
<dc:creator>Sayegh, M. N.</dc:creator>
<dc:creator>Wang, L.</dc:creator>
<dc:creator>Weiskopf, D.</dc:creator>
<dc:creator>Filev, P. D.</dc:creator>
<dc:creator>Waggoner, J.</dc:creator>
<dc:creator>Piantadosi, A.</dc:creator>
<dc:creator>Kasturi, S. P.</dc:creator>
<dc:creator>Al-Shakhshir, H.</dc:creator>
<dc:creator>Ribeiro, S. P.</dc:creator>
<dc:creator>Sekaly, R. P.</dc:creator>
<dc:creator>Levit, R. D.</dc:creator>
<dc:creator>Estes, J. D.</dc:creator>
<dc:creator>Vanderford, T. H.</dc:creator>
<dc:creator>Schinazi, R. F.</dc:creator>
<dc:creator>Bosinger,</dc:creator>
<dc:date>2020-09-16</dc:date>
<dc:identifier>doi:10.1101/2020.09.16.300277</dc:identifier>
<dc:title><![CDATA[Baricitinib treatment resolves lower airway inflammation and neutrophil recruitment in SARS-CoV-2-infected rhesus macaques]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-09-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.09.18.304048v1?rss=1">
<title>
<![CDATA[
Chronic Psychosocial Stress and Experimental Pubertal Delay Affect Socioemotional Behavior and Amygdala Functional Connectivity in Adolescent Female Rhesus Macaques 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.09.18.304048v1?rss=1</link>
<description><![CDATA[
In females, pubertal onset appears to signal the opening of a window of increased vulnerability to the effects of stress on neurobehavioral development. What is the impact of pubertal timing on this process? We assessed the effects of pubertal timing and stress on behavior and amygdala functional connectivity (FC) in adolescent female macaques, whose social hierarchy provides an ethologically valid model of chronic psychosocial stress. Monkeys experienced puberty spontaneously (n=34) or pubertal delay via Lupron treatment from age 16-33 months (n=36). We examined the effects of stress (continuous dimension spanning dominant/low-stress to subordinate/high-stress) and experimental pubertal delay (Lupron-treated vs. Control) on socioemotional behavior and FC at 43-46 months, after all animals had begun puberty. Regardless of treatment, subordinate monkeys were more submissive and less affiliative, and exhibited weaker FC between amygdala and dorsolateral prefrontal cortex and stronger FC between amygdala and temporal pole. Regardless of social rank, Lupron-treated monkeys were also more submissive, less affiliative, and explored less in a "Human Intruder" task but were less anxious than untreated monkeys; they exhibited stronger FC between amygdala and orbitofrontal cortex. No interactions between rank and Lupron treatment were observed. These data suggest that some of the effects of chronic subordination stress and delayed puberty overlap behaviorally, such that late-onset puberty-linked exposure to female hormones mimics chronic stress. In the brain, however, delayed puberty and subordination stress had separable effects, suggesting that the overlapping socioemotional outcomes may be mediated by distinct neuroplastic mechanisms. To gain further insights, additional longitudinal studies are required.
]]></description>
<dc:creator>Pincus, M.</dc:creator>
<dc:creator>Godfrey, J. S.</dc:creator>
<dc:creator>Feczko, E.</dc:creator>
<dc:creator>Earl, E.</dc:creator>
<dc:creator>Miranda Dominguez, O.</dc:creator>
<dc:creator>Fair, D.</dc:creator>
<dc:creator>Wilson, M. E.</dc:creator>
<dc:creator>Sanchez, M. M.</dc:creator>
<dc:creator>Kelly, C.</dc:creator>
<dc:date>2020-09-19</dc:date>
<dc:identifier>doi:10.1101/2020.09.18.304048</dc:identifier>
<dc:title><![CDATA[Chronic Psychosocial Stress and Experimental Pubertal Delay Affect Socioemotional Behavior and Amygdala Functional Connectivity in Adolescent Female Rhesus Macaques]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-09-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.09.21.306837v1?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/10.1101/2020.09.24.312702v1?rss=1">
<title>
<![CDATA[
Increased oxidative stress tolerance of a spontaneously-occurring perR gene mutation in Streptococcus mutans UA159 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.09.24.312702v1?rss=1</link>
<description><![CDATA[
The ability of bacteria such as the dental pathogen Streptococcus mutans to coordinate a response against damage-inducing oxidants is a critical aspect of their pathogenicity. The oxidative stress regulator SpxA1 has been proven to be a major player in the ability of S. mutans to withstand both disulfide and peroxide stresses. While studying spontaneously-occurring variants of an S. mutans {Delta}spxA1 strain, we serendipitously discovered that our S. mutans UA159 host strain bore a single nucleotide deletion within the coding region ofperR, resulting in a premature truncation of the encoded protein. PerR is a metal-dependent transcriptional repressor that senses and responds to peroxide stress such that loss of PerR activity results in activation of oxidative stress responses. To determine the impact of loss of PerR regulation, we obtained a UA159 isolated bearing an intact perR copy and created a clean perR deletion mutant. Our findings indicate that loss of PerR activity results in a strain that is primed to tolerate oxidative stresses in the laboratory setting. Interestingly, RNA-Seq and targeted transcriptional expression analyses reveal that PerR has a minor contribution to the ability of S. mutans to orchestrate a transcriptional response to peroxide stress. Furthermore, we detected loss-of-function perR mutations in two other commonly used laboratory strains of S. mutans suggesting that this may be not be an uncommon occurrence. This report serves as a cautionary warning regarding the so-called domestication of laboratory strains and advocates for the implementation of more stringent strain authentication practices.

ImportanceA resident of the human oral biofilm, Streptococcus mutans is one of the major bacterial pathogens associated with dental caries. This report highlights a spontaneously-occurring mutation within the laboratory strain S. mutans UA159, found in the coding region of perR, a gene encoding a transcriptional repressor associated with peroxide tolerance. Though perR mutant strains of S. mutans showed a distinct growth advantage and enhanced tolerance toward H2O2, a {Delta}perR deletion strain showed a small number of differentially expressed genes as compared to the parent strain, suggesting few direct regulatory targets. In addition to characterizing the role of PerR in S. mutans, our findings serve as a warning to laboratory researchers regarding bacterial adaptation to in vitro growth conditions.
]]></description>
<dc:creator>Kajfasz, J. K.</dc:creator>
<dc:creator>Zuber, P.</dc:creator>
<dc:creator>Ganguly, T.</dc:creator>
<dc:creator>Abranches, J.</dc:creator>
<dc:creator>Lemos, J. A.</dc:creator>
<dc:date>2020-09-25</dc:date>
<dc:identifier>doi:10.1101/2020.09.24.312702</dc:identifier>
<dc:title><![CDATA[Increased oxidative stress tolerance of a spontaneously-occurring perR gene mutation in Streptococcus mutans UA159]]></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/10.1101/2020.09.28.317859v1?rss=1">
<title>
<![CDATA[
Human cytomegalovirus UL7, miR-US5-1 and miR-UL112-3p inactivation of FOXO3a protects CD34+ hematopoietic progenitor cells from apoptosis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.09.28.317859v1?rss=1</link>
<description><![CDATA[
Human cytomegalovirus (HCMV) infection of myeloid-lineage cells, such as CD34+ hematopoietic progenitor cells (HPCs) or monocytes results in the upregulation of anti-apoptotic cellular proteins that protect the newly infected cells from programmed cell death. The mechanisms used by HCMV to regulate pro-apoptotic cellular proteins upon infection of CD34+ HPCs has not been fully explored. Here we show that HCMV utilizes pUL7, a secreted protein that signals through the FLT3 receptor, and miR-US5-1 and miR-UL112-3p to reduce the abundance and activity of the pro-apoptotic transcription factor FOXO3a at early times after infection of CD34+ HPCs. Regulation of FOXO3a by pUL7, miR-US5-1 and miR-UL112 results in reduced expression of the pro-apoptotic BCL2L11 transcript and protection of CD34+ HPCs from virus-induced apoptosis. These data highlight the importance of both viral proteins and miRNAs in protecting CD34+ HPCs from apoptosis at early times post-infection, allowing for the establishment of latency and maintenance of viral genome-containing cells.

ImportanceHuman cytomegalovirus (HCMV) causes serious disease in immunocompromised individuals and is a significant problem during transplantation. The virus can establish a latent infection in CD34+ hematopoietic progenitor cells (HPCs) and periodically reactivate to cause disease in the absence of an intact immune system. What viral gene products are required for successful establishment of latency are still not fully understood. Here we show that both a viral protein and viral miRNAs are required to prevent apoptosis after infection of CD34+ HPCs. HCMV pUL7 and miRNAs miR-US5-1 and miR-UL112-3p act to limit the expression and activation of the transcription factor FOXO3a which in turn reduces expression of pro-apoptotic gene BCL2L11 and prevents virus-induced apoptosis in CD34+ HPCs.
]]></description>
<dc:creator>Hancock, M. H.</dc:creator>
<dc:creator>Crawford, L. B.</dc:creator>
<dc:creator>Perez, W.</dc:creator>
<dc:creator>Struthers, H. M.</dc:creator>
<dc:creator>Mitchell, J.</dc:creator>
<dc:creator>Caposio, P.</dc:creator>
<dc:date>2020-09-29</dc:date>
<dc:identifier>doi:10.1101/2020.09.28.317859</dc:identifier>
<dc:title><![CDATA[Human cytomegalovirus UL7, miR-US5-1 and miR-UL112-3p inactivation of FOXO3a protects CD34+ hematopoietic progenitor cells from apoptosis]]></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/10.1101/2020.10.06.323063v1?rss=1">
<title>
<![CDATA[
The macaque brain ONPRC18 template with combined gray and white matter labelmap for multimodal neuroimaging studies of nonhuman primates 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.10.06.323063v1?rss=1</link>
<description><![CDATA[
Macaques are the most common nonhuman primate (NHP) species used in neuroscience research. With the advancement of many neuroimaging techniques, new studies are beginning to apply multiple types of in vivo magnetic resonance imaging (MRI), such as structural imaging (sMRI) with T1 and T2 weighted contrasts alongside diffusion weighed (DW) imaging. In studies involving rhesus macaques, this approach can be used to better understand micro-structural changes that occur during development, in various disease states or with normative aging. However, many of the available rhesus brain atlases have been designed for only one imaging modality, making it difficult to consistently define the same brain regions across multiple imaging modalities in the same subject. To address this, we created a brain atlas from 18 adult rhesus macaques that includes co-registered templates constructed from images frequently used to characterize macroscopic brain structure (T2/SPACE and T1/MP-RAGE), and a diffusion tensor imaging (DTI) template. The DTI template was up-sampled from 1 mm isotropic resolution to resolution match to the T1 and T2-weighted images (0.5 mm isotropic), and the parameter map was derived for fractional anisotropy (FA). The labelmap volumes delineate 57 gray matter regions of interest (ROIs; 36 cortical regions and 21 subcortical structures), as well as 74 white matter tracts. Importantly, the labelmap overlays both the structural and diffusion templates, enabling the same regions to be consistently identified across imaging modalities. A specialized condensed version of the labelmap ROIs are also included to further extend the usefulness of this tool for imaging data with lower spatial resolution, such as functional MRI (fMRI) or positron emission tomography (PET). O_FIG_DISPLAY_L [Figure 1] M_FIG_DISPLAY Graphical Abstract C_FIG_DISPLAY
]]></description>
<dc:creator>Weiss, A. R.</dc:creator>
<dc:creator>Liu, Z.</dc:creator>
<dc:creator>Wang, X.</dc:creator>
<dc:creator>Liguore, W. A.</dc:creator>
<dc:creator>Kroenke, C.</dc:creator>
<dc:creator>McBride, J.</dc:creator>
<dc:date>2020-10-08</dc:date>
<dc:identifier>doi:10.1101/2020.10.06.323063</dc:identifier>
<dc:title><![CDATA[The macaque brain ONPRC18 template with combined gray and white matter labelmap for multimodal neuroimaging studies of nonhuman primates]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-10-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.10.07.330316v1?rss=1">
<title>
<![CDATA[
The Trypanosoma brucei Cytoskeletal Protein KHARON Associates with Partner Proteins to Mediate Both Cytokinesis and Trafficking of Flagellar Membrane Proteins 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.10.07.330316v1?rss=1</link>
<description><![CDATA[
In the African trypanosome Trypanosoma brucei, the cytoskeletal protein TbKHARON is required for trafficking of a putative Ca2+ channel to the flagellar membrane, and it is essential for parasite viability in both the mammalian stage bloodstream forms and the tsetse fly procyclic forms. This protein is located at the base of the flagellum, in the pellicular cytoskeleton, and in the mitotic spindle in both life cycle forms, and it likely serves multiple functions for these parasites. To begin to deconvolve the functions of KHARON, we have investigated partners associated with this protein and their roles in parasite biology. One KHARON associated protein, TbKHAP1, is a close interaction partner that can be crosslinked to KHARON by formaldehyde and pulled down in a molecular complex, and it colocalizes with TbKHARON in the basal body at the base of the flagellum. Knockdown of TbKHAP1 mRNA has similar phenotypes to knockdown of its partner TbKHARON, impairing trafficking of the Ca2+ channel to the flagellar membrane and blocking cytokinesis, implying that the TbKHARON/TbKHAP1 complex mediates trafficking of flagellar membrane proteins. Two other KHAPs, TbKHAP2 and TbKHAP3, are in close proximity to TbKHARON, but knockdown of their mRNAs does not affect trafficking of the Ca2+ channel. Two different flagellar membrane proteins, which are extruded from the flagellar membrane into extracellular vesicles, are also dependent upon TbKHARON for flagellar trafficking. These studies confirm that TbKHARON acts in complexes with other proteins to carry out various biological functions, and that some partners are involved in the core activity of targeting membrane proteins to the flagellum.
]]></description>
<dc:creator>Landfear, S. M.</dc:creator>
<dc:creator>Sanchez, M. A.</dc:creator>
<dc:date>2020-10-07</dc:date>
<dc:identifier>doi:10.1101/2020.10.07.330316</dc:identifier>
<dc:title><![CDATA[The Trypanosoma brucei Cytoskeletal Protein KHARON Associates with Partner Proteins to Mediate Both Cytokinesis and Trafficking of Flagellar Membrane Proteins]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-10-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.10.08.330241v1?rss=1">
<title>
<![CDATA[
Atypical molecular basis for drug resistance to mitochondrial function inhibitors in Plasmodium falciparum 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.10.08.330241v1?rss=1</link>
<description><![CDATA[
The continued emergence of drug-resistant Plasmodium falciparum parasites hinders global attempts to eradicate malaria, emphasizing the need to identify new antimalarial drugs. Attractive targets for chemotherapeutic intervention are the cytochrome (cyt) bc1 complex, which is an essential component of the mitochondrial electron transport chain (mtETC) necessary for ubiquinone recycling and mitochondrially localized dihydroorotate dehydrogenase (DHODH) critical for de novo pyrimidine synthesis. Despite the essentiality of this complex, resistance to a novel acridone class of compounds targeting cyt bc1 was readily attained, resulting in a parasite strain (SB1-A6) that was pan-resistant to both mtETC and DHODH inhibitors. Here we describe the molecular mechanism behind the resistance of the SB1-A6 parasite line which lacks the common cyt bc1 point mutations characteristic of resistance to mtETC inhibitors. Using Illumina whole-genome sequencing, we have identified both a copy number variation ([~]2x) and a single-nucleotide polymorphism (C276F) associated with pfdhodh in SB1-A6. We have characterized the role of both genetic lesions by mimicking the copy number variation via episomal expression of pfdhodh and introducing the identified SNP using CRISPR/Cas9 and assessed their contributions to drug resistance. Although both of these genetic polymorphisms have been previously identified as contributing to both DSM-1 (1) and atovaquone resistance (2, 3), SB1-A6 represents a unique genotype in which both alterations are present in a single line, suggesting that the combination contributes to the pan-resistant phenotype. This novel mechanism of resistance to mtETC inhibition has critical implications for the development of future drugs targeting the bc1 complex or de novo pyrimidine synthesis that could help guide future anti-malarial combination therapies and reduce the rapid development of drug resistance in the field.
]]></description>
<dc:creator>Painter, H. J.</dc:creator>
<dc:creator>Morrisey, J. M.</dc:creator>
<dc:creator>Mather, M. W.</dc:creator>
<dc:creator>Orchard, L. M.</dc:creator>
<dc:creator>Luck, C.</dc:creator>
<dc:creator>Smilkstein, M. J.</dc:creator>
<dc:creator>Riscoe, M. K.</dc:creator>
<dc:creator>Vaidya, A. B.</dc:creator>
<dc:creator>Llinas, M.</dc:creator>
<dc:date>2020-10-08</dc:date>
<dc:identifier>doi:10.1101/2020.10.08.330241</dc:identifier>
<dc:title><![CDATA[Atypical molecular basis for drug resistance to mitochondrial function inhibitors in Plasmodium falciparum]]></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/10.1101/2020.10.15.336420v1?rss=1">
<title>
<![CDATA[
Distinct cis-acting elements govern purine-responsive regulation of the Leishmania donovani nucleoside transporters, LdNT1 and LdNT2 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.10.15.336420v1?rss=1</link>
<description><![CDATA[
Purine salvage from the host is an obligatory process for all protozoan parasites. In Leishmania donovani, this is accomplished by four membrane nucleoside and nucleobase transporters, or LdNTs. Previously, we demonstrated that purine starvation invokes a robust stress response in Leishmania and characterized the proteomic changes involved. However, because Leishmania do not control the transcription of individual genes, the underlying mechanisms responsible for these changes were ill-defined. LdNT1 and LdNT2 are among the most rapidly and significantly upregulated genes in purine-starved L. donovani parasites. Thus, to better understand post-transcriptional mechanisms of purine-responsive gene expression, we have examined regulation of these genes in molecular detail. Here we report that LdNT1 and LdNT2 are controlled by distinct cis-acting elements. In the case of LdNT2, mRNA abundance and translational enhancement under purine stress depend on a 76 nt-long polypyrimidine tract encoded in the LdNT2 mRNA 3-UTR. Transcripts containing the LdNT2 polypyrimidine tract were found to localize to discrete cytoplasmic foci in purine-replete cells, suggesting that the LdNT2 message may be stored in RNA granules at steady-state. In the case of LdNT1, we found that purine-responsiveness is conferred by a 48 nt-long polypyrimidine tract and additional upstream element, termed UE1. Both features are independently required for regulation, with the polypyrimidine tract and UE1 controlling mRNA abundance and translation, respectively. Together, these results highlight a remarkable degree of complexity in the regulation of the Leishmania purine stress response and set the stage for future investigations to identify the larger network of RNA-protein and protein-protein interactions involved.
]]></description>
<dc:creator>Licon, M. H.</dc:creator>
<dc:creator>Goodstein, F.</dc:creator>
<dc:creator>Ortiz, D.</dc:creator>
<dc:creator>Landfear, S. M.</dc:creator>
<dc:creator>Yates, P. A.</dc:creator>
<dc:date>2020-10-15</dc:date>
<dc:identifier>doi:10.1101/2020.10.15.336420</dc:identifier>
<dc:title><![CDATA[Distinct cis-acting elements govern purine-responsive regulation of the Leishmania donovani nucleoside transporters, LdNT1 and LdNT2]]></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/10.1101/2020.10.19.346346v1?rss=1">
<title>
<![CDATA[
Large, stable spikes exhibit differential broadening in excitatory and inhibitory neocortical boutons 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.10.19.346346v1?rss=1</link>
<description><![CDATA[
Presynaptic action potential spikes control neurotransmitter release and thus interneuronal communication. However, the properties and the dynamics of presynaptic spikes in the neocortex remain enigmatic because boutons in the neocortex are small and direct patch-clamp recordings have not been performed. Here we report direct recordings from boutons of neocortical pyramidal neurons and interneurons. Our data reveal rapid and large presynaptic action potentials in layer 5 neurons and fast-spiking interneurons reliably propagating into axon collaterals. For in-depth analyses we validate boutons of mature cultured neurons as models for excitatory neocortical boutons, demonstrating that the presynaptic spike amplitude was unaffected by potassium channels, homeostatic long-term plasticity, and high-frequency firing. In contrast to the stable amplitude, presynaptic spikes profoundly broadened for example during high-frequency firing in layer 5 pyramidal neurons but not in fast-spiking interneurons. Thus, our data demonstrate large presynaptic spikes and fundamental differences between excitatory and inhibitory boutons in the neocortex.
]]></description>
<dc:creator>Ritzau-Jost, A.</dc:creator>
<dc:creator>Tsintsadze, T.</dc:creator>
<dc:creator>Krueger, M.</dc:creator>
<dc:creator>Ader, J.</dc:creator>
<dc:creator>Bechmann, I.</dc:creator>
<dc:creator>Eilers, J.</dc:creator>
<dc:creator>Barbour, B.</dc:creator>
<dc:creator>Smith, S. M.</dc:creator>
<dc:creator>Hallermann, S.</dc:creator>
<dc:date>2020-10-20</dc:date>
<dc:identifier>doi:10.1101/2020.10.19.346346</dc:identifier>
<dc:title><![CDATA[Large, stable spikes exhibit differential broadening in excitatory and inhibitory neocortical boutons]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-10-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.10.21.349191v1?rss=1">
<title>
<![CDATA[
Antisense oligonucleotides targeting alpha-synuclein reduce pre-formed fibril-induced Lewy pathology and improve some domains of cognitive and motor performance 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.10.21.349191v1?rss=1</link>
<description><![CDATA[
Alpha-synuclein (syn) is a small protein involved in neurodegenerative diseases known as synucleinopathies. The phosphorylated form (psyn) is the primary component of protein aggregates known as Lewy bodies (LBs), which are the hallmark of diseases such as Parkinsons disease (PD) and Dementia with Lewy bodies (DLB). Synucleinopathies might spread in a prion-like fashion, leading to a progressive emergence of symptoms over time. syn pre-formed fibrils (PFFs) induce LB-like pathology in wild-type (WT) mice, but there are remaining questions about the progressive "spreading" of pathology and the cognitive and behavioral effects. Here, we induced LB-like pathology in the bilateral motor cortex of WT mice and assessed behavioral and cognitive performance. As there are no long-term effective treatments for synucleinopathies, and no therapies slow or reduce the spreading of LBs, we also assessed the effects of a mouse syn-targeted antisense oligonucleotides (ASOs) on pathology and behavioral and cognitive performance starting 5 weeks after ASO treatment. At 3 months post-PFF injection (mpi), mice injected with PFFs showed cognitive impairments and mild motor impairments. At 6 mpi, PFF-injected mice showed further cognitive and motor impairments that were partially ameliorated by the ASO. ASO treatment also reduced LB-like pathology, and pathology was significantly correlated with cognitive measures. However, the particular mouse ASO used in these assays was also associated with some possible off-target effects, defined as effects not involving lowering of syn, such as a decline in body weight. These results add to what is known about the progressive nature of the PFF model of synucleinopathies. These data also support the therapeutic potential of ASOs to improve Lewy pathology and associated behavioral and cognitive phenotypes.
]]></description>
<dc:creator>Boutros, S. J.</dc:creator>
<dc:creator>Raber, J.</dc:creator>
<dc:creator>Unni, V. K.</dc:creator>
<dc:date>2020-10-21</dc:date>
<dc:identifier>doi:10.1101/2020.10.21.349191</dc:identifier>
<dc:title><![CDATA[Antisense oligonucleotides targeting alpha-synuclein reduce pre-formed fibril-induced Lewy pathology and improve some domains of cognitive and motor performance]]></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/10.1101/2020.10.23.352120v1?rss=1">
<title>
<![CDATA[
Incomplete removal of extracellular glutamate controls synaptic transmission and integration at a cerebellar synapse 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.10.23.352120v1?rss=1</link>
<description><![CDATA[
Synapses of glutamatergic mossy fiber onto cerebellar unipolar brush cells (UBCs) generate slow excitatory (ON) or inhibitory (OFF) postsynaptic responses dependent on the complement of glutamate receptors expressed on the UBCs large dendritic brush. Using brain slice recording and computational modeling of synaptic transmission, we found that substantial glutamate is maintained in the UBC synaptic cleft, sufficient to modify spontaneous firing in OFF UBCs and tonically desensitize AMPARs of ON UBC. The source of ambient glutamate was spontaneous, spike-independent exocytosis from the mossy fiber terminal, and its level was dependent on activity of glutamate transporters EAAT1-2. Changing levels of ambient glutamate shifted the polarity of evoked synaptic responses in ON UBCs and altered the phase of responses to in vivo-like synaptic activity. Unlike classical fast synapses, receptors at the UBC synapse are virtually always exposed to a significant level of glutamate, which varies in a graded manner during transmission.
]]></description>
<dc:creator>Balmer, T. S.</dc:creator>
<dc:creator>Borges-Merjane, C.</dc:creator>
<dc:creator>Trussell, L. O.</dc:creator>
<dc:date>2020-10-23</dc:date>
<dc:identifier>doi:10.1101/2020.10.23.352120</dc:identifier>
<dc:title><![CDATA[Incomplete removal of extracellular glutamate controls synaptic transmission and integration at a cerebellar synapse]]></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/10.1101/2020.10.25.354134v1?rss=1">
<title>
<![CDATA[
Diurnal changes in perineuronal nets and parvalbumin neurons in the rat medial prefrontal cortex 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.10.25.354134v1?rss=1</link>
<description><![CDATA[
Perineuronal nets (PNNs) surrounding fast-spiking, parvalbumin (PV) inhibitory interneurons are vital for providing excitatory:inhibitory balance within cortical circuits, and this balance is impaired in disorders such as schizophrenia, autism spectrum disorder, and substance use disorders. These disorders are also associated with altered diurnal rhythms, yet few studies have examined the diurnal rhythms of PNNs or PV cells. We measured the intensity and number of PV cells and PNNs labeled with Wisteria floribunda agglutinin (WFA) in the rat prelimbic medial prefrontal cortex (mPFC) at Zeitgeber times (ZT) ZT0, 6, 12, and 18. We also measured the oxidative stress marker 8-oxo-deoxyguanosine (8-oxo-dG). Relative to ZT0, the intensities of PNN and PV staining were increased in the dark (active) phase compared with the light (inactive) phase. The intensity of 8-oxo-dG was decreased from ZT0 at all time points (ZT6,12,18), in both PV cells and non-PV cells. To examine corresponding changes in inhibitory and excitatory inputs, we measured GAD 65/67 and vGlut1 puncta apposed to PV cells with and without PNNs. Relative to ZT6, there were more excitatory puncta on PV cells surrounded by PNNs at ZT18, but no changes in PV cells devoid of PNNs. No changes in inhibitory puncta were observed. Whole-cell slice recordings in fast-spiking (PV) cells with PNNs showed an increased ratio of -amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptor:N-methyl-D-aspartate receptor (AMPA:NMDA) at ZT18 vs. ZT6. The number of PV cells and co-labeled PV/PNN cells containing the transcription factor orthodenticle homeobox 2 (OTX2), which maintains PNNs, showed a strong trend toward an increase from ZT6 to ZT18. These diurnal fluctuations in PNNs and PV cells are expected to alter cortical excitatory:inhibitory balance and provide new insights into treatment approaches for diseases impacted by imbalances in sleep and circadian rhythms.
]]></description>
<dc:creator>Harkness, J. H.</dc:creator>
<dc:creator>Gonzalez, A. E.</dc:creator>
<dc:creator>Bushana, P. N.</dc:creator>
<dc:creator>Jorgensen, E. T.</dc:creator>
<dc:creator>Hegarty, D. M.</dc:creator>
<dc:creator>Di Nardo, A. A.</dc:creator>
<dc:creator>Prochiantz, A.</dc:creator>
<dc:creator>Wisor, J. P.</dc:creator>
<dc:creator>Aicher, S. A.</dc:creator>
<dc:creator>Brown, T. E.</dc:creator>
<dc:creator>Sorg, B. A.</dc:creator>
<dc:date>2020-10-25</dc:date>
<dc:identifier>doi:10.1101/2020.10.25.354134</dc:identifier>
<dc:title><![CDATA[Diurnal changes in perineuronal nets and parvalbumin neurons in the rat medial prefrontal cortex]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-10-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.10.27.358408v1?rss=1">
<title>
<![CDATA[
Species level resolution of female bladder microbiota from marker gene surveys 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.10.27.358408v1?rss=1</link>
<description><![CDATA[
The human bladder contains bacteria in the absence of infection. Interest in studying these bacteria and their association with bladder conditions is increasing. However, the chosen experimental method can limit the resolution of the taxonomy that can be assigned to the bacteria found in the bladder. 16S rRNA amplicon sequencing is commonly used to identify bacteria in urinary specimens, but is typically restricted to genus-level identification. Our primary aim was to determine if accurate species-level identification of bladder bacteria is possible using 16S rRNA amplicon sequencing. We evaluated the ability of different classification schemes, each consisting of combinations of a reference database, a 16S rRNA gene variable region and a taxonomic classification algorithm to correctly classify bladder bacteria. We show that species-level identification is possible, and that the reference database chosen is the most important component, followed by the 16S variable region sequenced.

ImportanceSpecies-level information may deepen our understanding of associations between bladder microbiota and bladder conditions, such as lower urinary tract symptoms and urinary tract infections. The capability to identify bacterial species depends on large databases of sequences, algorithms that leverage statistics and available computer hardware, and knowledge of bacterial genetics and classification. Taken together, this is a daunting body of knowledge to become familiar with before the simple question of bacterial identity can be answered. Our results show the choice of taxonomic database and variable region of the 16S rRNA gene sequence makes species level identification possible. We also show this improvement can be achieved through the more careful application of existing methods and use of existing resources.
]]></description>
<dc:creator>Hoffman, C.</dc:creator>
<dc:creator>Siddiqui, N. Y.</dc:creator>
<dc:creator>Fields, I.</dc:creator>
<dc:creator>Gregory, W. T.</dc:creator>
<dc:creator>SImon, H.</dc:creator>
<dc:creator>Mooney, M. A.</dc:creator>
<dc:creator>Wolfe, A. J.</dc:creator>
<dc:creator>Karstens, L.</dc:creator>
<dc:date>2020-10-28</dc:date>
<dc:identifier>doi:10.1101/2020.10.27.358408</dc:identifier>
<dc:title><![CDATA[Species level resolution of female bladder microbiota from marker gene surveys]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-10-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.10.30.361949v1?rss=1">
<title>
<![CDATA[
Enhancer of Zeste Homolog 2 (Ezh2) is essential for patterning of multiple musculoskeletal tissues but dispensable for tendon differentiation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.10.30.361949v1?rss=1</link>
<description><![CDATA[
An efficient musculoskeletal system depends on the precise assembly and coordinated growth and function of muscles, skeleton and tendons. However, the mechanisms that drive integrated musculoskeletal development and coordinated growth and differentiation of each of these tissues are still being uncovered. Epigenetic modifiers have emerged as critical regulators of cell fate differentiation, but so far almost nothing is known about their roles in tendon biology. Previous studies have shown that epigenetic modifications driven by Enhancer of zeste homolog 2 (EZH2), a major histone methyltransferase, have significant roles in vertebrate development including skeletal patterning and bone formation. We now find that targeting Ezh2 through the limb mesenchyme also has significant effects on tendon and muscle patterning, likely reflecting the essential roles of early mesenchymal cues mediated by Ezh2 for coordinated patterning and development of all tissues of the musculoskeletal system. Conversely, loss of Ezh2 in the tendon cells did not disrupt the tendon cell fate suggesting that tenocyte differentiation and tendon maturation are independent of Ezh2 signaling.
]]></description>
<dc:creator>Pal, D.</dc:creator>
<dc:creator>Riester, S. M.</dc:creator>
<dc:creator>Hasan, B.</dc:creator>
<dc:creator>Tufa, S. F.</dc:creator>
<dc:creator>Dudakovic, A.</dc:creator>
<dc:creator>Keene, D. R.</dc:creator>
<dc:creator>van Wijnen, A.</dc:creator>
<dc:creator>Schweitzer, R.</dc:creator>
<dc:date>2020-10-30</dc:date>
<dc:identifier>doi:10.1101/2020.10.30.361949</dc:identifier>
<dc:title><![CDATA[Enhancer of Zeste Homolog 2 (Ezh2) is essential for patterning of multiple musculoskeletal tissues but dispensable for tendon differentiation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-10-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.11.01.364232v1?rss=1">
<title>
<![CDATA[
Regulatory diversity contributes to a divergent transcriptional response to dietary changes in mammals 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.11.01.364232v1?rss=1</link>
<description><![CDATA[
BACKGROUNDRegulatory innovation is central to the evolution of species. Different nutritional sources are one environmental pressure that can lead to selection for novel regulatory elements. Dietary composition changes, including exposure to "western" diets with excess fat and sugar content, can lead to transcriptional regulatory changes in the liver. In order to investigate how transcriptional regulatory changes in response to a high fat diet diverge across species, we profiled chromatin accessibility, histone modifications and the transcriptome in livers of rhesus macaques and mice fed high fat and normal diets.

RESULTSWhile the majority of elements exhibiting changes in chromatin accessibility in response to a high fat diet are enriched for similar transcription factors across species, the loci that change are mostly species specific. These unique responsive regulatory elements are largely derived from transposable elements and are enriched for liver-specific transcription factors, such as HNF4. Furthermore, the majority of genes that respond to a high fat diet in rhesus macaques do not have a shared response in mice and are proximal to regulatory elements that display changes in chromatin accessibility only in rhesus macaques.

CONCLUSIONSOur study demonstrates that most of the liver regulatory elements that exhibit changes in chromatin accessibility in response to a high fat diet do so in a species-specific manner. These findings illustrate how a similar environmental stimulus can drive a divergent chromatin and transcriptional responses in evolutionary distinct mammalian species.
]]></description>
<dc:creator>Costello, K. R.</dc:creator>
<dc:creator>Shin, H.</dc:creator>
<dc:creator>Trac, C.</dc:creator>
<dc:creator>Varlamov, O.</dc:creator>
<dc:creator>Schones, D. E.</dc:creator>
<dc:date>2020-11-02</dc:date>
<dc:identifier>doi:10.1101/2020.11.01.364232</dc:identifier>
<dc:title><![CDATA[Regulatory diversity contributes to a divergent transcriptional response to dietary changes in mammals]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-11-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.11.09.373407v1?rss=1">
<title>
<![CDATA[
A Simple and Efficient Pipeline for Construction, Merging, Expansion, and Simulation of Large-Scale, Single-Cell Mechanistic Models 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.11.09.373407v1?rss=1</link>
<description><![CDATA[
Mechanistic models of how single cells respond to different perturbagens can help integrate disparate big data sets or predict response to varied drug combinations. However, the construction and simulation of such models have proved challenging. Our lab previously constructed one of the largest mechanistic models for single mammalian cell regulation of proliferation and death (774 species, 141 genes, 8 ligands, 2400 reactions). However, this, as many other large-scale models, was written using licensed software (MATLAB) with intricate programming structure, impeding alteration, expansion, and sharing. Here, we generated a new foundation for this model, which includes a python-based creation and simulation pipeline converting a few structured text files into an SBML-compatible format. This new open-source model (named SPARCED) is high-performance- and cloud-computing compatible and enables the study of virtual cell population responses at the single-cell level. We applied this new model to a subset of the LINCS MCF10A Data Cube, which observed that IFN{gamma} acts as an anti-proliferative factor, but the reasons why were unknown. After expanding the SPARCED model with an IFN{gamma} signaling module (to 950 species, 150 genes, 9 ligands, 2500 reactions), we ran stochastic single-cell simulations for two different putative crosstalk mechanisms and looked at the number of cycling cells in each case. Our model-based analysis suggested, and experiments support that these observations are better explained by IFN{gamma}-induced SOCS1 expression sequestering activated EGF receptors, thereby downregulating AKT activity, as opposed to direct IFN{gamma}-induced upregulation of p21 expression. This work forms a foundation for increased mechanistic model-based data integration on a single-cell level, an important building block for clinically predictive mechanistic models.
]]></description>
<dc:creator>Erdem, C.</dc:creator>
<dc:creator>Bensman, E. M.</dc:creator>
<dc:creator>Mutsuddy, A.</dc:creator>
<dc:creator>Saint-Antoine, M. M.</dc:creator>
<dc:creator>Bouhaddou, M.</dc:creator>
<dc:creator>Blake, R. C.</dc:creator>
<dc:creator>Dodd, W.</dc:creator>
<dc:creator>Gross, S. M.</dc:creator>
<dc:creator>Heiser, L. M.</dc:creator>
<dc:creator>Feltus, F. A.</dc:creator>
<dc:creator>Birtwistle, M. R.</dc:creator>
<dc:date>2020-11-10</dc:date>
<dc:identifier>doi:10.1101/2020.11.09.373407</dc:identifier>
<dc:title><![CDATA[A Simple and Efficient Pipeline for Construction, Merging, Expansion, and Simulation of Large-Scale, Single-Cell Mechanistic Models]]></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/10.1101/2020.11.09.374496v1?rss=1">
<title>
<![CDATA[
What Markov state models can and cannot do: Correlation versus path-based observables in protein folding models 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.11.09.374496v1?rss=1</link>
<description><![CDATA[
Markov state models (MSMs) have been widely applied to study the kinetics and pathways of protein conformational dynamics based on statistical analysis of molecular dynamics (MD) simulations. These MSMs coarse-grain both configuration space and time in ways that limit what kinds of observables they can reproduce with high fidelity over different spatial and temporal resolutions. Despite their popularity, there is still limited understanding of which biophysical observables can be computed from these MSMs in a robust and unbiased manner, and which suffer from the space-time coarse-graining intrinsic in the MSM model. Most theoretical arguments and practical validity tests for MSMs rely on long-time equilibrium kinetics, such as the slowest relaxation timescales and experimentally observable time-correlation functions. Here, we perform an extensive assessment of the ability of well-validated protein folding MSMs to accuractely reproduce path-based observable such as mean first-passage times (MFPTs) and transition path mechanisms compared to a direct trajectory analysis. We also assess a recently proposed class of history-augmented MSMs (haMSMs) that exploit additional information not accounted for in standard MSMs. We conclude with some practical guidance on the use of MSMs to study various problems in conformational dynamics of biomolecules. In brief, MSMs can accurately reproduce correlation functions slower than the lag time, but path-based observables can only be reliably reproduced if the lifetimes of states exceed the lag time, which is a much stricter requirement. Even in the presence of short-lived states, we find that haMSMs reproduce path-based observables more reliably.
]]></description>
<dc:creator>Suarez, E.</dc:creator>
<dc:creator>Wiewiora, R. P.</dc:creator>
<dc:creator>Wehmeyer, C.</dc:creator>
<dc:creator>Noe, F.</dc:creator>
<dc:creator>Chodera, J. D.</dc:creator>
<dc:creator>Zuckerman, D. M.</dc:creator>
<dc:date>2020-11-09</dc:date>
<dc:identifier>doi:10.1101/2020.11.09.374496</dc:identifier>
<dc:title><![CDATA[What Markov state models can and cannot do: Correlation versus path-based observables in protein folding models]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-11-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.11.11.378505v1?rss=1">
<title>
<![CDATA[
Cell autonomous TGF-beta signaling is essential for cell recruitment into degenerating tendons 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.11.11.378505v1?rss=1</link>
<description><![CDATA[
Understanding the role of cell recruitment in tendon disorders is critical for improvements in regenerative therapy. We recently reported that targeted disruption of TGF{beta} type II receptor in the tendon cell lineage (Tgfbr2ScxCre) resulted in tenocyte dedifferentiation and tendon degradation in post-natal stages. Here we extend the analysis and identify direct recruitment of stem/progenitor cells into the degenerative mutant tendons. Cre-lineage tracing indicates that these cells are not derived from tendon ensheathing tissues or from a Scleraxis-lineage, and they turned on tendon markers only upon entering the mutant tendons. Through immunohistochemistry and inducible gene deletion, we further find that the recruited cells originated from a Sox9-expressing lineage and their recruitment was dependent on cell-autonomous TGF{beta} signaling. These results thus differ from previous reports of cell recruitment into injured tendons, and suggest a critical role for TGF{beta} signaling and cell recruitment in the etiology and treatment of tendon degeneration.
]]></description>
<dc:creator>Tan, G.-K.</dc:creator>
<dc:creator>Pryce, B. A.</dc:creator>
<dc:creator>Stabio, A.</dc:creator>
<dc:creator>Keene, D. R.</dc:creator>
<dc:creator>Tufa, S. F.</dc:creator>
<dc:creator>Schweitzer, R.</dc:creator>
<dc:date>2020-11-11</dc:date>
<dc:identifier>doi:10.1101/2020.11.11.378505</dc:identifier>
<dc:title><![CDATA[Cell autonomous TGF-beta signaling is essential for cell recruitment into degenerating tendons]]></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/10.1101/2020.11.16.385211v1?rss=1">
<title>
<![CDATA[
Adding software to package management systems can increase their citation by 280% 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.11.16.385211v1?rss=1</link>
<description><![CDATA[
A growing number of biomedical methods and protocols are being disseminated as open-source software packages. When put in concert with other packages, they can execute in-depth and comprehensive computational pipelines. Therefore, their integration with other software packages plays a prominent role in their adoption in addition to their availability. Accordingly, package management systems are developed to standardize the discovery and integration of software packages. Here we study the impact of package management systems on software dissemination and their scholarly recognition. We study the citation pattern of more than 18,000 scholarly papers referenced by more than 23,000 software packages hosted by Bioconda, Bioconductor, BioTools, and ToolShed--the package management systems primarily used by the Bioinformatics community. Our results suggest that there is significant evidence that the scholarly papers citation count increases after their respective software was published to package management systems. Additionally, our results show that the impact of different package management systems on the scholarly papers recognition is of the same magnitude. These results may motivate scientists to distribute their software via package management systems, facilitating the composition of computational pipelines and helping reduce redundancy in package development.

Significance StatementSoftware packages are the building blocks of computational pipelines. A myriad of packages are developed; however, the lack of integration and discovery standards hinders their adoption, leaving most scientists scholarly contributions unrecognized. Package management systems are developed to facilitate software dissemination and integration. However, developing software to meet their code and packaging standards is an involved process. Therefore, our study results on the significant impact of the package management systems on scholarly papers recognition can motivate scientists to invest in disseminating their software via package management systems. Dissemination of more software via package management systems will lead to a more straightforward composition of computational pipelines and less redundancy in software packages.
]]></description>
<dc:creator>Jalili, V.</dc:creator>
<dc:creator>Clements, D.</dc:creator>
<dc:creator>Gruning, B.</dc:creator>
<dc:creator>Blankenberg, D.</dc:creator>
<dc:creator>Goecks, J.</dc:creator>
<dc:date>2020-11-17</dc:date>
<dc:identifier>doi:10.1101/2020.11.16.385211</dc:identifier>
<dc:title><![CDATA[Adding software to package management systems can increase their citation by 280%]]></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/10.1101/2020.11.17.387548v1?rss=1">
<title>
<![CDATA[
Human methylome variation across Infinium 450K data on the Gene Expression Omnibus 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.11.17.387548v1?rss=1</link>
<description><![CDATA[
While DNA methylation (DNAm) is the most-studied epigenetic mark, few recent studies probe the breadth of publicly available DNAm array samples. We collectively analyzed 35,360 Illumina Infinium HumanMethylation450K DNAm array samples published on the Gene Expression Omnibus (GEO). We learned a controlled vocabulary of sample labels by applying regular expressions to metadata and used existing models to predict various sample properties including epigenetic age. We found approximately two-thirds of samples were from blood, one-quarter were from brain, and one-third were from cancer patients. 19% of samples failed at least one of Illuminas 17 prescribed quality assessments; signal distributions across samples suggest modifying manufacturer-recommended thresholds for failure would make these assessments more informative. We further analyzed DNAm variances in seven tissues (adipose, nasal, blood, brain, buccal, sperm, and liver) and characterized specific probes distinguishing them. Finally, we compiled DNAm array data and metadata, including our learned and predicted sample labels, into database files accessible via the recountmethylation R/Bioconductor companion package. Its vignettes walk the user through some analyses contained in this paper.
]]></description>
<dc:creator>Maden, S. K.</dc:creator>
<dc:creator>Thompson, R. F.</dc:creator>
<dc:creator>Hansen, K. D.</dc:creator>
<dc:creator>Nellore, A.</dc:creator>
<dc:date>2020-11-19</dc:date>
<dc:identifier>doi:10.1101/2020.11.17.387548</dc:identifier>
<dc:title><![CDATA[Human methylome variation across Infinium 450K data on the Gene Expression Omnibus]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-11-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.11.19.390633v1?rss=1">
<title>
<![CDATA[
Cancer cells are sensitive to wild-type IDH1 inhibition under nutrient limitation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.11.19.390633v1?rss=1</link>
<description><![CDATA[
Pancreatic cancer cells alter their metabolism to survive cancer-associated stress (1-4). For example, cancer cells must adapt to steep nutrient gradients that characterize the natural tumor microenvironment (TME) (5-7). In the absence of adaptive strategies, harsh metabolic conditions promote the generation of free radicals (8) and impair energy production in tumor cells. Towards this end, wild-type isocitrate dehydrogenase 1 (IDH1) activity is a metabolic requirement for cancer cells living in a harsh metabolic milieu. The cytosolic enzyme interconverts isocitrate and alpha-ketoglutarate, and uses NADP(H) as a cofactor. We show that under low nutrient conditions, the enzymatic reaction favors oxidative decarboxylation to yield NADPH and alpha-ketoglutarate. Metabolic studies showed that the IDH1 products directly support antioxidant defense and mitochondrial function in stressed cancer cells. Genetic IDH1 suppression reduced growth of pancreatic cancer cells in vitro under low nutrient conditions and in mouse models of pancreatic cancer. Surprisingly, allosteric inhibitors of mutant IDH1 proved to be potent wild-type IDH1 inhibitors under conditions specific to the TME, highlighting a natural therapeutic window. The presence of low magnesium enhanced allosteric inhibition by the drug, and ambient low glucose levels enhanced cancer cells dependence on wild-type IDH1. Thus, intrinsic TME conditions sensitized wild-type IDH1 to FDA-approved AG-120 (ivosidenib), and revealed the drug to be a potent single-agent therapeutic in cell culture and diverse in vivo cancer models. This work identified a potentially new repertoire of safe cancer therapies, including a clinically available compound, for the treatment of multiple wild-type IDH1 cancers (e.g., pancreatic).
]]></description>
<dc:creator>Vaziri-Gohar, A.</dc:creator>
<dc:creator>Hue, J. J.</dc:creator>
<dc:creator>Graor, H. J.</dc:creator>
<dc:creator>Prendergast, E.</dc:creator>
<dc:creator>Chen, V.</dc:creator>
<dc:creator>Cassel, J.</dc:creator>
<dc:creator>Mohammed, F. S.</dc:creator>
<dc:creator>Abbas, A.</dc:creator>
<dc:creator>Dukleska, K.</dc:creator>
<dc:creator>Khokhar, I.</dc:creator>
<dc:creator>Hajhassani, O.</dc:creator>
<dc:creator>Zarei, M.</dc:creator>
<dc:creator>Wang, R.</dc:creator>
<dc:creator>Rothermel, L. D.</dc:creator>
<dc:creator>Bederman, I.</dc:creator>
<dc:creator>Browers, J.</dc:creator>
<dc:creator>Getts, R.</dc:creator>
<dc:creator>Brunengraber, H.</dc:creator>
<dc:creator>Salvino, J. M.</dc:creator>
<dc:creator>Brody, J. R.</dc:creator>
<dc:creator>Winter, J. M.</dc:creator>
<dc:date>2020-11-20</dc:date>
<dc:identifier>doi:10.1101/2020.11.19.390633</dc:identifier>
<dc:title><![CDATA[Cancer cells are sensitive to wild-type IDH1 inhibition under nutrient limitation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-11-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.11.25.397059v1?rss=1">
<title>
<![CDATA[
AKAP79/150 coordinates leptin-induced PKA activation to regulate KATP channel trafficking in pancreatic β-cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.11.25.397059v1?rss=1</link>
<description><![CDATA[
The adipocyte hormone leptin regulates glucose homeostasis both centrally and peripherally. A key peripheral target is the pancreatic {beta}-cell, which secretes insulin upon glucose stimulation. Leptin suppresses glucose-stimulated insulin secretion by promoting trafficking of KATP channels to the {beta}-cell surface, which increases K+ conductance and causes {beta}-cell hyperpolarization. Here we investigate the signaling mechanism underlying leptin-induced KATP channel translocation with a focus on protein kinase A (PKA). Using FRET-based PKA activity reporters, we show that leptin increases PKA activity at the cell membrane via a signaling pathway involving NMDA receptors, CaMKK{beta} and AMPK. Genetic knockdown and rescue experiments reveal that leptin activation of PKA requires tethering of PKA to the membrane-targeted PKA-anchoring protein AKAP79/150. Interestingly, disrupting protein phosphatase 2B (PP2B) anchoring to AKAP79/150, known to elevate basal PKA signaling, increases surface KATP channels. Our findings uncover a novel role of AKAP79/150 in coordinating leptin and PKA signaling to regulate {beta}-cell function.
]]></description>
<dc:creator>Cochrane, V.</dc:creator>
<dc:creator>Yang, Z.</dc:creator>
<dc:creator>Dell'Acqua, M.</dc:creator>
<dc:creator>Shyng, S.-L.</dc:creator>
<dc:date>2020-11-26</dc:date>
<dc:identifier>doi:10.1101/2020.11.25.397059</dc:identifier>
<dc:title><![CDATA[AKAP79/150 coordinates leptin-induced PKA activation to regulate KATP channel trafficking in pancreatic β-cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-11-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.11.25.399006v1?rss=1">
<title>
<![CDATA[
Eosinophil peroxidase induces inflammation in an animal model of dermatitis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.11.25.399006v1?rss=1</link>
<description><![CDATA[
Eosinophils play an important role in mediating itch and inflammation in dermatitis. The role of the eosinophil granule protein eosinophil peroxidase (EPX) in mediating inflammation and itch was tested in a dermatitis mouse model. Mice were sensitized to trimellitic anhydride (TMA) and subsequently challenged chronically on the ear to establish dermatitis. Loss of EPX (in EPX (-/-) mice) or blocking EPX with the drug resorcinol significantly reduced dermatitis in mice exposed to TMA. Resorcinol also reduced levels of thymic stromal lymphopoietin protein (TSLP) in skin. Further studies showed that EPX increased different cytokines in keratinocytes in cell culture via two distinct mechanisms. EPX induced TSLP expression requires lysophosphatidic acid signaling while EPX induced expression of TNF-, CSF2, CSF3, and IL1 required IL-1 signaling. We also showed that blocking IL-1 reduced inflammation in skin following TMA exposure in mice. Thus, EPX is an important mediator of inflammation and itch, that are mediated via at least two pathways. This suggests that both EPX and its signaling pathways may provide novel therapeutic strategies in dermatitis.
]]></description>
<dc:creator>Roth-Carter, Q.</dc:creator>
<dc:creator>Luo, H.</dc:creator>
<dc:creator>Ochkur, S. I.</dc:creator>
<dc:creator>Jacobsen, E.</dc:creator>
<dc:creator>Datena, M.</dc:creator>
<dc:creator>Fryer, A. D.</dc:creator>
<dc:creator>Lee, J. J.</dc:creator>
<dc:creator>Jacoby, D. B.</dc:creator>
<dc:date>2020-11-27</dc:date>
<dc:identifier>doi:10.1101/2020.11.25.399006</dc:identifier>
<dc:title><![CDATA[Eosinophil peroxidase induces inflammation in an animal model of dermatitis]]></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/10.1101/2020.11.30.405670v1?rss=1">
<title>
<![CDATA[
African-lineage Zika virus replication dynamics and maternal-fetal interface infection in pregnant rhesus macaques 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.11.30.405670v1?rss=1</link>
<description><![CDATA[
Following the Zika virus (ZIKV) outbreak in the Americas, ZIKV was causally associated with microcephaly and a range of neurological and developmental symptoms, termed congenital Zika syndrome (CZS). The isolates responsible for this outbreak belonged to the Asian lineage of ZIKV. However, in-vitro and in-vivo studies assessing the pathogenesis of African-lineage ZIKV demonstrated that African-lineage isolates often replicated to high titer and caused more severe pathology than Asian-lineage isolates. To date, the pathogenesis of African-lineage ZIKV in a translational model, particularly during pregnancy, has not been rigorously characterized. Here we infected four pregnant rhesus macaques with a low-passage strain of African-lineage ZIKV and compared its pathogenesis to a cohort of four pregnant rhesus macaques infected with an Asian-lineage isolate and a cohort of mock-infected controls. Viral replication kinetics were not significantly different between the two experimental groups and both groups developed robust neutralizing antibody titers above levels considered to be protective. There was no evidence of significant fetal head growth restriction or gross fetal harm at delivery in either group. However, a significantly higher burden of ZIKV vRNA was found in maternal-fetal interface tissues in the macaques exposed to an African-lineage isolate. Our findings suggest that ZIKV isolates of any genetic lineage pose a threat to women and their infants.

IMPORTANCEZIKV was first identified over 70 years ago in Africa, but most of our knowledge of ZIKV is based on studies of the distinct Asian genetic lineage, which caused the outbreak in the Americas in 2015-16. In its most recent update, the WHO stated that improved understanding of African-lineage pathogenesis during pregnancy must be a priority. Recent detection of African-lineage isolates in Brazil underscores the need to understand the impact of these viruses. Here we provide the first comprehensive assessment of African-lineage ZIKV infection during pregnancy in a translational non-human primate model. We show African-lineage isolates replicate with similar kinetics to Asian-lineage isolates and are capable of infecting the placenta. However, there was no evidence of more severe outcomes with African-lineage isolates. Our results highlight both the threat that African-lineage ZIKV poses to women and their infants and the need for future epidemiological and translational in-vivo studies with African-lineage ZIKV.
]]></description>
<dc:creator>Crooks, C. M.</dc:creator>
<dc:creator>Weiler, A. M.</dc:creator>
<dc:creator>Rybarczyk, S. L.</dc:creator>
<dc:creator>Bliss, M. I.</dc:creator>
<dc:creator>Jaeger, A. S.</dc:creator>
<dc:creator>Murphy, M. E.</dc:creator>
<dc:creator>Simmons, H. A.</dc:creator>
<dc:creator>Mejia, A.</dc:creator>
<dc:creator>Fritsch, M. K.</dc:creator>
<dc:creator>Hayes, J. M.</dc:creator>
<dc:creator>Eickhoff, J. C.</dc:creator>
<dc:creator>Mitzey, A. M.</dc:creator>
<dc:creator>Razo, E.</dc:creator>
<dc:creator>Braun, K.</dc:creator>
<dc:creator>Brown, E. A.</dc:creator>
<dc:creator>Yamamoto, K.</dc:creator>
<dc:creator>Shepard, P. M.</dc:creator>
<dc:creator>Possell, A.</dc:creator>
<dc:creator>Weaver, K.</dc:creator>
<dc:creator>Antony, K. M.</dc:creator>
<dc:creator>Morgan, T. K.</dc:creator>
<dc:creator>Dudley, D.</dc:creator>
<dc:creator>Peterson, E.</dc:creator>
<dc:creator>Schultz-Darken, N.</dc:creator>
<dc:creator>O'Connor, D. H.</dc:creator>
<dc:creator>Mohr, E. L.</dc:creator>
<dc:creator>Golos, T. G.</dc:creator>
<dc:creator>Aliota, M.</dc:creator>
<dc:creator>Friedrich, T. C.</dc:creator>
<dc:date>2020-12-01</dc:date>
<dc:identifier>doi:10.1101/2020.11.30.405670</dc:identifier>
<dc:title><![CDATA[African-lineage Zika virus replication dynamics and maternal-fetal interface infection in pregnant rhesus macaques]]></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/10.1101/2020.12.01.406066v1?rss=1">
<title>
<![CDATA[
HLA-E restricted, HIV-1 suppressing, Gag specific CD8+ T cells offer universal vaccine opportunities 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.12.01.406066v1?rss=1</link>
<description><![CDATA[
Human leukocyte antigen-E (HLA-E) normally presents a HLA class Ia signal peptide to the NKG2A/C-CD94 regulatory receptors on natural killer (NK) cells and T cell subsets. Rhesus macaques immunized with a cytomegalovirus vectored simian immunodeficiency virus (SIV) vaccine, generated Mamu-E (HLA-E homolog) restricted T cell responses that mediated post-challenge SIV replication arrest in >50% of animals. However, human immunodeficiency virus type 1 (HIV-1) specific HLA-E restricted T cells have not been observed in HIV-1-infected individuals. Here we primed HLA-E restricted HIV-1 specific CD8+ T cells in vitro. These T cell clones, and allogeneic CD8+ T cells transduced with their T cell receptors, suppressed HIV-1 replication in CD4+ T cells in vitro. Vaccine induction of efficacious HLA-E restricted HIV-1 specific T cells should therefore be possible.

One Sentence SummaryCD8+ T cells that recognize a Gag peptide presented by HLA-E suppress HIV-1 replication in vitro.
]]></description>
<dc:creator>Yang, H.</dc:creator>
<dc:creator>Rei, M.</dc:creator>
<dc:creator>Brackenridge, S.</dc:creator>
<dc:creator>Brenna, E.</dc:creator>
<dc:creator>Sun, H.</dc:creator>
<dc:creator>abdulhaqq, s.</dc:creator>
<dc:creator>Liu, M. K.</dc:creator>
<dc:creator>Ma, W.</dc:creator>
<dc:creator>Kurupati, P.</dc:creator>
<dc:creator>Xu, x.</dc:creator>
<dc:creator>Cerundolo, V.</dc:creator>
<dc:creator>Jenkins, E.</dc:creator>
<dc:creator>Davis, S. J.</dc:creator>
<dc:creator>Sacha, J. B.</dc:creator>
<dc:creator>Frueh, K.</dc:creator>
<dc:creator>Picker, L. J.</dc:creator>
<dc:creator>Borrow, P.</dc:creator>
<dc:creator>Gillespie, G.</dc:creator>
<dc:creator>McMichael, A. J.</dc:creator>
<dc:date>2020-12-02</dc:date>
<dc:identifier>doi:10.1101/2020.12.01.406066</dc:identifier>
<dc:title><![CDATA[HLA-E restricted, HIV-1 suppressing, Gag specific CD8+ T cells offer universal vaccine opportunities]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-12-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.12.02.407601v1?rss=1">
<title>
<![CDATA[
Illuminating endosomal escape of polymorphic lipid nanoparticles that boost mRNA delivery 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.12.02.407601v1?rss=1</link>
<description><![CDATA[
Lipid-based nanoparticles (LNPs) for the delivery of mRNA have jumped to the forefront of non-viral gene delivery. Despite this exciting development, poor endosomal escape after LNP cell entry remains an unsolved, rate-limiting bottleneck. Here we report the use of galectin 8-GFP (Gal8-GFP) cell reporter system to visualize the endosomal escaping capabilities of LNPs encapsulating mRNA. LNPs substituted with phytosterols in place of cholesterol exhibited various levels of Gal8 recruitment in the Gal8-GFP reporter system. In live-cell imaging, the results of the LNPs containing {beta}-sitosterol (LNP-Sito) showed a 10-fold increase in detectable endosomal perturbation events when compared to the standard cholesterol LNPs (LNP-Chol), suggesting the superior capability of LNP-Sito to escape from endosomal entrapment. Trafficking studies of these LNPs showed strong localization with late endosomes. This highly sensitive and robust Gal8-GFP reporter system can be a valuable tool to elucidate intricacies of LNP trafficking and ephemeral endosomal escape events, enabling the advancement in gene delivery.
]]></description>
<dc:creator>Herrera, M.</dc:creator>
<dc:creator>Kim, J.</dc:creator>
<dc:creator>Eygeris, Y.</dc:creator>
<dc:creator>Jozic, A.</dc:creator>
<dc:creator>Sahay, G.</dc:creator>
<dc:date>2020-12-03</dc:date>
<dc:identifier>doi:10.1101/2020.12.02.407601</dc:identifier>
<dc:title><![CDATA[Illuminating endosomal escape of polymorphic lipid nanoparticles that boost mRNA delivery]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-12-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.12.02.409185v1?rss=1">
<title>
<![CDATA[
Functional and genomic adaptations of blood monocytes to pregravid obesity during pregnancy 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.12.02.409185v1?rss=1</link>
<description><![CDATA[
Pre-pregnancy obesity is associated with several adverse maternal health outcomes, notably increased risk of infection as well as the incidence of gestational diabetes, preeclampsia, and preterm birth. However, the mechanisms by which pregravid obesity disrupts the pregnancy associated "immune clock" are still unknown. To address this question, we collected blood samples from women during the first and third trimesters and determined the impact of both pregnancy and pregravid obesity on circulating immune mediators, immune cell subset frequencies, and peripheral immune responses. While regardless of BMI, pregnancy was associated with an elevation in both Th1 and Th2 cytokines, pregravid obesity was associated with a dysregulation in circulating myeloid factors at term. Moreover, pregnancy in lean subjects was associated with enhanced monocyte activation, augmented chromatin accessibility at inflammatory loci, and heightened responses to LPS. Pregravid obesity disrupted this trajectory and was accompanied by a lack of transcriptional and epigenetic changes and alterations in metabolic status strongly suggesting a skewing towards immunotolerance. These findings provide novel insight into the increased susceptibility to infections observed with obesity during pregnancy.

SUMMARYA healthy pregnancy is associated with progressive innate immune activation. Maternal factors such as obesity compromise this myeloid cell activation trajectory at genomic, epigenomic, functional, and metabolic levels, resulting in stagnant immune responses, suggestive of a state of tolerance.
]]></description>
<dc:creator>Messaoudi, I.</dc:creator>
<dc:creator>Sureshchandra, S.</dc:creator>
<dc:creator>Marshall, N. E.</dc:creator>
<dc:creator>Mendoza, N.</dc:creator>
<dc:creator>Jankeel, A.</dc:creator>
<dc:creator>Zulu, M. Z.</dc:creator>
<dc:date>2020-12-03</dc:date>
<dc:identifier>doi:10.1101/2020.12.02.409185</dc:identifier>
<dc:title><![CDATA[Functional and genomic adaptations of blood monocytes to pregravid obesity during pregnancy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-12-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.12.03.393306v1?rss=1">
<title>
<![CDATA[
Multiple viral microRNAs regulate interferon release and signaling early during infection with Epstein-Barr virus 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.12.03.393306v1?rss=1</link>
<description><![CDATA[
Epstein-Barr virus (EBV), a human herpes virus, encodes 44 microRNAs (miRNAs), which regulate many genes with various functions in EBV-infected cells. Multiple target genes of the EBV miRNAs have been identified, some of which play important roles in adaptive antiviral immune responses. Using EBV mutant derivatives, we identified additional roles of viral miRNAs in governing versatile type I interferon (IFN) responses upon infection of human primary mature B cells. We also found that Epstein-Barr virus-encoded small RNAs (EBERs) and LF2, viral genes with previously reported functions in inducing or regulating IFN-I pathways, had negligible or even contrary effects on secreted IFN- in our model. Data mining and Ago PAR-CLIP experiments uncovered more than a dozen of previously uncharacterized, direct cellular targets of EBV miRNA associated with type I IFN pathways. We also identified indirect targets of EBV miRNAs in B cells, such as TRL7 and TLR9, in the pre-latent phase of infection. The presence of epigenetically naive, non-CpG methylated viral DNA was essential to induce IFN- secretion during EBV infection in a TLR9-dependent manner. In a newly established fusion assay, we verified that EBV virions enter a subset of plasmacytoid dendritic cells (pDCs) and determined that these infected pDCs are the primary producers of IFN- in EBV-infected peripheral blood mononuclear cells. Our findings document that many EBV-encoded miRNAs regulate type I IFN response in newly EBV infected primary human B cells in the pre-latent phase of infection and dampen the acute release of IFN- in pDCs upon their encounter with EBV.

Author summaryAcute antiviral functions of all nucleated cells rely on type I interferon (IFN-I) pathways triggered upon viral infection. Host responses encompass the sensing of incoming viruses, the activation of specific transcription factors which induce transcription of IFN-I genes, the secretion of different IFN-I types and their recognition by the heterodimeric IFN-/{beta} receptor, the subsequent activation of JAK/STAT signaling pathways and, finally, the transcription of many IFN-stimulated genes (ISGs). In sum, these cellular functions establish a so-called antiviral state in infected and neighboring cells. To counteract these cellular defense mechanisms, viruses have evolved diverse strategies and encode gene products that target antiviral responses. Among such immune evasive factors are viral microRNAs (miRNAs) that can interfere with host gene expression. We discovered that multiple miRNAs encoded by Epstein-Barr virus (EBV) control over a dozen cellular genes that contribute to the anti-viral states of immune cells, specifically B cells and plasmacytoid dendritic cells (pDCs). We identified the viral DNA genome as the activator of IFN- and question the role of abundant EBV EBERs, that, contrary to previous reports, do not have an apparent inducing function in the IFN-I pathway early after infection.
]]></description>
<dc:creator>Bouvet, M.</dc:creator>
<dc:creator>Voigt, S.</dc:creator>
<dc:creator>Tagawa, T.</dc:creator>
<dc:creator>Albanese, M.</dc:creator>
<dc:creator>Chen, Y.-F. A.</dc:creator>
<dc:creator>Chen, Y.</dc:creator>
<dc:creator>Fachko, D. N.</dc:creator>
<dc:creator>Pich, D.</dc:creator>
<dc:creator>Goebel, C.</dc:creator>
<dc:creator>Skalsky, R.</dc:creator>
<dc:creator>Hammerschmidt, W.</dc:creator>
<dc:date>2020-12-03</dc:date>
<dc:identifier>doi:10.1101/2020.12.03.393306</dc:identifier>
<dc:title><![CDATA[Multiple viral microRNAs regulate interferon release and signaling early during infection with Epstein-Barr virus]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-12-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.12.03.408500v1?rss=1">
<title>
<![CDATA[
An Integrated Clinical, Omic, and Image Atlas of an Evolving Metastatic Breast Cancer 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.12.03.408500v1?rss=1</link>
<description><![CDATA[
Mechanisms of therapeutic resistance manifest in metastatic cancers as tumor cell intrinsic alterations and extrinsic microenvironmental influences that can change during treatment. To support the development of methods for the identification of these mechanisms in individual patients, we present here an Omic and Multidimensional Spatial (OMS) Atlas generated from four serial biopsies of a metastatic breast cancer patient during 3.5 years of therapy. This resource links detailed, longitudinal clinical metadata including treatment times and doses, anatomic imaging, and blood-based response measurements to exploratory analytics including comprehensive DNA, RNA, and protein profiles, images of multiplexed immunostaining, and 2- and 3-dimensional scanning electron micrographs. These data reveal aspects of therapy-associated heterogeneity and evolution of the cancers genome, signaling pathways, immune microenvironment, cellular composition and organization, and ultrastructure. We present illustrative examples showing how integrative analyses of these data provide insights into potential mechanisms of response and resistance, and suggest novel therapeutic vulnerabilities.
]]></description>
<dc:creator>Johnson, B. E.</dc:creator>
<dc:creator>Creason, A. L.</dc:creator>
<dc:creator>Stommel, J. M.</dc:creator>
<dc:creator>Keck, J.</dc:creator>
<dc:creator>Parmar, S.</dc:creator>
<dc:creator>Betts, C. B.</dc:creator>
<dc:creator>Blucher, A.</dc:creator>
<dc:creator>Boniface, C.</dc:creator>
<dc:creator>Bucher, E.</dc:creator>
<dc:creator>Burlingame, E. A.</dc:creator>
<dc:creator>Chin, K.</dc:creator>
<dc:creator>Eng, J.</dc:creator>
<dc:creator>Feiler, H. S.</dc:creator>
<dc:creator>Kolodzie, A.</dc:creator>
<dc:creator>Kong, B.</dc:creator>
<dc:creator>Labrie, M.</dc:creator>
<dc:creator>Leyshock, P.</dc:creator>
<dc:creator>Mitri, S.</dc:creator>
<dc:creator>Patterson, J.</dc:creator>
<dc:creator>Riesterer, J. L.</dc:creator>
<dc:creator>Sivagnanam, S.</dc:creator>
<dc:creator>Sudar, D.</dc:creator>
<dc:creator>Thibault, G.</dc:creator>
<dc:creator>Zheng, C.</dc:creator>
<dc:creator>Nan, X.</dc:creator>
<dc:creator>Heiser, L. M.</dc:creator>
<dc:creator>Spellman, P. T.</dc:creator>
<dc:creator>Thomas, G. V.</dc:creator>
<dc:creator>Demir, E.</dc:creator>
<dc:creator>Chang, Y. H.</dc:creator>
<dc:creator>Coussens, L. M.</dc:creator>
<dc:creator>Guimaraes, A. R.</dc:creator>
<dc:creator>Corless, C.</dc:creator>
<dc:creator>Goecks, J.</dc:creator>
<dc:creator>Bergan, R.</dc:creator>
<dc:creator>Mitri, Z.</dc:creator>
<dc:creator>Mills, G. B.</dc:creator>
<dc:creator>Gray, J. W.</dc:creator>
<dc:date>2020-12-03</dc:date>
<dc:identifier>doi:10.1101/2020.12.03.408500</dc:identifier>
<dc:title><![CDATA[An Integrated Clinical, Omic, and Image Atlas of an Evolving Metastatic Breast Cancer]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-12-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.12.03.409417v1?rss=1">
<title>
<![CDATA[
High-fidelity, efficient, and reversible labeling of endogenous proteins using CRISPR-based designer exon insertion 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.12.03.409417v1?rss=1</link>
<description><![CDATA[
Precise and efficient insertion of large DNA fragments into somatic cells using gene editing technologies to label or modify endogenous proteins remains challenging. Non-specific insertions/deletions (INDELs) resulting from the non-homologous end joining pathway make the process error-prone. Further, the insert is not readily removable. Here, we describe a method called CRISPR-mediated insertion of exon (CRISPIE) that can precisely and reversibly label endogenous proteins using CRISPR/Cas9-based editing. CRISPIE inserts a designer donor module, which consists of an exon encoding the protein sequence flanked by intron sequences, into an intronic location in the target gene. INDELs at the insertion junction will be spliced out, leaving mRNAs nearly error-free. We used CRISPIE to fluorescently label endogenous proteins in neurons in vivo with previously unachieved efficiency. We demonstrate that this method is broadly applicable, and that the insert can be readily removed later. CRISPIE permits protein sequence insertion with high fidelity, efficiency, and flexibility.
]]></description>
<dc:creator>Zhong, H.</dc:creator>
<dc:creator>Massengill, C. I.</dc:creator>
<dc:creator>Muniak, M. A.</dc:creator>
<dc:creator>Ma, L.</dc:creator>
<dc:creator>Qin, M.</dc:creator>
<dc:creator>Petrie, S. K.</dc:creator>
<dc:creator>Mao, T.</dc:creator>
<dc:date>2020-12-03</dc:date>
<dc:identifier>doi:10.1101/2020.12.03.409417</dc:identifier>
<dc:title><![CDATA[High-fidelity, efficient, and reversible labeling of endogenous proteins using CRISPR-based designer exon insertion]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-12-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.12.04.410092v1?rss=1">
<title>
<![CDATA[
Ribavirin shows antiviral activity against SARS-CoV-2 and downregulates the activity of TMPRSS2 and the expression of ACE2 in vitro 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.12.04.410092v1?rss=1</link>
<description><![CDATA[
Ribavirin is a guanosine analog and has a broad-spectrum antiviral activity against RNA viruses. Based on this, we aimed to show the anti-SARS-CoV-2 activity of this drug molecule via in vitro, in silico and molecular techniques. Ribavirin showed antiviral activity in Vero E6 cells following SARS-CoV-2 infection. In silico analysis suggested that Ribarivin has a broad-spectrum impact on Vero E6 cells. According to the detailed molecular techniques, Ribavirin was shown to decrease TMPRSS2 expression both at mRNA and protein level 48 hours after treatment. The suppressive effect of Ribavirin in ACE2 protein expression was shown to be dependent on cell types. Finally, proteolytic activity assays showed that Ribavirin also showed an inhibitory effect on TMPRSS2 enzyme. As a conclusion, Ribavirin is a potential antiviral drug for the treatment against SARS-CoV-2, and it interferes with the effect of TMPRSS2 and ACE2 expression.
]]></description>
<dc:creator>Unal, M. A.</dc:creator>
<dc:creator>Bitirim, C. V.</dc:creator>
<dc:creator>Summak, G. Y.</dc:creator>
<dc:creator>Bereketoglu, S.</dc:creator>
<dc:creator>Cevher Zeytin, I.</dc:creator>
<dc:creator>Besbinar, O.</dc:creator>
<dc:creator>Gurcan, C.</dc:creator>
<dc:creator>Aydos, D.</dc:creator>
<dc:creator>Goksoy, E.</dc:creator>
<dc:creator>Kocakaya, E.</dc:creator>
<dc:creator>Eran, Z.</dc:creator>
<dc:creator>Murat, M.</dc:creator>
<dc:creator>Demir, N.</dc:creator>
<dc:creator>Somers, J.</dc:creator>
<dc:creator>Demir, E.</dc:creator>
<dc:creator>Nazir, H.</dc:creator>
<dc:creator>Ozkan, S. A.</dc:creator>
<dc:creator>Ozkul, A.</dc:creator>
<dc:creator>Azap, A.</dc:creator>
<dc:creator>Yilmazer, A.</dc:creator>
<dc:creator>Akcali, K. C.</dc:creator>
<dc:date>2020-12-05</dc:date>
<dc:identifier>doi:10.1101/2020.12.04.410092</dc:identifier>
<dc:title><![CDATA[Ribavirin shows antiviral activity against SARS-CoV-2 and downregulates the activity of TMPRSS2 and the expression of ACE2 in vitro]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-12-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.12.10.420331v1?rss=1">
<title>
<![CDATA[
A pragmatic reevaluation of the efficacy of nonhuman primate optogenetics 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.12.10.420331v1?rss=1</link>
<description><![CDATA[
Translational neuroscience is committed to generating discoveries in the laboratory that ultimately can improve human lives. Optogenetics has received considerable attention because of its demonstrated promise in rodent brains to manipulate cells and circuits. In a recent report, Tremblay and colleagues (2020) introduce an open resource detailing optogenetic studies of the nonhuman primate (NHP) brain and make robust claims about the translatability of the technology. We propose that their quantitative (e.g., a 91% success rate) and theoretical claims are questionable because the data were analyzed at a level relevant to the rodent but not NHP brain, injections were clustered within a few monkeys in a few studies in a few brain regions, and their definitions of success was not clearly relevant to human neuropsychiatric disease. A reanalysis of the data with a modified definition of success that included a behavioral and biological effect revealed an 62.5% success rate that was lower when considering only strong outcomes (53.1%). This calls into question the current efficacy of optogenetic techniques in the NHP brain and suggests that we are a long way from being able to leverage them in "the service of patients with neurological or psychiatric conditions" as the Tremblay report claims.
]]></description>
<dc:creator>Bliss-Moreau, E.</dc:creator>
<dc:creator>Costa, V. D.</dc:creator>
<dc:creator>Baxter, M. G.</dc:creator>
<dc:date>2020-12-11</dc:date>
<dc:identifier>doi:10.1101/2020.12.10.420331</dc:identifier>
<dc:title><![CDATA[A pragmatic reevaluation of the efficacy of nonhuman primate optogenetics]]></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/10.1101/2020.12.11.422048v1?rss=1">
<title>
<![CDATA[
Toward reproducible, scalable, and robust data analysis across multiplex tissue imaging platforms 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.12.11.422048v1?rss=1</link>
<description><![CDATA[
The emergence of megascale single-cell multiplex tissue imaging (MTI) datasets necessitates reproducible, scalable, and robust tools for cell phenotyping and spatial analysis. We developed open-source, graphics processing unit (GPU)-accelerated tools for intensity normalization, phenotyping, and microenvironment characterization. We deploy the toolkit on a human breast cancer (BC) tissue microarray stained by cyclic immunofluorescence and benchmark our cell phenotypes against a published MTI dataset. Finally, we demonstrate an integrative analysis revealing BC subtype-specific features.
]]></description>
<dc:creator>Burlingame, E. A.</dc:creator>
<dc:creator>Eng, J.</dc:creator>
<dc:creator>Thibault, G.</dc:creator>
<dc:creator>Chin, K.</dc:creator>
<dc:creator>Gray, J. W.</dc:creator>
<dc:creator>Chang, Y. H.</dc:creator>
<dc:date>2020-12-11</dc:date>
<dc:identifier>doi:10.1101/2020.12.11.422048</dc:identifier>
<dc:title><![CDATA[Toward reproducible, scalable, and robust data analysis across multiplex tissue imaging platforms]]></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/10.1101/2020.12.14.422805v1?rss=1">
<title>
<![CDATA[
Longitudinal profiling of the macaque vaginal microbiome reveals similarities to diverse human vaginal communities: implications for use as a pre-clinical model for bacterial vaginosis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.12.14.422805v1?rss=1</link>
<description><![CDATA[
The vaginal microbiota plays an important role in womens reproductive and urogenital health. Disturbances in this microbial community can lead to several adverse outcomes including pelvic inflammatory disease, bacterial vaginosis (BV) as well as increased susceptibility to sexually transmitted infections, miscarriage, and pre-term births. It is now well accepted that while the microbiome of healthy women in the developed world is dominated by Lactobacillus species, vaginal communities in asymptomatic women, especially those in the developing world, can be comprised of a diverse set of micro-organisms. The presence of a diverse vaginal microbiome has been associated with increased susceptibility to HIV infection but their implications for womens health remain poorly understood. Rhesus macaques are an excellent translational animal model due to significant physiological and genetic homology with humans. In this study, we performed a longitudinal analysis of clinical and microbiome data from 16 reproductive age female rhesus macaques. Many animals showed hallmarks of BV, including Nugent scores above 7 and high vaginal pH. At both the taxonomic and functional level, the rhesus macaque vaginal microbiome was most similar to that of women who harbor a diverse vaginal community associated with asymptomatic/symptomatic bacterial vaginosis. Specifically, rhesus macaque vaginal microbiomes harbored a diverse set of anaerobic gram-negative bacteria, including; Snethia, Prevotella, Porphyromonas, and Mobilluncus. Interestingly, some animals were transiently colonized by Lactobacillus and some with Gardnerella. Our in-depth and comprehensive analysis highlights the importance of the model to test interventions for manipulating the vaginal microbiome.

IMPORTANCEIt is widely accepted that the "healthy" vaginal microbiome of the majority of women in the developed world is dominated by Lactobacillus species. However, in the developing world, a majority of women are colonized by diverse microbial communities, typically associated with bacterial vaginosis, but remain asymptomatic. Many questions remain about the drivers of this disparity and potential interventions to alter the vaginal microbiome. Rhesus macaques provide an excellent translational model due to significant physiological and genetic homology with humans. In this study, we performed a longitudinal analysis of clinical and microbiome data from a large cohort of reproductive age rhesus macaques. At the taxonomic, genomic, and functional level, the rhesus macaque vaginal microbiome was most similar to that of humans who harbor a diverse vaginal community associated with asymptomatic/symptomatic bacterial vaginosis. Our in-depth and comprehensive analysis highlights the utility of macaques to test interventions for manipulating the vaginal microbiome.
]]></description>
<dc:creator>Rhoades, N. S.</dc:creator>
<dc:creator>Hendrickson, S. M.</dc:creator>
<dc:creator>Gerken, D. R.</dc:creator>
<dc:creator>Martinez, K.</dc:creator>
<dc:creator>Slayden, O. D.</dc:creator>
<dc:creator>Slifka, M. K.</dc:creator>
<dc:creator>Messaoudi, I.</dc:creator>
<dc:date>2020-12-15</dc:date>
<dc:identifier>doi:10.1101/2020.12.14.422805</dc:identifier>
<dc:title><![CDATA[Longitudinal profiling of the macaque vaginal microbiome reveals similarities to diverse human vaginal communities: implications for use as a pre-clinical model for bacterial vaginosis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-12-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.12.15.422915v1?rss=1">
<title>
<![CDATA[
Stretch-activated ion channels identified in the touch-sensitive structures of carnivorous Droseraceae plants 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.12.15.422915v1?rss=1</link>
<description><![CDATA[
In response to touch, some carnivorous plants such as the Venus flytrap have evolved spectacular movements to capture animals for nutrient acquisition. However, the molecules that confer this sensitivity remain unknown. We used comparative transcriptomics to show that expression of three genes encoding homologs of the MscS-Like (MSL) and OSCA/TMEM63 family of mechanosensitive ion channels are localized to touch-sensitive trigger hairs of Venus flytrap. We focus here on the candidate with the most enriched expression in trigger hairs, the MSL homolog FLYCATCHER1 (FLYC1). We show that FLYC1 transcripts are localized to mechanosensory cells within the trigger hair, transfecting FLYC1 induces chloride-permeable stretch-activated currents in naive cells, and transcripts coding for FLYC1 homologs are expressed in touch-sensing cells of Cape sundew, a related carnivorous plant of the Droseraceae family. Our data suggest that the mechanism of prey recognition in carnivorous Droseraceae evolved by co-opting ancestral mechanosensitive ion channels to sense touch.
]]></description>
<dc:creator>Procko, C.</dc:creator>
<dc:creator>Murthy, S.</dc:creator>
<dc:creator>Keenan, W. T.</dc:creator>
<dc:creator>Mousavi, S. A. R.</dc:creator>
<dc:creator>Dabi, T.</dc:creator>
<dc:creator>Coombs, A.</dc:creator>
<dc:creator>Procko, E.</dc:creator>
<dc:creator>Baird, L.</dc:creator>
<dc:creator>Patapoutian, A.</dc:creator>
<dc:creator>Chory, J.</dc:creator>
<dc:date>2020-12-15</dc:date>
<dc:identifier>doi:10.1101/2020.12.15.422915</dc:identifier>
<dc:title><![CDATA[Stretch-activated ion channels identified in the touch-sensitive structures of carnivorous Droseraceae plants]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-12-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.12.15.422929v1?rss=1">
<title>
<![CDATA[
A neural crest stem cell-like state drives nongenetic resistance to targeted therapy in melanoma 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.12.15.422929v1?rss=1</link>
<description><![CDATA[
The ability to predict the future behaviour of an individual cancer is crucial for precision cancer medicine and, in particular, for the development of strategies that prevent acquisition of resistance to anti-cancer drugs. Therapy resistance, which often develops from a heterogeneous pool of drug-tolerant cells known as minimal residual disease (MRD), is thought to mainly occur through acquisition of genetic alterations. Increasing evidence, however, indicates that drug resistance might also be acquired though nongenetic mechanisms. A key emerging question is therefore whether specific molecular and/or cellular features of the MRD ecosystem determine which of these two distinct resistance trajectories will eventually prevail. We show herein that, in melanoma exposed to MAPK-therapeutics, the presence of a neural crest stem cell (NCSC) subpopulation in MRD concurred with the rapid development of resistance through nongenetic mechanisms. Emergence of this drug-tolerant population in MRD relies on a GDNF-dependent autocrine and paracrine signalling cascade, which activates the AKT survival pathway in a Focal-adhesion kinase-(FAK) dependent manner. Ablation of this subpopulation through inhibition of FAK/SRC-signalling delayed relapse in patient-derived tumour xenografts. Strikingly, all tumours that eventually escaped this treatment exhibited resistance-conferring genetic alterations and increased sensitivity to ERK-inhibition. These findings firmly establish that nongenetic reprogramming events contribute to therapy resistance in melanoma and identify a clinically-compatible approach that abrogates such a trajectory. Importantly, these data demonstrate that the cellular composition of MRD deterministically imposes distinct drug resistance evolutionary paths and highlight key principles that may permit more effective pre-emptive therapeutic interventions.
]]></description>
<dc:creator>Marin-Bejar, O.</dc:creator>
<dc:creator>Rogiers, A.</dc:creator>
<dc:creator>Dewaele, M.</dc:creator>
<dc:creator>Femel, J.</dc:creator>
<dc:creator>Karras, P.</dc:creator>
<dc:creator>Pozniak, J.</dc:creator>
<dc:creator>Bervoets, G.</dc:creator>
<dc:creator>Van Raemdonck, N.</dc:creator>
<dc:creator>Pedri, D.</dc:creator>
<dc:creator>Swings, T.</dc:creator>
<dc:creator>Demeulemeester, J.</dc:creator>
<dc:creator>Vander Borght, S.</dc:creator>
<dc:creator>Bosisio, F.</dc:creator>
<dc:creator>van den Oord, J. J.</dc:creator>
<dc:creator>Vanden Bempt, I.</dc:creator>
<dc:creator>Lambrechts, D.</dc:creator>
<dc:creator>Voet, T.</dc:creator>
<dc:creator>Bechter, O.</dc:creator>
<dc:creator>Rizos, H.</dc:creator>
<dc:creator>Levesque, M.</dc:creator>
<dc:creator>Leucci, E.</dc:creator>
<dc:creator>Lund, A. W.</dc:creator>
<dc:creator>Rambow, F.</dc:creator>
<dc:creator>Marine, J.-C.</dc:creator>
<dc:date>2020-12-16</dc:date>
<dc:identifier>doi:10.1101/2020.12.15.422929</dc:identifier>
<dc:title><![CDATA[A neural crest stem cell-like state drives nongenetic resistance to targeted therapy in melanoma]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-12-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.12.18.423492v1?rss=1">
<title>
<![CDATA[
TRPM channels mediate learned pathogen avoidance following intestinal distention 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.12.18.423492v1?rss=1</link>
<description><![CDATA[
Upon exposure to harmful microorganisms, hosts engage in protective molecular and behavioral immune responses, both of which are ultimately regulated by the nervous system. Using the nematode Caenorhabditis elegans, we show that ingestion of E. faecalis leads to a fast pathogen avoidance behavior that results in aversive learning. We have identified multiple sensory mechanisms involved in the regulation of avoidance of E. faecalis, including the GPCR NPR-1-dependent oxygen-sensing pathway, an ASE neuron-dependent pathway, and an AWB and AWC neuron-dependent pathway. Colonization of the anterior part of the intestine by E. faecalis leads to AWB and AWC mediated olfactory aversive learning. Finally, two transient receptor potential melastatin (TRPM) channels, GON-2 and GTL-2, mediate this newly described rapid pathogen avoidance. These results suggest a mechanism by which TRPM channels may sense the intestinal distension caused by bacterial colonization to elicit pathogen avoidance and aversive learning by detecting changes in host physiology.
]]></description>
<dc:creator>Filipowicz, A.</dc:creator>
<dc:creator>Aballay, A.</dc:creator>
<dc:date>2020-12-19</dc:date>
<dc:identifier>doi:10.1101/2020.12.18.423492</dc:identifier>
<dc:title><![CDATA[TRPM channels mediate learned pathogen avoidance following intestinal distention]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-12-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.12.21.423901v1?rss=1">
<title>
<![CDATA[
THE URINARY MICROBIOME IN POSTMENOPAUSAL WOMEN WITH RECURRENT URINARY TRACT INFECTIONS 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.12.21.423901v1?rss=1</link>
<description><![CDATA[
Recurrent urinary tract infections (UTI) are highly prevalent in postmenopausal women, where vaginal estrogen and prophylactic antibiotics are used for treatment. The etiology of recurrent UTIs is not completely known, but the urinary microbiome is thought to be implicated. Thus, we aimed to compare the "steady state" urinary microbiome in three groups of menopausal women who were all using topically-applied vaginal estrogen: 1) women with recurrent UTIs on daily antibiotic prophylaxis; 2) women with recurrent UTIs not on antibiotic prophylaxis; and 3) age-matched controls without recurrent UTIs. Here we present a cross-sectional analysis of baseline data from 64 women enrolled in a longitudinal cohort study. Catheterized urine samples were collected > 4 weeks after last treatment for UTI. Samples were evaluated using expanded quantitative urine culture (EQUC) and 16S rRNA gene sequencing. With EQUC techniques, there were no significant differences in the median numbers of microbial species isolated among groups (p=0.96), even when considering Lactobacilli (p=0.72). However, there were trends towards different Lactobacillus species between groups. With sequencing the overwhelming majority of urinary samples contained Lactobacilli, with non-significant trends in relative abundance of Lactobacilli among groups. Using a Bayesian regression analysis for compositional data, we identified significant differences in anaerobic taxa that were associated with phenotypic groups. Most of these differences centered on Bacteroidales and the family Prevotellaceae, though differences were also noted in Actinobacteria and certain genera of Clostridiales. Associations between anaerobes within the urinary microbiome and recurrent UTI warrants further investigation.

IMPORTANCEIn menopausal women with recurrent urinary tract infections (UTIs) compared to those without, the abundance of Lactobacillus within the urinary microbiome is not significantly different when vaginal estrogen is regularly used. In this population, Lactobacillaceae were identified in 97% of urine samples using culture-independent techniques. However, with expanded urine cultures, women with recurrent UTIs taking daily antibiotics had a disproportionately low amount of L. gasseri/L. acidophilus compared to the other phenotypic groups. These findings support the theory that certain Lactobacillus species may be more important than others in the pathophysiology of postmenopausal recurrent UTIs. Furthermore, when using culture-independent techniques to explore urinary microbiota across phenotypic groups, we identified differences in multiple anaerobic taxa. Taken together, these results suggest that altered ratios of anaerobes and certain Lactobacillus species within the urinary microbiome may be implicated in postmenopausal recurrent UTI.
]]></description>
<dc:creator>Vaughan, M. H.</dc:creator>
<dc:creator>Mao, J.</dc:creator>
<dc:creator>Karstens, L.</dc:creator>
<dc:creator>Ma, L.</dc:creator>
<dc:creator>Amundsen, C. L.</dc:creator>
<dc:creator>Schmader, K. E.</dc:creator>
<dc:creator>Siddiqui, N. Y.</dc:creator>
<dc:date>2020-12-22</dc:date>
<dc:identifier>doi:10.1101/2020.12.21.423901</dc:identifier>
<dc:title><![CDATA[THE URINARY MICROBIOME IN POSTMENOPAUSAL WOMEN WITH RECURRENT URINARY TRACT INFECTIONS]]></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/10.1101/2020.12.25.424393v1?rss=1">
<title>
<![CDATA[
A cancer cell-intrinsic GOT2-PPARδ axis suppresses antitumor immunity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.12.25.424393v1?rss=1</link>
<description><![CDATA[
Despite significant recent advances in precision medicine1,2, pancreatic ductal adenocarcinoma (PDAC) remains near-uniformly lethal. While the most frequent genomic alterations in PDAC are not presently druggable3 and conventional therapies are often ineffective in this disease4, immune-modulatory therapies hold promise to meaningfully improve outcomes for PDAC patients. Development of such therapies requires an improved understanding of the immune evasion mechanisms that characterize the PDAC microenvironment, including frequent exclusion of antineoplastic T cells and abundance of immune-suppressive myeloid cells5-9. Here we show that cancer cell-intrinsic glutamic-oxaloacetic transaminase 2 (GOT2) shapes the immune microenvironment to suppress antitumor immunity. Mechanistically, we find that GOT2 functions beyond its established role in the malate-aspartate shuttle10-13 and promotes the transcriptional activity of nuclear receptor peroxisome proliferator-activated receptor delta (PPAR{delta}), facilitated by direct fatty acid binding. While GOT2 in PDAC cells is dispensable for cancer cell proliferation in vivo, GOT2 loss results in T cell-dependent suppression of tumor growth, and genetic or pharmacologic activation of PPAR{delta} restores PDAC progression in the GOT2-null context. This cancer cell-intrinsic GOT2-PPAR{delta} axis promotes spatial restriction of both CD4+ and CD8+ T cells from the tumor microenvironment, and fosters the immune-suppressive phenotype of tumor-infiltrating myeloid cells. Our results demonstrate a non-canonical function for an established mitochondrial enzyme in transcriptional regulation of immune evasion, which may be exploitable to promote a productive antitumor immune response.
]]></description>
<dc:creator>Sanford-Crane, H.</dc:creator>
<dc:creator>Abrego, J.</dc:creator>
<dc:creator>Oon, C.</dc:creator>
<dc:creator>Xiao, X.</dc:creator>
<dc:creator>Nagarajan, S.</dc:creator>
<dc:creator>Bhattacharyya, S.</dc:creator>
<dc:creator>Tontonoz, P.</dc:creator>
<dc:creator>Sherman, M. H.</dc:creator>
<dc:date>2020-12-26</dc:date>
<dc:identifier>doi:10.1101/2020.12.25.424393</dc:identifier>
<dc:title><![CDATA[A cancer cell-intrinsic GOT2-PPARδ axis suppresses antitumor immunity]]></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/10.1101/2020.12.30.424365v1?rss=1">
<title>
<![CDATA[
Activity-Dependent Modulation of Synapse-Regulating Genes in Astrocytes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.12.30.424365v1?rss=1</link>
<description><![CDATA[
Astrocytes regulate the formation and function of neuronal synapses via multiple signals, however, what controls regional and temporal expression of these signals during development is unknown. We determined the expression profile of astrocyte synapse-regulating genes in the developing mouse visual cortex, identifying astrocyte signals that show differential temporal and layer-enriched expression. These patterns are not intrinsic to astrocytes, but regulated by visually-evoked neuronal activity, as they are absent in mice lacking glutamate release from thalamocortical terminals. Consequently, synapses remain immature. Expression of synapse-regulating genes and synaptic development are also altered when astrocyte signaling is blunted by diminishing calcium release from astrocyte stores. Single nucleus RNA sequencing identified groups of astrocytic genes regulated by neuronal and astrocyte activity, and a cassette of genes that show layer-specific enrichment. Thus, the development of cortical circuits requires coordinated signaling between astrocytes and neurons, identifying astrocytes as a target to manipulate in neurodevelopmental disorders.
]]></description>
<dc:creator>Farhy-Tselnicker, I.</dc:creator>
<dc:creator>Boisvert, M. M.</dc:creator>
<dc:creator>Liu, H.</dc:creator>
<dc:creator>Dowling, C.</dc:creator>
<dc:creator>Erikson, G. A.</dc:creator>
<dc:creator>Blanco-Suarez, E.</dc:creator>
<dc:creator>Farhy, C.</dc:creator>
<dc:creator>Shokhirev, M.</dc:creator>
<dc:creator>Ecker, J. R.</dc:creator>
<dc:creator>Allen, N. J.</dc:creator>
<dc:date>2020-12-30</dc:date>
<dc:identifier>doi:10.1101/2020.12.30.424365</dc:identifier>
<dc:title><![CDATA[Activity-Dependent Modulation of Synapse-Regulating Genes in Astrocytes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-12-30</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/10.1101/2021.01.11.425995v1?rss=1">
<title>
<![CDATA[
High-content single-cell combinatorial indexing 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.01.11.425995v1?rss=1</link>
<description><![CDATA[
Single-cell genomics assays have emerged as a dominant platform for interrogating complex biological systems. Methods to capture various properties at the single-cell level typically suffer a tradeoff between cell count and information content, which is defined by the number of unique and usable reads acquired per cell. We and others have described workflows that utilize single-cell combinatorial indexing (sci)1, leveraging transposase-based library construction2 to assess a variety of genomic properties in high throughput; however, these techniques often produce sparse coverage for the property of interest. Here, we describe a novel adaptor-switching strategy,  s3, capable of producing one-to-two order-of-magnitude improvements in usable reads obtained per cell for chromatin accessibility (s3-ATAC), whole genome sequencing (s3-WGS), and whole genome plus chromatin conformation (s3-GCC), while retaining the same high-throughput capabilities of predecessor  sci technologies. We apply s3 to produce high-coverage single-cell ATAC-seq profiles of mouse brain and human cortex tissue; and whole genome and chromatin contact maps for two low-passage patient-derived cell lines from a primary pancreatic tumor.
]]></description>
<dc:creator>Mulqueen, R. M.</dc:creator>
<dc:creator>Pokholok, D.</dc:creator>
<dc:creator>O'Connell, B. L.</dc:creator>
<dc:creator>Thornton, C. A.</dc:creator>
<dc:creator>Zhang, F.</dc:creator>
<dc:creator>O'Roak, B. J.</dc:creator>
<dc:creator>Link, J.</dc:creator>
<dc:creator>Yardimci, G. G.</dc:creator>
<dc:creator>Sears, R. C.</dc:creator>
<dc:creator>Steemers, F. J.</dc:creator>
<dc:creator>Adey, A. C.</dc:creator>
<dc:date>2021-01-12</dc:date>
<dc:identifier>doi:10.1101/2021.01.11.425995</dc:identifier>
<dc:title><![CDATA[High-content single-cell combinatorial indexing]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-01-12</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/10.1101/2021.01.14.425337v1?rss=1">
<title>
<![CDATA[
Gene expression profiling of lymph node sub-capsular sinus macrophages in cancer 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.01.14.425337v1?rss=1</link>
<description><![CDATA[
Lymph nodes are key lymphoid organs collecting lymph fluid and migratory cells from the tissue area they survey. When cancerous cells arise within a tissue, the sentinel lymph node is the first immunological organ to mount an immune response. Sub-capsular sinus macrophages (SSMs) are specialized macrophages residing in the lymph nodes that play important roles as gatekeepers against particulate antigenic material. In the context of cancer, SSMs capture tumor-derived extracellular vesicles (tEVs), a form of particulate antigen released in high amounts by tumor cells. We have recently demonstrated that SSMs possess anti-tumor activity because in their absence tumors grow faster. A comprehensive profiling of SSMs represents an important first step to identify the cellular and molecular mechanisms responsible for SSM anti-tumor activity. Unfortunately, the isolation of SSMs for molecular analyses is very challenging. Here, we combined an optimized dissociation protocol, careful marker selection and stringent gating strategies to highly purify SSMs. We provide evidence of decreased T and B cell contamination, which allowed us to reveal the gene expression profile of this elusive macrophage subset. Squamous cell carcinomas induced an increase in the expression of Fc receptors, lysosomal and proteasomal enzymes in SSMs. These results suggest that SSMs may be able to capture immune complexes for antigen processing and presentation to B and T cells on both MHC class I and II.
]]></description>
<dc:creator>Pucci, F.</dc:creator>
<dc:creator>Claudio, N.</dc:creator>
<dc:creator>Danilo, P.</dc:creator>
<dc:date>2021-01-15</dc:date>
<dc:identifier>doi:10.1101/2021.01.14.425337</dc:identifier>
<dc:title><![CDATA[Gene expression profiling of lymph node sub-capsular sinus macrophages in cancer]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-01-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.01.15.426843v1?rss=1">
<title>
<![CDATA[
The GA4GH Variation Representation Specification (VRS): a Computational Framework for the Precise Representation and Federated Identification of Molecular Variation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.01.15.426843v1?rss=1</link>
<description><![CDATA[
Maximizing the personal, public, research, and clinical value of genomic information will require that clinicians, researchers, and testing laboratories exchange genetic variation data reliably. Developed by a partnership among national information resource providers, public initiatives, and diagnostic testing laboratories under the auspices of the Global Alliance for Genomics and Health (GA4GH), the Variation Representation Specification (VRS, pronounced "verse") is an extensible framework for the semantically precise and computable representation of variation that complements contemporary human-readable and flat file standards for variation representation. VRS objects are designed to be semantically precise representations of variation, and leverage this design to enable unique, federated identification of molecular variation. We describe the components of this framework, including the terminology and information model, schema, data sharing conventions, and a reference implementation, each of which is intended to be broadly useful and freely available for community use. The specification, documentation, examples, and community links are available at https://vrs.ga4gh.org/.
]]></description>
<dc:creator>Wagner, A. H.</dc:creator>
<dc:creator>Babb, L.</dc:creator>
<dc:creator>Alterovitz, G.</dc:creator>
<dc:creator>Baudis, M.</dc:creator>
<dc:creator>Brush, M.</dc:creator>
<dc:creator>Cameron, D. L.</dc:creator>
<dc:creator>Cline, M.</dc:creator>
<dc:creator>Griffith, M.</dc:creator>
<dc:creator>Griffith, O.</dc:creator>
<dc:creator>Hunt, S.</dc:creator>
<dc:creator>Kreda, D.</dc:creator>
<dc:creator>Lee, J.</dc:creator>
<dc:creator>Lopez, J.</dc:creator>
<dc:creator>Moyer, E.</dc:creator>
<dc:creator>Nelson, T.</dc:creator>
<dc:creator>Patel, R. Y.</dc:creator>
<dc:creator>Riehle, K.</dc:creator>
<dc:creator>Robinson, P. N.</dc:creator>
<dc:creator>Rynearson, S.</dc:creator>
<dc:creator>Schuilenburg, H.</dc:creator>
<dc:creator>Tsukanov, K.</dc:creator>
<dc:creator>Walsh, B.</dc:creator>
<dc:creator>Konopko, M.</dc:creator>
<dc:creator>Rehm, H.</dc:creator>
<dc:creator>Yates, A. D.</dc:creator>
<dc:creator>Freimuth, R. R.</dc:creator>
<dc:creator>Hart, R. K.</dc:creator>
<dc:date>2021-01-17</dc:date>
<dc:identifier>doi:10.1101/2021.01.15.426843</dc:identifier>
<dc:title><![CDATA[The GA4GH Variation Representation Specification (VRS): a Computational Framework for the Precise Representation and Federated Identification of Molecular Variation]]></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/10.1101/2021.01.18.427078v1?rss=1">
<title>
<![CDATA[
Human Neural Stem Cells Differentiate and Integrate, Innervating Implanted zQ175 Huntington's Disease Mouse Striatum 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.01.18.427078v1?rss=1</link>
<description><![CDATA[
Huntingtons disease (HD), a genetic neurodegenerative disorder, primarily impacts the striatum and cortex with progressive loss of medium-sized spiny neurons (MSNs) and pyramidal neurons, disrupting cortico-striatal circuitry. A promising regenerative therapeutic strategy of transplanting human neural stem cells (hNSCs) is challenged by the need for long-term functional integration. We previously described that hNSCs transplanted into the striatum of HD mouse models differentiated into electrophysiologically active immature neurons, improving behavior and biochemical deficits. Here we show that 8-month implantation of hNSCs into the striatum of zQ175 HD mice ameliorates behavioral deficits, increases brain-derived neurotrophic factor (BDNF) and reduces mutant Huntingtin (mHTT) accumulation. Patch clamp recordings, immunohistochemistry and electron microscopy demonstrates that hNSCs differentiate into diverse neuronal populations, including MSN- and interneuron-like cells. Remarkably, hNSCs receive synaptic inputs, innervate host neurons, and improve membrane and synaptic properties. Overall, the findings support hNSC transplantation for further evaluation and clinical development for HD.
]]></description>
<dc:creator>Thompson, L. M.</dc:creator>
<dc:creator>Holley, S. M.</dc:creator>
<dc:creator>Reidling, J. C.</dc:creator>
<dc:creator>Cepeda, C.</dc:creator>
<dc:creator>Lau, A.</dc:creator>
<dc:creator>Moore, C.</dc:creator>
<dc:creator>Orellana, I.</dc:creator>
<dc:creator>Fury, B.</dc:creator>
<dc:creator>Kopan, L.</dc:creator>
<dc:creator>Yeung, S.</dc:creator>
<dc:creator>Neel, M.</dc:creator>
<dc:creator>Coleal-Bergum, D.</dc:creator>
<dc:creator>Monuki, E.</dc:creator>
<dc:creator>K. Meshul, C.</dc:creator>
<dc:creator>Bauer, G.</dc:creator>
<dc:creator>Levine, M. S.</dc:creator>
<dc:date>2021-01-19</dc:date>
<dc:identifier>doi:10.1101/2021.01.18.427078</dc:identifier>
<dc:title><![CDATA[Human Neural Stem Cells Differentiate and Integrate, Innervating Implanted zQ175 Huntington's Disease Mouse Striatum]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-01-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.01.25.428149v1?rss=1">
<title>
<![CDATA[
TREM2 is thyroid hormone regulated making the TREM2 pathway druggable with ligands for thyroid hormone receptor 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.01.25.428149v1?rss=1</link>
<description><![CDATA[
Triggering receptor expressed on myeloid cells-2 (TREM2) is a cell surface receptor on macrophages and microglia that senses and responds to disease associated signals to regulate the phenotype of these innate immune cells. The TREM2 signaling pathway has been implicated in a variety of diseases ranging from neurodegeneration in the central nervous system to metabolic disease in the periphery. We report here that TREM2 is a thyroid hormone regulated gene and its expression in macrophages and microglia is stimulated by thyroid hormone. Both endogenous thyroid hormone and sobetirome, a synthetic thyroid hormone agonist drug, suppress pro-inflammatory cytokine production from myeloid cells including macrophages that have been treated with the SARS-CoV-2 spike protein which produces a strong, pro-inflammatory phenotype. Thyroid hormone agonism was also found to induce phagocytic behavior in microglia, a phenotype consistent with activation of the TREM2 pathway. The thyroid hormone antagonist NH-3 blocks the anti-inflammatory effects of thyroid hormone agonists and suppresses microglia phagocytosis. Finally, in a murine experimental autoimmune encephalomyelitis (EAE) multiple sclerosis model, treatment with Sob-AM2, a CNS-penetrating sobetirome prodrug, results in increased Trem2 expression in disease lesion resident myeloid cells which correlates with therapeutic benefit in the EAE clinical score and reduced damage to myelin. Our findings represent the first report of endocrine regulation of TREM2 and provide a unique opportunity to drug the TREM2 signaling pathway with orally active small molecule therapeutic agents.
]]></description>
<dc:creator>Ferrara, S. J.</dc:creator>
<dc:creator>Chaudhary, P.</dc:creator>
<dc:creator>DeBell, M. J.</dc:creator>
<dc:creator>Marracci, G.</dc:creator>
<dc:creator>Miller, H.</dc:creator>
<dc:creator>Calkins, E.</dc:creator>
<dc:creator>Pocius, E.</dc:creator>
<dc:creator>Napier, B. A.</dc:creator>
<dc:creator>Emery, B.</dc:creator>
<dc:creator>Bourdette, D.</dc:creator>
<dc:creator>Scanlan, T. S.</dc:creator>
<dc:date>2021-01-27</dc:date>
<dc:identifier>doi:10.1101/2021.01.25.428149</dc:identifier>
<dc:title><![CDATA[TREM2 is thyroid hormone regulated making the TREM2 pathway druggable with ligands for thyroid hormone receptor]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-01-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.01.25.428184v1?rss=1">
<title>
<![CDATA[
Prior dengue immunity enhances Zika virus infection of the maternal-fetal interface in rhesus macaques 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.01.25.428184v1?rss=1</link>
<description><![CDATA[
Concerns have arisen that pre-existing immunity to dengue virus (DENV) could enhance Zika virus (ZIKV) disease, due to the homology between ZIKV and DENV and the observation of antibody-dependent enhancement (ADE) among DENV serotypes. To date, no study has examined the impact of pre-existing DENV immunity on ZIKV pathogenesis during pregnancy in a translational non-human primate model. Here we show that prior DENV-2 exposure enhanced ZIKV infection of maternal-fetal interface tissues in macaques. However, pre-existing DENV immunity had no detectable impact on ZIKV replication kinetics in maternal plasma, and all pregnancies progressed to term without adverse outcomes or gross fetal abnormalities detectable at delivery. Understanding the risks of ADE to pregnant women worldwide is critical as vaccines against DENV and ZIKV are developed and licensed and as DENV and ZIKV continue to circulate.
]]></description>
<dc:creator>Crooks, C. M.</dc:creator>
<dc:creator>Weiler, A. M.</dc:creator>
<dc:creator>Rybarczyk, S. L.</dc:creator>
<dc:creator>Bliss, M. I.</dc:creator>
<dc:creator>Jaeger, A. S.</dc:creator>
<dc:creator>Murphy, M. E.</dc:creator>
<dc:creator>Simmons, H. A.</dc:creator>
<dc:creator>Mejia, A.</dc:creator>
<dc:creator>Fritsch, M. K.</dc:creator>
<dc:creator>Hayes, J. M.</dc:creator>
<dc:creator>Eickhoff, J. C.</dc:creator>
<dc:creator>Mitzey, A. M.</dc:creator>
<dc:creator>Razo, E.</dc:creator>
<dc:creator>Braun, K. M.</dc:creator>
<dc:creator>Brown, E. A.</dc:creator>
<dc:creator>Yamamoto, K.</dc:creator>
<dc:creator>Shepherd, P. M.</dc:creator>
<dc:creator>Possell, A.</dc:creator>
<dc:creator>Weaver, K.</dc:creator>
<dc:creator>Antony, K. M.</dc:creator>
<dc:creator>Morgan, T. K.</dc:creator>
<dc:creator>Newman, C. M.</dc:creator>
<dc:creator>Dudley, D. M.</dc:creator>
<dc:creator>Schultz-Darken, N.</dc:creator>
<dc:creator>Peterson, E.</dc:creator>
<dc:creator>Katzelnick, L. C.</dc:creator>
<dc:creator>Balmaseda, A.</dc:creator>
<dc:creator>Harris, E.</dc:creator>
<dc:creator>OConnor, D. H.</dc:creator>
<dc:creator>Mohr, E. L.</dc:creator>
<dc:creator>Golos, T. G.</dc:creator>
<dc:creator>Friedrich, T. C.</dc:creator>
<dc:creator>Aliota, M. T.</dc:creator>
<dc:date>2021-01-26</dc:date>
<dc:identifier>doi:10.1101/2021.01.25.428184</dc:identifier>
<dc:title><![CDATA[Prior dengue immunity enhances Zika virus infection of the maternal-fetal interface in rhesus macaques]]></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/10.1101/2021.01.27.428336v1?rss=1">
<title>
<![CDATA[
A noncanonical FLT3 gatekeeper mutation disrupts gilteritinib binding and confers resistance 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.01.27.428336v1?rss=1</link>
<description><![CDATA[
The recent FDA approval of the FLT3 inhibitor, gilteritinib, for AML represents a major breakthrough for treatment of FLT3 mutated AML. However, patients only respond to gilteritinib for 6-7 months due to the emergence of drug resistance. Clinical resistance to gilteritinib is often associated with expansion of NRAS mutations, and less commonly via gatekeeper mutations in FLT3, with F691L being the most common. We developed an in vitro model that charts the temporal evolution of resistance to gilteritinib from early microenvironmental-mediated resistance to late intrinsic resistance mutations. Our model system accurately recapitulates the expansion of NRAS mutations and the F691L gatekeeper mutations found in AML patients. As part of this study, we also identified a novel FLT3N701K mutation that also appeared to promote resistance to gilteritinib. Using the Ba/F3 system, we demonstrate that N701K mutations effectively act like a gatekeeper mutation and block gilteritinib from binding to FLT3, thereby promoting resistance. Structural modeling of FLT3 reveals how N701K, and other reported gilteritinib resistance mutations, obstruct the gilteritinib binding pocket on FLT3. Interestingly, FLT3N701K does not block quizartinib binding, suggesting that FLT3N701K mutations are more specific for type 1 FLT3 inhibitors (gilteritinib, midostaurin, and crenolanib). Thus, our data suggests that for the FLT3N701K mutation, switching classes of FLT3 inhibitors may restore clinical response. As the use of gilteritinib expands in the clinic, this information will become critical to define clinical strategies to manage gilteritinib resistance.
]]></description>
<dc:creator>Joshi, S. K.</dc:creator>
<dc:creator>Sharzehi, S.</dc:creator>
<dc:creator>Pittsenbarger, J.</dc:creator>
<dc:creator>Bottomly, D.</dc:creator>
<dc:creator>Tognon, C. E.</dc:creator>
<dc:creator>McWeeney, S.</dc:creator>
<dc:creator>Druker, B.</dc:creator>
<dc:creator>Traer, E.</dc:creator>
<dc:date>2021-01-28</dc:date>
<dc:identifier>doi:10.1101/2021.01.27.428336</dc:identifier>
<dc:title><![CDATA[A noncanonical FLT3 gatekeeper mutation disrupts gilteritinib binding and confers resistance]]></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/10.1101/2021.01.29.428748v1?rss=1">
<title>
<![CDATA[
Natural loss of function of ephrin-B3 shapes spinal flight circuitry in birds 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.01.29.428748v1?rss=1</link>
<description><![CDATA[
Flight in birds evolved through patterning of the wings from forelimbs and transition from alternating gait to synchronous flapping. In mammals, the spinal midline guidance molecule ephrin-B3 instructs the wiring that enables limb alternation, and its deletion leads to synchronous hopping gait. Here we show that the ephrin-B3 protein in birds lacks several motifs present in other vertebrates, diminishing its affinity for the EphA4 receptor. The avian ephrin-B3 gene lacks an enhancer that drives midline expression, and is missing in Galliformes. The morphology and wiring at brachial levels of the chick spinal cord resemble those of ephrin-B3 null mice. Importantly, dorsal midline decussation, evident in the mutant mouse, is apparent at the chick brachial level, and is prevented by expression of exogenous ephrin-B3 at the roof plate. Our findings support a role for loss of ephrin-B3 function in shaping the avian brachial spinal cord circuitry and facilitating synchronous wing flapping.

TeaserWalking vs flying: Deciphering the organization and evolution of the neuronal network that controls wing flapping in birds.
]]></description>
<dc:creator>Haimson, B.</dc:creator>
<dc:creator>Meir, O.</dc:creator>
<dc:creator>Sudakevitz-Merzbach, R.</dc:creator>
<dc:creator>Elberg, G.</dc:creator>
<dc:creator>Friedrich, S.</dc:creator>
<dc:creator>Lovell, p.</dc:creator>
<dc:creator>Paixao, s.</dc:creator>
<dc:creator>Klen, R.</dc:creator>
<dc:creator>Mello, C. V.</dc:creator>
<dc:creator>Klar, A.</dc:creator>
<dc:date>2021-01-29</dc:date>
<dc:identifier>doi:10.1101/2021.01.29.428748</dc:identifier>
<dc:title><![CDATA[Natural loss of function of ephrin-B3 shapes spinal flight circuitry in birds]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-01-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.02.01.429244v1?rss=1">
<title>
<![CDATA[
Mutant SETBP1 enhances NRAS-driven MAPK pathway activation to promote aggressive leukemia 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.02.01.429244v1?rss=1</link>
<description><![CDATA[
Mutations in SET binding protein 1 (SETBP1) are associated with poor outcomes in myeloid leukemias. In the Ras-driven leukemia, juvenile myelomonocytic leukemia, SETBP1 mutations are enriched in relapsed disease. While some mechanisms for SETBP1-driven oncogenesis have been established, it remains unclear how SETBP1 specifically modulates the biology of Ras-driven leukemias. In this study, we found that when co-expressed with Ras pathway mutations, SETBP1 promoted oncogenic transformation of murine bone marrow in vitro and aggressive myeloid leukemia in vivo. We demonstrate that SETBP1 enhances the NRAS gene expression signature, driving upregulation of mitogen-activated protein kinase (MAPK) signaling and downregulation of differentiation pathways. SETBP1 also enhances NRAS-driven phosphorylation of MAPK proteins. Cells expressing NRAS and SETBP1 are sensitive to inhibitors of the MAPK pathway, and treatment with the MEK inhibitor trametinib conferred a survival benefit in a mouse model of NRAS/SETBP1-mutant disease. Our data demonstrate that despite driving enhanced MAPK signaling, SETBP1-mutant cells remain susceptible to trametinib in vitro and in vivo, providing encouraging pre-clinical data for the use of trametinib in SETBP1-mutant disease.
]]></description>
<dc:creator>Carratt, S. A.</dc:creator>
<dc:creator>Braun, T. P.</dc:creator>
<dc:creator>Coblentz, C.</dc:creator>
<dc:creator>Schonrock, Z.</dc:creator>
<dc:creator>Callahan, R.</dc:creator>
<dc:creator>Smith, B. M.</dc:creator>
<dc:creator>Maloney, L.</dc:creator>
<dc:creator>Foley, A.</dc:creator>
<dc:creator>Maxson, J. E.</dc:creator>
<dc:date>2021-02-02</dc:date>
<dc:identifier>doi:10.1101/2021.02.01.429244</dc:identifier>
<dc:title><![CDATA[Mutant SETBP1 enhances NRAS-driven MAPK pathway activation to promote aggressive leukemia]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-02-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.02.03.429523v1?rss=1">
<title>
<![CDATA[
The impact of genetic background and sex on the phenotype of IL-23 induced murine arthritis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.02.03.429523v1?rss=1</link>
<description><![CDATA[
BackgroundOverexpression of IL-23 in adult mice by means of hydrodynamic tail vein injection of IL-23 minicircles has been reported to result in spondyloarthritis-like disease. The impact of genetic background and sex on the disease phenotype in this model has not been investigated.

MethodsWe compared male B10.RIII mice with male C57BL/6 mice, and male with female B10.RIII mice after hydrodynamic injection of IL-23 enhanced episomal vector (EEV) at 8-12 weeks of age. We monitored clinical arthritis scores, paw swelling, and body weight. Animals were euthanized after two weeks and tissues were harvested for histology, flow cytometry and gene expression analysis. Serum cytokine levels were determined by ELISA.

FindingsMale B10.RIII mice developed arthritis in the forepaws and feet within 6 days after IL-23 EEV injection; they also exhibited psoriasis-like skin disease, colitis, weight loss, and osteopenia. In contrast to previous reports, we did not observe spondylitis or uveitis. Male C57BL/6 mice injected with IL-23 EEV had serum IL-23 levels comparable with B10.RIII mice and developed skin inflammation, colitis, weight loss, and osteopenia but failed to develop arthritis. Female B10.RIII mice had more severe arthritis than male B10.RIII mice but did not lose weight.

ConclusionsSystemic IL-23 overexpression results in spondyloarthritis-like disease in B10.RIII mice. The development of extra-articular manifestations but absence of arthritis in C57BL/6 mice suggests organ-specific genetic control mechanisms of IL-23 driven inflammation. Discrepancies regarding the phenotype of IL-23 induced disease in different labs and the sexual dimorphism observed in this study warrant further exploration.
]]></description>
<dc:creator>Haley, E.</dc:creator>
<dc:creator>Matmusaev, M.</dc:creator>
<dc:creator>Hossain, I. N.</dc:creator>
<dc:creator>Davin, S.</dc:creator>
<dc:creator>Martin, T. M.</dc:creator>
<dc:creator>Ermann, J.</dc:creator>
<dc:date>2021-02-03</dc:date>
<dc:identifier>doi:10.1101/2021.02.03.429523</dc:identifier>
<dc:title><![CDATA[The impact of genetic background and sex on the phenotype of IL-23 induced murine arthritis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-02-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.02.12.431000v1?rss=1">
<title>
<![CDATA[
Single-cell Visualization and Quantification of Trace Metals in Chlamydomonas Lysosome-Related Organelles 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.02.12.431000v1?rss=1</link>
<description><![CDATA[
The acidocalcisome is an acidic organelle in the cytosol of eukaryotes, defined by its low pH and high calcium and polyphosphate content. It is visualized as an electron-dense object by transmission electron microscopy (TEM) or described with mass-spectrometry (MS)-based imaging techniques or multimodal X-ray fluorescence microscopy (XFM) based on its unique elemental composition. Compared to MS-based imaging techniques, XFM offers the advantage of absolute quantification of trace metal content, since sectioning of the cell is not required and metabolic states can be preserved rapidly by either vitrification or chemical fixation. We employed XFM in Chlamydomonas reinhardtii, to determine single-cell and organelle trace metal quotas within algal cells in situations of trace metal over-accumulation (Fe, Cu). We found up to 70% of the cellular Cu and 80% of Fe sequestered in acidocalcisomes in these conditions, and identified two distinct populations of acidocalcisomes, defined by their unique trace elemental makeup. We utilized the vtc1 mutant, defective in polyphosphate synthesis and failing to accumulate Ca to show that Fe sequestration is not dependent on either. Finally, quantitation of the Fe and Cu contents of individual cells and compartments via XFM, over a range of cellular metal quotas created by nutritional and genetic perturbations, indicated excellent correlation with bulk data from corresponding cell cultures, establishing a framework to distinguish the nutritional status of single cells.

Significance statementTransition metals are of crucial importance for primary productivity; their scarcity limits crop yield in agriculture and carbon sequestration at global scale. Copper (Cu), iron (Fe) and manganese (Mn) are among the most important trace elements that enable the redox chemistry in oxygenic photosynthesis. The single-celled, eukaryotic green alga Chlamydomonas reinhardtii is a choice experimental system for studying trace metal homeostasis in the context of phototrophy, offering all the advantages of a classical microbial system with a well-characterized photosystem and trace metal metabolism machinery of relevance to plants. This project identifies and differentiates different trace metal storage sites in Chlamydomonas and uncovers the dynamics of trace metal storage and mobilization in situations of fluctuating resources.
]]></description>
<dc:creator>Schmollinger, S.</dc:creator>
<dc:creator>Chen, S.</dc:creator>
<dc:creator>Strenkert, D.</dc:creator>
<dc:creator>Hui, C.</dc:creator>
<dc:creator>Ralle, M.</dc:creator>
<dc:creator>Merchant, S. S.</dc:creator>
<dc:date>2021-02-13</dc:date>
<dc:identifier>doi:10.1101/2021.02.12.431000</dc:identifier>
<dc:title><![CDATA[Single-cell Visualization and Quantification of Trace Metals in Chlamydomonas Lysosome-Related Organelles]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-02-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.02.16.431411v1?rss=1">
<title>
<![CDATA[
Human cytomegalovirus blocks canonical TGFβ signaling during lytic infection to limit induction of type I interferons 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.02.16.431411v1?rss=1</link>
<description><![CDATA[
Human cytomegalovirus (HCMV) microRNAs (miRNAs) significantly rewire host signaling pathways to support the viral lifecycle and regulate host cell responses. Here we show that SMAD3 expression is regulated by HCMV miR-UL22A and contributes to the IRF7-mediated induction of type I IFNs and IFN-stimulated genes (ISGs) in human fibroblasts. Addition of exogenous TGF{beta} interferes with the replication of a miR-UL22A mutant virus in a SMAD3-dependent manner in wild type fibroblasts, but not in cells lacking IRF7, indicating that downregulation of SMAD3 expression to limit IFN induction is important for efficient lytic replication. These findings uncover a novel interplay between SMAD3 and innate immunity during HCMV infection and highlight the role of viral miRNAs in modulating these responses.

Author SummaryCells trigger the interferon (IFN) response to induce the expression of cellular genes that limit virus replication. In turn, viruses have evolved numerous countermeasures to avoid the effects of IFN signaling. Using a microRNA (miRNA) mutant virus we have uncovered a novel means of regulating the IFN response during human cytomegalovirus (HCMV) infection. Lytic HCMV infection induces the production of TGF{beta}, which binds to the TGF{beta} receptor and activates the receptor-associated SMAD SMAD3. SMAD3, together with IRF7, induces the expression of IFN{beta} and downstream IFN-stimulated genes in human fibroblasts. To counteract this, HCMV miR-UL22A, along with other HCMV gene products, directly targets SMAD3 for downregulation. Infection of fibroblasts with a miR-UL22A mutant virus results in enhanced type I IFN production in a SMAD3-dependent manner and the virus is impaired for growth in the presence of TGF{beta}, but only when both SMAD3 and IRF7 are present, highlighting the unique interaction between TGF{beta} and innate immune signaling.
]]></description>
<dc:creator>Pham, A. H.</dc:creator>
<dc:creator>Mitchell, J.</dc:creator>
<dc:creator>Botto, S.</dc:creator>
<dc:creator>Pryke, K. M.</dc:creator>
<dc:creator>Defilippis, V. R.</dc:creator>
<dc:creator>Hancock, M.</dc:creator>
<dc:date>2021-02-16</dc:date>
<dc:identifier>doi:10.1101/2021.02.16.431411</dc:identifier>
<dc:title><![CDATA[Human cytomegalovirus blocks canonical TGFβ signaling during lytic infection to limit induction of type I interferons]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-02-16</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/10.1101/2021.02.23.432582v1?rss=1">
<title>
<![CDATA[
Widespread ripples synchronize human cortical activity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.02.23.432582v1?rss=1</link>
<description><![CDATA[
Declarative memory encoding, consolidation, and retrieval require the integration of elements encoded in widespread cortical locations. The mechanism whereby such  binding of different components of mental events into unified representations occurs is unknown. The  binding-bysynchrony theory proposes that distributed encoding areas are bound by synchronous oscillations enabling enhanced communication. However, evidence for such oscillations is sparse. Brief high-frequency oscillations ( ripples) occur in the hippocampus and cortex, and help organize memory recall and consolidation. Here, using intracranial recordings in humans, we report that these ~70ms duration 90Hz ripples often couple (within {+/-}500ms), co-occur ([&ge;]25ms overlap), and crucially, phase-lock (have consistent phase-lags) between widely distributed focal cortical locations during both sleep and waking, even between hemispheres. Cortical ripple co-occurrence is facilitated through activation across multiple sites, and phaselocking increases with more cortical sites co-rippling. Ripples in all cortical areas co-occur with hippocampal ripples but do not phase-lock with them, further suggesting that cortico-cortical synchrony is mediated by cortico-cortical connections. Ripple phase-lags vary across sleep nights, consistent with participation in different networks. During waking, we show that hippocampo-cortical and cortico-cortical co-ripples increase preceding successful delayed memory recall, when binding between the cue and response is essential. Ripples increase and phase-modulate unit firing, and co-ripples increase high-frequency correlations between areas, suggesting synchronized unit-spiking facilitating information exchange. Co-occurrence, phasesynchrony, and high-frequency correlation are maintained with little decrement over very long distances (25cm). Hippocampo-cortico-cortical co-ripples appear to possess the essential properties necessary to support binding-by-synchrony during memory retrieval, and perhaps generally in cognition.

Significance StatementDifferent elements of a memory, or any mental event, are encoded in locations distributed across the cortex. A prominent hypothesis proposes that widespread networks are integrated with bursts of synchronized high-frequency oscillations called  ripples, but evidence is limited. Here, using recordings inside the human brain, we show that ripples occur simultaneously in multiple lobes in both cortical hemispheres, and the hippocampus, generally during sleep and waking, and especially during memory recall. Ripples phase-lock local cell firing, and phase-synchronize with little decay between locations separated by up to 25cm, enabling long-distance integration. Indeed, co-rippling sites have increased correlation of very high-frequency activity which reflects cell firing. Thus, ripples may help bind information across the cortex in memory and other mental events.
]]></description>
<dc:creator>Dickey, C. W.</dc:creator>
<dc:creator>Verzhbinsky, I. A.</dc:creator>
<dc:creator>Jiang, X.</dc:creator>
<dc:creator>Rosen, B. Q.</dc:creator>
<dc:creator>Kajfez, S.</dc:creator>
<dc:creator>Stedelin, B.</dc:creator>
<dc:creator>Shih, J. J.</dc:creator>
<dc:creator>Ben-Haim, S.</dc:creator>
<dc:creator>Raslan, A. M.</dc:creator>
<dc:creator>Madsen, J. R.</dc:creator>
<dc:creator>Eskandar, E. N.</dc:creator>
<dc:creator>Gonzalez-Martinez, J.</dc:creator>
<dc:creator>Cash, S. S.</dc:creator>
<dc:creator>Halgren, E.</dc:creator>
<dc:date>2021-02-24</dc:date>
<dc:identifier>doi:10.1101/2021.02.23.432582</dc:identifier>
<dc:title><![CDATA[Widespread ripples synchronize human cortical activity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-02-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.02.25.432801v1?rss=1">
<title>
<![CDATA[
Immunity-longevity tradeoff neurally controlled by GABAergic transcription factor PITX1/UNC-30 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.02.25.432801v1?rss=1</link>
<description><![CDATA[
A body of evidence indicates that metazoan immune and aging pathways are largely interconnected, but the mechanisms involved in their homeostatic control remain unclear. In this study, we found that the PITX (paired like homeodomain) transcription factor UNC-30 controls the tradeoff between immunity and longevity from the nervous system in Caenorhabditis elegans. PITX/UNC-30 functional loss enhanced immunity in a GATA/ELT-2- and p38 MAPK/PMK-1-dependent manner and reduced longevity by activating MXD/MDL-1 and the C2H2-type zinc finger transcription factor PQM-1. The immune inhibitory and longevity stimulatory functions of PITX/UNC-30 required the sensory neuron ASG and a neurotransmitter signaling pathway controlled by NPR-1, which is a G protein-coupled receptor related to mammalian neuropeptide Y receptors. Our findings uncovered a suppressive role of GABAergic signaling in the neural control of a biological tradeoff where energy is allocated towards immunity at the expense of longevity.
]]></description>
<dc:creator>Otarigho, B.</dc:creator>
<dc:creator>Aballay, A.</dc:creator>
<dc:date>2021-02-26</dc:date>
<dc:identifier>doi:10.1101/2021.02.25.432801</dc:identifier>
<dc:title><![CDATA[Immunity-longevity tradeoff neurally controlled by GABAergic transcription factor PITX1/UNC-30]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-02-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.02.26.432955v1?rss=1">
<title>
<![CDATA[
IL-10 driven memory T cell survival and Tfh differentiation promote HIV persistence 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.02.26.432955v1?rss=1</link>
<description><![CDATA[
Mechanisms regulating HIV persistence are complex and not well understood. Increased IL-10 levels were positively associated with HIV reservoir in blood and lymph nodes (LN) of treated HIV aviremic individuals. In LNs, B cells, regulatory T cells, follicular T helper cells (Tfh), monocytes and macrophages contributed to the frequencies of IL10+ cells. Cells with HIV DNA in LNs were in close proximity to IL-10+ cells and/or had the active form of STAT3, the transcription downstream of IL-10. Gene signatures and proteins associated to cell survival, Co-inhibitory receptors expression, maintenance of memory T cells, immune metabolism and Tfh frequencies were all modulated by IL-10 and associated with HIV reservoir persistence. In vitro, STAT3 knockout or neutralization of IL-10, reverted all the aforementioned pathways and resulted in 10-fold decay in HIV reservoir. Collectively, these results provide strong evidence for a pivotal role of IL-10 in HIV persistence, and a potential therapeutic strategy for HIV cure.
]]></description>
<dc:creator>Pereira Ribeiro, S.</dc:creator>
<dc:creator>Aid, M.</dc:creator>
<dc:creator>Dupuy, F. P.</dc:creator>
<dc:creator>Ngai Chan, C.</dc:creator>
<dc:creator>Hultquist, J.</dc:creator>
<dc:creator>Deleage, C.</dc:creator>
<dc:creator>Moysi, E.</dc:creator>
<dc:creator>Kulpa, D. A.</dc:creator>
<dc:creator>Li, L.</dc:creator>
<dc:creator>Xu, X.</dc:creator>
<dc:creator>Tamilselvan, B.</dc:creator>
<dc:creator>Tomalka, J. A.</dc:creator>
<dc:creator>Nekorchuk, M.</dc:creator>
<dc:creator>Busman-Sahay, K.</dc:creator>
<dc:creator>Bordi, R.</dc:creator>
<dc:creator>Simoneau, C.</dc:creator>
<dc:creator>Goulet, J. P.</dc:creator>
<dc:creator>Marconi, V.</dc:creator>
<dc:creator>Routy, J. P.</dc:creator>
<dc:creator>Balderas, R.</dc:creator>
<dc:creator>Micci, L.</dc:creator>
<dc:creator>J Howell, B.</dc:creator>
<dc:creator>H Barouch, D.</dc:creator>
<dc:creator>Krogan, N. J.</dc:creator>
<dc:creator>Petrovas, C.</dc:creator>
<dc:creator>Paiardini, M.</dc:creator>
<dc:creator>Deeks, S.</dc:creator>
<dc:creator>Estes, J. D.</dc:creator>
<dc:creator>Gorman, D.</dc:creator>
<dc:creator>J Hazuda, D.</dc:creator>
<dc:creator>Sekaly, R.-P.</dc:creator>
<dc:date>2021-02-26</dc:date>
<dc:identifier>doi:10.1101/2021.02.26.432955</dc:identifier>
<dc:title><![CDATA[IL-10 driven memory T cell survival and Tfh differentiation promote HIV persistence]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-02-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.02.27.433155v1?rss=1">
<title>
<![CDATA[
A de novo paradigm for male infertility 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.02.27.433155v1?rss=1</link>
<description><![CDATA[
IntroductionDe novo mutations (DNMs) are known to play a prominent role in sporadic disorders with reduced fitness1. We hypothesize that DNMs play an important role in male infertility and explain a significant fraction of the genetic causes of this understudied disorder. To test this hypothesis, we performed trio-based exome-sequencing in a unique cohort of 185 infertile males and their unaffected parents. Following a systematic analysis, 29 of 145 rare protein altering DNMs were classified as possibly causative of the male infertility phenotype. We observed a significant enrichment of Loss-of-Function (LoF) DNMs in LoF-intolerant genes (p-value=1.00x10-5) as well as predicted pathogenic missense DNMs in missense-intolerant genes (p-value=5.01x10-4). One DNM gene identified, RBM5, is an essential regulator of male germ cell pre-mRNA splicing2. In a follow-up study, 5 rare pathogenic missense mutations affecting this gene were observed in a cohort of 2,279 infertile patients, with no such mutations found in a cohort of 5,784 fertile men (p-value=0.009). Our results provide the first evidence for the role of DNMs in severe male infertility and point to many new candidate genes affecting fertility.
]]></description>
<dc:creator>Oud, M. S.</dc:creator>
<dc:creator>Smits, R. M.</dc:creator>
<dc:creator>Smith, H. E.</dc:creator>
<dc:creator>Mastrorosa, F. K.</dc:creator>
<dc:creator>Holt, G. S.</dc:creator>
<dc:creator>Houston, B. J.</dc:creator>
<dc:creator>de Vries, P. F.</dc:creator>
<dc:creator>Alobaidi, B. K.</dc:creator>
<dc:creator>Batty, L. E.</dc:creator>
<dc:creator>Ismail, H.</dc:creator>
<dc:creator>Greenwood, J.</dc:creator>
<dc:creator>Sheth, H.</dc:creator>
<dc:creator>Mikulasova, A.</dc:creator>
<dc:creator>Astuti, G.</dc:creator>
<dc:creator>Gilissen, C. S.</dc:creator>
<dc:creator>McEleny, K.</dc:creator>
<dc:creator>Turner, H.</dc:creator>
<dc:creator>Coxhead, J.</dc:creator>
<dc:creator>Cockell, S. J.</dc:creator>
<dc:creator>Braat, D.</dc:creator>
<dc:creator>Fleischer, K.</dc:creator>
<dc:creator>D'Hauwers, K.</dc:creator>
<dc:creator>Schaafsma, E.</dc:creator>
<dc:creator>GEMINI Consortium,</dc:creator>
<dc:creator>Nagirnaja, L.</dc:creator>
<dc:creator>Conrad, D.</dc:creator>
<dc:creator>Friedrich, C.</dc:creator>
<dc:creator>Kliesch, S.</dc:creator>
<dc:creator>Aston, K. I.</dc:creator>
<dc:creator>Riera-Escamilla, A.</dc:creator>
<dc:creator>Krausz, C. G.</dc:creator>
<dc:creator>Gonzaga-Jauregui, C.</dc:creator>
<dc:creator>Santibanez-Koref, M.</dc:creator>
<dc:creator>Elliott, D.</dc:creator>
<dc:creator>Vissers, L.</dc:creator>
<dc:creator>Tüttelmann, F.</dc:creator>
<dc:creator>O'Bryan, M.</dc:creator>
<dc:creator>Ramos, L.</dc:creator>
<dc:creator>Xavier, M. J.</dc:creator>
<dc:creator>van der Heijden, G</dc:creator>
<dc:date>2021-02-27</dc:date>
<dc:identifier>doi:10.1101/2021.02.27.433155</dc:identifier>
<dc:title><![CDATA[A de novo paradigm for male infertility]]></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/10.1101/2021.03.02.433557v1?rss=1">
<title>
<![CDATA[
Distinct forms of synaptic plasticity during ascending vs. descending control of medial olivocochlear efferent neurons 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.03.02.433557v1?rss=1</link>
<description><![CDATA[
Activity in each brain region is shaped by the convergence of ascending and descending axonal pathways, and the balance and characteristics of these determine neural output. The medial olivocochlear (MOC) efferent system is part of a reflex arc that critically controls auditory sensitivity. Multiple central pathways contact MOC neurons, raising the question of how a reflex arc could be engaged by diverse inputs. We examined functional properties of synapses onto brainstem MOC neurons from ascending (ventral cochlear nucleus, VCN), and descending (inferior colliculus, IC) sources in mice using an optogenetic approach. We found that these pathways exhibited opposing forms of short-term plasticity, with VCN input showing depression and IC input showing marked facilitation. By using a conductance clamp approach, we found that combinations of facilitating and depressing inputs enabled firing of MOC neurons over a surprisingly wide dynamic range, suggesting an essential role for descending signaling to a brainstem nucleus.
]]></description>
<dc:creator>Romero, G. E.</dc:creator>
<dc:creator>Trussell, L. O.</dc:creator>
<dc:date>2021-03-02</dc:date>
<dc:identifier>doi:10.1101/2021.03.02.433557</dc:identifier>
<dc:title><![CDATA[Distinct forms of synaptic plasticity during ascending vs. descending control of medial olivocochlear efferent neurons]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-03-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.03.03.433701v1?rss=1">
<title>
<![CDATA[
CaSR modulates sodium channel-mediated Ca 2+ -dependent excitability 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.03.03.433701v1?rss=1</link>
<description><![CDATA[
Increasing extracellular [Ca2+] ([Ca2+]o) strongly decreases intrinsic excitability in neurons but the mechanism is unclear. By one hypothesis, [Ca2+]o screens surface charge reducing voltage-dependent sodium channel (VGSC) activation and by another [Ca2+]o activates Calcium-sensing receptor (CaSR) closing the sodium-leak channel (NALCN). Here we report that action potential (AP) firing rates increased in wild-type (WT), but not CaSR null mutant (Casr-/-) neocortical neurons, following the switch from physiological to reduced Ca2+-containing Tyrode. However, after membrane potential correction, AP firing increased similarly in both genotypes inconsistent with CaSR regulation of NALCN. Activation of VGSCs was the dominant contributor to the increase in excitability after the [Ca2+]o change. VGSC conductance-voltage relationships were hyperpolarized by decreasing [Ca2+]o for Casr-/- neurons indicating CaSR contributes to [Ca2+]o-dependent excitability via VGSCs. Regulation of VGSC gating by [Ca2+]o is the key mechanism mediating [Ca2+]o-dependent changes in neocortical neuron excitability and CaSR influences neuronal excitability by its effects on VGSC gating.
]]></description>
<dc:creator>Martiszus, B. J.</dc:creator>
<dc:creator>Tsintsadze, T.</dc:creator>
<dc:creator>Smith, S. M.</dc:creator>
<dc:creator>Chang, W.</dc:creator>
<dc:date>2021-03-04</dc:date>
<dc:identifier>doi:10.1101/2021.03.03.433701</dc:identifier>
<dc:title><![CDATA[CaSR modulates sodium channel-mediated Ca 2+ -dependent excitability]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-03-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.03.10.382333v1?rss=1">
<title>
<![CDATA[
Capturing scientific knowledge in computable form 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.03.10.382333v1?rss=1</link>
<description><![CDATA[
Technological advances in computing provide major opportunities to accelerate scientific discovery. The wide availability of structured knowledge would allow us to take full advantage of these by enabling efficient human-computer interaction. Traditionally, biological knowledge is captured in publications and knowledge bases, however, the information in articles is not directly accessible to computers, and knowledge bases are constrained by finite resources available for manual curation. To accelerate knowledge capture and communication and to keep pace with the rapid growth of scientific reports, we developed the Biofactoid (biofactoid.org) software suite, which crowdsources structured knowledge in articles from authors. Biofactoid is a web-based system that lets scientists draw a network of interactions between genes, their products, and chemical compounds and employs smart-automation to translate user input into a structured language using the expressive power of a formal ontology. The resulting data is shared via public information resources, enabling author-curated knowledge to be appreciated in the context of all existing computable knowledge. Authors of recently published papers across a range of journals have already contributed their pathway information, much of which is novel and extends existing pathway databases into new biological areas. We envision the adoption of Biofactoid for crowdsourced curation by scientists and publishers as part of an ecosystem of tools that accelerate scientific communication and discovery.

AvailabilityBiofactoid server at https://biofactoid.org
]]></description>
<dc:creator>Wong, J. V.</dc:creator>
<dc:creator>Franz, M.</dc:creator>
<dc:creator>Siper, M. C.</dc:creator>
<dc:creator>Fong, D.</dc:creator>
<dc:creator>Durupinar, F.</dc:creator>
<dc:creator>Dallago, C.</dc:creator>
<dc:creator>Luna, A.</dc:creator>
<dc:creator>Giorgi, J. M.</dc:creator>
<dc:creator>Rodchenkov, I.</dc:creator>
<dc:creator>Babur, O.</dc:creator>
<dc:creator>Gyori, B. M.</dc:creator>
<dc:creator>Bachman, J. A.</dc:creator>
<dc:creator>Demir, E.</dc:creator>
<dc:creator>Bader, G. D.</dc:creator>
<dc:creator>Sander, C.</dc:creator>
<dc:date>2021-03-11</dc:date>
<dc:identifier>doi:10.1101/2021.03.10.382333</dc:identifier>
<dc:title><![CDATA[Capturing scientific knowledge in computable form]]></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/10.1101/2021.03.10.434649v1?rss=1">
<title>
<![CDATA[
Single-Cell Spatial Proteomics Analyses of Head and Neck Squamous Cell Carcinoma Reveal Tumor Heterogeneity and Immune Architectures Associated with Clinical Outcome 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.03.10.434649v1?rss=1</link>
<description><![CDATA[
There is increasing evidence that the spatial organization of cells within the tumor-immune microenvironment (TiME) of solid tumors influences survival and response to therapy in numerous cancer types. Here, we report results and demonstrate the applicability of quantitative single-cell spatial proteomics analyses in the TiME of primary and recurrent human papillomavirus (HPV)-negative head and neck squamous cell carcinoma (HNSCC) tumors. Single-cell compositions of a nine patient, primary and recurrent (n=18), HNSCC cohort is presented, followed by deeper investigation into the spatial architecture of the TiME and its relationship with clinical variables and progression free survival (PFS). Multiple spatial algorithms were used to quantify the spatial landscapes of immune cells within TiMEs and demonstrate that neoplastic tumor-immune cell spatial compartmentalization, rather than mixing, is associated with longer PFS. Mesenchymal (SMA+) cellular neighborhoods describe distinct immune landscapes associated with neoplastic tumor-immune compartmentalization and improved patient outcomes. Results from this investigation are concordant with studies in other tumor types, suggesting that trends in TiME cellular heterogeneity and spatial organization may be shared across cancers and may provide prognostic value in multiple cancer types.
]]></description>
<dc:creator>Blise, K. E.</dc:creator>
<dc:creator>Sivagnanam, S.</dc:creator>
<dc:creator>Banik, G. L.</dc:creator>
<dc:creator>Coussens, L. M.</dc:creator>
<dc:creator>Goecks, J.</dc:creator>
<dc:date>2021-03-11</dc:date>
<dc:identifier>doi:10.1101/2021.03.10.434649</dc:identifier>
<dc:title><![CDATA[Single-Cell Spatial Proteomics Analyses of Head and Neck Squamous Cell Carcinoma Reveal Tumor Heterogeneity and Immune Architectures Associated with Clinical Outcome]]></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/10.1101/2021.03.11.434896v1?rss=1">
<title>
<![CDATA[
Relevance of Circulating Hybrid Cells as a Non-Invasive Biomarker for Myriad Solid Tumors 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.03.11.434896v1?rss=1</link>
<description><![CDATA[
AbstractMetastatic progression defines the final stages of tumor evolution and underlies the majority of cancer-related deaths. The heterogeneity in disseminated tumor cell populations capable of seeding and growing in distant organ sites contributes to the development of treatment resistant disease. We recently reported the identification of a novel tumor-derived cell population, circulating hybrid cells (CHCs), harboring attributes from both macrophages and neoplastic cells, including functional characteristics important to metastatic spread. These disseminated hybrids outnumber conventionally defined circulating tumor cells (CTCs) in cancer patients. It is unknown if CHCs represent a generalized cancer mechanism for cell dissemination, or if this population is relevant to the metastatic cascade. Herein, we detect CHCs in the peripheral blood of patients with cancer in myriad disease sites encompassing epithelial and non-epithelial malignancies. Further, we demonstrate that in vivo-derived hybrid cells harbor tumor-initiating capacity in murine cancer models and that CHCs from human breast cancer patients express stem cell antigens, features consistent with the ability to seed and grow at metastatic sites. Finally, we reveal heterogeneity of CHC phenotypes reflect key tumor features, including oncogenic mutations and functional protein expression. Importantly, this novel population of disseminated neoplastic cells opens a new area in cancer biology and renewed opportunity for battling metastatic disease.

Simple SummaryThere is an incomplete understanding of circulating neoplastic cell populations and the fundamental mechanisms that drive dissemination, immune evasion, and growth --all critical information to more effectively prevent and treat cancer progression. A novel disseminated tumor cell population, circulating hybrid cells, are detected across many cancer types and carry functional tumor-initiating properties. Additionally, circulating hybrid cells are found at significantly higher levels than conventionally defined circulating tumor cells. Our study demonstrates that neoplastic hybrid cells harbor phenotypic and genetic characteristics of tumor and immune cells, display stem features, and are a generalizable phenomenon in solid tumors. Circulating hybrid cells therefore have relevance as a novel biomarker and open a new field of study in malignancy.
]]></description>
<dc:creator>Dietz, M. S.</dc:creator>
<dc:creator>Sutton, T. L.</dc:creator>
<dc:creator>Walker, B. S.</dc:creator>
<dc:creator>Gast, C. E.</dc:creator>
<dc:creator>Zarour, L.</dc:creator>
<dc:creator>Sengupta, S. K.</dc:creator>
<dc:creator>Swain, J. R.</dc:creator>
<dc:creator>Eng, J.</dc:creator>
<dc:creator>Parappilly, M.</dc:creator>
<dc:creator>Limbach, K.</dc:creator>
<dc:creator>Sattler, A.</dc:creator>
<dc:creator>Burlingame, E.</dc:creator>
<dc:creator>Chin, Y.</dc:creator>
<dc:creator>Gower, A.</dc:creator>
<dc:creator>Montoya Mira, J. L.</dc:creator>
<dc:creator>Sapre, A.</dc:creator>
<dc:creator>Chiu, Y.-J.</dc:creator>
<dc:creator>Clayburgh, D. R.</dc:creator>
<dc:creator>Pommier, S. J.</dc:creator>
<dc:creator>Cetnar, J. P.</dc:creator>
<dc:creator>Fischer, J. M.</dc:creator>
<dc:creator>Jaboin, J. J.</dc:creator>
<dc:creator>Pommier, R. F.</dc:creator>
<dc:creator>Sheppard, B. C.</dc:creator>
<dc:creator>Tsikitis, V. L.</dc:creator>
<dc:creator>Skalet, A. H.</dc:creator>
<dc:creator>Mayo, S. C.</dc:creator>
<dc:creator>Lopez, C. D.</dc:creator>
<dc:creator>Gray, J. W.</dc:creator>
<dc:creator>Mills, G. B.</dc:creator>
<dc:creator>Mitri, Z.</dc:creator>
<dc:creator>Chang, Y. H.</dc:creator>
<dc:creator>Chin, K.</dc:creator>
<dc:creator>Wong, M. H.</dc:creator>
<dc:date>2021-03-12</dc:date>
<dc:identifier>doi:10.1101/2021.03.11.434896</dc:identifier>
<dc:title><![CDATA[Relevance of Circulating Hybrid Cells as a Non-Invasive Biomarker for Myriad Solid Tumors]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-03-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.03.13.435270v1?rss=1">
<title>
<![CDATA[
Using synthetic MR images for field map-less distortion correction 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.03.13.435270v1?rss=1</link>
<description><![CDATA[
Functional MRI (fMRI) data acquired using echo-planar imaging (EPI) are highly distorted by magnetic field inhomogeneities. Distortion combined with underlying differences in image contrast between EPI and T1-weighted and T2-weighted (T1w/T2w) structural images makes the alignment of functional and anatomical images a challenge. Typically, separately acquired field map data are used to correct fMRI distortions and a flexible cost function insensitive to cross-modal differences in image contrast and intensity is used for aligning fMRI and anatomical images. The quality of alignment achieved with this approach can vary greatly and depends on the quality of field map data. In addition, many publicly available datasets lack field map data entirely. To address this issue, we developed Synth, a software package for distortion correction and cross-modal image registration that does not require separately acquired field map data. Synth combines information from T1w and T2w anatomical images to construct an idealized undistorted synthetic image that has similar contrast properties to fMRI data. The undistorted synthetic image then serves as an effective reference for individual-specific nonlinear unwarping to correct fMRI distortions. We demonstrate, in both pediatric (ABCD: Adolescent Brain Cognitive Development) and adult (MSC: Midnight Scan Club) data that Synth performs comparably well to other leading distortion correction approaches that utilize field map data, and often outperforms them. Field map-less distortion correction with Synth allows accurate and precise registration of fMRI data with missing or corrupted field map information.
]]></description>
<dc:creator>Montez, D. F.</dc:creator>
<dc:creator>Van, A. N.</dc:creator>
<dc:creator>Miller, R. L.</dc:creator>
<dc:creator>Seider, N. A.</dc:creator>
<dc:creator>Marek, S.</dc:creator>
<dc:creator>Zheng, A.</dc:creator>
<dc:creator>Newbold, D. J.</dc:creator>
<dc:creator>Scheidter, K.</dc:creator>
<dc:creator>Feczko, E.</dc:creator>
<dc:creator>Perrone, A. J.</dc:creator>
<dc:creator>Miranda Dominguez, O.</dc:creator>
<dc:creator>Earl, E. A.</dc:creator>
<dc:creator>Kay, B. P.</dc:creator>
<dc:creator>Jha, A. K.</dc:creator>
<dc:creator>Sotiras, A.</dc:creator>
<dc:creator>Laumann, T. O.</dc:creator>
<dc:creator>Greene, D. J.</dc:creator>
<dc:creator>Gordon, E. M.</dc:creator>
<dc:creator>Tisdall, M. D.</dc:creator>
<dc:creator>van der Kouwe, A.</dc:creator>
<dc:creator>Fair, D. A.</dc:creator>
<dc:creator>Dosenbach, N. U. F.</dc:creator>
<dc:date>2021-03-15</dc:date>
<dc:identifier>doi:10.1101/2021.03.13.435270</dc:identifier>
<dc:title><![CDATA[Using synthetic MR images for field map-less distortion correction]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-03-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.03.14.435295v1?rss=1">
<title>
<![CDATA[
3D genomic capture of regulatory immuno-genetic profiles in COVID-19 patients for prognosis of severe COVID disease outcome 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.03.14.435295v1?rss=1</link>
<description><![CDATA[
Human infection with the SARS-CoV-2 virus leads to coronavirus disease (COVID-19). A striking characteristic of COVID-19 infection in humans is the highly variable host response and the diverse clinical outcomes, ranging from clinically asymptomatic to severe immune reactions leading to hospitalization and death. Here we used a 3D genomic approach to analyse blood samples at the time of COVID diagnosis, from a global cohort of 80 COVID-19 patients, with different degrees of clinical disease outcomes. Using 3D whole genome EpiSwitch(R) arrays to generate over 1 million data points per patient, we identified a distinct and measurable set of differences in genomic organization at immune-related loci that demonstrated prognostic power at baseline to stratify patients with mild forms of illness and those with severe forms that required hospitalization and intensive care unit (ICU) support. Further analysis revealed both well established and new COVID-related dysregulated pathways and loci, including innate and adaptive immunity; ACE2; olfactory, G{beta}{psi}, Ca2+ and nitric oxide (NO) signalling; prostaglandin E2 (PGE2), the acute inflammatory cytokine CCL3, and the T-cell derived chemotactic cytokine CCL5. We identified potential therapeutic agents for mitigation of severe disease outcome, with several already being tested independently, including mTOR inhibitors (rapamycin and tacrolimus) and general immunosuppressants (dexamethasone and hydrocortisone). Machine learning algorithms based on established EpiSwitch(R) methodology further identified a subset of 3D genomic changes that could be used as prognostic molecular biomarker leads for the development of a COVID-19 disease severity test.
]]></description>
<dc:creator>Hunter, E.</dc:creator>
<dc:creator>Koutsothanasi, C.</dc:creator>
<dc:creator>Wilson, A.</dc:creator>
<dc:creator>Santos, F. C.</dc:creator>
<dc:creator>Salter, M.</dc:creator>
<dc:creator>Powell, R.</dc:creator>
<dc:creator>Dring, A.</dc:creator>
<dc:creator>Brajer, P.</dc:creator>
<dc:creator>Egan, B.</dc:creator>
<dc:creator>Westra, J.</dc:creator>
<dc:creator>Ramadass, A.</dc:creator>
<dc:creator>Messner, W.</dc:creator>
<dc:creator>Brunton, A.</dc:creator>
<dc:creator>Lyski, Z.</dc:creator>
<dc:creator>Vancheeswaran, R.</dc:creator>
<dc:creator>Barlow, A.</dc:creator>
<dc:creator>Pchejetski, D.</dc:creator>
<dc:creator>Robbins, P.</dc:creator>
<dc:creator>Mellor, J.</dc:creator>
<dc:creator>Akoulitchev, A.</dc:creator>
<dc:date>2021-03-16</dc:date>
<dc:identifier>doi:10.1101/2021.03.14.435295</dc:identifier>
<dc:title><![CDATA[3D genomic capture of regulatory immuno-genetic profiles in COVID-19 patients for prognosis of severe COVID disease outcome]]></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/10.1101/2021.03.15.435473v1?rss=1">
<title>
<![CDATA[
MCMICRO: A scalable, modular image-processing pipeline for multiplexed tissue imaging 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.03.15.435473v1?rss=1</link>
<description><![CDATA[
Highly multiplexed tissue imaging makes molecular analysis of single cells possible in a preserved spatial context. However, reproducible analysis of the underlying data poses a substantial computational challenge. Here we describe a modular and open-source computational pipeline (MCMICRO) for performing the sequential steps needed to transform large, multi-channel whole slide images into single-cell data. We demonstrate use of MCMICRO on images of different tissues and tumors acquired using multiple imaging platforms, thereby providing a solid foundation for the continued development of tissue imaging software.
]]></description>
<dc:creator>Schapiro, D.</dc:creator>
<dc:creator>Sokolov, A.</dc:creator>
<dc:creator>Yapp, C.</dc:creator>
<dc:creator>Muhlich, J. L.</dc:creator>
<dc:creator>Hess, J.</dc:creator>
<dc:creator>Lin, J.-R.</dc:creator>
<dc:creator>Chen, Y.-A.</dc:creator>
<dc:creator>Nariya, M. K.</dc:creator>
<dc:creator>Baker, G. J.</dc:creator>
<dc:creator>Ruokonen, J.</dc:creator>
<dc:creator>Maliga, Z.</dc:creator>
<dc:creator>Jacobson, C. A.</dc:creator>
<dc:creator>Farhi, S. L.</dc:creator>
<dc:creator>Abbondanza, D.</dc:creator>
<dc:creator>McKinley, E. T.</dc:creator>
<dc:creator>Betts, C.</dc:creator>
<dc:creator>Regev, A.</dc:creator>
<dc:creator>Coffey, R. J.</dc:creator>
<dc:creator>Coussens, L. M.</dc:creator>
<dc:creator>Santagata, S.</dc:creator>
<dc:creator>Sorger, P. K.</dc:creator>
<dc:date>2021-03-16</dc:date>
<dc:identifier>doi:10.1101/2021.03.15.435473</dc:identifier>
<dc:title><![CDATA[MCMICRO: A scalable, modular image-processing pipeline for multiplexed tissue imaging]]></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/10.1101/2021.03.16.435600v1?rss=1">
<title>
<![CDATA[
BCL-XL blockage in TNBC models confers vulnerability to inhibition of specific cell cycle regulators 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.03.16.435600v1?rss=1</link>
<description><![CDATA[
Cell cycle regulators are frequently altered in Triple-Negative Breast Cancer (TNBC). Emerging agents targeting these signals offer the possibility to design new combinatorial therapies. However, preclinical models that recapitulate TNBC primary resistance and heterogeneity are essential to evaluate the potency of these combined treatments.

MethodsBioinformatic processing of human breast cancer datasets was used to analyse correlations between expression levels of cell cycle regulators and patient survival outcome. The MMTV-R26Met mouse model of TNBC resistance and heterogeneity was employed to analyse expression and targeting vulnerability of cell cycle regulators in the presence of BCL-XL blockage. Robustness of outcomes and selectivity was further explored using a panel of human breast cancer cells. Alterations of protein expression, phosphorylation, and/or cellular localisation were analysed by western blots, reverse phase protein array, and immunocytochemistry. Bioinformatics was performed to highlight drugs mechanisms of action.

ResultsWe report that high expression levels of BCL-XL and specific cell cycle regulators correlate with poor survival outcomes of TNBC patients. Blockage of BCL-XL confers vulnerability to drugs targeting CDK1/2/4, but not FOXM1, CDK4/6, Aurora A and Aurora B, to all MMTV-R26Met and human TNBC cell lines tested. Mechanistically, we show that, co-targeting of BCL-XL and CDK1/2/4 synergistically inhibited cell growth by combinatorial depletion of survival and RTK/AKT signals, and concomitantly restoring FOXO3a tumour suppression actions. This was accompanied by an accumulation of DNA damage and consequently apoptosis.

ConclusionsOur studies illustrate the possibility to exploit the vulnerability of TNBC cells to CDK1/2/4 inhibition by targeting BCL-XL. Moreover, they underline that specificity matters in targeting cell cycle regulators for combinatorial anticancer therapies.
]]></description>
<dc:creator>Castellanet, O.</dc:creator>
<dc:creator>Ahmad, F.</dc:creator>
<dc:creator>Vinik, Y.</dc:creator>
<dc:creator>Mills, G. B.</dc:creator>
<dc:creator>Habermann, B. H.</dc:creator>
<dc:creator>Borg, J.-P. H.</dc:creator>
<dc:creator>Lev, S.</dc:creator>
<dc:creator>Lamballe, F.</dc:creator>
<dc:creator>Maina, F.</dc:creator>
<dc:date>2021-03-17</dc:date>
<dc:identifier>doi:10.1101/2021.03.16.435600</dc:identifier>
<dc:title><![CDATA[BCL-XL blockage in TNBC models confers vulnerability to inhibition of specific cell cycle regulators]]></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/10.1101/2021.03.23.436530v1?rss=1">
<title>
<![CDATA[
Generation of functional ciliates cholangiocytes from human pluripotent stem cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.03.23.436530v1?rss=1</link>
<description><![CDATA[
The derivation of mature functional cholangiocytes from human pluripotent stem cells (hPSCs) would provide a model for studying the pathogenesis of cholangiopathies and for developing novel therapies to treat them. Current differentiation protocols are not efficient and give rise to cholangiocytes that are not fully mature, limiting their therapeutic applications. Here, we describe a new strategy to generate functional hPSC-derived cholangiocytes that display many characteristics of mature bile duct cells including high levels of CFTR and the presence of primary cilia capable of sensing flow. With this level of maturation, these cholangiocytes are amenable for testing the efficacy of new cystic fibrosis drugs and for studying the role of cilia in cholangiocyte development and function. Transplantation studies showed that the mature cholangiocytes generate ductal structures in the liver of immunocompromised mice providing the first indication that it may be possible to develop cell-based therapies to restore bile duct function in patients with biliary disease.
]]></description>
<dc:creator>Ogawa, M.</dc:creator>
<dc:creator>Jiang, J.-X.</dc:creator>
<dc:creator>Xia, S.</dc:creator>
<dc:creator>Yang, D.</dc:creator>
<dc:creator>Ding, A.</dc:creator>
<dc:creator>Laselva, O.</dc:creator>
<dc:creator>Chin, S.</dc:creator>
<dc:creator>Hernandez, M.</dc:creator>
<dc:creator>Cui, C.</dc:creator>
<dc:creator>Higuchi, Y.</dc:creator>
<dc:creator>Suemizu, H.</dc:creator>
<dc:creator>Dorrell, C.</dc:creator>
<dc:creator>Grompe, M.</dc:creator>
<dc:creator>Bear, C.</dc:creator>
<dc:creator>Keller, G.</dc:creator>
<dc:creator>Ogawa, S.</dc:creator>
<dc:date>2021-03-23</dc:date>
<dc:identifier>doi:10.1101/2021.03.23.436530</dc:identifier>
<dc:title><![CDATA[Generation of functional ciliates cholangiocytes from human pluripotent stem cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-03-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.03.25.437080v1?rss=1">
<title>
<![CDATA[
α2δ-2 is Required for Functional Postsynaptic Calcium Channel Nanodomain Signaling 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.03.25.437080v1?rss=1</link>
<description><![CDATA[
2{delta} proteins (CACNA2D1-4) are required for normal neurological function, although how they control neuronal output remains unclear. Using whole-cell recordings of mouse Purkinje cells, we show 2{delta}-2 is required for functional coupling of postsynaptic voltage-dependent calcium entry with effector mechanisms controlling two different outputs, depolarization-induced suppression of excitation mediated by endocannabinoid signaling, and spike afterhyperpolarization generated by calcium-dependent potassium channels. Our findings indicate an important role for 2{delta}-2 proteins in regulating functional postsynaptic calcium channel-coupling in neurons.

Significance StatementCalcium influx via membrane voltage-dependent calcium channels drives numerous neuronal processes by signaling through calcium-dependent effector molecules. Signal precision is achieved in part by calcium channel-effector coupling. In mouse Purkinje cell neurons, we show that neuronal 2{delta}-2 protein functionally couples calcium entry to two different postsynaptic calcium-dependent signals, retrograde endocannabinoid signaling and the action potential afterhyperpolarization. Our findings provide new insights about the control of calcium channel-effector coupling as well as new roles for 2{delta}-2 proteins in neurons.
]]></description>
<dc:creator>Beeson, K.</dc:creator>
<dc:creator>Westbrook, G.</dc:creator>
<dc:creator>Schnell, E.</dc:creator>
<dc:date>2021-03-26</dc:date>
<dc:identifier>doi:10.1101/2021.03.25.437080</dc:identifier>
<dc:title><![CDATA[α2δ-2 is Required for Functional Postsynaptic Calcium Channel Nanodomain Signaling]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-03-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.03.30.437775v1?rss=1">
<title>
<![CDATA[
Novel insights in the pathophysiology of α-synuclein dysregulation on D2 receptor activity contributing to the vulnerability of dopamine neurons 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.03.30.437775v1?rss=1</link>
<description><![CDATA[
Pathophysiological damages and loss of function of dopamine neurons precedes their demise and contributes to the early phases of Parkinsons disease. The presence of aberrant intercellular pathological inclusions of the protein -synuclein within ventral midbrain dopaminergic neurons is one of the cardinal features of Parkinsons disease. We employed multiple complementary approaches in molecular biology, electrophysiology, and live-cell imaging to investigate how excessive -synuclein levels alters multiple characteristics of dopaminergic neuronal dynamics and dopamine transmission prior to neuronal demise. These studies demonstrate that -synuclein dysregulation of D2 receptor autoinhibition contributes to the vulnerability of dopaminergic neurons, and that modulation thereof can ameliorate the resulting pathophysiology. These novel findings provide mechanistic insights in the insidious loss of dopaminergic function and neurons that characterize Parkinsons disease progression with significant therapeutic implications.
]]></description>
<dc:creator>Dagra, A.</dc:creator>
<dc:creator>Miller, D. R.</dc:creator>
<dc:creator>Shaerzadeh, F.</dc:creator>
<dc:creator>Lin, M.</dc:creator>
<dc:creator>Gopinath, A.</dc:creator>
<dc:creator>Harris, S.</dc:creator>
<dc:creator>Sorrentino, Z. A.</dc:creator>
<dc:creator>Velasco, S.</dc:creator>
<dc:creator>Alonge, A. R.</dc:creator>
<dc:creator>Azar, J.</dc:creator>
<dc:creator>Lebowitz, J. J.</dc:creator>
<dc:creator>Ulm, B.</dc:creator>
<dc:creator>Bu, A.-M.</dc:creator>
<dc:creator>Hansen, C. A.</dc:creator>
<dc:creator>Urs, N. M.</dc:creator>
<dc:creator>Giasson, B. I.</dc:creator>
<dc:creator>Khoshbouei, H.</dc:creator>
<dc:date>2021-03-30</dc:date>
<dc:identifier>doi:10.1101/2021.03.30.437775</dc:identifier>
<dc:title><![CDATA[Novel insights in the pathophysiology of α-synuclein dysregulation on D2 receptor activity contributing to the vulnerability of dopamine neurons]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-03-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.03.31.438003v1?rss=1">
<title>
<![CDATA[
Effective Therapy Targeting Cytochrome bc1 Prevents Babesia Erythrocytic Development and Protects from Lethal Infection 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.03.31.438003v1?rss=1</link>
<description><![CDATA[
Targeting conserved metabolic processes that are essential for viability of pathogens, such as Plasmodium and Babesia that cause blood-borne diseases, is an effective strategy to eliminate malaria and babesiosis infections with no recrudescence. One interesting target is the mitochondrial cytochrome bc1 complex, which could be inhibited by drugs such as endochin-like quinolones (ELQ) and atovaquone. We used the tick-transmitted and culturable blood-borne pathogen Babesia duncani to evaluate the structure-activity relationship, safety, efficacy and mode of action of ELQs. We identified a potent and highly selective ELQ prodrug (ELQ-502), which alone or in combination with atovaquone eliminates B. microti and B. duncani infections in vitro and in mouse models of parasitemia and lethal infection. The strong efficacy at low dose, excellent safety, bioavailability and long half-life of this experimental therapy makes it an ideal clinical candidate for the treatment of human infections caused by Babesia and its closely related apicomplexan parasites.
]]></description>
<dc:creator>Chiu, J. E.</dc:creator>
<dc:creator>Renard, I.</dc:creator>
<dc:creator>Chattopadhyay Pal, A.</dc:creator>
<dc:creator>Singh, P.</dc:creator>
<dc:creator>Vydyam, P.</dc:creator>
<dc:creator>Thekkiniath, J.</dc:creator>
<dc:creator>Kumar, M.</dc:creator>
<dc:creator>Gihaz, S.</dc:creator>
<dc:creator>Pou, S.</dc:creator>
<dc:creator>Winter, R. W.</dc:creator>
<dc:creator>Dodean, R.</dc:creator>
<dc:creator>Nilsen, A.</dc:creator>
<dc:creator>Riscoe, M. K.</dc:creator>
<dc:creator>Doggett, J. S.</dc:creator>
<dc:creator>Ben Mamoun, C.</dc:creator>
<dc:date>2021-04-01</dc:date>
<dc:identifier>doi:10.1101/2021.03.31.438003</dc:identifier>
<dc:title><![CDATA[Effective Therapy Targeting Cytochrome bc1 Prevents Babesia Erythrocytic Development and Protects from Lethal Infection]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-04-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.04.01.438108v1?rss=1">
<title>
<![CDATA[
Acetylated α-tubulin residue K394 regulates microtubule stability to shape the growth of axon terminals 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.04.01.438108v1?rss=1</link>
<description><![CDATA[
Microtubules are essential to neuron shape and function. Therefore, the stability of the microtubule cytoskeleton must be carefully regulated. Acetylation of tubulin has the potential to directly tune microtubule stability, and proteomic studies have identified several acetylation sites in -tubulin. This includes the highly conserved residue lysine 394 (K394), which is located at the {beta}-tubulin dimer interface. Using a fly model, we show that -tubulin K394 is acetylated in the nervous system and is an essential residue. We found that an acetylation-blocking mutation in endogenous -tubulin, K394R, perturbs the synaptic morphogenesis of motoneurons by reducing microtubule stability. Intriguingly, the K394R mutation has opposite effects on the growth of two functionally and morphologically distinct motoneurons, revealing neuron-type-specific responses when microtubule stability is altered. Eliminating the deacetylase HDAC6 increases K394 acetylation, and the over-expression of HDAC6 reduces microtubule stability similar to the K394 mutant. Thus, our findings implicate -tubulin K394 and its acetylation in the regulation of microtubule stability and suggest that HDAC6 regulates K394 acetylation during synaptic morphogenesis.
]]></description>
<dc:creator>Saunders, H. A. J.</dc:creator>
<dc:creator>Johnson-Schlitz, D. M.</dc:creator>
<dc:creator>Jenkins, B. V.</dc:creator>
<dc:creator>Volkert, P. J.</dc:creator>
<dc:creator>Yang, S. Z.</dc:creator>
<dc:creator>Wildonger, J.</dc:creator>
<dc:date>2021-04-02</dc:date>
<dc:identifier>doi:10.1101/2021.04.01.438108</dc:identifier>
<dc:title><![CDATA[Acetylated α-tubulin residue K394 regulates microtubule stability to shape the growth of axon terminals]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-04-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.04.03.437254v1?rss=1">
<title>
<![CDATA[
Early embryonic loss following intravaginal Zika virus challenge in rhesus macaques 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.04.03.437254v1?rss=1</link>
<description><![CDATA[
Zika virus (ZIKV) is an arthropod-borne virus (arbovirus) and is primarily transmitted by Aedes species mosquitoes; however, ZIKV can also be sexually transmitted. During the initial epidemic and in places where ZIKV is now considered endemic, it is difficult to disentangle the risks and contributions of sexual versus vector-borne transmission to adverse pregnancy outcomes. To examine the potential impact of sexual transmission of ZIKV on pregnancy outcome, we challenged three rhesus macaques (Macaca mulatta) three times intravaginally with 1 x 107 PFU of a low passage, African lineage ZIKV isolate (ZIKV-DAK) in the first trimester ([~]30 days gestational age). Samples were collected from all animals initially on days 3 through 10 post challenge, followed by twice, and then once weekly sample collection; ultrasound examinations were performed every 3-4 days then weekly as pregnancies progressed. All three dams had ZIKV RNA detectable in plasma on day 3 post-ZIKV challenge. At approximately 45 days gestation (17-18 days post-challenge), two of the three dams were found to have nonviable embryos by ultrasound. Viral RNA was detected in recovered tissues and at the maternal-fetal interface (MFI) in both cases. The remaining viable pregnancy proceeded to near term ([~]155 days gestational age) and ZIKV RNA was detected at the MFI but not in fetal tissues. These results suggest that sexual transmission of ZIKV may represent an underappreciated risk of pregnancy loss during early gestation.
]]></description>
<dc:creator>Newman, C. M.</dc:creator>
<dc:creator>Tarantal, A. F.</dc:creator>
<dc:creator>Martinez, M. L.</dc:creator>
<dc:creator>Simmons, H. A.</dc:creator>
<dc:creator>Morgan, T. K.</dc:creator>
<dc:creator>Zeng, X.</dc:creator>
<dc:creator>Rosinski, J. R.</dc:creator>
<dc:creator>Bliss, M. I.</dc:creator>
<dc:creator>Bohm, E. K.</dc:creator>
<dc:creator>Dudley, D. M.</dc:creator>
<dc:creator>Aliota, M.</dc:creator>
<dc:creator>Friedrich, T.</dc:creator>
<dc:creator>Miller, C. J.</dc:creator>
<dc:creator>O'Connor, D.</dc:creator>
<dc:date>2021-04-03</dc:date>
<dc:identifier>doi:10.1101/2021.04.03.437254</dc:identifier>
<dc:title><![CDATA[Early embryonic loss following intravaginal Zika virus challenge in rhesus macaques]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-04-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.04.04.438397v1?rss=1">
<title>
<![CDATA[
An aging clock using metabolomic CSF 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.04.04.438397v1?rss=1</link>
<description><![CDATA[
Quantifying the physiology of aging is essential for improving our understanding of age-related disease and the heterogeneity of healthy aging. Recent studies have shown that in regression models using "-omic" platforms to predict chronological age, residual variation in predicted age is correlated with health outcomes, and suggest that these "omic clocks" provide measures of biological age. This paper presents predictive models for age using metabolomic profiles of cerebrospinal fluid from healthy human subjects, and finds that metabolite and lipid data are generally able to predict chronological age within 10 years. We use these models to predict the age of a cohort of subjects with Alzheimers and Parkinsons disease and find an increase in prediction error, potentially indicating that the relationship between the metabolome and chronological age differs with these diseases. In our analysis of control subjects, we find the carnitine shuttle, sucrose, biopterin, vitamin E metabolism, tryptophan, and tyrosine to be the most associated with age. We showcase the potential usefulness of age prediction models in a small dataset (n = 85), and discuss techniques for drift correction, missing data imputation, and regularized regression which can be used to help mitigate the statistical challenges that commonly arise in this setting. To our knowledge, this work presents the first multivariate predictive metabolomic and lipidomic models for age using mass spectrometry analysis of cerebrospinal fluid.
]]></description>
<dc:creator>Hwangbo, N.</dc:creator>
<dc:creator>Zhang, X.</dc:creator>
<dc:creator>Raftery, D.</dc:creator>
<dc:creator>Gu, H.</dc:creator>
<dc:creator>Hu, S.-C.</dc:creator>
<dc:creator>Montine, T. J.</dc:creator>
<dc:creator>Quinn, J. F.</dc:creator>
<dc:creator>Chung, K. A.</dc:creator>
<dc:creator>Hiller, A. L.</dc:creator>
<dc:creator>Wang, D.</dc:creator>
<dc:creator>Fei, Q.</dc:creator>
<dc:creator>Bettcher, L.</dc:creator>
<dc:creator>Zabetian, C. P.</dc:creator>
<dc:creator>Peskind, E.</dc:creator>
<dc:creator>Li, G.</dc:creator>
<dc:creator>Promislow, D. E. L.</dc:creator>
<dc:creator>Franks, A.</dc:creator>
<dc:date>2021-04-05</dc:date>
<dc:identifier>doi:10.1101/2021.04.04.438397</dc:identifier>
<dc:title><![CDATA[An aging clock using metabolomic CSF]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-04-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.04.04.438418v1?rss=1">
<title>
<![CDATA[
Distinct in vivo dynamics of excitatory synapses onto cortical pyramidal neurons and inhibitory interneurons 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.04.04.438418v1?rss=1</link>
<description><![CDATA[
Cortical function relies on the balanced activation of excitatory and inhibitory neurons. However, little is known about the organization and dynamics of shaft excitatory synapses onto cortical inhibitory interneurons, which cannot be easily identified morphologically. Here, we fluorescently visualize the excitatory postsynaptic marker PSD-95 at endogenous levels as a proxy for excitatory synapses onto layer 2/3 pyramidal neurons and parvalbumin-positive (PV+) inhibitory interneurons in the mouse barrel cortex. Longitudinal in vivo imaging reveals that, while synaptic weights in both neuronal types are log-normally distributed, synapses onto PV+ neurons are less heterogeneous and more stable. Markov-model analyses suggest that the synaptic weight distribution is set intrinsically by ongoing cell type-specific dynamics, and substantial changes are due to accumulated gradual changes. Synaptic weight dynamics are multiplicative, i.e., changes scale with weights, though PV+ synapses also exhibit an additive component. These results reveal that cell type-specific processes govern cortical synaptic strengths and dynamics.
]]></description>
<dc:creator>Melander, J. B.</dc:creator>
<dc:creator>Nayebi, A.</dc:creator>
<dc:creator>Jongbloets, B. C.</dc:creator>
<dc:creator>Fortin, D. A.</dc:creator>
<dc:creator>Qin, M.</dc:creator>
<dc:creator>Ganguli, S.</dc:creator>
<dc:creator>Mao, T.</dc:creator>
<dc:creator>Zhong, H.</dc:creator>
<dc:date>2021-04-05</dc:date>
<dc:identifier>doi:10.1101/2021.04.04.438418</dc:identifier>
<dc:title><![CDATA[Distinct in vivo dynamics of excitatory synapses onto cortical pyramidal neurons and inhibitory interneurons]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-04-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.04.09.438906v1?rss=1">
<title>
<![CDATA[
False gene and chromosome losses affected by assembly and sequence errors 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.04.09.438906v1?rss=1</link>
<description><![CDATA[
Many genome assemblies have been found to be incomplete and contain misassemblies. The Vertebrate Genomes Project (VGP) has been producing assemblies with an emphasis on being as complete and error-free as possible, utilizing long reads, long-range scaffolding data, new assembly algorithms, and manual curation. Here we evaluate these new vertebrate genome assemblies relative to the previous references for the same species, including a mammal (platypus), two birds (zebra finch, Annas hummingbird), and a fish (climbing perch). We found that 3 to 11% of genomic sequence was entirely missing in the previous reference assemblies, which included nearly entire GC-rich and repeat-rich microchromosomes with high gene density. Genome-wide, between 25 to 60% of the genes were either completely or partially missing in the previous assemblies, and this was in part due to a bias in GC-rich 5-proximal promoters and 5 exon regions. Our findings reveal novel regulatory landscapes and protein coding sequences that have been greatly underestimated in previous assemblies and are now present in the VGP assemblies.
]]></description>
<dc:creator>Kim, J.</dc:creator>
<dc:creator>Lee, C.</dc:creator>
<dc:creator>Ko, B. J.</dc:creator>
<dc:creator>Yoo, D.</dc:creator>
<dc:creator>Won, S.</dc:creator>
<dc:creator>Phillippy, A.</dc:creator>
<dc:creator>Fedrigo, O.</dc:creator>
<dc:creator>Zhang, G.</dc:creator>
<dc:creator>Howe, K.</dc:creator>
<dc:creator>Wood, J.</dc:creator>
<dc:creator>Durbin, R.</dc:creator>
<dc:creator>Formenti, G.</dc:creator>
<dc:creator>Brown, S.</dc:creator>
<dc:creator>Cantin, L.</dc:creator>
<dc:creator>Mello, C. V.</dc:creator>
<dc:creator>Cho, S.</dc:creator>
<dc:creator>Rhie, A.</dc:creator>
<dc:creator>Kim, H.</dc:creator>
<dc:creator>Jarvis, E. D.</dc:creator>
<dc:date>2021-04-09</dc:date>
<dc:identifier>doi:10.1101/2021.04.09.438906</dc:identifier>
<dc:title><![CDATA[False gene and chromosome losses affected by assembly and sequence errors]]></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/10.1101/2021.04.10.439295v1?rss=1">
<title>
<![CDATA[
High-speed widefield handheld swept-source OCT angiography with a VCSEL light source 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.04.10.439295v1?rss=1</link>
<description><![CDATA[
Optical coherence tomography (OCT) and OCT angiography (OCTA) enable noninvasive structural and angiographic imaging of the eye. Portable handheld OCT/OCTA systems are required for imaging patients in the supine position. Examples include infants in the neonatal intensive care unit (NICU) and operating room (OR). The speed of image acquisition plays a pivotal role in acquiring high quality OCT/OCTA images, particularly with the handheld system, since both the operator hand tremor and subject motion can cause significant motion artifacts. In addition, having a large field of view and the ability of real-time data visualization are critical elements in rapid disease screening, reducing imaging time, and detecting peripheral retinal pathologies. The arrangement of optical components is less flexible in the handheld system due to the limitation of size and weight. In this paper, we introduce a 400-kHz, 55-degree field of view handheld OCT/OCTA system that has overcome many technical challenges as a portable OCT system as well as a high-speed OCTA system. We demonstrate imaging premature infants with retinopathy of prematurity (ROP) in the NICU, and patients with incontinentia pigmenti (IP) in the OR using our handheld OCT system. Our design may have potential for improving the diagnosis of retinal diseases and help provide a practical guideline for designing a flexible and portable OCT system.
]]></description>
<dc:creator>Ni, S.</dc:creator>
<dc:creator>Wei, X.</dc:creator>
<dc:creator>Ng, R.</dc:creator>
<dc:creator>Ostmo, S.</dc:creator>
<dc:creator>Chiang, M.</dc:creator>
<dc:creator>Huang, D.</dc:creator>
<dc:creator>Jia, Y.</dc:creator>
<dc:creator>Campbell, P.</dc:creator>
<dc:creator>Jian, Y.</dc:creator>
<dc:date>2021-04-11</dc:date>
<dc:identifier>doi:10.1101/2021.04.10.439295</dc:identifier>
<dc:title><![CDATA[High-speed widefield handheld swept-source OCT angiography with a VCSEL light source]]></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/10.1101/2021.04.11.439331v1?rss=1">
<title>
<![CDATA[
A complex feature-based representation of vocalizations emerges in the superficial layers of primary auditory cortex 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.04.11.439331v1?rss=1</link>
<description><![CDATA[
Early in auditory processing, neural responses faithfully reflect acoustic input. At higher stages of auditory processing, however, neurons become selective for particular call types, eventually leading to specialized regions of cortex that preferentially process calls at the highest auditory processing stages. We previously proposed that an intermediate step in how non-selective responses are transformed into call-selective responses is the detection of informative call features. But how neural selectivity for informative call features emerges from non-selective inputs, whether feature selectivity gradually emerges over the processing hierarchy, and how stimulus information is represented in non-selective and feature-selective populations remain open questions. In this study, using unanesthetized guinea pigs, a highly vocal and social rodent, as an animal model, we characterized the neural representation of calls in three auditory processing stages - the thalamus (vMGB), and thalamorecipient (L4) and superficial layers (L2/3) of primary auditory cortex (A1). We found that neurons in vMGB and A1 L4 did not exhibit call-selective responses and responded throughout the call durations. However, A1 L2/3 neurons showed high call-selectivity with about a third of neurons responding to only one or two call types. These A1 L2/3 neurons only responded to restricted portions of calls suggesting that they were highly selective for call features. Receptive fields of these A1 L2/3 neurons showed complex spectrotemporal structures that could underlie their high call feature selectivity. Information theoretic analysis revealed that in A1 L4 stimulus information was distributed over the population and was spread out over the call durations. In contrast, in A1 L2/3, individual neurons showed brief bursts of high stimulus-specific information, and conveyed high levels of information per spike. These data demonstrate that a transformation in the neural representation of calls occurs between A1 L4 and A1 L2/3, leading to the emergence of a feature-based representation of calls in A1 L2/3. Our data thus suggest that observed cortical specializations for call processing emerge in A1, and set the stage for further mechanistic studies.
]]></description>
<dc:creator>Montes-Lourido, P.</dc:creator>
<dc:creator>Kar, M.</dc:creator>
<dc:creator>David, S. V.</dc:creator>
<dc:creator>Sadagopan, S.</dc:creator>
<dc:date>2021-04-11</dc:date>
<dc:identifier>doi:10.1101/2021.04.11.439331</dc:identifier>
<dc:title><![CDATA[A complex feature-based representation of vocalizations emerges in the superficial layers of primary auditory cortex]]></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/10.1101/2021.04.12.439511v1?rss=1">
<title>
<![CDATA[
Adaptive mossy cell circuit plasticity after status epilepticus 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.04.12.439511v1?rss=1</link>
<description><![CDATA[
Hilar mossy cells control network function in the hippocampus through both direct excitation and di-synaptic inhibition of dentate granule cells (DGCs). Substantial mossy cell loss occurs after epileptic seizures; however the contribution of surviving mossy cells to network activity in the reorganized dentate gyrus is unknown. To examine functional circuit changes after pilocarpine-induced status epilepticus, we optogenetically stimulated mossy cells in acute hippocampal slices. In control mice, activation of mossy cells produced monosynaptic excitatory and di-synaptic GABAergic currents in DGCs. In pilocarpine-treated mice, mossy cell density and excitation of DGCs were reduced in parallel, with only a minimal reduction in feedforward inhibition, enhancing the inhibition:excitation ratio. Surprisingly, mossy cell-driven excitation of parvalbumin-positive basket cells, the primary mediators of feed-forward inhibition, was maintained, indicating increased connectivity between surviving mossy cells and these targets. Our results suggest that mossy cell outputs reorganize following seizures, increasing their net inhibitory effect in the hippocampus.
]]></description>
<dc:creator>Butler, C. R.</dc:creator>
<dc:creator>Westbrook, G. L.</dc:creator>
<dc:creator>Schnell, E.</dc:creator>
<dc:date>2021-04-14</dc:date>
<dc:identifier>doi:10.1101/2021.04.12.439511</dc:identifier>
<dc:title><![CDATA[Adaptive mossy cell circuit plasticity after status epilepticus]]></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/10.1101/2021.04.14.439861v1?rss=1">
<title>
<![CDATA[
Targeting Adaptation to Cancer Treatment by Drug Combinations 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.04.14.439861v1?rss=1</link>
<description><![CDATA[
Adaptation of tumors to therapeutic interventions contributes to dismal long-term patient outcomes. Adaptation to therapy involves co-action of functionally related proteins that together activate cell survival programs and compensate for the therapeutic impact. Oncogenic dependencies to such adaptive events, however, can generate new therapeutic vulnerabilities that can be targeted with drug combinations. The precision medicine approaches in which targeted drugs are matched to pre-existing genomic aberrations fail to address the adaptive responses and resulting vulnerabilities. Here, we provide the mathematical formulation, implementation and validation of the TargetScore method. The TargetScore identifies collective adaptive responses to targeted interventions as concurrent changes of phospho-proteins that are connected within a signaling network. Based on the adaptive responses, the method predicts drug-induced vulnerabilities. Using TargetScore, we inferred the adaptive responses with short-term (i.e., days) stress and long-term (i.e., months) acquired resistance to inhibitors of anti-apoptotic mediators, MCL1 and BCL2. With experiments guided by the predictions, we identified synergistic interactions between inhibitors of PARP, SHP2, and MCL1 in breast cancer cells. TargetScore is readily applicable to existing precision oncology efforts by matching targeted drug combinations to emerging molecular signatures under therapeutic stress.
]]></description>
<dc:creator>Wang, H.</dc:creator>
<dc:creator>Luna, A.</dc:creator>
<dc:creator>Yan, G.</dc:creator>
<dc:creator>Li, X.</dc:creator>
<dc:creator>Babur, O.</dc:creator>
<dc:creator>Mills, G. B.</dc:creator>
<dc:creator>Sander, C.</dc:creator>
<dc:creator>Korkut, A.</dc:creator>
<dc:date>2021-04-14</dc:date>
<dc:identifier>doi:10.1101/2021.04.14.439861</dc:identifier>
<dc:title><![CDATA[Targeting Adaptation to Cancer Treatment by Drug Combinations]]></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/10.1101/2021.04.14.439910v1?rss=1">
<title>
<![CDATA[
Speech-related auditory salience detection in the posterior superior temporal region 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.04.14.439910v1?rss=1</link>
<description><![CDATA[
Processing auditory human speech requires both detection (early and transient) and analysis (sustained). We analyzed high gamma (70-110Hz) activity of intracranial electroencephalography waveforms acquired during an auditory task that paired forward speech, reverse speech, and signal correlated noise. We identified widespread superior temporal sites with sustained activity responding only to forward and reverse speech regardless of paired order. More localized superior temporal auditory onset sites responded to all stimulus types when presented first in a pair and responded in recurrent fashion to the second paired stimulus in select conditions even in the absence of interstimulus silence; a novel finding. Auditory onset activity to a second paired sound recurred according to relative salience, with evidence of partial suppression during linguistic processing. We propose that temporal lobe auditory onset sites facilitate a salience detector function with hysteresis of 200ms and are influenced by cortico-cortical feedback loops involving linguistic processing and articulation.
]]></description>
<dc:creator>Brown, E. C.</dc:creator>
<dc:creator>Stedelin, B.</dc:creator>
<dc:creator>Raslan, A. M.</dc:creator>
<dc:creator>Selden, N. R.</dc:creator>
<dc:date>2021-04-14</dc:date>
<dc:identifier>doi:10.1101/2021.04.14.439910</dc:identifier>
<dc:title><![CDATA[Speech-related auditory salience detection in the posterior superior temporal region]]></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/10.1101/2021.04.16.439833v1?rss=1">
<title>
<![CDATA[
Cross-Species Analysis Defines the Conservation of Anatomically-Segregated VMH Neuron Populations 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.04.16.439833v1?rss=1</link>
<description><![CDATA[
The ventromedial hypothalamic nucleus (VMH) controls diverse behaviors and physiologic functions, suggesting the existence of multiple VMH neural subtypes with distinct functions. Combing Translating Ribosome Affinity Purification with RNA sequencing (TRAP-seq) data with snRNA-seq data, we identified 24 mouse VMH neuron clusters. Further analysis, including snRNA-seq data from macaque tissue, defined a more tractable VMH parceling scheme consisting of 6 major genetically- and anatomically-differentiated VMH neuron classes with good cross-species conservation. In addition to two major ventrolateral classes, we identified three distinct classes of dorsomedial VMH neurons. Consistent with previously-suggested unique roles for leptin receptor (Lepr)-expressing VMH neurons, Lepr expression marked a single dorsomedial class. We also identified a class of glutamatergic VMH neurons that resides in the tuberal region, anterolateral to the neuroanatomical core of the VMH. This atlas of conserved VMH neuron populations provides an unbiased starting point for the analysis of VMH circuitry and function.
]]></description>
<dc:creator>Affinati, A. H.</dc:creator>
<dc:creator>Sabatini, P. V.</dc:creator>
<dc:creator>True, C.</dc:creator>
<dc:creator>Tomlinson, A. J.</dc:creator>
<dc:creator>Kirigiti, M.</dc:creator>
<dc:creator>Lindsley, S. R.</dc:creator>
<dc:creator>Li, C.</dc:creator>
<dc:creator>Olson, D. P.</dc:creator>
<dc:creator>Kievit, P.</dc:creator>
<dc:creator>Myers, M. G.</dc:creator>
<dc:creator>Rupp, A. C.</dc:creator>
<dc:date>2021-04-18</dc:date>
<dc:identifier>doi:10.1101/2021.04.16.439833</dc:identifier>
<dc:title><![CDATA[Cross-Species Analysis Defines the Conservation of Anatomically-Segregated VMH Neuron Populations]]></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/10.1101/2021.04.16.440238v1?rss=1">
<title>
<![CDATA[
Single-cell analysis of localized low- and high-grade prostate cancers 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.04.16.440238v1?rss=1</link>
<description><![CDATA[
Approximately, 30% of early-stage localized prostate cancer cases reoccur within 5 to 10 years [1, 2]. However, identifying precise molecular subtypes attributable to specific stages of prostate cancer has proven difficult due to high heterogeneity within localized tumors [3-5]. Bulk assays represent a population average, which is a result of the heterogeneity that exists at the individual prostate cancer cell level [6]. Here, we sequenced the accessible chromatin regions of 14,424 single-cells collected from 18 fresh-frozen prostate tumors using sci-ATAC-seq [7, 8]. We observed that shared chromatin features among low-grade prostate cancer epithelial cells were lost in high-grade tumors. Despite this loss, all high-grade tumors exhibited an enrichment for FOXA1, HOXB13 and CDX2 transcription factor binding sites within their accessible chromatin regions, indicating a shared trans-regulatory program. Single-cell analysis of the differentially accessible regions in high- versus low-grade prostate tumors identified two unique genes encoding neuronal adhesion molecules, NRXN1 and NLGN1. We found that NRXN1 and NLGN1 are expressed in the epithelial luminal, basal and neuroendocrine cells, as well as the immune, endothelial and neuronal cell types in all prostate tumors. Overall, these results provide a deeper understanding of the active gene regulatory networks in low- and high-grade prostate tumors at a striking resolution and provide critical insights for molecular stratification of the disease.
]]></description>
<dc:creator>Eksi, S. E.</dc:creator>
<dc:creator>Chitsazan, A.</dc:creator>
<dc:creator>Sayar, Z.</dc:creator>
<dc:creator>Fields, A.</dc:creator>
<dc:creator>Kopp, R.</dc:creator>
<dc:creator>Thomas, G. V.</dc:creator>
<dc:creator>Spellman, P. T.</dc:creator>
<dc:creator>Adey, A.</dc:creator>
<dc:date>2021-04-17</dc:date>
<dc:identifier>doi:10.1101/2021.04.16.440238</dc:identifier>
<dc:title><![CDATA[Single-cell analysis of localized low- and high-grade prostate cancers]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-04-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.04.18.440336v1?rss=1">
<title>
<![CDATA[
Dimensionality reduction for neural population decoding 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.04.18.440336v1?rss=1</link>
<description><![CDATA[
Rapidly developing technology for large scale neural recordings has allowed researchers to measure the activity of hundreds to thousands of neurons at single cell resolution in vivo. Neural decoding analyses are a widely used tool used for investigating what information is represented in this complex, high-dimensional neural population activity. Most population decoding methods assume that correlated activity between neurons has been estimated accurately. In practice, this requires large amounts of data, both across observations and across neurons. Unfortunately, most experiments are fundamentally constrained by practical variables that limit the number of times the neural population can be observed under a single stimulus and/or behavior condition. Therefore, new analytical tools are required to study neural population coding while taking into account these limitations. Here, we present a simple and interpretable method for dimensionality reduction that allows neural decoding metrics to be calculated reliably, even when experimental trial numbers are limited. We illustrate the method using simulations and compare its performance to standard approaches for dimensionality reduction and decoding by applying it to single-unit electrophysiological data collected from auditory cortex.
]]></description>
<dc:creator>Heller, C. R.</dc:creator>
<dc:creator>David, S. V.</dc:creator>
<dc:date>2021-04-20</dc:date>
<dc:identifier>doi:10.1101/2021.04.18.440336</dc:identifier>
<dc:title><![CDATA[Dimensionality reduction for neural population decoding]]></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/10.1101/2021.04.21.440847v1?rss=1">
<title>
<![CDATA[
An ELISA-based method for rapid genetic screens in Drosophila 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.04.21.440847v1?rss=1</link>
<description><![CDATA[
Drosophila is a powerful model in which to perform genetic screens, but screening assays that are both rapid and can be used to examine a wide variety of cellular and molecular pathways are limited. Drosophila offer an extensive toolbox of GFP-based transcriptional reporters, GFP-tagged proteins, and driver lines which can be used to express GFP in numerous subpopulations of cells. Thus, a tool that can rapidly and quantitatively evaluate GFP levels in Drosophila tissue would provide a broadly applicable screening platform. To quantify GFP levels from Drosophila lysates, we developed a GFP-based ELISA assay. We demonstrate that this assay can detect membrane localized GFP in a variety of neuronal and glial cell populations and validate that it can identify genes that change the morphology of these cells. This assay was also able to detect STAT transcriptional activity after injury. We found that this assay can detect endogenously GFP-tagged proteins, including Draper and Cryptochrome, and it is able to report developmental and circadian changes in the expression of these proteins. Finally, we validated that the assay can be used to detect changes in synapse elimination upon genetic manipulation of astrocytes. We then used the assay to perform a small-scale screen, which identified Syntaxins as novel regulators of astrocyte-mediated synapse elimination. Together, these studies establish an ELISA as a rapid, easy and quantitative in vivo screening method to assay a wide breadth of fundamental questions in neurobiology.

Significance StatementForward genetic screens in Drosophila have played an integral role in elucidating the cellular and molecular pathways that govern almost every facet of biology. However, current screening methods in Drosophila are either fast, but limited in their specificity for particular pathways or processes, or rely on imaging, which requires substantial expertise, time, and cost. We have developed a rapid GFP-based ELISA screening method that, when paired with the wealth of GFP-based genetic tools already available in Drosophila, can be used to screen for regulators of many subpopulations of cells, transcriptional programs and levels of thousands of different proteins. Using this assay, we have identified a novel family of genes required for astrocytes to mediate developmental synapse elimination. This technique provides a screening platform that is fast, accessible, and broadly applicable to many pathways and processes, making Drosophila an even more powerful screening tool.
]]></description>
<dc:creator>Jay, T. R.</dc:creator>
<dc:creator>Kang, Y.</dc:creator>
<dc:creator>Jefferson, A.</dc:creator>
<dc:creator>Freeman, M.</dc:creator>
<dc:date>2021-04-22</dc:date>
<dc:identifier>doi:10.1101/2021.04.21.440847</dc:identifier>
<dc:title><![CDATA[An ELISA-based method for rapid genetic screens in Drosophila]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-04-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.04.21.440864v1?rss=1">
<title>
<![CDATA[
Transfer Learning for Inference of Metastatic Origin from Whole Slide Histology 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.04.21.440864v1?rss=1</link>
<description><![CDATA[
Accurate diagnosis of metastatic cancer is essential for prescribing optimal control strategies to halt further spread of metastasizing disease. While pathological inspection aided by immunohistochemistry staining provides a valuable gold standard for clinical diagnostics, deep learning methods have emerged as powerful tools for identifying clinically relevant features of whole slide histology relevant to a tumors metastatic origin. Although deep learning models require significant training data to learn effectively, transfer learning paradigms provide mechanisms to circumvent limited training data by first training a model on related data prior to fine-tuning on smaller data sets of interest. In this work we propose a transfer learning approach that trains a convolutional neural network to infer the metastatic origin of tumor tissue from whole slide images of hematoxylin and eosin (H&E) stained tissue sections and illustrate the advantages of pre-training network on whole slide images of primary tumor morphology. We further characterize statistical dissimilarity between primary and metastatic tumors of various indications on patch-level images to highlight limitations of our indication-specific transfer learning approach. Using a primary-to-metastatic transfer learning approach, we achieved mean class-specific areas under receiver operator characteristics curve (AUROC) of 0.779, which outperformed comparable models trained on only images of primary tumor (mean AUROC of 0.691) or trained on only images of metastatic tumor (mean AUROC of 0.675), supporting the use of large scale primary tumor imaging data in developing computer vision models to characterize metastatic origin of tumor lesions.
]]></description>
<dc:creator>Schau, G.</dc:creator>
<dc:creator>Ghani, H.</dc:creator>
<dc:creator>Burlingame, E. A.</dc:creator>
<dc:creator>Thibault, G.</dc:creator>
<dc:creator>Gray, J. W.</dc:creator>
<dc:creator>Corless, C.</dc:creator>
<dc:creator>Chang, Y. H.</dc:creator>
<dc:date>2021-04-22</dc:date>
<dc:identifier>doi:10.1101/2021.04.21.440864</dc:identifier>
<dc:title><![CDATA[Transfer Learning for Inference of Metastatic Origin from Whole Slide Histology]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-04-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.04.22.436044v1?rss=1">
<title>
<![CDATA[
Inverting the model of genomics data sharing with the NHGRI Genomic Data Science Analysis, Visualization, and Informatics Lab-space (AnVIL) 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.04.22.436044v1?rss=1</link>
<description><![CDATA[
The traditional model of genomic data analysis - downloading data from centralized warehouses for analysis with local computing resources - is increasingly unsustainable. Not only are transfers slow and cost prohibitive, but this approach also leads to redundant and siloed compute infrastructure that makes it difficult to ensure security and compliance of protected data. The NHGRI Genomic Data Science Analysis, Visualization, and Informatics Lab-space (AnVIL; https://anvilproject.org) inverts this model, providing a unified cloud computing environment for data storage, management, and analysis. AnVIL eliminates the need for data movement, allows for active threat detection and monitoring, and provides scalable, shared computing resources that can be acquired by researchers as needed. This presents many new opportunities for collaboration and data sharing that will ultimately lead to scientific discoveries at scales not previously possible.
]]></description>
<dc:creator>Schatz, M. C.</dc:creator>
<dc:creator>Philippakis, A. A.</dc:creator>
<dc:creator>Afgan, E.</dc:creator>
<dc:creator>Banks, E.</dc:creator>
<dc:creator>Carey, V. J.</dc:creator>
<dc:creator>Carroll, R. J.</dc:creator>
<dc:creator>Culotti, A.</dc:creator>
<dc:creator>Ellrott, K.</dc:creator>
<dc:creator>Goecks, J.</dc:creator>
<dc:creator>Grossman, R. L.</dc:creator>
<dc:creator>Hall, I.</dc:creator>
<dc:creator>Hansen, K. D.</dc:creator>
<dc:creator>Lawson, J.</dc:creator>
<dc:creator>Leek, J. T.</dc:creator>
<dc:creator>O'Donnell Luria, A.</dc:creator>
<dc:creator>Mosher, S.</dc:creator>
<dc:creator>Morgan, M.</dc:creator>
<dc:creator>Nekrutenko, A.</dc:creator>
<dc:creator>O'Connor, B. D.</dc:creator>
<dc:creator>Osborn, K.</dc:creator>
<dc:creator>Paten, B.</dc:creator>
<dc:creator>Patterson, C.</dc:creator>
<dc:creator>Tan, F. J.</dc:creator>
<dc:creator>Overby Taylor, C.</dc:creator>
<dc:creator>Vessio, J.</dc:creator>
<dc:creator>Waldron, L. D.</dc:creator>
<dc:creator>Wang, T.</dc:creator>
<dc:creator>Wuichet, K.</dc:creator>
<dc:creator>AnVIL Team,</dc:creator>
<dc:date>2021-04-23</dc:date>
<dc:identifier>doi:10.1101/2021.04.22.436044</dc:identifier>
<dc:title><![CDATA[Inverting the model of genomics data sharing with the NHGRI Genomic Data Science Analysis, Visualization, and Informatics Lab-space (AnVIL)]]></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/10.1101/2021.04.22.441005v1?rss=1">
<title>
<![CDATA[
ME-VAE: Multi-Encoder Variational AutoEncoder for Controlling Multiple Transformational Features in Single Cell Image Analysis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.04.22.441005v1?rss=1</link>
<description><![CDATA[
Image-based cell phenotyping relies on quantitative measurements as encoded representations of cells; however, defining suitable representations that capture complex imaging features is challenged by the lack of robust methods to segment cells, identify subcellular compartments, and extract relevant features. Variational autoencoder (VAE) approaches produce encouraging results by mapping an image to a representative descriptor, and outperform classical hand-crafted features for morphology, intensity, and texture at differentiating data. Although VAEs show promising results for capturing morphological and organizational features in tissue, single cell image analyses based on VAEs often fail to identify biologically informative features due to uninformative technical variation. Herein, we propose a multi-encoder VAE (ME-VAE) in single cell image analysis using transformed images as a self-supervised signal to extract transform-invariant biologically meaningful features, including emergent features not obvious from prior knowledge. We show that the proposed architecture improves analysis by making distinct cell populations more separable compared to traditional VAEs and intensity measurements by enhancing phenotypic differences between cells and by improving correlations to other analytic modalities.
]]></description>
<dc:creator>Ternes, L.</dc:creator>
<dc:creator>Dane, M.</dc:creator>
<dc:creator>Labrie, M.</dc:creator>
<dc:creator>Mills, G. B.</dc:creator>
<dc:creator>Gray, J. W.</dc:creator>
<dc:creator>Heiser, L.</dc:creator>
<dc:creator>Chang, Y. H.</dc:creator>
<dc:date>2021-04-23</dc:date>
<dc:identifier>doi:10.1101/2021.04.22.441005</dc:identifier>
<dc:title><![CDATA[ME-VAE: Multi-Encoder Variational AutoEncoder for Controlling Multiple Transformational Features in Single Cell Image Analysis]]></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/10.1101/2021.04.23.441175v1?rss=1">
<title>
<![CDATA[
Functional independence of endogenous mu- and delta-opioid receptors co-expressed in cholinergic interneurons 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.04.23.441175v1?rss=1</link>
<description><![CDATA[
Class A G protein-coupled receptors (GPCRs) normally function as monomers, although evidence from heterologous expression systems suggests they may form homodimers and/or heterodimers. Detection of GPCR dimers in native tissues has however been challenging due to the lack of suitable tools. - and {delta}-Opioid receptors (MORs and DORs) co-expressed in transfected cells has been reported to form heterodimers. The co-localization of MORs and DORs in neurons has been studied in knock-in mice expressing genetically engineered receptors fused to fluorescent proteins. Here we report that single cholinergic neurons in the mouse striatum endogenously express both MORs and DORs. The receptors were fluorescently labeled in live brain slices with a ligand-directed labeling reagent, NAI-A594. The selective activation of MORs and DORs, with DAMGO (-agonist) and deltorphin ({delta}-agonist) inhibited spontaneous firing in all cells examined. In the continued presence of agonist, the firing rate returned to baseline with the application of deltorphin but was persistently inhibited with the application of DAMGO. In addition, agonist-induced internalization of DORs but not MORs was detected. When MORs and DORs were activated simultaneously with [Met5]enkephalin, desensitization of MORs was facilitated but internalization was not increased. Together, these results indicate that while MORs and DORs are expressed in single striatal cholinergic interneurons, the two receptors function independently.
]]></description>
<dc:creator>Arttamangkul, S.</dc:creator>
<dc:creator>Platt, E. J.</dc:creator>
<dc:creator>Carroll, J.</dc:creator>
<dc:creator>Farrens, D. L.</dc:creator>
<dc:date>2021-04-24</dc:date>
<dc:identifier>doi:10.1101/2021.04.23.441175</dc:identifier>
<dc:title><![CDATA[Functional independence of endogenous mu- and delta-opioid receptors co-expressed in cholinergic interneurons]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-04-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.04.26.441492v1?rss=1">
<title>
<![CDATA[
Neuronal Dystroglycan regulates postnatal development of CCK/cannabinoid receptor-1 interneurons 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.04.26.441492v1?rss=1</link>
<description><![CDATA[
The development of functional neural circuits requires the precise formation of synaptic connections between diverse neuronal populations. The molecular pathways that allow GABAergic interneuron subtypes in the mammalian brain to recognize their postsynaptic partners remain largely unknown. The transmembrane glycoprotein Dystroglycan is localized to inhibitory synapses in pyramidal neurons, where it is required for the proper function of CCK+ interneurons. We show that deletion of Dystroglycan from pyramidal neurons selectively impairs CCK+ interneuron development during the first postnatal week. In the absence of postsynaptic Dystroglycan, presynaptic CCK+ interneurons fail to elaborate their axons and largely disappear from the cortex, hippocampus, amygdala, and olfactory bulb. Bax deletion did not rescue CCK+ interneurons, suggesting that they are not eliminated by canonical apoptosis in Dystroglycan mutants. Rather, we observed an increase in CCK+ interneuron innervation of the striatum, suggesting that the remaining CCK+ interneurons re-directed their axons to neighboring areas where Dystroglycan expression remained intact. Together these findings identify Dystroglycan as a critical regulator of CCK+ interneuron development.
]]></description>
<dc:creator>Miller, D. S.</dc:creator>
<dc:creator>Wright, K. M.</dc:creator>
<dc:date>2021-04-26</dc:date>
<dc:identifier>doi:10.1101/2021.04.26.441492</dc:identifier>
<dc:title><![CDATA[Neuronal Dystroglycan regulates postnatal development of CCK/cannabinoid receptor-1 interneurons]]></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/10.1101/2021.04.26.441556v1?rss=1">
<title>
<![CDATA[
Maternal Obesity Dysregulates Fetal Hematopoietic Stem and Progenitor Cell Development in Rhesus Macaques 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.04.26.441556v1?rss=1</link>
<description><![CDATA[
BackgroundMaternal obesity adversely impacts the in utero metabolic environment and offsprings health, but its effect on fetal hematopoiesis and immune cell development remains incompletely understood, particularly in models that resemble human development.

MethodsWe studied gestational day 130-135 fetuses derived from rhesus macaque dams chronically exposed to a high-fat Western-style diet (WSD) or a low-fat control diet. Fetal immune cell phenotypes and fetal bone marrow architecture and hematopoietic stem and progenitor cell (FBM HSPC) function were examined using bone computed tomography, histology, flow cytometry, single-cell RNA-sequencing, and HSPC transplantation assays.

FindingsMaternal WSD induced premature FBM cavity opening and a codominant increase in the number of FBM adipocytes. Furthermore, a maternal WSD induced a proinflammatory transcriptional response in FBM HSPCs. FBM macrophages from the WSD group exhibited heightened proinflammatory responses to toll-like receptor agonist stimulation. Maternal WSD exposure suppressed the expression of genes required for B-cell development and decreased the frequencies of FBM B-cells. Finally, maternal WSD led to poor engraftment of FBM HSPCs in nonlethally irradiated immunodeficient NOD/SCID/IL2r{gamma}-/- mice.

InterpretationsMaternal WSD impairs FBM development, drives a hyperinflammatory phenotype, and induces functional and differentiation impairment in FBM HSPCs in a translationally relevant nonhuman primate model.

FundingNational Institute of Health

RESEARCH IN CONTEXTO_ST_ABSEvidence before this studyC_ST_ABSMaternal obesity is associated with increased risk of infections and proinflammatory disease in offspring. The translationally-relevant rhesus macaque model was utilized to address the effects of maternal obesogenic diet on fetal hematopoietic and immune cell development.

Added value of this studyWe assessed changes in fetal immune cell phenotypes and fetal hematopoietic stem and progenitor cell function using immunohistochemistry, flow cytometry, single-cell RNA sequencing, and transplantation assays. We determined that chronic consumption of a maternal obesogenic diet induced the development of adipogenic and proinflammatory environments in the fetal bone marrow. Additionally, we detected the impairment in B-cell differentiation program in fetal hematopoietic stem and progenitor cells.

Implications of all the available evidenceThese data demonstrate that maternal obesogenic diet modulates fetal hematopoietic development and could impact the offsprings immune system, including proinflammatory phenotype and a decline in B-cell function.
]]></description>
<dc:creator>Sureshchandra, S.</dc:creator>
<dc:creator>Robino, J. J.</dc:creator>
<dc:creator>Goldman, D.</dc:creator>
<dc:creator>Takahashi, D.</dc:creator>
<dc:creator>Shen, W.</dc:creator>
<dc:creator>Hennebold, J. D.</dc:creator>
<dc:creator>Roberts, C. T.</dc:creator>
<dc:creator>Burwitz, B. J.</dc:creator>
<dc:creator>Messaoudi, I.</dc:creator>
<dc:creator>Varlamov, O.</dc:creator>
<dc:date>2021-04-27</dc:date>
<dc:identifier>doi:10.1101/2021.04.26.441556</dc:identifier>
<dc:title><![CDATA[Maternal Obesity Dysregulates Fetal Hematopoietic Stem and Progenitor Cell Development in Rhesus Macaques]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-04-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.04.27.441638v1?rss=1">
<title>
<![CDATA[
Astrocytic GABA Transporter controls sleep by modulating GABAergic signaling in Drosophila circadian neurons 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.04.27.441638v1?rss=1</link>
<description><![CDATA[
A precise balance between sleep and wakefulness is essential to sustain a good quality of life and optimal brain function. GABA is known to play a key and conserved role in sleep control, and GABAergic tone should therefore be tightly controlled in sleep circuits. Here we examined the role of the astrocytic GABA transporter (GAT) in sleep regulation using Drosophila melanogaster. We found that a hypomorphic gat mutation (gat33-1) increased sleep amount, decreased sleep latency, and increased sleep consolidation. Interestingly, sleep defects were suppressed when gat33-1 was combined with a mutation disrupting wide-awake (wake), a gene that regulates the cell-surface levels of the GABAA receptor Resistance to Dieldrin (RDL) in the wake-promoting large ventral lateral neurons (l-LNvs). Moreover, RNAi knockdown of rdl and its modulator dnlg4 in these circadian neurons also suppressed gat33-1 sleep phenotypes. Brain immunohistochemistry showed that GAT-expressing astrocytes were located near RDL-positive l-LNvs cell bodies and dendritic processes. We conclude that astrocytic GAT decreases GABAergic tone and RDL activation in arousal promoting LNvs, thus determining proper sleep amount and quality in Drosophila
]]></description>
<dc:creator>Chaturvedi, R.</dc:creator>
<dc:creator>Stork, T.</dc:creator>
<dc:creator>Yuan, C.</dc:creator>
<dc:creator>Freeman, M. R.</dc:creator>
<dc:creator>Emery, P.</dc:creator>
<dc:date>2021-04-27</dc:date>
<dc:identifier>doi:10.1101/2021.04.27.441638</dc:identifier>
<dc:title><![CDATA[Astrocytic GABA Transporter controls sleep by modulating GABAergic signaling in Drosophila circadian neurons]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-04-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.04.29.441927v1?rss=1">
<title>
<![CDATA[
Effects of BCG vaccination on donor unrestricted T cells in humans 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.04.29.441927v1?rss=1</link>
<description><![CDATA[
Antigen classes other than proteins can be presented to T cells by near-monomorphic antigen-presenting molecules such as CD1, MR1, and butyrophilin 3A1. We sought to define the roles of donor unrestricted T (DURT) cells, including MR1-reactive MAIT cells, CD1b-reactive glucose monomycolate (GMM)-specific T cells, CD1d-reactive NKT cells, and {gamma}{delta} T cells, in vaccination against Mycobacterium tuberculosis. We characterized DURT cells following primary bacille Calmette-Guerin (BCG) vaccination in infants or BCG-revaccination in adults. BCG (re)vaccination did not modulate peripheral blood frequencies, T cell activation or memory profiles of MAIT cells, CD1b-restricted GMM-specific and germline-encoded mycolyl-reactive (GEM) cells or CD1d- restricted NKT cells. By contrast, BCG vaccination was associated with increased frequencies of {gamma}{delta} T cells as well as a novel subset of IFN-{gamma}-expressing CD4+ T cells with a CD26+CD161+TRAV1-2- phenotype in infants. More studies are required to understand the full potential of DURT cells in new TB vaccine strategies.
]]></description>
<dc:creator>Gela, A.</dc:creator>
<dc:creator>Murphy, M.</dc:creator>
<dc:creator>Hadley, K.</dc:creator>
<dc:creator>Hanekom, W.</dc:creator>
<dc:creator>Boom, H.</dc:creator>
<dc:creator>Johnson, J.</dc:creator>
<dc:creator>Hoft, D.</dc:creator>
<dc:creator>Joosten, S.</dc:creator>
<dc:creator>Ottenhoff, T.</dc:creator>
<dc:creator>Suliman, S.</dc:creator>
<dc:creator>Moody, B.</dc:creator>
<dc:creator>Lewinsohn, D.</dc:creator>
<dc:creator>Hatherill, M.</dc:creator>
<dc:creator>Seshadri, C.</dc:creator>
<dc:creator>Nemes, E.</dc:creator>
<dc:creator>Scriba, T.</dc:creator>
<dc:date>2021-04-29</dc:date>
<dc:identifier>doi:10.1101/2021.04.29.441927</dc:identifier>
<dc:title><![CDATA[Effects of BCG vaccination on donor unrestricted T cells in humans]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-04-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.05.01.442252v1?rss=1">
<title>
<![CDATA[
Mesenchymal Lineage Heterogeneity Underlies Non-Redundant Functions of Pancreatic Cancer-Associated Fibroblasts 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.05.01.442252v1?rss=1</link>
<description><![CDATA[
Cancer-associated fibroblast (CAF) heterogeneity is increasingly appreciated, but the origins and functions of distinct CAF subtypes remain poorly understood. The abundant and transcriptionally diverse CAF population in pancreatic ductal adenocarcinoma (PDAC) is thought to arise from a common cell of origin, pancreatic stellate cells (PSCs), with diversification resulting from cytokine and growth factor gradients within the tumor microenvironment. Here we analyzed the differentiation and function of PSCs during tumor progression in vivo. Contrary to expectations, we found that PSCs give rise to a numerically minor subset of PDAC CAFs. Targeted ablation of PSC-derived CAFs within their host tissue revealed non-redundant functions for this defined CAF population in shaping the PDAC microenvironment, including production of specific components of the extracellular matrix. Together, these findings link stromal evolution from distinct cells of origin to transcriptional heterogeneity among PDAC CAFs, and demonstrate unique functions for CAFs of a defined cellular origin.

Statement of significanceBy tracking and ablating a specific CAF population, we find that a numerically minor CAF subtype from a defined cell of origin plays unique roles in establishing the pancreatic tumor microenvironment. Together with prior studies, this work suggests that mesenchymal lineage heterogeneity as well as signaling gradients diversify PDAC CAFs.
]]></description>
<dc:creator>Helms, E. J.</dc:creator>
<dc:creator>Berry, M. W.</dc:creator>
<dc:creator>Chaw, R. C.</dc:creator>
<dc:creator>DuFort, C. C.</dc:creator>
<dc:creator>Sun, D.</dc:creator>
<dc:creator>Onate, M. K.</dc:creator>
<dc:creator>Oon, C.</dc:creator>
<dc:creator>Bhattacharyya, S.</dc:creator>
<dc:creator>Sanford-Crane, H.</dc:creator>
<dc:creator>Horton, W.</dc:creator>
<dc:creator>Finan, J. M.</dc:creator>
<dc:creator>Sattler, A.</dc:creator>
<dc:creator>Makar, R.</dc:creator>
<dc:creator>Dawson, D. W.</dc:creator>
<dc:creator>Xia, Z.</dc:creator>
<dc:creator>Hingorani, S. R.</dc:creator>
<dc:creator>Sherman, M. H.</dc:creator>
<dc:date>2021-05-02</dc:date>
<dc:identifier>doi:10.1101/2021.05.01.442252</dc:identifier>
<dc:title><![CDATA[Mesenchymal Lineage Heterogeneity Underlies Non-Redundant Functions of Pancreatic Cancer-Associated Fibroblasts]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-05-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.05.12.443856v1?rss=1">
<title>
<![CDATA[
Transcriptional, epigenetic, and functional reprogramming of blood monocytes in non-human primates following chronic alcohol drinking 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.05.12.443856v1?rss=1</link>
<description><![CDATA[
Chronic heavy drinking (CHD) of alcohol is a known risk factor for increased susceptibility to bacterial and viral infection as well as impaired wound healing. Evidence suggests that these defects are mediated by a dysregulated inflammatory response originating from myeloid cells, notably monocytes and macrophages, but the mechanisms remain poorly understood. Our ability to study CHD is impacted by the complexities of human drinking patterns and behavior as well as comorbidities and confounding risk factors for patients with alcohol use disorders. To overcome these challenges, we utilize a translational rhesus macaque model of voluntary ethanol self-administration that closely recapitulates human drinking patterns and chronicity. In this study, we examined the effects of CHD on blood monocytes and alveolar macrophages in control and CHD female macaques after 12 months of daily ethanol consumption. While monocytes from CHD female macaques generated a hyper-inflammatory response to ex vivo LPS stimulation, their response to E.Coli was dampened. In depth scRNA-Seq analysis of purified monocytes revealed significant shifts in classical monocyte subsets with accumulation of cells expressing markers of hypoxia (HIF1A) and inflammation (NFkB signaling pathway) in CHD macaques. The increased presence of monocyte subsets poised to generate a hyperinflammatory response was confirmed by the epigenetic analysis which revealed higher accessibility of promoter regions that regulate genes involved in cytokine signaling pathways. Finally, alveolar macrophages (AM) from the same animals produced higher levels of inflammatory mediators in response to LPS stimulation, but reduced ability to phagocytose bacteria. Collectively, data presented in this manuscript demonstrate that CHD primes monocytes and tissue-resident macrophages towards a more hyper-inflammatory immune response with compromised functional abilities, which could be used in diagnostic purposes or preventative measures for patients with alcohol use disorders.
]]></description>
<dc:creator>Lewis, S. A.</dc:creator>
<dc:creator>Sureshchandra, S.</dc:creator>
<dc:creator>Doratt, B.</dc:creator>
<dc:creator>Jimenez, V.</dc:creator>
<dc:creator>Stull, C.</dc:creator>
<dc:creator>Grant, K. A.</dc:creator>
<dc:creator>Messaoudi, I.</dc:creator>
<dc:date>2021-05-13</dc:date>
<dc:identifier>doi:10.1101/2021.05.12.443856</dc:identifier>
<dc:title><![CDATA[Transcriptional, epigenetic, and functional reprogramming of blood monocytes in non-human primates following chronic alcohol drinking]]></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/10.1101/2021.05.13.444033v1?rss=1">
<title>
<![CDATA[
The FDA-approved drug apremilast suppresses alcohol intake: clinical and pre-clinical validation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.05.13.444033v1?rss=1</link>
<description><![CDATA[
Treatment options for Alcohol Use Disorders (AUD) have minimally advanced since 2004, while the annual deaths and economic toll have become alarmingly high. Bringing potential therapeutics beyond the bench and into the clinic for AUD requires rigorous pharmacological screening across molecular, behavioral, pre-clinical, and clinical studies in neuroscience. The repurposing of FDA-approved compounds is an effective and expedited means of screening pharmacotherapies for AUD. Here, we demonstrate that apremilast, a phosphodiesterase type 4 inhibitor that is FDA approved for psoriasis and psoriatic arthritis, reduces binge-like alcohol intake and behavioral measures of motivation in unique, preclinical genetic risk models for drinking to intoxication and reduces excessive alcohol drinking in models of stress-facilitated drinking and alcohol dependence. In a double blind, placebo-controlled human laboratory study in non-treatment seeking individuals with AUD, apremilast significantly reduced the number of drinks per day. Lastly, using site-directed drug infusions and electrophysiology we determined that apremilast may act by increasing neural activity in the nucleus accumbens, an important alcohol-related brain region, to reduce alcohol intake in mice. These results demonstrate that apremilast reduces excessive alcohol drinking across a spectrum of AUD severity and support its importance as a potential therapeutic for AUD.
]]></description>
<dc:creator>Grigsby, K. B.</dc:creator>
<dc:creator>Mangieri, R. A.</dc:creator>
<dc:creator>Roberts, A. J.</dc:creator>
<dc:creator>Lopez, M. J.</dc:creator>
<dc:creator>Tran, A.</dc:creator>
<dc:creator>Firsick, E. J.</dc:creator>
<dc:creator>Townsley, K. G.</dc:creator>
<dc:creator>Beneze, A.</dc:creator>
<dc:creator>Bess, J.</dc:creator>
<dc:creator>Eisenstein, T. K.</dc:creator>
<dc:creator>Meissler, J. J.</dc:creator>
<dc:creator>Light, J. M.</dc:creator>
<dc:creator>Miller, J.</dc:creator>
<dc:creator>Quello, S.</dc:creator>
<dc:creator>Shadan, F.</dc:creator>
<dc:creator>Skinner, M.</dc:creator>
<dc:creator>Aziz, H. C.</dc:creator>
<dc:creator>Metten, P.</dc:creator>
<dc:creator>Morissett, R. A.</dc:creator>
<dc:creator>Crabbe, J. C.</dc:creator>
<dc:creator>Roberto, M.</dc:creator>
<dc:creator>Becker, H. C.</dc:creator>
<dc:creator>Mason, B. J.</dc:creator>
<dc:creator>Ozburn, A. R.</dc:creator>
<dc:date>2021-05-15</dc:date>
<dc:identifier>doi:10.1101/2021.05.13.444033</dc:identifier>
<dc:title><![CDATA[The FDA-approved drug apremilast suppresses alcohol intake: clinical and pre-clinical validation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-05-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.05.14.444244v1?rss=1">
<title>
<![CDATA[
Pepsickle rapidly and accurately predicts proteasomal cleavage sites for improved neoantigen identification 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.05.14.444244v1?rss=1</link>
<description><![CDATA[
Proteasomal cleavage is a key component in protein turnover, as well as antigen presentation and subsequent immune response. Herein we present pepsickle, an open-source tool for proteasomal cleavage prediction with better in vivo prediction performance (AUC) and computational speed than current models available in the field, and with the ability to predict sites based on both constitutive and immunoproteasome profiles. Post-hoc filtering of predicted patient neoepitopes using pepsickle significantly enriches for immune-responsive epitopes and may represent a significant opportunity to improve current epitope prediction and vaccine development pipelines.
]]></description>
<dc:creator>Weeder, B.</dc:creator>
<dc:creator>Wood, M.</dc:creator>
<dc:creator>Li, E.</dc:creator>
<dc:creator>Nellore, A.</dc:creator>
<dc:creator>Thompson, R. F.</dc:creator>
<dc:date>2021-05-17</dc:date>
<dc:identifier>doi:10.1101/2021.05.14.444244</dc:identifier>
<dc:title><![CDATA[Pepsickle rapidly and accurately predicts proteasomal cleavage sites for improved neoantigen identification]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-05-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.05.15.444267v1?rss=1">
<title>
<![CDATA[
Vascular KATP channel structural dynamics reveal regulatory mechanism by Mg-nucleotides 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.05.15.444267v1?rss=1</link>
<description><![CDATA[
Vascular tone is dependent on smooth muscle KATP channels comprising pore-forming Kir6.1 and regulatory SUR2B subunits, in which mutations cause Cantu syndrome. Unique among KATP isoforms, they lack spontaneous activity and require Mg-nucleotides for activation. Structural mechanisms underlying these properties are unknown. Here, we determined the first cryoEM structures of vascular KATP channels bound to inhibitory ATP and glibenclamide, which differ informatively from similarly determined pancreatic KATP channel isoform (Kir6.2/SUR1). Unlike SUR1, SUR2B subunits adopt distinct rotational "propeller" and "quatrefoil" geometries surrounding their Kir6.1 core. The previously unseen ED-rich linker connecting the two halves of the SUR-ABC core is observed in a quatrefoil-like conformation. MD simulations reveal MgADP-dependent dynamic tripartite interactions between this linker, SUR2B and Kir6.1. The structures captured implicate a progression of intermediate states between MgADP-free inactivated and MgADP-bound activated conformations wherein the ED-rich linker participates as mobile autoinhibitory domain, suggesting a conformational pathway toward KATP channel activation.
]]></description>
<dc:creator>Sung, M. W.</dc:creator>
<dc:creator>Yang, Z.</dc:creator>
<dc:creator>Patton, B. L.</dc:creator>
<dc:creator>Mostofian, B.</dc:creator>
<dc:creator>Russo, J.</dc:creator>
<dc:creator>Zuckerman, D. M.</dc:creator>
<dc:creator>Shyng, S.-L.</dc:creator>
<dc:date>2021-05-16</dc:date>
<dc:identifier>doi:10.1101/2021.05.15.444267</dc:identifier>
<dc:title><![CDATA[Vascular KATP channel structural dynamics reveal regulatory mechanism by Mg-nucleotides]]></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/10.1101/2021.05.21.444548v1?rss=1">
<title>
<![CDATA[
Virus-Dependent Immune Conditioning of Tissue Microenvironments 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.05.21.444548v1?rss=1</link>
<description><![CDATA[
A thorough understanding of complex spatial host-disease interactions in situ is necessary in order to develop effective preventative measures and therapeutic strategies. Here, we developed Protein And Nucleic acid IN situ Imaging (PANINI) and coupled it with Multiplexed Ion Beam Imaging (MIBI) to sensitively and simultaneously quantify DNA, RNA, and protein levels within the microenvironments of tissue compartments. The PANINI-MIBI approach was used to measure over 30 parameters simultaneously across large sections of archival lymphoid tissues from non-human primates that were healthy or infected with simian immunodeficiency virus (SIV), a model that accurately recapitulates human immunodeficiency virus infection (HIV). This enabled multiplexed dissection of cellular phenotypes, functional markers, viral DNA integration events, and viral RNA transcripts as resulting from viral infection. The results demonstrated immune coordination from an unexpected upregulation of IL10 in B cells in response to SIV infection that correlated with macrophage M2 polarization, thus conditioning a potential immunosuppressive environment that allows for viral production. This multiplexed imaging strategy also allowed characterization of the coordinated microenvironment around latently or actively infected cells to provide mechanistic insights into the process of viral latency. The spatial multi-modal framework presented here is applicable to deciphering tissue responses in other infectious diseases and tumor biology.
]]></description>
<dc:creator>Jiang, S.</dc:creator>
<dc:creator>Chan, C. N.</dc:creator>
<dc:creator>Rovira-Clave, X.</dc:creator>
<dc:creator>Chen, H.</dc:creator>
<dc:creator>Bai, Y.</dc:creator>
<dc:creator>Zhu, B.</dc:creator>
<dc:creator>McCaffrey, E.</dc:creator>
<dc:creator>Greenwald, N. F.</dc:creator>
<dc:creator>Liu, C.</dc:creator>
<dc:creator>Barlow, G. L.</dc:creator>
<dc:creator>Weirather, J. L.</dc:creator>
<dc:creator>Busman-Sahay, K.</dc:creator>
<dc:creator>Nekorchuk, M.</dc:creator>
<dc:creator>Terry, M.</dc:creator>
<dc:creator>Younger, S.</dc:creator>
<dc:creator>Oliveria, J. P.</dc:creator>
<dc:creator>Bosse, M.</dc:creator>
<dc:creator>Demeter, J.</dc:creator>
<dc:creator>Philips, D.</dc:creator>
<dc:creator>Mukherjee, N.</dc:creator>
<dc:creator>Golstev, Y.</dc:creator>
<dc:creator>McIlwain, D. R.</dc:creator>
<dc:creator>Angelo, M.</dc:creator>
<dc:creator>Estes, J. D.</dc:creator>
<dc:creator>Nolan, G. P.</dc:creator>
<dc:date>2021-05-23</dc:date>
<dc:identifier>doi:10.1101/2021.05.21.444548</dc:identifier>
<dc:title><![CDATA[Virus-Dependent Immune Conditioning of Tissue Microenvironments]]></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/10.1101/2021.05.24.445493v1?rss=1">
<title>
<![CDATA[
Non-neutralizing antibodies targeting the immunogenic regions of HIV-1 envelope reduce mucosal infection and virus burden in humanized mice 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.05.24.445493v1?rss=1</link>
<description><![CDATA[
Antibodies are principal immune components elicited by vaccines to induce protection from microbial pathogens. In the Thai RV144 HIV-1 vaccine trial, vaccine efficacy was 31% and the sole primary correlate of reduced risk was shown to be vigorous antibody response targeting the V1V2 region of HIV-1 envelope. Antibodies against V3 also were inversely correlated with infection risk in subsets of vaccinees. Antibodies recognizing these regions, however, do not exhibit potent neutralizing activity. Therefore, we examined the antiviral potential of poorly neutralizing monoclonal antibodies (mAbs) against immunodominant V1V2 and V3 sites by passive administration of human mAbs to humanized mice engrafted with CD34+ hematopoietic stem cells, followed by mucosal challenge with an HIV-1 infectious molecular clone (IMC) expressing the envelope of a tier 2 resistant HIV-1 strain. Treatment with anti-V1V2 mAb 2158 or anti-V3 mAb 2219 did not prevent infection, but both reduced the virus burden, and V3 mAb 2219 displayed a superior potency compared to V1V2 mAb 2158. While these mAbs had no or weak neutralizing activity and elicited undetectable levels of antibody-dependent cellular cytotoxicity (ADCC), V3 mAb 2219 displayed a greater capacity to bind virus- and cell-associated HIV-1 envelope and to mediate antibody-dependent cellular phagocytosis (ADCP) and C1q complement binding as compared to V1V2 mAb 2158. Mutations in the Fc region of 2219 abolished these effector activities and abrogated virus control in humanized mice. These results demonstrate the importance of Fc functions other than ADCC for antibodies without potent neutralizing activity.
]]></description>
<dc:creator>Hioe, C. E.</dc:creator>
<dc:creator>Li, G.</dc:creator>
<dc:creator>Liu, X.</dc:creator>
<dc:creator>Tsahouridis, O.</dc:creator>
<dc:creator>He, X.</dc:creator>
<dc:creator>Funaki, M.</dc:creator>
<dc:creator>Klingler, J.</dc:creator>
<dc:creator>Tang, A. F.</dc:creator>
<dc:creator>Feyznezhad, R.</dc:creator>
<dc:creator>Heindel, D. W.</dc:creator>
<dc:creator>Wang, X.-H.</dc:creator>
<dc:creator>Spencer, D. A.</dc:creator>
<dc:creator>Hu, G.</dc:creator>
<dc:creator>Satija, N.</dc:creator>
<dc:creator>Prevost, J.</dc:creator>
<dc:creator>Finzi, A.</dc:creator>
<dc:creator>Hessell, A.</dc:creator>
<dc:creator>Wang, S.</dc:creator>
<dc:creator>Lu, S.</dc:creator>
<dc:creator>Chen, B. K.</dc:creator>
<dc:creator>Zolla-Pazner, S.</dc:creator>
<dc:creator>Upadhyay, C.</dc:creator>
<dc:creator>Alvarez, R. A.</dc:creator>
<dc:creator>Su, L.</dc:creator>
<dc:date>2021-05-24</dc:date>
<dc:identifier>doi:10.1101/2021.05.24.445493</dc:identifier>
<dc:title><![CDATA[Non-neutralizing antibodies targeting the immunogenic regions of HIV-1 envelope reduce mucosal infection and virus burden in humanized mice]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-05-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.05.25.445648v1?rss=1">
<title>
<![CDATA[
Mu opioid receptors acutely regulate adenosine signaling in a thalamo-striatal circuit. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.05.25.445648v1?rss=1</link>
<description><![CDATA[
Endogenous adenosine plays a crucial role in maintaining energy homeostasis and adenosine levels are tightly regulated across neural circuits. In the dorsal medial striatum (DMS) adenosine inhibits neurotransmitter release, but the source and mechanism underlying its accumulation are largely unknown. Opioids also inhibit neurotransmitter release in the DMS and influences adenosine accumulation after prolonged exposure. However, how these two neurotransmitter systems interact acutely is also largely unknown. This study demonstrates that activation of opioid receptors (MORs), but not {delta} opioid receptors (DORs) or {kappa} opioid receptors (KORs), inhibits tonic activation of adenosine A1Rs via a cyclic adenosine monophosphate (cAMP) dependent mechanism in both male and female mice. Further, selectively knocking-out MORs from presynaptic terminals and postsynaptic medium spiny neurons (MSNs) revealed that activation of MORs on D1R positive MSNs, but not D2R positive MSNs, is necessary to inhibit tonic adenosine signaling on presynaptic terminals. Given the role of D1R positive MSNs in movement and motivated behaviors, these findings reveal a novel mechanism by which these neurons regulate their own synaptic inputs.

Significance StatementUnderstanding interactions between neuromodulatory systems within brain circuits is a fundamental question in neuroscience. The present work uncovers a novel role of opioids in acutely inhibiting adenosine accumulation and subsequent adenosine receptor signaling in the striatum by inhibiting the production of cAMP. Adenosine receptor signaling regulates striatal neurotransmitters including glutamate, GABA, dopamine and acetylcholine. Furthermore, interactions between adenosine2A receptors and numerous other GPCRs, including D2 dopamine and CB1 cannabinoid receptors, suggest that endogenous adenosine broadly modulates striatal GPCR signaling. Additionally, this work discovered that resting endogenous adenosine is released by D1, but not D2 receptor positive MSNs, suggesting that opioid signaling and manipulation of D1R-expressing MSN cAMP activity can broadly affect striatal function and behavior.
]]></description>
<dc:creator>Adhikary, S.</dc:creator>
<dc:creator>Birdsong, W. T.</dc:creator>
<dc:date>2021-05-25</dc:date>
<dc:identifier>doi:10.1101/2021.05.25.445648</dc:identifier>
<dc:title><![CDATA[Mu opioid receptors acutely regulate adenosine signaling in a thalamo-striatal circuit.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-05-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.05.25.445671v1?rss=1">
<title>
<![CDATA[
Sex Differences in Functional Topography of Association Networks 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.05.25.445671v1?rss=1</link>
<description><![CDATA[
Prior work has shown that there is substantial interindividual variation in the spatial distribution of functional networks across the cerebral cortex, or functional topography. However, it remains unknown whether there are sex differences in the topography of individualized networks in youth. Here we leveraged an advanced machine learning method (sparsity-regularized nonnegative matrix factorization) to define individualized functional networks in 693 youth (ages 8-23 years) who underwent functional magnetic resonance imaging as part of the Philadelphia Neurodevelopmental Cohort. Multivariate pattern analysis using support vector machines classified participant sex based on functional topography with 83% accuracy (p<0.0001). Brain regions most effective in classifying participant sex belonged to association networks, including the ventral attention and default mode networks. Mass-univariate analyses using generalized additive models with penalized splines provided convergent results. Comparative analysis using transcriptomic data from the Allen Human Brain Atlas revealed that sex differences in multivariate patterns of functional topography correlated with the expression of genes on the X-chromosome. These results identify normative developmental sex differences in the functional topography of association networks and highlight the role of sex as a biological variable in shaping brain development in youth.

SIGNIFICANCE STATEMENTWe identify normative developmental sex differences in the functional topography of personalized association networks including the ventral attention network and default mode network. Furthermore, chromosomal enrichment analyses revealed that sex differences in multivariate patterns of functional topography were spatially coupled to the expression of X-linked genes as well as astrocytic and excitatory neuronal cell-type signatures. These results highlight the role of sex as a biological variable in shaping functional brain development in youth.
]]></description>
<dc:creator>Shanmugan, S.</dc:creator>
<dc:creator>Seidlitz, J.</dc:creator>
<dc:creator>Cui, Z.</dc:creator>
<dc:creator>Adebimpe, A.</dc:creator>
<dc:creator>Bassett, D. S.</dc:creator>
<dc:creator>Bertolero, M. A.</dc:creator>
<dc:creator>Davatzikos, C.</dc:creator>
<dc:creator>Fair, D. A.</dc:creator>
<dc:creator>Gur, R. E.</dc:creator>
<dc:creator>Gur, R. C.</dc:creator>
<dc:creator>Larsen, B.</dc:creator>
<dc:creator>Li, H.</dc:creator>
<dc:creator>Pines, A. R.</dc:creator>
<dc:creator>Raznahan, A.</dc:creator>
<dc:creator>Roalf, D. R.</dc:creator>
<dc:creator>Shinohara, R. T.</dc:creator>
<dc:creator>Vogel, J.</dc:creator>
<dc:creator>Wolf, D. H.</dc:creator>
<dc:creator>Fan, Y.</dc:creator>
<dc:creator>Alexander-Bloch, A.</dc:creator>
<dc:creator>Satterthwaite, T. D.</dc:creator>
<dc:date>2021-05-26</dc:date>
<dc:identifier>doi:10.1101/2021.05.25.445671</dc:identifier>
<dc:title><![CDATA[Sex Differences in Functional Topography of Association Networks]]></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/10.1101/2021.05.26.445708v1?rss=1">
<title>
<![CDATA[
The P5-ATPase ATP13A1 modulates MR1-mediated antigen presentation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.05.26.445708v1?rss=1</link>
<description><![CDATA[
The monomorphic antigen presenting molecule MHC-I-related protein 1 (MR1) presents small molecule metabolites to mucosal-associated invariant T (MAIT) cells. The MR1-MAIT cell axis has been implicated in a variety of infectious and non-communicable diseases and recent studies have begun to develop an understanding of the molecular mechanisms underlying this specialised antigen presentation pathway. Yet, the proteins regulating MR1 folding, loading, stability, and surface expression remain to be identified. Here, we performed a gene trap screen to discover novel modulators of MR1 surface expression through insertional mutagenesis of an MR1-overexpressing clone derived from the near-haploid human cell line HAP1 (HAP1.MR1). The most significant positive regulators identified included {beta}2-microglobulin, a known regulator of MR1 surface expression, and ATP13A1, a P5-ATPase in the endoplasmic reticulum (ER) with putative transporter function not previously associated with MR1-mediated antigen presentation. CRISPR/Cas9-mediated knock-out of ATP13A1 in both HAP1.MR1 and THP-1 cell lines revealed a profound reduction in MR1 protein levels and a concomitant functional defect specific to MR1-mediated antigen presentation. Collectively, these data are consistent with the ER-resident ATP13A1 as a key post-transcriptional determinant of MR1 surface expression.
]]></description>
<dc:creator>Kulicke, C. A.</dc:creator>
<dc:creator>De Zan, E.</dc:creator>
<dc:creator>Hein, Z.</dc:creator>
<dc:creator>Gonzalez-Lopez, C.</dc:creator>
<dc:creator>Ghanwat, S.</dc:creator>
<dc:creator>Veerapen, N.</dc:creator>
<dc:creator>Besra, G.</dc:creator>
<dc:creator>Klenerman, P.</dc:creator>
<dc:creator>Christianson, J. C.</dc:creator>
<dc:creator>Springer, S.</dc:creator>
<dc:creator>Nijman, S.</dc:creator>
<dc:creator>Cerundolo, V.</dc:creator>
<dc:creator>Salio, M.</dc:creator>
<dc:date>2021-05-27</dc:date>
<dc:identifier>doi:10.1101/2021.05.26.445708</dc:identifier>
<dc:title><![CDATA[The P5-ATPase ATP13A1 modulates MR1-mediated antigen presentation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-05-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.05.27.445749v1?rss=1">
<title>
<![CDATA[
Novel aspects of iron homeostasis in pathogenic bloodstream form Trypanosoma brucei 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.05.27.445749v1?rss=1</link>
<description><![CDATA[
Iron is an essential regulatory signal for virulence factors in many pathogens. Mammals and bloodstream form (BSF) Trypanosoma brucei obtain iron by receptor-mediated endocytosis of transferrin bound to receptors (TfR) but the mechanisms by which T. brucei subsequently handles iron remains enigmatic. Here, we analyse the transcriptome of T. brucei cultured in iron-rich and iron-poor conditions. We show that adaptation to iron-deprivation induces upregulation of TfR, a cohort of parasite-specific genes (ESAG3, PAGS), genes involved in glucose uptake and glycolysis (THT1 and hexokinase), endocytosis (Phosphatidic Acid Phosphatase, PAP2), and most notably a divergent RNA binding protein RBP5, indicative of a non-canonical mechanism for regulating intracellular iron levels. We show that cells depleted of TfR by RNA silencing import free iron as a compensatory survival strategy. The TfR and RBP5 iron response are reversible by genetic complementation, the response kinetics are similar, but the regulatory mechanisms are distinct. Increased TfR protein is due to increased mRNA. Increased RBP5 expression, however, occurs by a post-transcriptional feedback mechanism whereby RBP5 interacts with its own, and with PAP2 mRNAs. Further observations suggest that increased RBP5 expression in iron-deprived cells has a maximum threshold as ectopic overexpression above this threshold disrupts normal cell cycle progression resulting in an accumulation of anucleate cells and cells in G2/M phase. This phenotype is not observed with overexpression of RPB5 containing a point mutation (F61A) in its single RNA Recognition Motif. Our experiments shed new light on how T. brucei BSFs reorganise their transcriptome to deal with iron stress revealing the first iron responsive RNA binding protein that is co-regulated with TfR, is important for cell viability and iron homeostasis; two essential processes for successful proliferation.

Author SummaryAfrican trypanosomes are single-celled extracellular parasites of humans and animals relying on essential host nutrients for survival. They satisfy their iron needs by capturing host transferrin-bound iron using a surface-localised transferrin receptor (TfR) that is structurally distinct from its host counterpart. Little is known about the trypanosome response to fluctuations in host iron availability, with the exception of modulated TfR expression. We show that unlike other eukaryotes, at the transcriptome level, trypanosomes do not regulate iron-dependent enzymes as a mechanism to cope with iron deprivation.

Instead, we identify a group of novel iron responsive trypanosome-specific genes, particularly an RNA Binding Protein RBP5 that is responsive to iron levels, albeit mediated by a distinct mechanism from TfR. We show that although RBP5 expression is elevated at the mRNA and protein levels, increased abundance above a maximum threshold is toxic. The trypanosome TfR has been suggested as a therapeutic target, but whether it is essential for optimal host colonisation is unclear. Our data demonstrate that trypanosomes efficiently import free iron from their environment independent of TfR suggesting that alternative iron uptake pathways exist, and that any therapeutic interventions targeting TfR must be evaluated with caution.
]]></description>
<dc:creator>Carbajo, C. G.</dc:creator>
<dc:creator>Cornell, L. J.</dc:creator>
<dc:creator>Madbouly, Y.</dc:creator>
<dc:creator>Lai, Z.</dc:creator>
<dc:creator>Yates, P. A.</dc:creator>
<dc:creator>Tinti, M.</dc:creator>
<dc:creator>Tiengwe, C.</dc:creator>
<dc:date>2021-05-27</dc:date>
<dc:identifier>doi:10.1101/2021.05.27.445749</dc:identifier>
<dc:title><![CDATA[Novel aspects of iron homeostasis in pathogenic bloodstream form Trypanosoma brucei]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-05-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.05.27.446019v1?rss=1">
<title>
<![CDATA[
Robust Segmentation of Cellular Ultrastructure on Sparsely Labeled 3D Electron Microscopy Images using Deep Learning 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.05.27.446019v1?rss=1</link>
<description><![CDATA[
A deeper understanding of the cellular and subcellular organization of tumor cells and their interactions with the tumor microenvironment will shed light on how cancer evolves and guide effective therapy choices. Electron microscopy (EM) images can provide detailed view of the cellular ultrastructure and are being generated at an ever-increasing rate. However, the bottleneck in their analysis is the delineation of the cellular structures to enable interpretable rendering. We have mitigated this limitation by using deep learning, specifically, the ResUNet architecture, to segment cells and subcellular ultrastructure. Our initial prototype focuses on segmenting nuclei and nucleoli in 3D FIB-SEM images of tumor biopsies obtained from patients with metastatic breast and pancreatic cancers. Trained with sparse manual labels, our method results in accurate segmentation of nuclei and nucleoli with best Dice score of 0.99 and 0.98 respectively. This method can be extended to other cellular structures, enabling deeper analysis of inter- and intracellular state and interactions.
]]></description>
<dc:creator>Machireddy, A.</dc:creator>
<dc:creator>Thibault, G.</dc:creator>
<dc:creator>Loftis, K. G.</dc:creator>
<dc:creator>Stoltz, K.</dc:creator>
<dc:creator>Bueno, C. E.</dc:creator>
<dc:creator>Smith, H. R.</dc:creator>
<dc:creator>Riesterer, J. L.</dc:creator>
<dc:creator>Gray, J. W.</dc:creator>
<dc:creator>Song, X.</dc:creator>
<dc:date>2021-05-28</dc:date>
<dc:identifier>doi:10.1101/2021.05.27.446019</dc:identifier>
<dc:title><![CDATA[Robust Segmentation of Cellular Ultrastructure on Sparsely Labeled 3D Electron Microscopy Images using Deep Learning]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-05-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.05.27.446051v1?rss=1">
<title>
<![CDATA[
Noise Reconstruction & Removal Network: A New Architecture to Denoise FIB-SEM Images 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.05.27.446051v1?rss=1</link>
<description><![CDATA[
Recent advances in Focused Ion Beam-Scanning Electron Microscopy (FIB-SEM) allows the imaging and analysis of cellular ultrastructure at nanoscale resolution, but the collection of labels and/or noise-free data sets has several challenges, often immutable. Reasons range from time consuming manual annotations, requiring highly trained specialists, to introducing imaging artifacts from the prolonged scanning during acquisition. We propose a fully unsupervised Noise Reconstruction and Removal Network for denoising scanning electron microscopy images.

The architecture, inspired by gated recurrent units, reconstructs and removes the noise by synthesizing the sequential data. At the same time the fully unsupervised training guides the network in distinguishing true signal from noise and gives comparable results to supervised architectures. We demonstrate that this new network specialized on 3D electron microscopy data sets, achieves comparable and even better results than supervised networks.
]]></description>
<dc:creator>Giannios, K.</dc:creator>
<dc:creator>Chaurasia, A.</dc:creator>
<dc:creator>Thibault, G.</dc:creator>
<dc:creator>Riesterer, J. L.</dc:creator>
<dc:creator>Stempinski, E.</dc:creator>
<dc:creator>Lo, T.</dc:creator>
<dc:creator>DeLaRosa, B.</dc:creator>
<dc:creator>Gray, J. W.</dc:creator>
<dc:date>2021-05-28</dc:date>
<dc:identifier>doi:10.1101/2021.05.27.446051</dc:identifier>
<dc:title><![CDATA[Noise Reconstruction & Removal Network: A New Architecture to Denoise FIB-SEM Images]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-05-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.06.02.446828v1?rss=1">
<title>
<![CDATA[
A homeostatic interferon-lambda response to bacterial microbiota stimulates preemptive antiviral defense within discrete pockets of intestinal epithelium 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.06.02.446828v1?rss=1</link>
<description><![CDATA[
Interferon-lambda (IFN-{lambda}) protects intestinal epithelial cells (IECs) from enteric viruses by inducing expression of antiviral IFN-stimulated genes (ISGs). Here, we find that bacterial microbiota stimulate a homeostatic ISG signature in the intestine of specific pathogen-free mice. This homeostatic ISG expression is restricted to IECs, depends on IEC-intrinsic expression of IFN-{lambda} receptor (Ifnlr1), and is associated with IFN-{lambda} production by leukocytes. Strikingly, imaging of these homeostatic ISGs reveals localization to pockets of the epithelium and concentration in mature IECs. Correspondingly, a minority of mature IECs express these ISGs in public single-cell RNA sequencing datasets from mice and humans. Furthermore, we assessed the ability of orally-administered bacterial components to restore localized ISGs in mice lacking bacterial microbiota. Lastly, we find that IECs lacking Ifnlr1 are hyper-susceptible to initiation of murine rotavirus infection. These observations indicate that bacterial microbiota stimulate ISGs in localized regions of the intestinal epithelium at homeostasis, thereby preemptively activating antiviral defenses in vulnerable IECs to improve host defense against enteric viruses.
]]></description>
<dc:creator>Van Winkle, J. A.</dc:creator>
<dc:creator>Peterson, S. T.</dc:creator>
<dc:creator>Kennedy, E. A.</dc:creator>
<dc:creator>Wheadon, M. J.</dc:creator>
<dc:creator>Ingle, H.</dc:creator>
<dc:creator>Desai, C.</dc:creator>
<dc:creator>Constant, D. A.</dc:creator>
<dc:creator>Wright, A. P.</dc:creator>
<dc:creator>Li, L.</dc:creator>
<dc:creator>Artyomov, M.</dc:creator>
<dc:creator>Lee, S.</dc:creator>
<dc:creator>Baldridge, M. T.</dc:creator>
<dc:creator>Nice, T. J.</dc:creator>
<dc:date>2021-06-02</dc:date>
<dc:identifier>doi:10.1101/2021.06.02.446828</dc:identifier>
<dc:title><![CDATA[A homeostatic interferon-lambda response to bacterial microbiota stimulates preemptive antiviral defense within discrete pockets of intestinal epithelium]]></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/10.1101/2021.06.09.447758v1?rss=1">
<title>
<![CDATA[
Incorporation of unnatural amino acid for the tagging of cannabinoid receptors 1 and 2 reveals receptor roles in regulating cAMP levels 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.06.09.447758v1?rss=1</link>
<description><![CDATA[
The role of CB1/CB2 co-expression in cell signaling remains elusive. We established a simplified mammalian cell model system in which expression of CB1 or CB2 can be easily monitored under a confocal microscope. For this, we applied amber codon suppression in live cells to incorporate a single trans-cyclooctene (TCO) bearing amino acid in one of the extracellular loops of CB1 or CB2, followed by fluorescent labeling via click chemistry. We employed genetically encoded biosensors to measure the roles of CB1 and/or CB2 in regulating intracellular calcium ([Ca2+]i) and cAMP ([cAMP]i) levels. We show that the agonist-mediated activation of tagged-CB1 or -CB2 can transiently elevate [Ca2+]i levels. However, when the two receptors were co-expressed in the same cell, CB2 no longer signaled through calcium although CB1-mediated transient elevation of [Ca2+]i levels was unaffected. Because of the existence of crosstalk between calcium and cAMP signaling, we measured the effects of CB1 and/or CB2 in regulating adenylate cyclase activity. We found that the expression of CB1 increased forskolin-induced [cAMP]i levels compared to non-transfected cells. Conversely, CB2 expression decreased stimulated [cAMP]i levels under the same conditions. Finally, co-expressed CB1 and CB2 receptors showed additive yet opposing effects on stimulated [cAMP]i levels. These observations suggest that co-expressed CB1/CB2 act locally as a pair in regulating cell excitability by modulating stimulated [cAMP]i levels.
]]></description>
<dc:creator>Thomas, A.</dc:creator>
<dc:creator>Schultz, C.</dc:creator>
<dc:creator>Laguerre, A.</dc:creator>
<dc:date>2021-06-09</dc:date>
<dc:identifier>doi:10.1101/2021.06.09.447758</dc:identifier>
<dc:title><![CDATA[Incorporation of unnatural amino acid for the tagging of cannabinoid receptors 1 and 2 reveals receptor roles in regulating cAMP levels]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-06-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.06.10.447598v1?rss=1">
<title>
<![CDATA[
Encapsulated allografts preclude host sensitization and promote ovarian endocrine function in ovariectomized young rhesus monkeys and sensitized mice 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.06.10.447598v1?rss=1</link>
<description><![CDATA[
Transplantation of allogeneic donor ovarian tissue holds great potential for female cancer survivors who often experience premature ovarian insufficiency. To avoid complications associated with immune suppression and to protect transplanted ovarian allografts from immune-mediated injury, we have developed an immuno-isolating hydrogel-based capsule that supports the function of ovarian allografts without triggering an immune response. Encapsulated ovarian allografts implanted in naive ovariectomized BALB/c mice responded to the circulating gonadotropins without direct revascularization and maintained function for 4 months, as evident by regular estrous cycles and presence of antral follicles in the retrieved grafts. Repeated implantations of encapsulated mouse ovarian allografts did not sensitize naive BALB/c mice in contrast to non-encapsulated controls, which was confirmed with undetectable levels of allo-antibody. Further, encapsulated allografts implanted in hosts previously sensitized by implantation of non-encapsulated allografts restored estrous cycles similarly to our results in naive recipients. Next, we tested the translational potential and efficiency of the immune-isolating capsule in a rhesus monkey model by implanting encapsulated ovarian auto- and allografts in young ovariectomized animals. The encapsulated ovarian grafts survived and restored basal levels of urinary estrone conjugate and pregnanediol 3-glucuronide during the approximate 4-5 month observation period. We demonstrate, for the first time, that encapsulation of ovarian allografts prevents sensitization and protects the allograft from rejection in young rhesus monkeys and in sensitized mice.
]]></description>
<dc:creator>Day, J. R.</dc:creator>
<dc:creator>Flanagan, C. L.</dc:creator>
<dc:creator>David, A.</dc:creator>
<dc:creator>Hartigan-O'Connor, D. J.</dc:creator>
<dc:creator>de Mattos Barbosa, M. G.</dc:creator>
<dc:creator>Martinez, M. L.</dc:creator>
<dc:creator>Lee, C.</dc:creator>
<dc:creator>Barnes, J.</dc:creator>
<dc:creator>Farkash, E.</dc:creator>
<dc:creator>Zelinski, M.</dc:creator>
<dc:creator>Tarantal, A.</dc:creator>
<dc:creator>Cascalho, M.</dc:creator>
<dc:creator>Shikanov, A.</dc:creator>
<dc:date>2021-06-13</dc:date>
<dc:identifier>doi:10.1101/2021.06.10.447598</dc:identifier>
<dc:title><![CDATA[Encapsulated allografts preclude host sensitization and promote ovarian endocrine function in ovariectomized young rhesus monkeys and sensitized mice]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-06-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.06.11.448065v1?rss=1">
<title>
<![CDATA[
Mechanical stress in pancreatic cancer: Signaling pathway adaptation activates cytoskeletal remodeling and enhances cell migration 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.06.11.448065v1?rss=1</link>
<description><![CDATA[
New treatments for patients with advanced or metastatic pancreatic cancers are urgently needed due to their resistance to all current therapies. Current studies focus on alternative treatment approaches that target or normalize the abnormal microenvironment of pancreatic tumors, which among others, is responsible for elevated mechanical stress in the tumor interior. Nevertheless, the underlying mechanisms by which mechanical stress regulates pancreatic cancer metastatic potential remain elusive. Herein, we used a large-scale proteomic assay to profile mechanical stress-induced signaling cascades that drive the motility of pancreatic cancer cells. Proteomic analysis, together with selective protein inhibition and siRNA treatments, revealed that mechanical stress enhances cell migration through activation of the p38 MAPK/HSP27 and JNK/c-Jun signaling axes, and activation of the actin cytoskeleton remodelers: Rac1, cdc42, and Myosin II. Our results highlight targeting aberrant signaling in cancer cells that are adapted to the mechanical tumor microenvironment as a novel approach to effectively limit pancreatic cancer cell migration.
]]></description>
<dc:creator>Kalli, M.</dc:creator>
<dc:creator>Li, R.</dc:creator>
<dc:creator>Mills, G. B.</dc:creator>
<dc:creator>Stylianopoulos, T.</dc:creator>
<dc:creator>Zervantonakis, I. K.</dc:creator>
<dc:date>2021-06-11</dc:date>
<dc:identifier>doi:10.1101/2021.06.11.448065</dc:identifier>
<dc:title><![CDATA[Mechanical stress in pancreatic cancer: Signaling pathway adaptation activates cytoskeletal remodeling and enhances cell migration]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-06-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.06.15.448579v1?rss=1">
<title>
<![CDATA[
Dietary palmitic acid induces trained immunity that controls inflammation and infection 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.06.15.448579v1?rss=1</link>
<description><![CDATA[
Trained immunity is an innate immune memory response that is induced by primary microbial or sterile stimuli that sensitizes monocytes and macrophages to a secondary pathogenic challenge, reprogramming the host response to infection and inflammatory disease. Nutritional components, such as dietary fatty acids, can act as inflammatory stimuli, but it is unknown if they can act as the primary stimuli in the context of innate immune memory. Here we find mice fed diets enriched in saturated fatty acids (SFAs) confer a hyper-inflammatory response to systemic lipopolysaccharide (LPS) and increased mortality, independent of diet-induced microbiome and glycemic modulation. Ex vivo, we show monocytes and splenocytes from mice fed enriched SFAs do not have altered baseline inflammation, but enhanced responses to a secondary inflammatory challenge. Lipidomics identified enhanced free palmitic acid (PA) and PA-associated lipids in SFA-fed mice serum. We found pre-treatment with physiologically relevant concentrations of PA alone reprograms macrophages to induce a hyper-inflammatory response to secondary challenge with LPS. This response was found to be dependent on the synthesis of ceramide, and reversible when treated with oleic acid, a mono-unsaturated FA that depletes intracellular ceramide. In vivo, we found systemic PA confers enhanced inflammation and mortality during an acute septic response to systemic LPS, which was not reversible for up to 7 days post-PA-exposure. While PA-treatment is harmful for acute septic shock outcome, we find PA exposure enhanced clearance of Candida albicans in RAG-/- mice. These are the first data to implicate enriched dietary SFAs, and specifically PA, in the induction of long-lived innate immune memory that is detrimental during an acute septic response, but beneficial for clearance of pathogens.
]]></description>
<dc:creator>Seufert, A. L.</dc:creator>
<dc:creator>Hickman, J. W.</dc:creator>
<dc:creator>Traxler, S. K.</dc:creator>
<dc:creator>Peterson, R. M.</dc:creator>
<dc:creator>Lashley, S. J.</dc:creator>
<dc:creator>Shulzhenko, N.</dc:creator>
<dc:creator>Napier, R. J.</dc:creator>
<dc:creator>Napier, B. A.</dc:creator>
<dc:date>2021-06-16</dc:date>
<dc:identifier>doi:10.1101/2021.06.15.448579</dc:identifier>
<dc:title><![CDATA[Dietary palmitic acid induces trained immunity that controls inflammation and infection]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-06-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.06.16.448597v1?rss=1">
<title>
<![CDATA[
Physiologically distinct neurons within the ventrolateral periaqueductal gray are not defined by mu-opioid receptor expression but are differentially activated by persistent inflammation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.06.16.448597v1?rss=1</link>
<description><![CDATA[
The ventrolateral periaqueductal gray (vlPAG) is a key structure within the descending pain modulatory pathway and an important target for opioid-induced analgesia. This area contains heterogeneous neurons with respect to neurotransmitter and receptor expression so it is difficult to define vlPAG neurons that contribute to pain and analgesia. Characterization of intrinsic membrane properties of 371 vlPAG neurons from female and male Long-Evans rats identified 4 neuron types with distinct intrinsic firing patterns: Phasic, Tonic, Onset, and Random. Phasic and Tonic neurons comprise the majority of the neurons sampled. Mu-opioid receptor (MOR) expression was determined by the ability of the selective MOR agonist DAMGO to activate G protein-coupled inwardly-rectifying potassium channel (GIRK) currents. Opioid-sensitive and -insensitive neurons were observed within each neuron type in naive rats and in rats pretreated with Complete Freunds adjuvant in a hindpaw to produce persistent inflammation. The presence of low threshold spikes (LTS) did not correlate with MOR-mediated GIRK currents indicating that MOR expression alone does not define a physiologically distinct neuron type in the vlPAG. MOR activation inhibited firing in nearly all spontaneously active neurons, both in naive and persistent inflammation conditions. CFA-induced inflammation increased Fos expression at both acute (2 h) and persistent inflammation (5-7 d) time points. However, persistent, but not acute, inflammation selectively enhanced spontaneous firing and lowered firing thresholds of Phasic neurons which was maintained in the absence of synaptic inputs. Taken together, persistent inflammation selectively activates Phasic neurons, of which only a subset was opioid-sensitive.

Significance statementIntrinsic membrane properties define separate vlPAG neurons types that are functionally important. Persistent, and not acute, inflammation selectively activates Phasic firing vlPAG neurons that are not defined by MOR expression. Although the vlPAG is known to contribute to the descending inhibition of pain, the activation of a single physiologically-defined neuron type in the presence of inflammation may represent a mechanism by which the vlPAG participates in descending facilitation of pain.
]]></description>
<dc:creator>McPherson, K. B.</dc:creator>
<dc:creator>Bouchet, C. A.</dc:creator>
<dc:creator>Ingram, S. L.</dc:creator>
<dc:date>2021-06-17</dc:date>
<dc:identifier>doi:10.1101/2021.06.16.448597</dc:identifier>
<dc:title><![CDATA[Physiologically distinct neurons within the ventrolateral periaqueductal gray are not defined by mu-opioid receptor expression but are differentially activated by persistent inflammation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-06-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.06.21.449128v1?rss=1">
<title>
<![CDATA[
The neurocomputational architecture of explore-exploit decision making 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.06.21.449128v1?rss=1</link>
<description><![CDATA[
Humans and other animals often make the difficult decision to try new options (exploration) and forego immediate rewards (exploitation). Novelty-seeking is an adaptive solution to this explore-exploit dilemma, but our understanding of the neural computations supporting novelty-seeking in humans is limited. Here, we presented the same explore-exploit decision making task to monkeys and humans and found evidence that the computational basis for novelty-seeking is conserved across primate species. Critically, through computational model-based decomposition of event-related functional magnetic resonance imaging (fMRI) in humans, these findings reveal a previously unidentified cortico-subcortical architecture mediating explore-exploit behavior in humans.
]]></description>
<dc:creator>Hogeveen, J.</dc:creator>
<dc:creator>Mullins, T. S.</dc:creator>
<dc:creator>Romero, J.</dc:creator>
<dc:creator>Eversole, E.</dc:creator>
<dc:creator>Rogge-Obando, K.</dc:creator>
<dc:creator>Mayer, A. R.</dc:creator>
<dc:creator>Costa, V. D.</dc:creator>
<dc:date>2021-06-21</dc:date>
<dc:identifier>doi:10.1101/2021.06.21.449128</dc:identifier>
<dc:title><![CDATA[The neurocomputational architecture of explore-exploit decision making]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-06-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.06.25.449905v1?rss=1">
<title>
<![CDATA[
Persistence of SARS CoV-2 S1 Protein in CD16+ Monocytes in Post-Acute Sequelae of COVID-19 (PASC) Up to 15 Months Post-Infection 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.06.25.449905v1?rss=1</link>
<description><![CDATA[
The recent COVID-19 pandemic is a treatment challenge in the acute infection stage but the recognition of chronic COVID-19 symptoms termed post-acute sequelae SARS-CoV-2 infection (PASC) may affect up to 30% of all infected individuals. The underlying mechanism and source of this distinct immunologic condition three months or more after initial infection remains elusive. Here, we investigated the presence of SARS-CoV-2 S1 protein in 46 individuals. We analyzed T-cell, B-cell, and monocytic subsets in both severe COVID-19 patients and in patients with post-acute sequelae of COVID-19 (PASC). The levels of both intermediate (CD14+, CD16+) and non-classical monocyte (CD14Lo, CD16+) were significantly elevated in PASC patients up to 15 months post-acute infection compared to healthy controls (P=0.002 and P=0.01, respectively). A statistically significant number of non-classical monocytes contained SARS-CoV-2 S1 protein in both severe (P=0.004) and PASC patients (P=0.02) out to 15 months post-infection. Non-classical monocytes were sorted from PASC patients using flow cytometric sorting and the SARS-CoV-2 S1 protein was confirmed by mass spectrometry. Cells from 4 out of 11 severe COVID-19 patients and 1 out of 26 PASC patients contained ddPCR+ peripheral blood mononuclear cells, however, only fragmented SARS-CoV-2 RNA was found in PASC patients. No full length sequences were identified, and no sequences that could account for the observed S1 protein were identified in any patient. Non-classical monocytes are capable of causing inflammation throughout the body in response to fractalkine/CX3CL1 and RANTES/CCR5.
]]></description>
<dc:creator>Patterson, B.</dc:creator>
<dc:creator>Francisco, E. B.</dc:creator>
<dc:creator>Yogendra, R.</dc:creator>
<dc:creator>Long, E.</dc:creator>
<dc:creator>Pise, A.</dc:creator>
<dc:creator>Rodrigues, H.</dc:creator>
<dc:creator>Hall, E.</dc:creator>
<dc:creator>Herrera, o.</dc:creator>
<dc:creator>Parikh, P.</dc:creator>
<dc:creator>Guevara-Coto, J.</dc:creator>
<dc:creator>Chang, X.</dc:creator>
<dc:creator>Sacha, J. B.</dc:creator>
<dc:creator>Rodriguez, R.</dc:creator>
<dc:creator>Mora, J.</dc:creator>
<dc:date>2021-06-25</dc:date>
<dc:identifier>doi:10.1101/2021.06.25.449905</dc:identifier>
<dc:title><![CDATA[Persistence of SARS CoV-2 S1 Protein in CD16+ Monocytes in Post-Acute Sequelae of COVID-19 (PASC) Up to 15 Months Post-Infection]]></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/10.1101/2021.06.30.449980v1?rss=1">
<title>
<![CDATA[
Structural Insights into the Venus flytrap Mechanosensitive Ion Channel Flycatcher1 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.06.30.449980v1?rss=1</link>
<description><![CDATA[
Flycatcher1 (FLYC1), a MscS homolog, has recently been identified as a candidate mechanosensitive (MS) ion channel involved in Venus flytrap prey recognition. FLYC1 is larger and its sequence diverges from previously studied MscS homologs, suggesting it has unique structural features that contribute to its function. Here, we characterized FLYC1 by cryo-electron microscopy, molecular dynamics simulations, and electrophysiology. Akin to bacterial MscS and plant MSL1 channels, we find that FLYC1 central core includes side portals in the cytoplasmic cage that regulate ion conduction, by identifying critical residues that modulate channel conductance. Topologically unique cytoplasmic flanking regions can adopt  up or  down conformations, making the channel asymmetric. Disruption of an up conformation-specific interaction severely delays channel deactivation by 40-fold likely due to stabilization of the channel open state. Our results illustrate novel structural features and likely conformational transitions that regulate mechano-gating of FLYC1.
]]></description>
<dc:creator>Jojoa-Cruz, S.</dc:creator>
<dc:creator>Saotome, K.</dc:creator>
<dc:creator>Tsui, C. C.</dc:creator>
<dc:creator>Lee, W.-H.</dc:creator>
<dc:creator>Sansom, M. S. P.</dc:creator>
<dc:creator>Murthy, S. E.</dc:creator>
<dc:creator>Patapoutian, A.</dc:creator>
<dc:creator>Ward, A. B.</dc:creator>
<dc:date>2021-06-30</dc:date>
<dc:identifier>doi:10.1101/2021.06.30.449980</dc:identifier>
<dc:title><![CDATA[Structural Insights into the Venus flytrap Mechanosensitive Ion Channel Flycatcher1]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-06-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.07.05.451195v1?rss=1">
<title>
<![CDATA[
Adolescent dopamine neurons represent reward differently during action and state guided learning 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.07.05.451195v1?rss=1</link>
<description><![CDATA[
The neuronal underpinning of learning cause-and-effect associations in the adolescent brain remains poorly understood. Two fundamental forms of associative learning are Pavlovian (classical) conditioning, where a stimulus is followed by an outcome, and operant (instrumental) conditioning, where outcome is contingent on action execution. Both forms of learning, when associated with a rewarding outcome, rely on midbrain dopamine neurons in the ventral tegmental area (VTA) and substantia nigra (SN). We find that in adolescent male rats, reward-guided associative learning is encoded differently by midbrain dopamine neurons in each conditioning paradigm. Whereas simultaneously recorded VTA and SN adult neurons have a similar phasic response to reward delivery during both forms of conditioning, adolescent neurons display a muted reward response during operant but a profoundly larger reward response during Pavlovian conditioning suggesting that adolescent neurons assign a different value to reward when it is not gated by action. The learning rate of adolescents and adults during both forms of conditioning was similar further supporting the notion that differences in reward response in each paradigm are due to differences in motivation and independent of state versus action value learning. Static characteristics of dopamine neurons such as dopamine cell number and size were similar in the VTA and SN but there were age differences in baseline firing rate, stimulated release and correlated spike activity suggesting that differences in reward responsiveness by adolescent dopamine neurons are not due to differences in intrinsic properties of these neurons but engagement of different networks.
]]></description>
<dc:creator>McCane, A.</dc:creator>
<dc:creator>Wegener, M.</dc:creator>
<dc:creator>Faraji, M.</dc:creator>
<dc:creator>Garcia, M. R.</dc:creator>
<dc:creator>Wallin-Miller, K.</dc:creator>
<dc:creator>Costa, V.</dc:creator>
<dc:creator>Moghaddam, B.</dc:creator>
<dc:date>2021-07-06</dc:date>
<dc:identifier>doi:10.1101/2021.07.05.451195</dc:identifier>
<dc:title><![CDATA[Adolescent dopamine neurons represent reward differently during action and state guided learning]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-07-06</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/10.1101/2021.07.08.451636v1?rss=1">
<title>
<![CDATA[
Pyroptosis-dependent and -independent cross-priming of CD8+ T cells by intestinal epithelial cell-derived antigen 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.07.08.451636v1?rss=1</link>
<description><![CDATA[
The innate immune system detects pathogens and initiates adaptive immune responses. Inflammasomes are central components of the innate immune system, but whether inflammasomes provide sufficient signals to activate adaptive immunity is unclear. In intestinal epithelial cells (IECs), inflammasomes activate a lytic form of cell death called pyroptosis, leading to epithelial cell expulsion and the release of cytokines. Here we employed a genetic system to show that simultaneous antigen expression and inflammasome activation specifically in IECs is sufficient to activate CD8+ T cells. By genetic elimination of direct T cell priming by IECs, we found that IEC-derived antigens are cross-presented to CD8+ T cells. However, activation of CD8+ T cells by IEC-derived antigen only partially depended on IEC pyroptosis. In the absence of inflammasome activation, cross-priming of CD8+ T cells required Batf3+ dendritic cells (cDC1), whereas cross-priming in the presence of pyroptosis did not. These data suggest the existence of parallel pyroptosis-dependent and pyroptosis-independent but cDC1-dependent pathways for cross-presentation of IEC-derived antigens.
]]></description>
<dc:creator>Deets, K. A.</dc:creator>
<dc:creator>Nichols, R. D.</dc:creator>
<dc:creator>Rauch, I.</dc:creator>
<dc:creator>Vance, R. E.</dc:creator>
<dc:date>2021-07-08</dc:date>
<dc:identifier>doi:10.1101/2021.07.08.451636</dc:identifier>
<dc:title><![CDATA[Pyroptosis-dependent and -independent cross-priming of CD8+ T cells by intestinal epithelial cell-derived antigen]]></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/10.1101/2021.07.14.452355v1?rss=1">
<title>
<![CDATA[
Prolonged partner separation erodes nucleus accumbens transcriptional signatures of pair bonding in male prairie voles 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.07.14.452355v1?rss=1</link>
<description><![CDATA[
The loss of a spouse is often cited as the most traumatic event in a persons life. However, for most people, the severity of grief and its maladaptive effects subside over time via an understudied adaptive process. Like humans, socially monogamous prairie voles (Microtus ochrogaster) form opposite-sex pair bonds, and upon partner separation, show stress phenotypes that diminish over time. We test the hypothesis that extended partner separation diminishes pair bond-associated behaviors and causes pair bond transcriptional signatures to erode. Pairs were cohoused for 2 weeks and then either remained paired or were separated for 48hrs or 4wks before collecting fresh nucleus accumbens tissue for RNAseq. In a separate cohort, we assessed partner preference and selective aggression at these time points, finding that these behaviors persist despite prolonged separation in both same-sex and opposite-sex paired voles. Opposite-sex pair bonding led to changes in accumbal transcription that were stably maintained while animals remained paired but eroded following prolonged partner separation. Eroded genes are associated with gliogenesis and myelination, suggesting a previously undescribed role for glia in pair bonding and loss. Further, we pioneered neuron-specific translating ribosomal affinity purification in voles. Neuronally-enriched transcriptional changes revealed dopaminergic-, mitochondrial-, and steroid hormone signaling-associated gene clusters sensitive to acute pair bond disruption and loss adaptation. Our results suggest that partner separation erodes transcriptomic signatures of pair bonding despite core behavioral features of the bond remaining intact, revealing potential molecular processes priming a vole to be able to form a new bond.
]]></description>
<dc:creator>Sadino, J. M.</dc:creator>
<dc:creator>Bradeen, X. G.</dc:creator>
<dc:creator>Kelly, C. J.</dc:creator>
<dc:creator>Walker, D. M.</dc:creator>
<dc:creator>Donaldson, Z. R.</dc:creator>
<dc:date>2021-07-14</dc:date>
<dc:identifier>doi:10.1101/2021.07.14.452355</dc:identifier>
<dc:title><![CDATA[Prolonged partner separation erodes nucleus accumbens transcriptional signatures of pair bonding in male prairie voles]]></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/10.1101/2021.07.15.452525v1?rss=1">
<title>
<![CDATA[
GPR39 Localization in Aging Human Brain and Correlation of Expression and Polymorphism with Vascular Cognitive Impairment 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.07.15.452525v1?rss=1</link>
<description><![CDATA[
INTRODUCTIONThe pathogenesis of vascular cognitive impairment (VCI) is not fully understood. GPR39, an orphan G-protein coupled receptor, is implicated in neurological disorders but its role in VCI is unknown.

METHODSWe performed GPR39 immunohistochemical analysis in postmortem brain samples from mild cognitive impairment (MCI) and control subjects. DNA was analyzed for GPR39 SNPs, and correlated with white matter hyperintensity (WMH) burden on premortem MRI.

RESULTSGPR39 is expressed in aged human dorsolateral prefrontal cortex, localized to microglia and peri-capillary cells resembling pericytes. GPR39-capillary colocalization, and density of GPR39-expressing microglia was increased in aged brains compared to young. SNP distribution was equivalent between groups; however, homozygous SNP carriers were present only in the MCI group, and had higher WMH volume than WT or heterozygous SNP carriers.

DISCUSSIONGPR39 may play a role in aging-related VCI, and may serve as a therapeutic target and biomarker for the risk of developing VCI.
]]></description>
<dc:creator>Davis, C.</dc:creator>
<dc:creator>Bah, T.</dc:creator>
<dc:creator>Zhang, W.</dc:creator>
<dc:creator>Nelson, J.</dc:creator>
<dc:creator>Golgotiu, K.</dc:creator>
<dc:creator>Nie, X.</dc:creator>
<dc:creator>Alkayed, F.</dc:creator>
<dc:creator>Young, J.</dc:creator>
<dc:creator>Woltjer, R.</dc:creator>
<dc:creator>Silbert, L.</dc:creator>
<dc:creator>Grafe, M.</dc:creator>
<dc:creator>Alkayed, N. J.</dc:creator>
<dc:date>2021-07-15</dc:date>
<dc:identifier>doi:10.1101/2021.07.15.452525</dc:identifier>
<dc:title><![CDATA[GPR39 Localization in Aging Human Brain and Correlation of Expression and Polymorphism with Vascular Cognitive Impairment]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-07-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.07.15.452558v1?rss=1">
<title>
<![CDATA[
Axial spondyloarthritis patients have altered mucosal IgA response to oral and fecal microbiota 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.07.15.452558v1?rss=1</link>
<description><![CDATA[
ObjectiveTo investigate whether axial spondyloarthritis (AxSpA) patients have an altered immunoglobulin A (IgA) response in the gut and oral microbial communities.

MethodsWe performed 16S rRNA gene (16S) sequencing on IgA positive (IgA+) and IgA negative (IgA-) fractions (IgA-SEQ) from feces (n=17 AxSpA; n=14 healthy) and saliva (n=17 AxSpA; n=12 healthy), as well as on IgA-unsorted fecal and salivary samples. PICRUSt2 was used to predict microbial metabolic potential in AxSpA patients and healthy controls (HCs).

ResultsIgA-SEQ revealed enrichment of several microbes in the fecal (Akkermansia, Ruminococcaceae, Lachnospira) and salivary (Prevotellaceae, Actinobacillus) microbiome in AxSpA patients as compared with HCs. Fecal microbiome from AxSpA patients showed a trend towards increased alpha diversity of the IgA+ fraction and decreased diversity in the IgA- fraction in comparison with HCs, while the salivary microbiome exhibits a significant decrease in alpha diversity in both IgA+ and IgA- fractions. Increased IgA coating of Clostridiales Family XIII correlated with disease severity. Inferred metagenomic analysis suggests perturbation of metabolites and metabolic pathways for inflammation (oxidative phosphorylation, glutathione metabolism) and metabolism (propanoate and butanoate metabolism) in AxSpA patients.

ConclusionsAnalyses of fecal and salivary microbes from AxSpA patients reveal distinct populations of immunoreactive microbes using novel IgA-SEQ approach, which were not captured by comparing their relative abundance with HCs. Predictive metagenomic analysis revealed perturbation of metabolites/metabolic pathways in AxSpA patients. Future studies on these immunoreactive microbes may lead to better understanding of the functional role of IgA in maintaining microbial structure and human health.
]]></description>
<dc:creator>Gill, T.</dc:creator>
<dc:creator>Stauffer, P.</dc:creator>
<dc:creator>Asquith, M.</dc:creator>
<dc:creator>Laderas, T.</dc:creator>
<dc:creator>Martin, T. M.</dc:creator>
<dc:creator>Davin, S.</dc:creator>
<dc:creator>Schleisman, M.</dc:creator>
<dc:creator>Ramirez, C.</dc:creator>
<dc:creator>Ogle, K.</dc:creator>
<dc:creator>Lindquist, I.</dc:creator>
<dc:creator>Nguyen, J.</dc:creator>
<dc:creator>Planck, S. R.</dc:creator>
<dc:creator>Shaut, C.</dc:creator>
<dc:creator>Diamond, S.</dc:creator>
<dc:creator>Rosenbaum, J. T.</dc:creator>
<dc:creator>Karstens, L.</dc:creator>
<dc:date>2021-07-19</dc:date>
<dc:identifier>doi:10.1101/2021.07.15.452558</dc:identifier>
<dc:title><![CDATA[Axial spondyloarthritis patients have altered mucosal IgA response to oral and fecal microbiota]]></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/10.1101/2021.07.24.453572v1?rss=1">
<title>
<![CDATA[
Lateral habenula M2 muscarinic receptor control of neuronal activity and cocaine seeking behavior 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.07.24.453572v1?rss=1</link>
<description><![CDATA[
The lateral habenula (LHb) plays a central role in balancing reward and aversion by opposing the contributions of brain reward nuclei. Using a rat cocaine self-administration model, we previously found that LHb inhibition or non-selective blockade of LHb muscarinic acetylcholine receptors (mAChRs) led to persistent cocaine seeking despite its signaled unavailability. As understanding roles for the LHb and cholinergic signaling in behavioral control is important to psychiatric illness and addiction, we examine how mAChRs act on LHb neurons using in vitro electrophysiology. We find that different groups of LHb neurons are depolarized or hyperpolarized by the cholinergic agonist carbachol (CCh), and that CCh could inhibit GABAergic and glutamatergic synaptic inputs to these cells. Presynaptic CCh effects were reversed by the M2 mAChR (M2R) antagonist AFDX-116, but not by pirenzepine, an M1R antagonist. Contemporaneous measurement of CCh effects on synaptic inhibition and excitation in LHb neurons showed a smaller effect on inhibition, suggesting a net shift in synaptic integration toward greater inhibition by mAChRs. Synaptic currents elicited by light-activation of ventral tegmental area (VTA) axons in the LHb, following channelrhodopsin-2 transfection of VTA, were also inhibited by M2Rs, suggesting the VTA as at least one M2R-sensitive LHb afferent. Finally, Go-NoGo cocaine seeking studies showed that blockade of LHb M2Rs, and not M1Rs, triggered continued cocaine seeking. These data identify LHb M2Rs as a potential control point of LHb function that enables withholding responses for cocaine and define cellular mechanisms through which mAChRs modulate LHb activity.
]]></description>
<dc:creator>Wolfe, C. I. C.</dc:creator>
<dc:creator>Hwang, E.-K.</dc:creator>
<dc:creator>Zapata, A.</dc:creator>
<dc:creator>Hoffman, A. F.</dc:creator>
<dc:creator>Lupica, C. R.</dc:creator>
<dc:date>2021-07-25</dc:date>
<dc:identifier>doi:10.1101/2021.07.24.453572</dc:identifier>
<dc:title><![CDATA[Lateral habenula M2 muscarinic receptor control of neuronal activity and cocaine seeking behavior]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-07-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.07.26.453746v1?rss=1">
<title>
<![CDATA[
EXPRESSION OF A SECRETABLE, CELL-PENETRATING CDKL5 PROTEIN ENHANCES THE EFFICACY OF AAV VECTOR-MEDIATED GENE THERAPY FOR CDKL5 DEFICIENCY DISORDER. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.07.26.453746v1?rss=1</link>
<description><![CDATA[
No therapy is currently available for CDKL5 (cyclin-dependent kinase-like 5) deficiency disorder (CDD), a severe neurodevelopmental disorder caused by mutations in the CDKL5 gene. Although delivery of a wild-type copy of the mutated gene to cells represents the most curative approach for a monogenic disease, proof-of-concept studies highlight significant efficacy caveats for brain gene therapy. Herein, we used a secretable TATk-CDKL5 protein to enhance the efficiency of a gene therapy for CDD. We found that, although AAVPHP.B_Igk-TATk-CDKL5 and AAVPHP.B_CDKL5 vectors had similar brain infection efficiency, the AAVPHP.B_Igk-TATk-CDKL5 vector led to a higher CDKL5 protein replacement due to secretion and transduction of the TATk-CDKL5 protein into the neighboring cells. Importantly, Cdkl5 KO mice treated with the AAVPHP.B_Igk-TATk-CDKL5 vector showed a behavioral and neuroanatomical improvement in comparison with vehicle-treated Cdkl5 KO mice or Cdkl5 KO mice treated with the AAVPHP.B_CDKL5 vector, indicating that a gene therapy based on a secretable recombinant TATk-CDKL5 protein is more effective at compensating Cdkl5-null brain defects than gene therapy based on the expression of the native CDKL5.
]]></description>
<dc:creator>Medici, G.</dc:creator>
<dc:creator>Tassinari, M.</dc:creator>
<dc:creator>Galvani, G.</dc:creator>
<dc:creator>Bastianini, S.</dc:creator>
<dc:creator>Gennaccaro, L.</dc:creator>
<dc:creator>Loi, M.</dc:creator>
<dc:creator>Fuchs, C.</dc:creator>
<dc:creator>Mottolese, N.</dc:creator>
<dc:creator>Alvente, S.</dc:creator>
<dc:creator>Berteotti, C.</dc:creator>
<dc:creator>Bagget, H. R.</dc:creator>
<dc:creator>Zoccoli, G.</dc:creator>
<dc:creator>Giustetto, M.</dc:creator>
<dc:creator>Muotri, A. R.</dc:creator>
<dc:creator>Nakai, H.</dc:creator>
<dc:creator>Trazzi, S.</dc:creator>
<dc:creator>Ciani, E.</dc:creator>
<dc:date>2021-07-26</dc:date>
<dc:identifier>doi:10.1101/2021.07.26.453746</dc:identifier>
<dc:title><![CDATA[EXPRESSION OF A SECRETABLE, CELL-PENETRATING CDKL5 PROTEIN ENHANCES THE EFFICACY OF AAV VECTOR-MEDIATED GENE THERAPY FOR CDKL5 DEFICIENCY DISORDER.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-07-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.07.28.454223v1?rss=1">
<title>
<![CDATA[
Exploratory extracellular vesicle-bound miRNA profiling to identify candidate biomarkers of chronic alcohol drinking in non-human primates 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.07.28.454223v1?rss=1</link>
<description><![CDATA[
BackgroundLong-term alcohol drinking is associated with numerous health complications including susceptibility to infection, cancer, and organ damage. However, due to the complex nature of human drinking behavior, it is challenging to determine whether alcohol use should be considered a risk factor during diagnosis and treatment. We lack reliable biomarkers of alcohol use that could be used to determine drinking behavior prior to signs of overt organ damage. Recently, extracellular vesicle-bound microRNA (EV-miRNA) have been discovered to be consistent biomarkers of conditions including cancer and liver disease.

MethodsIn this study, we profiled the plasma EV-miRNA content by miRNA-Seq from 80 non-human primates after 12 months of voluntary ethanol drinking.

ResultsWe identified a list of up- and downregulated EV-miRNA candidate biomarkers of both heavy drinking as well as those positively correlated with ethanol dose. We further overexpressed these candidate miRNA in control primary peripheral immune cells to assess potential functional mechanisms of these EV-miRNA. We identified that overexpression of miR-155, miR-154, miR-34c, miR-450a, and miR-204 led to increased inflammatory TNF or IL-6 production in PBMC after stimulation.

ConclusionThis exploratory study identified several EV-miRNA that could serve as biomarkers of long-term alcohol drinking as well as provided a mechanism for alcohol-induced peripheral inflammation.
]]></description>
<dc:creator>Lewis, S. A.</dc:creator>
<dc:creator>Doratt, B.</dc:creator>
<dc:creator>Sureshchandra, S.</dc:creator>
<dc:creator>Pan, T.</dc:creator>
<dc:creator>Gonzalez, S. W.</dc:creator>
<dc:creator>Shen, W.</dc:creator>
<dc:creator>Grant, K. A.</dc:creator>
<dc:creator>Messaoudi, I.</dc:creator>
<dc:date>2021-07-29</dc:date>
<dc:identifier>doi:10.1101/2021.07.28.454223</dc:identifier>
<dc:title><![CDATA[Exploratory extracellular vesicle-bound miRNA profiling to identify candidate biomarkers of chronic alcohol drinking in non-human primates]]></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/10.1101/2021.07.30.454528v1?rss=1">
<title>
<![CDATA[
Chronic ethanol drinking in non-human primates induces inflammatory cathepsin gene expression in alveolar macrophages accompanied by functional defects 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.07.30.454528v1?rss=1</link>
<description><![CDATA[
Chronic alcohol drinking is associated with increased susceptibility to viral and bacterial respiratory pathogens. Investigating the effects of alcohol on the lung is challenging in humans because of the complexity of human drinking behavior and the challenge of obtaining samples. In this study, we utilize a rhesus macaque model of voluntary ethanol self-administration to study the effects of alcohol on the lung in a physiologically and genetically relevant model. We report a heightened activation and inflammatory state in alveolar macrophages (AM) obtained from ethanol drinking animals that is accompanied by increased chromatin accessibility in intergenic regions that regulate inflammatory genes and contain binding motifs for transcription factors AP-1, IRF8, and NFKB p-65. In line with these transcriptional and epigenetic changes at basal state, AM from ethanol drinking animals generate elevated inflammatory mediator responses to LPS and respiratory syncytial virus (RSV). Analysis using scRNA-Seq revealed heterogeneity in lung-resident macrophage and monocyte populations, including increased abundance of activated and cathepsin-expressing clusters and accelerated differentiation with ethanol. Finally, functional assays show increased mitochondrial content in AM from ethanol drinking animals, which is associated with observed increased ROS and decreased phagocytosis capacity. This comprehensive epigenomic, transcriptional and functional profiling of lung macrophages after ethanol drinking in macaques provides previously unidentified mechanisms of ethanol induced infection susceptibility in patients with alcohol use disorders.
]]></description>
<dc:creator>Lewis, S. A.</dc:creator>
<dc:creator>Doratt, B.</dc:creator>
<dc:creator>Sureshchandra, S.</dc:creator>
<dc:creator>Jankeel, A.</dc:creator>
<dc:creator>Newman, N.</dc:creator>
<dc:creator>Grant, K. A.</dc:creator>
<dc:creator>Messaoudi, I.</dc:creator>
<dc:date>2021-07-31</dc:date>
<dc:identifier>doi:10.1101/2021.07.30.454528</dc:identifier>
<dc:title><![CDATA[Chronic ethanol drinking in non-human primates induces inflammatory cathepsin gene expression in alveolar macrophages accompanied by functional defects]]></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/10.1101/2021.08.04.455138v1?rss=1">
<title>
<![CDATA[
Sex-dependent development of Kras-induced anal squamous cell carcinoma in mice 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.08.04.455138v1?rss=1</link>
<description><![CDATA[
Anal squamous cell carcinoma (SCC) will be diagnosed in an estimated 9,080 adults in the United States this year, and rates have been rising over the last several decades. Most people that develop anal SCC have associated human papillomavirus (HPV) infection ([~]85-95%), with approximately 5-15% of anal SCC cases occurring in HPV-negative patients from unknown etiology. This study identified and characterized a Kras-driven, female sex hormone-dependent development of anal squamous cell carcinoma (SCC) in the LSL-KrasG12D ; Pdx1-Cre (KC) mouse model that is not dependent on papillomavirus infection. One hundred percent of female KC mice develop anal SCC, while no male KC mice develop tumors. Both male and female KC anal tissue express Pdx1 and Cre-recombinase mRNA, and the activated mutant KrasG12D gene. Although the driver gene mutation KrasG12D is present in anus of both sexes, only female KC mice develop Kras-mutant induced anal SCC. To understand the sex-dependent differences, KC male mice were castrated and KC female mice were ovariectomized. Castrated KC males displayed an unchanged phenotype with no anal tumor formation. In contrast, ovariectomized KC females demonstrated a marked reduction in anal SCC development, with only 15% developing anal SCC. Finally, exogenous administration of estrogen rescued the tumor development in ovariectomized KC female mice and induced tumor development in castrated KC males. These results confirm that the anal SCC is estrogen mediated. The delineation of the role of female sex hormones in mediating mutant Kras to drive anal SCC pathogenesis highlights a subtype of anal SCC that is independent of papillomavirus infection. These findings may have clinical applicability for the papillomavirus-negative subset of anal SCC patients that typically respond poorly to standard of care chemoradiation.
]]></description>
<dc:creator>Walcheck, M.</dc:creator>
<dc:creator>Matkowskyj, K.</dc:creator>
<dc:creator>Turco, A. E.</dc:creator>
<dc:creator>Blaine-Sauer, S.</dc:creator>
<dc:creator>Nukaya, M.</dc:creator>
<dc:creator>Noel, J. L.</dc:creator>
<dc:creator>Ronnekleiv, O. K.</dc:creator>
<dc:creator>Ronnekleiv-Kelly, S.</dc:creator>
<dc:date>2021-08-06</dc:date>
<dc:identifier>doi:10.1101/2021.08.04.455138</dc:identifier>
<dc:title><![CDATA[Sex-dependent development of Kras-induced anal squamous cell carcinoma in mice]]></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/10.1101/2021.08.04.455147v1?rss=1">
<title>
<![CDATA[
BET inhibitors synergize with anti-PD1 by rescuing TCF1+ progenitor exhausted CD8+ T cells in Acute Myeloid Leukemia 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.08.04.455147v1?rss=1</link>
<description><![CDATA[
Many acute myeloid leukemia (AML) patients exhibit hallmarks of immune exhaustion, such as increased myeloid derived suppressor cells (MDSCs), suppressive regulatory T cells (Tregs) and dysfunctional T cells. We have developed a mouse model of AML driven by Flt3-ITD and Tet2 deficiency displays these immune-related features, including CD8+ T cells exhibiting a terminally exhausted phenotype (TEx). This T cell subset has been shown to be refractory to immune checkpoint blockade (ICB) monotherapy. Here we show that small molecule inhibitors which target bromodomain and extra-terminal domain (BET) proteins affect both tumor-intrinsic factors but also rescue T cell exhaustion and ICB resistance. Ex vivo treatment of cells from AML mice and AML patients with BET inhibitors (BETi) reversed CD8+ T cell exhaustion by restoring proliferative capacity and expansion of the more functional precursor exhausted T cells (TPEx). This reversal is enhanced by combined BETi and anti-PD1 treatment. Finally, we show that BETi synergizes with anti-PD1 in vivo, resulting in the reduction of circulating leukemia cells, enrichment of CD8+ T cells in the bone marrow, and increased expression of Tcf7, Slamf6, and Cxcr5 in CD8+ T cells. In total, we show the potential efficacy of combining BETi and ICB therapy in the treatment of AML.
]]></description>
<dc:creator>Romine, K. A.</dc:creator>
<dc:creator>Cho, H. J.</dc:creator>
<dc:creator>Kosaka, Y.</dc:creator>
<dc:creator>Byrd, K.</dc:creator>
<dc:creator>Coy, J. L.</dc:creator>
<dc:creator>Flynn, P. A.</dc:creator>
<dc:creator>Newman, M. T.</dc:creator>
<dc:creator>Loo, C.</dc:creator>
<dc:creator>Scott, J.</dc:creator>
<dc:creator>Lind, E. F.</dc:creator>
<dc:date>2021-08-04</dc:date>
<dc:identifier>doi:10.1101/2021.08.04.455147</dc:identifier>
<dc:title><![CDATA[BET inhibitors synergize with anti-PD1 by rescuing TCF1+ progenitor exhausted CD8+ T cells in Acute Myeloid 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/10.1101/2021.08.06.455429v1?rss=1">
<title>
<![CDATA[
A LINCS microenvironment perturbation resource for integrative assessment of ligand-mediated molecular and phenotypic responses 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.08.06.455429v1?rss=1</link>
<description><![CDATA[
The phenotype of a cell and its underlying molecular state is strongly influenced by extracellular signals, including growth factors, hormones, and extracellular matrix. While these signals are normally tightly controlled, their dysregulation leads to phenotypic and molecular states associated with diverse diseases. To develop a detailed understanding of the linkage between molecular and phenotypic changes, we generated a comprehensive dataset that catalogs the transcriptional, proteomic, epigenomic and phenotypic responses of MCF10A mammary epithelial cells after exposure to the ligands EGF, HGF, OSM, IFNG, TGFB and BMP2. Systematic assessment of the molecular and cellular phenotypes induced by these ligands comprise the LINCS Microenvironment (ME) perturbation dataset, which has been curated and made publicly available for community-wide analysis and development of novel computational methods (synapse.org/LINCS_MCF10A). In illustrative analyses, we demonstrate how this dataset can be used to discover functionally related molecular features linked to specific cellular phenotypes.
]]></description>
<dc:creator>Gross, S. M.</dc:creator>
<dc:creator>Dane, M. A.</dc:creator>
<dc:creator>Smith, R. L.</dc:creator>
<dc:creator>Devlin, K.</dc:creator>
<dc:creator>Mclean, I.</dc:creator>
<dc:creator>Derrick, D.</dc:creator>
<dc:creator>Mills, C.</dc:creator>
<dc:creator>Subramanian, K.</dc:creator>
<dc:creator>London, A.</dc:creator>
<dc:creator>Torre, D.</dc:creator>
<dc:creator>Erdem, C.</dc:creator>
<dc:creator>Lyons, N.</dc:creator>
<dc:creator>Natoli, T.</dc:creator>
<dc:creator>Pessa, S.</dc:creator>
<dc:creator>Lu, X.</dc:creator>
<dc:creator>Mullahoo, J.</dc:creator>
<dc:creator>Li, J.</dc:creator>
<dc:creator>Adam, M.</dc:creator>
<dc:creator>Wassie, B.</dc:creator>
<dc:creator>Liu, M.</dc:creator>
<dc:creator>Kilburn, D. F.</dc:creator>
<dc:creator>Liby, T.</dc:creator>
<dc:creator>Bucher, E.</dc:creator>
<dc:creator>Sanchez-Aguila, C.</dc:creator>
<dc:creator>Daily, K.</dc:creator>
<dc:creator>Omberg, L.</dc:creator>
<dc:creator>Wang, Y.</dc:creator>
<dc:creator>Jacobson, C.</dc:creator>
<dc:creator>Yapp, C.</dc:creator>
<dc:creator>Chung, M.</dc:creator>
<dc:creator>Vidovic, D.</dc:creator>
<dc:creator>Lu, Y.</dc:creator>
<dc:creator>Schurer, S.</dc:creator>
<dc:creator>Lee, A.</dc:creator>
<dc:creator>Pillai, A.</dc:creator>
<dc:creator>Subramanian, A.</dc:creator>
<dc:creator>Papanastasiou, M.</dc:creator>
<dc:creator>Fraenkel, E.</dc:creator>
<dc:creator>Feiler, H.</dc:creator>
<dc:creator>Mills, G. B.</dc:creator>
<dc:creator>Jaffe, J.</dc:creator>
<dc:creator>Ma'ayan, A.</dc:creator>
<dc:creator>Birtwistle, M. R.</dc:creator>
<dc:creator>Sorger, P. K.</dc:creator>
<dc:creator>Korkola, J. E.</dc:creator>
<dc:creator>Gray, J. W.</dc:creator>
<dc:creator>Heiser, L. M.</dc:creator>
<dc:date>2021-08-09</dc:date>
<dc:identifier>doi:10.1101/2021.08.06.455429</dc:identifier>
<dc:title><![CDATA[A LINCS microenvironment perturbation resource for integrative assessment of ligand-mediated molecular and phenotypic responses]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-08-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.08.09.455068v1?rss=1">
<title>
<![CDATA[
Immediate Impact of Yogic Breathing on Pulsatile Cerebrospinal Fluid Dynamics 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.08.09.455068v1?rss=1</link>
<description><![CDATA[
Cerebrospinal fluid (CSF), a clear fluid bathing the central nervous system (CNS), undergoes pulsatile movements, and plays a critical role for the removal of waste products from the brain including amyloid beta, a protein associated with Alzheimers disease. Regulation of CSF dynamics is critical for maintaining CNS health, and increased pulsatile CSF dynamics may alter brains waste clearance due to increased mixing and diffusion. As such, understanding the mechanisms driving CSF movement, and interventions that influence its resultant removal of wastes from the brain is of high scientific and clinical impact. Since pulsatile CSF dynamics is sensitive and synchronous to respiratory movements, we are interested in identifying potential integrative therapies such as yogic breathing to regulate and enhance CSF dynamics, which has not been reported before. Here, we investigated the pre-intervention baseline data from our ongoing randomized controlled trial, and examined whether yogic breathing immediately impacts pulsatile CSF dynamics compared to spontaneous breathing. We utilized our previously established non-invasive real-time phase contrast magnetic resonance imaging (RT-PCMRI) approach using a 3T MRI instrument, and computed and rigorously tested differences in CSF velocities (instantaneous, respiratory, cardiac 1st and 2nd harmonics) at the level of foramen magnum during spontaneous versus four yogic breathing patterns. In examinations of 18 healthy participants (eight females, ten males; mean age 34.9 {+/-} 14 (SD) years; age range: 18-61 years), we discovered immediate increase in cranially-directed velocities of instantaneous-CSF 16% - 28% and respiratory-CSF 60% - 118% during yogic versus spontaneous breathing, with most statistically significant changes during deep abdominal breathing (28%, p=0.0008, and 118%, p=0.0001, respectively). Further, cardiac pulsation was the primary source of pulsatile CSF during all breathing conditions except during deep abdominal breathing, when there was a comparable contribution of respiratory and cardiac 1st harmonic power [0.59 {+/-} 0.78], demonstrating respiration can be the primary regulator of CSF depending on individual differences in breath depth and location. Further work is needed to investigate the impact of sustained training yogic breathing on increased pulsatile CSF dynamics and brain waste clearance for CNS health.
]]></description>
<dc:creator>Yildiz, S.</dc:creator>
<dc:creator>Grinstead, J.</dc:creator>
<dc:creator>Hildebrand, A.</dc:creator>
<dc:creator>Oshinski, J.</dc:creator>
<dc:creator>Rooney, W. D.</dc:creator>
<dc:creator>Lim, M. M.</dc:creator>
<dc:creator>Oken, B.</dc:creator>
<dc:date>2021-08-10</dc:date>
<dc:identifier>doi:10.1101/2021.08.09.455068</dc:identifier>
<dc:title><![CDATA[Immediate Impact of Yogic Breathing on Pulsatile Cerebrospinal Fluid Dynamics]]></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/10.1101/2021.08.13.456301v1?rss=1">
<title>
<![CDATA[
FLARIM v2.0, an improved method to quantify transcript-ribosome interactions in vivo in the adult Drosophila brain 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.08.13.456301v1?rss=1</link>
<description><![CDATA[
Neural injury triggers striking immune reactions from glial cells, including significant transcriptional and morphological changes, but it is unclear how these events are coordinated to mount an effective immune response. Here, we present a new variant of the Fluorescence assay to detect ribosome interactions with mRNA (FLARIM), which we term FLARIM v2.0, to visualize single immune gene transcripts and association with ribosomes in glia responding to neurodegeneration. Specifically, using an in vivo axotomy assay in Drosophila, we show that matrix metalloproteinase-1 (Mmp-1) mRNAs and associated ribosomes are detected in distal processes of reactive glia where they are actively engulfing degenerating axonal material, suggesting that local translation is an important component of the glial immune response to axotomy. This work also validates our enhanced FLARIM assay as a promising tool to investigate mechanisms of mRNA transport and translation in a wide range of in vitro and in vivo paradigms.
]]></description>
<dc:creator>Richer, P.</dc:creator>
<dc:creator>Speese, S. D.</dc:creator>
<dc:creator>Logan, M. A.</dc:creator>
<dc:date>2021-08-14</dc:date>
<dc:identifier>doi:10.1101/2021.08.13.456301</dc:identifier>
<dc:title><![CDATA[FLARIM v2.0, an improved method to quantify transcript-ribosome interactions in vivo in the adult Drosophila brain]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-08-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.08.14.456365v1?rss=1">
<title>
<![CDATA[
Fanconi Anemia Pathway Deficiency Drives Copy Number Variation in Squamous Cell Carcinomas 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.08.14.456365v1?rss=1</link>
<description><![CDATA[
Fanconi anemia (FA), a model syndrome of genome instability, is caused by a deficiency in DNA interstrand crosslink (ICL) repair resulting in chromosome breakage1-3. The FA repair pathway comprises at least 22 FANC proteins including BRCA1 and BRCA24-6, and protects against carcinogenic endogenous and exogenous aldehydes7-10. Individuals with FA are hundreds to thousands-fold more likely to develop head and neck (HNSCC), esophageal and anogenital squamous cell carcinomas (SCCs) with a median onset age of 31 years11. The aggressive nature of these tumors and poor patient tolerance of platinum and radiation-based therapy have been associated with short survival in FA11-16. Molecular studies of SCCs from individuals with FA (FA SCCs) have been limited, and it is unclear how they relate to sporadic HNSCCs primarily driven by tobacco and alcohol exposure or human papillomavirus (HPV) infection17. Here, by sequencing FA SCCs, we demonstrate that the primary genomic signature of FA-deficiency is the presence of a high number of structural variants (SVs). SVs are enriched for small deletions, unbalanced translocations, and fold-back inversions that arise in the context of TP53 loss. The SV breakpoints preferentially localize to early replicating regions, common fragile sites, tandem repeats, and SINE elements. SVs are often connected forming complex rearrangements. Resultant genomic instability underlies elevated copy number alteration (CNA) rates of key HNSCC-associated genes, including PIK3CA, MYC, CSMD1, PTPRD, YAP1, MXD4, and EGFR. In contrast to sporadic HNSCC, we find no evidence of HPV infection in FA HNSCC, although positive cases were identified in gynecologic tumors. A murine allograft model of FA pathway-deficient SCC was enriched in SVs, exhibited dramatic tumor growth advantage, more rapid epithelial-to-mesenchymal transition (EMT), and enhanced autonomous inflammatory signaling when compared to an FA pathway-proficient model. In light of the protective role of the FA pathway against SV formation uncovered here, and recent findings of FA pathway insufficiency in the setting of increased formaldehyde load resulting in hematopoietic stem cell failure and carcinogenesis18-20, we propose that high copy-number instability in sporadic HNSCC may result from functional overload of the FA pathway by endogenous and exogenous DNA crosslinking agents. Our work lays the foundation for improved FA patient treatment and demonstrates that FA SCC is a powerful model to study tumorigenesis resulting from DNA crosslinking damage.
]]></description>
<dc:creator>Webster, A. L. H.</dc:creator>
<dc:creator>Sanders, M. A.</dc:creator>
<dc:creator>Patel, K.</dc:creator>
<dc:creator>Dietrich, R.</dc:creator>
<dc:creator>Noonan, R. J.</dc:creator>
<dc:creator>Lach, F. P.</dc:creator>
<dc:creator>White, R. R.</dc:creator>
<dc:creator>Goldfarb, A.</dc:creator>
<dc:creator>Hadi, K.</dc:creator>
<dc:creator>Edwards, M. M.</dc:creator>
<dc:creator>Donovan, F. X.</dc:creator>
<dc:creator>Jung, M.</dc:creator>
<dc:creator>Sridhar, S.</dc:creator>
<dc:creator>Fedrigo, O.</dc:creator>
<dc:creator>Tian, H.</dc:creator>
<dc:creator>Rosiene, J.</dc:creator>
<dc:creator>Heineman, T.</dc:creator>
<dc:creator>Kennedy, J. A.</dc:creator>
<dc:creator>Bean, L.</dc:creator>
<dc:creator>Rosti, O.</dc:creator>
<dc:creator>Tryon, R.</dc:creator>
<dc:creator>Gonzalez, A.-M.</dc:creator>
<dc:creator>Rosenberg, A.</dc:creator>
<dc:creator>Luo, J.-D.</dc:creator>
<dc:creator>Carrol, T.</dc:creator>
<dc:creator>Velleuer, E.</dc:creator>
<dc:creator>Rastatter, J. C.</dc:creator>
<dc:creator>Wells, S. I.</dc:creator>
<dc:creator>Surralles, J.</dc:creator>
<dc:creator>Bagby, G.</dc:creator>
<dc:creator>MacMillan, M. L.</dc:creator>
<dc:creator>Wagner, J. E.</dc:creator>
<dc:creator>Cancio, M.</dc:creator>
<dc:creator>Boulad, F.</dc:creator>
<dc:creator>Scognamiglio, T.</dc:creator>
<dc:creator>Vaughan, R.</dc:creator>
<dc:creator>Koren, A.</dc:creator>
<dc:creator>Imielinski, M.</dc:creator>
<dc:creator>Chandrasekharappa, S.</dc:creator>
<dc:creator>Auerbach, A. D.</dc:creator>
<dc:creator>Singh, B.</dc:creator>
<dc:creator>Kutler, D. I.</dc:creator>
<dc:creator>Campbell, P.</dc:creator>
<dc:date>2021-08-16</dc:date>
<dc:identifier>doi:10.1101/2021.08.14.456365</dc:identifier>
<dc:title><![CDATA[Fanconi Anemia Pathway Deficiency Drives Copy Number Variation in Squamous Cell Carcinomas]]></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/10.1101/2021.08.23.457408v1?rss=1">
<title>
<![CDATA[
Deep immune profiling of the maternal-fetal interface with mild SARS-CoV-2 infection 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.08.23.457408v1?rss=1</link>
<description><![CDATA[
Pregnant women are an at-risk group for severe COVID-19, though the majority experience mild/asymptomatic disease. Although severe COVID-19 has been shown to be associated with immune activation at the maternal-fetal interface even in the absence of active viral replication, the immune response to asymptomatic/mild COVID-19 remains unknown. Here, we assessed immunological adaptations in both blood and term decidua from 9 SARS-exposed pregnant women with asymptomatic/mild disease and 15 pregnant SARS-naive women. In addition to selective loss of tissue-resident decidual macrophages, we report attenuation of antigen presentation and type I IFN signaling but upregulation of inflammatory cytokines and chemokines in blood monocyte derived decidual macrophages. On the other hand, infection was associated with remodeling of the T cell compartment with increased frequencies of activated CD69+ tissue-resident T cells and decreased abundance of Tregs. Interestingly, frequencies of cytotoxic CD4 and CD8 T cells increased only in the blood, while CD8 effector memory T cells were expanded in the decidua. In contrast to decidual macrophages, signatures of type I IFN signaling were increased in decidual T cells. Finally, T cell receptor diversity was significantly reduced with infection in both compartments, albeit to a much greater extent in the blood. The resulting aberrant immune activation in the placenta, even with asymptomatic disease may alter the exquisitely sensitive developing fetal immune system, leading to long-term adverse outcomes for offspring.
]]></description>
<dc:creator>Sureshchandra, S.</dc:creator>
<dc:creator>Zulu, M. Z.</dc:creator>
<dc:creator>Doratt, B.</dc:creator>
<dc:creator>Jankeel, A.</dc:creator>
<dc:creator>Tifrea, D.</dc:creator>
<dc:creator>Edwards, R. A.</dc:creator>
<dc:creator>Rincon, M.</dc:creator>
<dc:creator>Marshall, N. E.</dc:creator>
<dc:creator>Messaoudi, I.</dc:creator>
<dc:date>2021-08-23</dc:date>
<dc:identifier>doi:10.1101/2021.08.23.457408</dc:identifier>
<dc:title><![CDATA[Deep immune profiling of the maternal-fetal interface with mild SARS-CoV-2 infection]]></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/10.1101/2021.08.24.456354v1?rss=1">
<title>
<![CDATA[
PU.1 and MYC transcriptional network defines synergistic drug responses to KIT and LSD1 inhibition in acute myeloid leukemia 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.08.24.456354v1?rss=1</link>
<description><![CDATA[
Activating mutations in the KIT tyrosine receptor kinase confer an adverse prognosis for patients with acute myeloid leukemia (AML). Outside of bone marrow transplantation, treatment options are limited. Here we demonstrate combined KIT and LSD1 inhibition produces synergistic cell death against KIT mutant AML cells. This combination suppresses MYC expression to drive cell cycle exit and apoptosis. This decreased MYC expression results from a loss of PU.1 binding at downstream MYC enhancers. The drug combination also inactivates PI3K/AKT/GSK3a/b signaling to decrease MYC protein abundance. KIT-mutant AML cells rapidly adapt to KIT inhibitor monotherapy by restoring PI3K/AKT activity, but cannot when treated with combined KIT and LSD1 inhibitor. In addition, we validate MYC suppression as a mechanism of synergy in KIT-mutant AML patient samples. Collectively, this work provides rational for a clinical trial to assess the efficacy of KIT and LSD1 inhibition in patients with KIT-mutant AML.

Statement of significanceEffective treatment options for AML are limited. We describe the synergistic response to combined KIT and LSD1 inhibition in KIT-mutant AML and identify key biomarkers of drug response. The specificity and efficacy of this combination in cell lines and patient samples provides rationale for investigation in early phase clinical trials.
]]></description>
<dc:creator>Smith, B. M.</dc:creator>
<dc:creator>VanCampen, J.</dc:creator>
<dc:creator>Kong, G. L.</dc:creator>
<dc:creator>Yashar, W.</dc:creator>
<dc:creator>Horton, W.</dc:creator>
<dc:creator>Coleman, D. J.</dc:creator>
<dc:creator>Estabrook, J.</dc:creator>
<dc:creator>Lusardi, T. A.</dc:creator>
<dc:creator>Druker, B. J.</dc:creator>
<dc:creator>Maxson, J. E.</dc:creator>
<dc:creator>Braun, T. P.</dc:creator>
<dc:date>2021-08-24</dc:date>
<dc:identifier>doi:10.1101/2021.08.24.456354</dc:identifier>
<dc:title><![CDATA[PU.1 and MYC transcriptional network defines synergistic drug responses to KIT and LSD1 inhibition in acute myeloid leukemia]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-08-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.08.25.457547v1?rss=1">
<title>
<![CDATA[
Increased 20-HETE signaling suppresses neurovascular coupling after ischemic stroke in regions beyond the infarct 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.08.25.457547v1?rss=1</link>
<description><![CDATA[
Neurovascular coupling, the process by which neuronal activity elicits increases in the local blood supply, is impaired in stroke patients in brain regions outside the infarct. Such impairment may contribute to neurological deterioration over time, but its mechanism is unknown. Using the middle cerebral artery occlusion (MCAO) model of stroke, we show that neuronal activity-evoked capillary dilation is reduced by [~]75% in the intact cortical tissue outside the infarct border. This decrease in capillary responsiveness was not explained by a decrease in local neuronal activity or a loss of vascular contractility. Inhibiting synthesis of the vasoconstrictive molecule 20-HETE, either by inhibiting its synthetic enzyme CYP450 {omega}-hydroxylases or by increasing nitric oxide (NO), which is a natural inhibitor of {omega}-hydroxylases, rescued activity-evoked capillary dilation. The capillary dilation unmasked by inhibiting 20-HETE was dependent on PGE2 activation of EP4 receptors, a vasodilatory pathway previously identified in healthy animals. Cortical 20-HETE levels were increased following MCAO, in agreement with data from stroke patients. Inhibition of {omega}-hydroxylases normalized 20-HETE levels in vivo and increased cerebral blood flow in the peri-infarct cortex. These data identify 20-HETE-dependent vasoconstriction as a mechanism underlying neurovascular coupling impairment after stroke. Our results suggest that the brains energy supply may be significantly reduced after stroke in regions previously believed to be asymptomatic and that {omega}-hydroxylase inhibition may restore healthy neurovascular coupling post-stroke.
]]></description>
<dc:creator>Li, Z.</dc:creator>
<dc:creator>McConnell, H. L.</dc:creator>
<dc:creator>Stackhouse, T. L.</dc:creator>
<dc:creator>Pike, M. M.</dc:creator>
<dc:creator>Zhang, W.</dc:creator>
<dc:creator>Mishra, A.</dc:creator>
<dc:date>2021-08-27</dc:date>
<dc:identifier>doi:10.1101/2021.08.25.457547</dc:identifier>
<dc:title><![CDATA[Increased 20-HETE signaling suppresses neurovascular coupling after ischemic stroke in regions beyond the infarct]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-08-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.08.26.457551v1?rss=1">
<title>
<![CDATA[
Infection-induced dermal lymphatic zippering restricts viral dissemination from skin and promotes anti-viral CD8+ T cell expansion. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.08.26.457551v1?rss=1</link>
<description><![CDATA[
Lymphatic vessels are often considered passive conduits that rapidly flush antigenic material, pathogens, and cells to draining lymph nodes. Recent evidence, however, suggests that lymphatic vessels actively regulate diverse processes from antigen transport to leukocyte trafficking and dietary lipid absorption. Here we tested the hypothesis that dermal lymphatic transport is dynamic and contributes to innate host defense during viral infection. We demonstrate that cutaneous vaccinia virus infection activates the tightening of lymphatic interendothelial junctions, termed zippering, in a VEGFA/VEGFR2-dependent manner. Both antibody-mediated blockade of VEGFA/VEGFR2 and lymphatic-specific deletion of Vegfr2 impaired lymphatic capillary zippering and increased fluid flux out of tissue. Strikingly, inhibition of lymphatic zippering allows viral dissemination to draining lymph nodes independent of dendritic cell migration and impairs CD8+ T cell priming. These data indicate that infection-induced dermal lymphatic capillary zippering is a context-dependent, active mechanism of innate host defense that limits interstitial fluid and virion flux and promotes protective, anti-viral CD8+ T cell responses.

SummaryCutaneous infection with vaccinia virus induces VEGFR2-dependent dermal lymphatic capillary zippering. This tightening of lymphatic junctions exacerbates tissue edema, sequesters virus, and promotes anti-viral CD8+ T cell responses. Dermal lymphatic capillaries are therefore an active component of innate host defense.
]]></description>
<dc:creator>Churchill, M. J.</dc:creator>
<dc:creator>du Bois, H.</dc:creator>
<dc:creator>Heim, T. A.</dc:creator>
<dc:creator>Mudianto, T.</dc:creator>
<dc:creator>Steele, M. M.</dc:creator>
<dc:creator>Nolz, J. C.</dc:creator>
<dc:creator>Lund, A. W.</dc:creator>
<dc:date>2021-08-27</dc:date>
<dc:identifier>doi:10.1101/2021.08.26.457551</dc:identifier>
<dc:title><![CDATA[Infection-induced dermal lymphatic zippering restricts viral dissemination from skin and promotes anti-viral CD8+ T cell expansion.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-08-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.08.26.457870v1?rss=1">
<title>
<![CDATA[
MRI-Visible Perivascular Space (PVS) Changes with Long-Duration Spaceflight 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.08.26.457870v1?rss=1</link>
<description><![CDATA[
Humans are exposed to extreme environmental stressors during spaceflight and return with alterations in brain structure and shifts in intracranial fluids. To date, no studies have evaluated the effects of spaceflight on perivascular spaces (PVSs) within the brain, which are believed to facilitate fluid drainage and brain homeostasis. Here, we examined how the number and morphology of magnetic resonance imaging (MRI)-visible PVSs are affected by spaceflight, including prior spaceflight experience. Fifteen astronauts underwent six T1-weighted 3T MRI scans, twice prior to launch and four times following their return to Earth after [~]6-month missions to the International Space Station. White matter MRI-visible PVS number and morphology were calculated using an established automated segmentation algorithm. We found that novice astronauts showed an increase in total PVS volume from pre- to post-flight, whereas experienced crewmembers did not (adjusted for age, sex, and time between landing and first MRI scan). Moreover, experienced astronauts exhibited a significant correlation between more previous flight days and greater PVS median length at baseline, suggesting that experienced astronauts exhibit holdover effects from prior spaceflight(s). There was also a significant positive correlation between pre- to post-flight increases in PVS median length and increases in right lateral ventricular volume. The presence of spaceflight associated neuro-ocular syndrome (SANS) was not associated with PVS number or morphology. Together, these findings demonstrate that spaceflight is associated with PVS morphological changes, and specifically that spaceflight experience is an important factor in determining PVS characteristics.
]]></description>
<dc:creator>Hupfeld, K.</dc:creator>
<dc:creator>Richmond, S.</dc:creator>
<dc:creator>McGregor, H.</dc:creator>
<dc:creator>Schwartz, D.</dc:creator>
<dc:creator>Luther, M.</dc:creator>
<dc:creator>Beltran, N.</dc:creator>
<dc:creator>Kofman, I.</dc:creator>
<dc:creator>De Dios, Y.</dc:creator>
<dc:creator>Riascos, R.</dc:creator>
<dc:creator>Wood, S.</dc:creator>
<dc:creator>Bloomberg, J.</dc:creator>
<dc:creator>Mulavara, A.</dc:creator>
<dc:creator>Silbert, L.</dc:creator>
<dc:creator>Iliff, J.</dc:creator>
<dc:creator>Seidler, R.</dc:creator>
<dc:creator>Piantino, J.</dc:creator>
<dc:date>2021-08-28</dc:date>
<dc:identifier>doi:10.1101/2021.08.26.457870</dc:identifier>
<dc:title><![CDATA[MRI-Visible Perivascular Space (PVS) Changes with Long-Duration Spaceflight]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-08-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.08.27.457999v1?rss=1">
<title>
<![CDATA[
Highly sensitive genetically-encoded sensors for population and subcellular imaging of cAMP in vivo 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.08.27.457999v1?rss=1</link>
<description><![CDATA[
Cyclic adenosine monophosphate (cAMP) integrates information from diverse G protein-coupled receptors, such as neuromodulator receptors, to regulate pivotal biological processes in a cellular- and subcellular-specific manner. However, in vivo cellular-resolution imaging of cAMP dynamics in neurons has not been demonstrated. Here, we screen existing genetically-encoded cAMP sensors, and further develop the best performer to derive three improved variants, called cAMPFIREs. These sensors exhibit up to ten-fold increased sensitivity to cAMP and a corrected, cytosolic distribution. cAMPFIREs are compatible with both ratiometric and fluorescence lifetime imaging, and can detect cAMP dynamics elicited by norepinephrine at physiologically-relevant, nanomolar concentrations. Imaging of cAMPFIREs in awake mice reveals tonic levels of cAMP in cortical neurons that are associated with wakefulness, and are differentially regulated in different subcellular compartments. Furthermore, enforced locomotion elicits neuron-specific, bidirectional cAMP dynamics, in part, mediated by norepinephrine. Finally, cAMPFIREs also function in Drosophila, suggesting that they have broad applicability for studying intracellular signaling in vivo.
]]></description>
<dc:creator>Massengill, C. I.</dc:creator>
<dc:creator>Bayless-Edwards, L.</dc:creator>
<dc:creator>Ceballos, C. C.</dc:creator>
<dc:creator>Cebul, E. R.</dc:creator>
<dc:creator>Qin, M.</dc:creator>
<dc:creator>Whorton, M. R.</dc:creator>
<dc:creator>Ye, B.</dc:creator>
<dc:creator>Mao, T.</dc:creator>
<dc:creator>Zhong, H.</dc:creator>
<dc:date>2021-08-28</dc:date>
<dc:identifier>doi:10.1101/2021.08.27.457999</dc:identifier>
<dc:title><![CDATA[Highly sensitive genetically-encoded sensors for population and subcellular imaging of cAMP in vivo]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-08-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.09.01.458631v1?rss=1">
<title>
<![CDATA[
Multiplex spatial systems analysis of local nanodose drug responses predicts effective treatment combinations of immunotherapies and targeted agents in mammary carcinoma 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.09.01.458631v1?rss=1</link>
<description><![CDATA[
Better methods are needed to identify effective combinations of immunotherapies with chemotherapies and targeted anti-cancer agents. Here we present a Multiplex Implantable Microdevice Assay (MIMA) system for rapid in vivo assessment of the effects of multiple, spatially separate anticancer drugs directly in the complex tumor microenvironment. In prototypic experiments, olaparib, lenvatinib, palbociclib, venetoclax, panobinostat, doxorubicin, and paclitaxel and combinations thereof were administered simultaneously to murine mammary tumor models. Quantitative multiplex immunohistochemistry and spatial systems analyses of each local drug response defined cellular relations of fibroblasts, endothelial cells, immune lineages, immunogenic cell death, tumor proliferation and/or cancer stem cells that were used to predict effective drug combinations. A predicted combination of panobinostat, venetoclax and anti-CD40 showed long-term anti-tumor efficacy in multiple mouse models with no observable toxicity when administered systemically. Future MIMA use promises to design effective drug combinations for tumor cell control and immune activation on a personalized basis.
]]></description>
<dc:creator>Tatarova, Z.</dc:creator>
<dc:creator>Blumberg, D. C.</dc:creator>
<dc:creator>Korkola, J. E.</dc:creator>
<dc:creator>Heiser, L. M.</dc:creator>
<dc:creator>Muschler, J. L.</dc:creator>
<dc:creator>Schedin, P. J.</dc:creator>
<dc:creator>Ahn, S. W.</dc:creator>
<dc:creator>Mills, G. B.</dc:creator>
<dc:creator>Coussens, L. M.</dc:creator>
<dc:creator>Jonas, O.</dc:creator>
<dc:creator>Gray, J. W.</dc:creator>
<dc:date>2021-09-02</dc:date>
<dc:identifier>doi:10.1101/2021.09.01.458631</dc:identifier>
<dc:title><![CDATA[Multiplex spatial systems analysis of local nanodose drug responses predicts effective treatment combinations of immunotherapies and targeted agents in mammary carcinoma]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-09-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.09.02.458753v1?rss=1">
<title>
<![CDATA[
Glial TGFβ activity promotes axon survival in peripheral nerves 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.09.02.458753v1?rss=1</link>
<description><![CDATA[
Axons can represent the majority of the volume of a neuron and are energetically very demanding. Specialized glia ensheathe axons and are believed to support axon function and maintenance throughout life, but molecular details of glia-neuron support mechanisms remain poorly defined. Here we identify a collection of secreted and transmembrane genes that are required in glia for long-term axon survival in vivo. We show that key components of the TGF{beta} superfamily are required cell-autonomously in glia for peripheral nerve maintenance, although their loss does not disrupt glial morphology. We observe age-dependent neurodegeneration in the absence of glial TGF{beta} signaling that can be rescued by genetic blockade of Wallerian degeneration. Our data argue that glial TGF{beta} signaling normally acts to promote axon survival and suppress neurodegeneration.

Significance StatementAxon maintenance is critical to preserving the functional integrity of the nervous system across animal lifespan. Glia contribute to axon maintenance, but their precise roles remain to be fully characterized. We identify glial genes that regulate axon support and provide new molecular insight into the means by which glia promote axon survival, which may help explain why neurodegeneration occurs when glia are lost in disease. We show that TGF{beta} signaling in mature glia is essential for long-term maintenance of axons, and that loss of TGF{beta} signaling activates an axon death signaling pathway.
]]></description>
<dc:creator>Lassetter, A.</dc:creator>
<dc:creator>Corty, M.</dc:creator>
<dc:creator>Barria, R.</dc:creator>
<dc:creator>Sheehan, A.</dc:creator>
<dc:creator>Aicher, S.</dc:creator>
<dc:creator>Fox, A. N.</dc:creator>
<dc:creator>Hill, J.</dc:creator>
<dc:creator>Freeman, M.</dc:creator>
<dc:date>2021-09-03</dc:date>
<dc:identifier>doi:10.1101/2021.09.02.458753</dc:identifier>
<dc:title><![CDATA[Glial TGFβ activity promotes axon survival in peripheral nerves]]></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/10.1101/2021.09.06.459091v1?rss=1">
<title>
<![CDATA[
Analysis of Rod/Cone Gap Junctions from the Reconstruction of Mouse Photoreceptor Terminals 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.09.06.459091v1?rss=1</link>
<description><![CDATA[
Using serial blockface-scanning electron microscopy (SBF-SEM) and focused ion beam-scanning electron microscopy (FIB-SEM), combined with confocal microscopy for the gap junction protein Cx36, we reconstructed mouse photoreceptor terminals and located the gap junctions between them. An exuberant spray of fine telodendria extends from each cone pedicle (including blue cones) to contact 40-50 nearby rod spherules where Cx36 clusters were located, close to the mouth of the synaptic opening. There were approximately 50 Cx36 clusters per cone pedicle and 2-3 per rod spherule. We were unable to detect rod/rod or cone/cone coupling. Thus, rod/cone coupling accounts for nearly all gap junctions between photoreceptors. Our calculations suggest a mean of 82 Cx36 channels between a rod/cone pair of which 25% are open at rest. Rod/cone gap junctions are modulated by dopamine. Comparing our morphological calculations of maximum coupling to previous physiological results suggests that dopamine antagonists can drive rod/cone gap junctions to a surprisingly high open probability, approaching 100%.
]]></description>
<dc:creator>Massey, S.</dc:creator>
<dc:creator>Ishibashi, M.</dc:creator>
<dc:creator>Keung, J.</dc:creator>
<dc:creator>Morgans, C. W.</dc:creator>
<dc:creator>Aicher, S.</dc:creator>
<dc:creator>Carroll, J.</dc:creator>
<dc:creator>Singer, J. H.</dc:creator>
<dc:creator>Jia, L.</dc:creator>
<dc:creator>Li, W.</dc:creator>
<dc:creator>Fahrenfort, I.</dc:creator>
<dc:creator>Ribelayga, C.</dc:creator>
<dc:date>2021-09-06</dc:date>
<dc:identifier>doi:10.1101/2021.09.06.459091</dc:identifier>
<dc:title><![CDATA[Analysis of Rod/Cone Gap Junctions from the Reconstruction of Mouse Photoreceptor Terminals]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-09-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.09.06.459177v1?rss=1">
<title>
<![CDATA[
Ascertaining cells' synaptic connections and RNA expression simultaneously with massively barcoded rabies virus libraries 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.09.06.459177v1?rss=1</link>
<description><![CDATA[
Brain function depends on forming and maintaining connections between neurons of specific types, ensuring neural function while allowing the plasticity necessary for cellular and behavioral dynamics. However, systematic descriptions of how brain cell types organize into synaptic networks and which molecules instruct these relationships are not readily available. Here, we introduce SBARRO (Synaptic Barcode Analysis by Retrograde Rabies ReadOut), a method that uses single-cell RNA sequencing to reveal directional, monosynaptic relationships based on the paths of a barcoded rabies virus from its "starter" postsynaptic cell to that cells presynaptic partners1. Thousands of these partner relationships can be ascertained in a single experiment, alongside genome-wide RNA profiles - and thus cell identities and molecular states - of each host cell. We used SBARRO to describe synaptic networks formed by diverse mouse brain cell types in vitro, leveraging a system similar to those used to identify synaptogenic molecules. We found that the molecular identity (cell type/subtype) of the starter cell predicted the number and types of cells that had synapsed onto it. Rabies transmission tended to occur into cells with RNA-expression signatures related to developmental maturation and synaptic transmission. The estimated size of a cells presynaptic network, relative to that of other cells of the same type, associated with increased expression of Arpp21 and Cdh13. By tracking individual virions and their clonal progeny as they travel among host cells, single-cell, single-virion genomic technologies offer new opportunities to map the synaptic organization of neural circuits in health and disease.
]]></description>
<dc:creator>Saunders, A.</dc:creator>
<dc:creator>Huang, K. W.</dc:creator>
<dc:creator>Vondrak, C.</dc:creator>
<dc:creator>Hughes, C.</dc:creator>
<dc:creator>Smolyar, K.</dc:creator>
<dc:creator>Sen, H.</dc:creator>
<dc:creator>Philson, A. C.</dc:creator>
<dc:creator>Nemesh, J.</dc:creator>
<dc:creator>Wysoker, A.</dc:creator>
<dc:creator>Kashin, S.</dc:creator>
<dc:creator>Sabatini, B. L.</dc:creator>
<dc:creator>McCarroll, S. A.</dc:creator>
<dc:date>2021-09-06</dc:date>
<dc:identifier>doi:10.1101/2021.09.06.459177</dc:identifier>
<dc:title><![CDATA[Ascertaining cells' synaptic connections and RNA expression simultaneously with massively barcoded rabies virus libraries]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-09-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.09.09.459677v1?rss=1">
<title>
<![CDATA[
Resurgent Na+ currents promote ultrafast spiking in projection neurons that drive fine motor control 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.09.09.459677v1?rss=1</link>
<description><![CDATA[
The underlying mechanisms that promote precise spiking in upper motor neurons controlling fine motor skills are not well understood. Here we report that projection neurons in the adult zebra finch song nucleus RA display: 1) robust high-frequency firing, 2) ultra-short half-width spike waveforms, 3) superfast Na+ current inactivation kinetics and 4) large resurgent Na+ currents (INaR). These spiking properties closely resemble those of specialized pyramidal neurons in mammalian motor cortex and are well suited for precise temporal coding. They emerge during the critical period for vocal learning in males but not females, coinciding with a complete switch of modulatory Na+ channel subunit expression from Nav{beta}3 to Nav{beta}4. Dynamic clamping and dialysis of Nav{beta}4s C-terminal peptide into juvenile RA neurons provide evidence that this subunit, and its associated INaR, promote neuronal excitability. We propose that Nav{beta}4 underpins INaR that facilitates precise, prolonged, and reliable high-frequency firing in upper motor neurons.
]]></description>
<dc:creator>Zemel, B.</dc:creator>
<dc:creator>Nevue, A. A.</dc:creator>
<dc:creator>Dagostin, A.</dc:creator>
<dc:creator>Lovell, P. V.</dc:creator>
<dc:creator>Mello, C. V.</dc:creator>
<dc:creator>von Gersdorff, H.</dc:creator>
<dc:date>2021-09-10</dc:date>
<dc:identifier>doi:10.1101/2021.09.09.459677</dc:identifier>
<dc:title><![CDATA[Resurgent Na+ currents promote ultrafast spiking in projection neurons that drive fine motor control]]></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/10.1101/2021.09.13.460106v1?rss=1">
<title>
<![CDATA[
c-Kit mediates cutaneous sensory axon innervation and multi-kinase inhibitor-induced neurotoxicity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.09.13.460106v1?rss=1</link>
<description><![CDATA[
Peripheral somatosensory neurons innervate the skin and sense the environment. Loss of skin innervation, often caused by the "dying back" of distal somatosensory axons, is a common side effect of drug-induced peripheral neuropathies (DIPNs) and results in pain and sensory dysfunction. Targeted cancer therapies frequently employ multi-kinase inhibitor (MKI) drugs that each block multiple receptor tyrosine kinases. Many MKIs produce DIPNs but the molecular targets and cellular mechanisms underlying these are unknown. We performed live-imaging of cutaneous somatosensory axons in larval zebrafish during treatment with several MKIs known to induce DIPNs, and observed axonal retraction consistent with a dying back pathology. These results were replicated in mouse somatosensory neurons. Genetic knockout of potential MKI targets identified c-Kit receptor as a regulator of sensory axon innervation and a major target of these MKIs mediating loss of axonal density. In both fish and mammals, Kit receptor is expressed in cutaneous somatosensory neurons and its ligand, Kitlg, is expressed in the skin. Mosaic misexpression of Kitlg in the skin induced dramatic increases in local sensory axon density, suggesting an important role for Kit signaling in cutaneous axon growth and maintenance. Immunostaining and structure-function analysis revealed Src, a downstream Kit target, mediates Kits role in cutaneous axon innervation and MKI neurotoxicity. Our data shows that the Kit-Src signaling pathway has a major role in cutaneous sensory axon innervation and is a potential therapeutic target to address DIPNs caused by MKIs and other compounds.
]]></description>
<dc:creator>Tuttle, A. M.</dc:creator>
<dc:creator>Pomaville, M. B.</dc:creator>
<dc:creator>Delgado, K. C.</dc:creator>
<dc:creator>Wright, K. M.</dc:creator>
<dc:creator>Nechiporuk, A. V.</dc:creator>
<dc:date>2021-09-15</dc:date>
<dc:identifier>doi:10.1101/2021.09.13.460106</dc:identifier>
<dc:title><![CDATA[c-Kit mediates cutaneous sensory axon innervation and multi-kinase inhibitor-induced neurotoxicity]]></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/10.1101/2021.09.21.461270v1?rss=1">
<title>
<![CDATA[
Discoidin domain receptor regulates ensheathment, survival, and caliber of peripheral axons 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.09.21.461270v1?rss=1</link>
<description><![CDATA[
Invertebrate axons and small caliber axons in mammalian peripheral nerves are unmyelinated but still ensheathed by glia. How this type of ensheathment is controlled and its roles in supporting neuronal function remain unclear. Here we use Drosophila wrapping glia, which ensheathe peripheral axons to study the function and development of non-myelinating ensheathment. We developed a new SplitGal4 intersectional driver to target wrapping glia for genetic ablation and found that loss of wrapping glia severely impaired larval locomotor behavior. We performed an in vivo RNAi screen in Drosophila to identify glial genes required for axon ensheathment during development and identified the conserved receptor tyrosine kinase Discoidin domain receptor (Ddr). In larval peripheral nerves, loss of Ddr resulted in severely reduced ensheathment of axons. We found a strong dominant genetic interaction between Ddr and the fly type XV/XVIII collagen Multiplexin (Mp), suggesting Ddr functions a collagen receptor to drive wrapping of axons during development. Surprisingly, despite severe impairment of ensheathment, the residual wrapping in Ddr mutants was sufficient to support basic circuit function during larval stages. In adult nerves, loss of Ddr from glia decreased long-term survival of sensory neurons and significantly reduced axon caliber in an identifiable neuron without overtly affecting ensheathment. Our data establish a crucial role for non-myelinating glia in peripheral nerve development and function across the lifespan, and identify Ddr as a key regulator of axon-glia interactions during ensheathment and nerve growth.
]]></description>
<dc:creator>Corty, M. M.</dc:creator>
<dc:creator>Lassetter, A. P.</dc:creator>
<dc:creator>Hill, J. Q.</dc:creator>
<dc:creator>Sheehan, A. E.</dc:creator>
<dc:creator>Bernardo-Garcia, F. J.</dc:creator>
<dc:creator>Davis, G. W.</dc:creator>
<dc:creator>Aicher, S. A.</dc:creator>
<dc:creator>Freeman, M. R.</dc:creator>
<dc:date>2021-09-23</dc:date>
<dc:identifier>doi:10.1101/2021.09.21.461270</dc:identifier>
<dc:title><![CDATA[Discoidin domain receptor regulates ensheathment, survival, and caliber of peripheral axons]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-09-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.09.23.461061v1?rss=1">
<title>
<![CDATA[
Aldh2 dependent formaldehyde metabolism fuels purine demands in melanocyte stem cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.09.23.461061v1?rss=1</link>
<description><![CDATA[
Melanocyte stem cells (McSCs) in zebrafish serve as an on-demand source of melanocytes during growth and regeneration, but metabolic programs associated with their activation and regenerative processes are not well known. Here, using live imaging coupled with scRNA-sequencing, we discovered that quiescent McSCs during regeneration activate a dormant embryonic neural crest transcriptional program followed by an aldehyde dehydrogenase (Aldh) 2 metabolic switch to generate progeny. Unexpectedly, while ALDH2 is well known for its aldehyde clearing mechanisms we find that in regenerating McSCs, Aldh2 activity is required to generate formate - the one-carbon (1C) building block for nucleotide biosynthesis - through formaldehyde metabolism. Consequently, we find that disrupting the 1C cycle with low-doses of methotrexate caused melanocyte regeneration defects. In the absence of Aldh2, we find that purines (but not pyrimidines) are the metabolic end product sufficient for activated McSCs to generate progeny. Together, our work reveals McSCs undergo a two-step cell state transition during regeneration, and that the reaction products of Aldh2 enzymes have tissue-specific stem cell functions that meet metabolic demands in regeneration.

SUMMARY STATEMENTIn melanocyte regeneration, quiescent McSCs respond by re-expressing a neural crest identity, followed by an Aldh2-dependent metabolic switch to generate progeny.
]]></description>
<dc:creator>Brunsdon, H.</dc:creator>
<dc:creator>Brombin, A.</dc:creator>
<dc:creator>Peterson, S.</dc:creator>
<dc:creator>Postlethwait, J. H.</dc:creator>
<dc:creator>Patton, E. E.</dc:creator>
<dc:date>2021-09-23</dc:date>
<dc:identifier>doi:10.1101/2021.09.23.461061</dc:identifier>
<dc:title><![CDATA[Aldh2 dependent formaldehyde metabolism fuels purine demands in melanocyte stem cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-09-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.09.23.461551v1?rss=1">
<title>
<![CDATA[
Vertex-wise characterization of Non-Human Primate cortical development with prenatal insights. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.09.23.461551v1?rss=1</link>
<description><![CDATA[
Characterization of the interwoven complexities of early cortical thickness development has been an ongoing undertaking in neuroscience research. Longitudinal studies of Non-Human Primates (NHP) offer unique advantages to categorizing the diverse patterns of cortical growth trajectories. Here, we used latent growth models to characterize the trajectories of typical cortical thickness development in Japanese macaques at each cortical surface vertex (i.e. grayordinate). Cortical thickness from 4 to 36 months showed regional specific linear and non-linear trajectories and distinct maturation timing across the cortex. Intriguingly, we revealed a "accumulation/ablation phenomenon" of cortical maturation where the most profound development changes in cortical thickness occur in the accumulation or ablation zones surrounding the focal points (i.e., a center of a delineated regions where cortical thickness is thickest or thinnest) throughout the brain. We further examined maternal diet and inflammation in the context of these typical brain trajectories and known network architecture. A well-controlled NHP model of a maternal "Western-style" diet was used alongside measures of inflammatory cytokine interleukin-6 (IL-6) in the mothers during gestation. We observed that these accumulation and ablation zones of variable change might be most susceptible to environmental effects. The maternal factors, diet and inflammation during pregnancy were distinctively associated with different aspects of offspring cortical development reflected in regions related to distinctive functional networks. Our findings characterize the versatile intricacies of typical cortical thickness development and highlight how the maternal environment plays a role in offspring cortical development.
]]></description>
<dc:creator>Ramirez, J. S. B.</dc:creator>
<dc:creator>Hermosillo, R.</dc:creator>
<dc:creator>Thomas, E.</dc:creator>
<dc:creator>Zhu, J. Y.</dc:creator>
<dc:creator>Sturgeon, D.</dc:creator>
<dc:creator>Schifsky, E.</dc:creator>
<dc:creator>Galassi, A.</dc:creator>
<dc:creator>Thompson, J. R.</dc:creator>
<dc:creator>Bagley, J. L.</dc:creator>
<dc:creator>Milham, M. P.</dc:creator>
<dc:creator>Miranda-Dominguez, O.</dc:creator>
<dc:creator>Papadakis, S.</dc:creator>
<dc:creator>Bah, M.</dc:creator>
<dc:creator>Mitchell, A.</dc:creator>
<dc:creator>Xu, T.</dc:creator>
<dc:creator>Graham, A. M.</dc:creator>
<dc:creator>Feczko, E.</dc:creator>
<dc:creator>Sullivan, E. L.</dc:creator>
<dc:creator>Fair, D. A.</dc:creator>
<dc:date>2021-09-24</dc:date>
<dc:identifier>doi:10.1101/2021.09.23.461551</dc:identifier>
<dc:title><![CDATA[Vertex-wise characterization of Non-Human Primate cortical development with prenatal insights.]]></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/10.1101/2021.09.27.462028v1?rss=1">
<title>
<![CDATA[
Fast resupply of synaptic vesicles requires Synaptotagmin-3 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.09.27.462028v1?rss=1</link>
<description><![CDATA[
Sustained neuronal activity demands quick resupply of synaptic vesicles in order to maintain reliable synaptic transmission. Such vesicle replenishment is accelerated by sub-micromolar presynaptic Ca2+ signals by an as yet unidentified high-affinity Ca2+ sensor1-4. Here we identify a novel presynaptic role for the high-affinity Ca2+ sensor Synaptotagmin-3 (SYT3)5 in driving vesicle replenishment and short-term synaptic plasticity. Synapses in Syt3 knockout mice exhibit enhanced short-term depression, and recovery is slower and insensitive to presynaptic residual Ca2+. During sustained neuronal firing, SYT3 speeds vesicle replenishment and increases the size of the readily releasable pool of vesicles. SYT3 also mediates a second form of short-term enhancement called facilitation, under conditions of low vesicle release probability. Models of vesicle trafficking suggest that SYT3 could combat synaptic depression by accelerating vesicle docking at active zones. Our results reveal a critical role for presynaptic SYT3 in maintaining reliable high-frequency synaptic transmission in neural circuits.
]]></description>
<dc:creator>Weingarten, D. J.</dc:creator>
<dc:creator>Shrestha, A.</dc:creator>
<dc:creator>Kissiwaa, S. A.</dc:creator>
<dc:creator>Spruston, E.</dc:creator>
<dc:creator>Jackman, S. L.</dc:creator>
<dc:date>2021-09-27</dc:date>
<dc:identifier>doi:10.1101/2021.09.27.462028</dc:identifier>
<dc:title><![CDATA[Fast resupply of synaptic vesicles requires Synaptotagmin-3]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-09-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.09.28.462211v1?rss=1">
<title>
<![CDATA[
A cell-based screen identifies HDAC inhibitors and PLK inhibitor as activators of RIG-I signaling 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.09.28.462211v1?rss=1</link>
<description><![CDATA[
Enhancing the immune microenvironment in cancer by targeting the nucleic acid sensors is becoming a potent therapeutic strategy. Among the nucleic acid sensors, activation of the RNA sensor Retinoic Acid-inducible Gene (RIG-I) using small hairpin RNAs has been shown to elicit powerful innate and adaptive immune responses. Given the challenges inherent in pharmacokinetics and delivery of RNA based agonists, we set out to discover small molecule agonists of RIG-I using a cell-based assay. To this end, we established and validated a robust high throughput screening assay based on a commercially available HEK293 reporter cell line with a luciferase reporter downstream of tandem interferon stimulated gene 54 (ISG54) promoter elements. We first confirmed that the luminescence in this cell line is dependent on RIG-I and the interferon receptor using a hairpin RNA RIG-I agonist. We established a 96-well and a 384-well format HTS based on this cell line and performed a proof-of-concept screen using an FDA approved drug library of 1200 compounds. Surprisingly, we found two HDAC inhibitors Entinostat, Mocetinostat and the PLK1 inhibitor Volasertib significantly enhanced ISG-luciferase activity. This luminescence was substantially diminished in the null reporter cell line indicating the increase in signaling was dependent on RIG-I expression. Combination treatment of tumor cell lines with Entinostat increased RIG-I induced cell death in a mammary carcinoma cell line that is resistant to either Entinostat or RIG-I agonist alone. Taken together, our data indicates an unexpected role for HDAC1,-3 inhibitors in enhancing RIG-I signaling and highlight potential opportunities for therapeutic combinations.
]]></description>
<dc:creator>Fraile-Bethencourt, E.</dc:creator>
<dc:creator>Foss, M. H.</dc:creator>
<dc:creator>Nelson, D.</dc:creator>
<dc:creator>Malhotra, S. V.</dc:creator>
<dc:creator>Anand, S.</dc:creator>
<dc:date>2021-09-30</dc:date>
<dc:identifier>doi:10.1101/2021.09.28.462211</dc:identifier>
<dc:title><![CDATA[A cell-based screen identifies HDAC inhibitors and PLK inhibitor as activators of RIG-I signaling]]></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/10.1101/2021.09.30.462514v1?rss=1">
<title>
<![CDATA[
Ad26.COV2.S Prevents SARS-CoV-2 Induced Pathways of Inflammation and Thrombosis in Hamsters 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.09.30.462514v1?rss=1</link>
<description><![CDATA[
Syrian golden hamsters exhibit features of severe disease after SARS-CoV-2 challenge and are therefore useful models of COVID-19 pathogenesis and prevention with vaccines. Recent studies have shown that SARS-CoV-2 infection stimulates type I interferon, myeloid, and inflammatory signatures similar to human disease, and that weight loss can be prevented with vaccines. However, the impact of vaccination on transcriptional programs associated with COVID-19 pathogenesis and protective adaptive immune responses is unknown. Here we show that SARS-CoV-2 challenge in hamsters stimulates antiviral, myeloid, and inflammatory programs as well as signatures of complement and thrombosis associated with human COVID-19. Notably, single dose immunization with Ad26.COV2.S, an adenovirus serotype 26 vector (Ad26)-based vaccine expressing a stabilized SARS-CoV-2 spike protein, prevents the upregulation of these pathways such that the gene expression profiles of vaccinated hamsters are comparable to uninfected animals. Finally, we show that Ad26.COV2.S vaccination induces T and B cell signatures that correlate with binding and neutralizing antibody responses. These data provide further insights into the mechanisms of Ad26.COV2.S based protection against severe COVID-19 in hamsters.

Author SummaryIn this study, we show that vaccination with Ad26.COV2.S protected SARS-CoV-2 challenged hamsters from developing severe COVID-19 disease by attenuating excessive proinflammatory responses, such as IL-6 and IL-1, macrophages and neutrophils signaling. Ad26 vaccination also prevented the upregulation of pathways associated with thrombosis such coagulation and clotting cascades associated with infection, and the transcriptomic profiles of vaccinated animals were largely comparable to control uninfected hamsters. In contrast, SARS-CoV-2 challenged unvaccinated hamsters showed significant increase of these proinflammatory and prothrombotic pathways and significant weight loss compared to vaccinated hamsters.
]]></description>
<dc:creator>Aid, M.</dc:creator>
<dc:creator>Vidal, S.</dc:creator>
<dc:creator>Piedra-Mora, C.</dc:creator>
<dc:creator>Ducat, S.</dc:creator>
<dc:creator>Chan, C.</dc:creator>
<dc:creator>Bondoc, S.</dc:creator>
<dc:creator>Colarusse, A.</dc:creator>
<dc:creator>Starke, C.</dc:creator>
<dc:creator>Nekorchuk, M.</dc:creator>
<dc:creator>Busman-Sahay, K.</dc:creator>
<dc:creator>Estes, J.</dc:creator>
<dc:creator>Martinot, A.</dc:creator>
<dc:creator>Barouch, D. H.</dc:creator>
<dc:date>2021-09-30</dc:date>
<dc:identifier>doi:10.1101/2021.09.30.462514</dc:identifier>
<dc:title><![CDATA[Ad26.COV2.S Prevents SARS-CoV-2 Induced Pathways of Inflammation and Thrombosis in Hamsters]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-09-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.10.01.462615v1?rss=1">
<title>
<![CDATA[
Structural basis for C-type inactivation in a Shaker family voltage gated K+ channel 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.10.01.462615v1?rss=1</link>
<description><![CDATA[
C-type inactivation is a process by which ion flux through a voltage-gated K+ (Kv) channel is regulated at the selectivity filter. While prior studies have indicated that C-type inactivation involves structural changes at the selectivity filter, the nature of the changes have not been resolved. Here we report the crystal structure of the Kv1.2 channel in a C-type inactivated state. The structure shows that C-type inactivation involves changes in the selectivity filter that disrupt the outer two ion binding sites in the filter. The changes at the selectivity filter propagate to the extracellular mouth and the turret regions of the channel pore. The structural changes observed are consistent with the functional hallmarks of C-type inactivation. This study highlights the intricate interplay between K+ occupancy at the ion binding sites and the interactions of the selectivity filter in determining the balance between the conductive and the inactivated conformations of the filter.
]]></description>
<dc:creator>Reddi, R.</dc:creator>
<dc:creator>Matulef, K.</dc:creator>
<dc:creator>Riederer, E.</dc:creator>
<dc:creator>Whorton, M.</dc:creator>
<dc:creator>Valiyaveetil, F.</dc:creator>
<dc:date>2021-10-01</dc:date>
<dc:identifier>doi:10.1101/2021.10.01.462615</dc:identifier>
<dc:title><![CDATA[Structural basis for C-type inactivation in a Shaker family voltage gated K+ channel]]></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/10.1101/2021.10.01.462800v1?rss=1">
<title>
<![CDATA[
Molecular structure and conformation of stereocilia tip-links elucidated by cryo-electron tomography 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.10.01.462800v1?rss=1</link>
<description><![CDATA[
Mechanosensory transduction (MT), the conversion of mechanical stimuli into electrical signals, underpins hearing and balance and is carried out within hair cells in the inner ear. Hair cells harbor actin-filled stereocilia, arranged in rows of descending heights, where the tips of stereocilia are connected to their taller neighbors by a filament composed of protocadherin 15 (PCDH15) and cadherin 23 (CDH23), deemed the  tip-link. Tension exerted on the tip-link opens an ion channel at the tip of the shorter stereocilia, thus converting mechanical force into an electrical signal. While biochemical and structural studies have provided insights into the molecular composition and structure of isolated portions of the tip-link, the architecture, location and conformational states of intact tip-links, on stereocilia, remains unknown. Here we report in situ cryo-electron microscopy imaging of the tip-link in mouse stereocilia. We observe individual PCDH15 molecules at the tip and shaft of stereocilia and determine their stoichiometry, conformational heterogeneity, and their complexes with CDH23. The PCDH15/CDH23 complexes occur in clusters, frequently with more than one copy of PCDH15 at the tip of stereocilia, suggesting that tip-links might consist of more than one copy of the PCDH15/CDH23 heterotetramer and by extension, might include multiple MT complexes.
]]></description>
<dc:creator>Elferich, J.</dc:creator>
<dc:creator>Clark, S.</dc:creator>
<dc:creator>Ge, J.</dc:creator>
<dc:creator>Goehring, A.</dc:creator>
<dc:creator>Matsui, A.</dc:creator>
<dc:creator>Gouaux, E.</dc:creator>
<dc:date>2021-10-01</dc:date>
<dc:identifier>doi:10.1101/2021.10.01.462800</dc:identifier>
<dc:title><![CDATA[Molecular structure and conformation of stereocilia tip-links elucidated by cryo-electron tomography]]></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/10.1101/2021.10.03.462422v1?rss=1">
<title>
<![CDATA[
Systematic discovery of mutation-directed neo-protein-protein interactions in cancer 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.10.03.462422v1?rss=1</link>
<description><![CDATA[
Comprehensive sequencing of patient tumors reveals numerous genomic mutations across tumor types that enable tumorigenesis and progression. A subset of oncogenic driver mutations results in neomorphic activity where the mutant protein mediates functions not engaged by the parental molecule. Here, we identify prevalent variant-enabled neomorph-protein-protein interactions (neoPPI) with a quantitative High Throughput differential Screening (qHT-dS) platform. Coupling of highly sensitive BRET biosensors with miniaturized co-expression in an ultra-HTS format allows large-scale monitoring of interactions of wild-type and mutant variant counterparts with a library of cancer-associated proteins in live cells. Screening of 13,392 interactions with 1,474,560 data points revealed a landscape of gain-of-interactions encompassing both oncogenic and tumor suppressor mutations. For example, the recurrent BRAF V600E lesion mediates KEAP1 neoPPI, rewiring a BRAFV600E-KEAP1 signaling axis and creating collateral vulnerability to NQO1 substrates, offering a combination therapeutic strategy. Thus, cancer genomic alterations can create neo-interactions, informing variant-directed therapeutic approaches for precision medicine.
]]></description>
<dc:creator>Mo, X.</dc:creator>
<dc:creator>Niu, Q.</dc:creator>
<dc:creator>Ivanov, A. A.</dc:creator>
<dc:creator>Tsang, Y. H.</dc:creator>
<dc:creator>Tang, C.</dc:creator>
<dc:creator>Shu, C.</dc:creator>
<dc:creator>Wahafu, A.</dc:creator>
<dc:creator>Doyle, S.</dc:creator>
<dc:creator>Cicka, D.</dc:creator>
<dc:creator>Yang, X.</dc:creator>
<dc:creator>Fan, D.</dc:creator>
<dc:creator>Reyna, M. A.</dc:creator>
<dc:creator>Cooper, L. A. D.</dc:creator>
<dc:creator>Moreno, C. S.</dc:creator>
<dc:creator>Zhou, W.</dc:creator>
<dc:creator>Owonikoko, T.</dc:creator>
<dc:creator>Lonial, S.</dc:creator>
<dc:creator>Khuri, F. R.</dc:creator>
<dc:creator>Du, Y.</dc:creator>
<dc:creator>Ramalingam, S. S.</dc:creator>
<dc:creator>Mill, G. B.</dc:creator>
<dc:creator>Fu, H.</dc:creator>
<dc:date>2021-10-03</dc:date>
<dc:identifier>doi:10.1101/2021.10.03.462422</dc:identifier>
<dc:title><![CDATA[Systematic discovery of mutation-directed neo-protein-protein interactions in cancer]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-10-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.10.06.463274v1?rss=1">
<title>
<![CDATA[
Cell Stiffening Contributes to Complement-mediated Injury of Choroidal Endothelial Cells in Early AMD 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.10.06.463274v1?rss=1</link>
<description><![CDATA[
Age-related macular degeneration (AMD) is the leading cause of blindness in the aging population. Yet, no therapies exist for [~]85% of all AMD patients who have the dry form that is marked by degeneration of the retinal pigment epithelium (RPE) and underlying choroidal vasculature. As the choroidal vessels are crucial for RPE development and maintenance, understanding how they degenerate may lead to effective therapies for dry AMD. One likely causative factor for choroidal vascular loss is the cytolytic membrane attack complex (MAC) of the complement pathway that is abundant on choroidal vessels of humans with early dry AMD. To examine this possibility, we studied the effect of complement activation on choroidal endothelial cells (ECs) isolated from a rhesus monkey model of early AMD that, we report, exhibits MAC deposition and choriocapillaris endothelial loss similar to that seen in human early AMD. Treatment of choroidal ECs from AMD eyes with complement-competent normal human serum caused extensive actin cytoskeletal injury that was significantly less pronounced in choroidal ECs from young normal monkey eyes. We further show that ECs from AMD eyes are significantly stiffer than their younger counterparts and exhibit peripheral actin organization that is distinct from the longitudinal stress fibers in young ECs. Finally, these differences in complement susceptibility and mechanostructural properties were found to be regulated by the differential activity of small GTPases Rac and Rho because Rac inhibition in AMD cells led to simultaneous reduction in stiffness and complement susceptibility while Rho inhibition in young cells exacerbated complement injury. Thus, by identifying cell stiffness and cytoskeletal regulators Rac and Rho as important determinants of complement susceptibility, the current findings offer a new mechanistic insight into choroidal vascular loss in early AMD that warrants further investigation for assessment of translational potential.
]]></description>
<dc:creator>Cabrera, A. P.</dc:creator>
<dc:creator>Stoddard, J.</dc:creator>
<dc:creator>Tierno, I. S.</dc:creator>
<dc:creator>Matisioudis, N.</dc:creator>
<dc:creator>Agarwal, M.</dc:creator>
<dc:creator>Renner, L.</dc:creator>
<dc:creator>Palegar, N.</dc:creator>
<dc:creator>Neuringer, M.</dc:creator>
<dc:creator>McGill, T. J.</dc:creator>
<dc:creator>Ghosh, K.</dc:creator>
<dc:date>2021-10-08</dc:date>
<dc:identifier>doi:10.1101/2021.10.06.463274</dc:identifier>
<dc:title><![CDATA[Cell Stiffening Contributes to Complement-mediated Injury of Choroidal Endothelial Cells in Early AMD]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-10-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.10.07.463498v1?rss=1">
<title>
<![CDATA[
Morphodynamical cell-state description via live-cell imaging trajectory embedding 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.10.07.463498v1?rss=1</link>
<description><![CDATA[
Time-lapse imaging is a powerful approach to gain insight into the dynamic responses of cells, but the quantitative analysis of morphological changes over time remains challenging. Here, we exploit the concept of "trajectory embedding" to analyze cellular behavior using morphological feature trajectory histories--that is, multiple time points simultaneously, rather than the more common practice of examining morphological feature time courses in single timepoint (snapshot) morphological features. We apply this approach to analyze live-cell images of MCF10A mammary epithelial cells after treatment with a panel of microenvironmental perturbagens that strongly modulate cell motility, morphology, and cell cycle behavior. Our morphodynamical trajectory embedding analysis constructs a shared cell state landscape revealing ligand-specific regulation of cell state transitions and enables quantitative and descriptive models of single-cell trajectories. Additionally, we show that incorporation of trajectories into single-cell morphological analysis enables (i) systematic characterization of cell state trajectories, (ii) better separation of phenotypes, and (iii) more descriptive models of ligand-induced differences as compared to snapshot-based analysis. This morphodynamical trajectory embedding is broadly applicable to the quantitative analysis of cell responses via live-cell imaging across many biological and biomedical applications.
]]></description>
<dc:creator>Copperman, J.</dc:creator>
<dc:creator>Gross, S. M.</dc:creator>
<dc:creator>Chang, Y. H.</dc:creator>
<dc:creator>Heiser, L. M.</dc:creator>
<dc:creator>Zuckerman, D. M.</dc:creator>
<dc:date>2021-10-09</dc:date>
<dc:identifier>doi:10.1101/2021.10.07.463498</dc:identifier>
<dc:title><![CDATA[Morphodynamical cell-state description via live-cell imaging trajectory embedding]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-10-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.10.08.463675v1?rss=1">
<title>
<![CDATA[
Quantitative multiplex immunohistochemistry reveals inter- and intra-patient lymphovascular and immune heterogeneity in primary cutaneous melanoma 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.10.08.463675v1?rss=1</link>
<description><![CDATA[
PurposeQuantitative, multiplexed imaging is revealing complex spatial relationships between phenotypically diverse tumor infiltrating leukocyte populations and their prognostic implications. The underlying mechanisms and tissue structures that determine leukocyte distribution within and around tumor nests, however, remain poorly understood. While presumed players in metastatic dissemination, new preclinical data demonstrates that blood and lymphatic vessels (lymphovasculature) also dictate leukocyte trafficking within tumor microenvironments and thereby impact anti-tumor immunity. Here we interrogate these relationships in primary human cutaneous melanoma.

Experimental DesignWe established a quantitative, multiplexed imaging platform to simultaneously detect immune infiltrates and tumor-associated vessels in formalin-fixed paraffin embedded patient samples. We performed a discovery, retrospective analysis of 28 treatment-naive, primary cutaneous melanomas.

ResultsHere we find that the lymphvasculature and immune infiltrate is heterogenous across patients in treatment naive, primary melanoma. We categorized five lymphovascular subtypes that differ by functionality and morphology and mapped their localization in and around primary tumors. Interestingly, the localization of specific vessel subtypes, but not overall vessel density, significantly associated with the presence of lymphoid aggregates, regional progression, and intratumoral T cell infiltrates.

ConclusionsWe describe a quantitative platform to enable simultaneous lymphovascular and immune infiltrate analysis and map their spatial relationships in primary melanoma. Our data indicate that tumor-associated vessels exist in different states and that their localization may determine potential for tumor cell exit (metastasis) or leukocyte trafficking (immune response).

This platform will support future efforts to map tumor-associated lymphovascular evolution across stage, assess its prognostic value, and stratify patients for adjuvant therapy.

TRANSLATIONAL RELEVANCEThis report describes a quantitative, image-based method to investigate the relationship between the tumor-associated lymphovasculature and immune landscape in treatment naive, primary human melanoma. The research shows that melanoma-associated blood and lymphatic vessels display context-dependent phenotypes that associate both with the risk of regional progression and immune infiltration. These findings indicate that stromal/vascular heterogeneity may underlie regional differences in immunogenicity and thus present opportunities for future biomarker development and therapeutic intervention.
]]></description>
<dc:creator>Femel, J.</dc:creator>
<dc:creator>Booth, J. L.</dc:creator>
<dc:creator>Asnaashari, T. G.</dc:creator>
<dc:creator>Leachman, S. A.</dc:creator>
<dc:creator>Tsujikawa, T.</dc:creator>
<dc:creator>White, K.</dc:creator>
<dc:creator>Chang, Y. H. W.</dc:creator>
<dc:creator>Lund, A. W.</dc:creator>
<dc:date>2021-10-09</dc:date>
<dc:identifier>doi:10.1101/2021.10.08.463675</dc:identifier>
<dc:title><![CDATA[Quantitative multiplex immunohistochemistry reveals inter- and intra-patient lymphovascular and immune heterogeneity in primary cutaneous melanoma]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-10-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.10.11.463883v1?rss=1">
<title>
<![CDATA[
Structural Diversity of Photoswitchable Sphingolipids for Optodynamic Control of Lipid Raft Microdomains 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.10.11.463883v1?rss=1</link>
<description><![CDATA[
Sphingolipids are a structurally diverse class of lipids predominantly found in the plasma membrane of eukaryotic cells. These lipids can laterally segregate with other saturated lipids and cholesterol into lipid rafts; liquid-ordered (Lo) microdomains that act as organizing centers within biomembranes. Owing the vital role of sphingolipids for lipid segregation, controlling their lateral localization is of utmost significance. Hence, we made use of the light-induced trans-cis isomerization of azobenzene-modified acyl chains, to develop a set of photoswitchable sphingolipids, with different headgroups (hydroxyl, galactosyl, phosphocholine) and backbones (sphingosine, phytosphingosine, tetrahydropyran (THP)-blocked sphingosine), able to shuttle between liquid-ordered (Lo) and liquid-disordered (Ld) regions of model membranes upon irradiation with UV-A ({lambda} = 365 nm) and blue ({lambda} = 470 nm) light, respectively. Using combined high-speed atomic force microscopy, fluorescence microscopy, and force spectroscopy, we investigated how these active sphingolipids laterally remodel supported bilayers upon photo-isomerization, notably in terms of domain area changes, height mismatch, line tension, and membrane piercing. Hereby, we show that all sphingosine-(Azo-{beta}-Gal-Cer, Azo-SM, Azo-Cer) and phytosphingosine-based (Azo--Gal-PhCer, Azo-PhCer) photolipids behave similarly, promoting a reduction in Lo domain area when in the UV-adapted cis-isoform. In contrast, azo-sphingolipids having THP groups that block H-bonding at the sphingosine backbone (Azo-THP-SM, Azo-THP-Cer) induce an increase in the Lo domain area when in cis, accompanied by a major rise in height mismatch and line tension. These changes were fully reversible upon blue light-triggered isomerization of the various lipids back to trans, pinpointing the role of interfacial interactions for the formation of stable Lo lipid raft domains.
]]></description>
<dc:creator>Hartrampf, N.</dc:creator>
<dc:creator>Leitao, S. M.</dc:creator>
<dc:creator>Winter, N.</dc:creator>
<dc:creator>Toombs-Ruane, H.</dc:creator>
<dc:creator>Frank, J. A.</dc:creator>
<dc:creator>Schwille, P.</dc:creator>
<dc:creator>Trauner, D.</dc:creator>
<dc:creator>Franquelim, H. G.</dc:creator>
<dc:date>2021-10-12</dc:date>
<dc:identifier>doi:10.1101/2021.10.11.463883</dc:identifier>
<dc:title><![CDATA[Structural Diversity of Photoswitchable Sphingolipids for Optodynamic Control of Lipid Raft Microdomains]]></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/10.1101/2021.10.17.464710v1?rss=1">
<title>
<![CDATA[
Synaptotagmin-7 enhances phasic dopamine release 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.10.17.464710v1?rss=1</link>
<description><![CDATA[
Dopamine released from substantia nigra pars compacta (SNc) neurons modulates movement, motivation, and reward. In addition to their tonic firing pattern, dopamine neurons also fire high-frequency bursts that cause superlinear increases in dopamine release. To study this poorly understood form of short-term plasticity, we used the fluorescent dopamine sensor dLight1.3b to examine the role of the calcium-binding protein synaptotagmin-7 (SYT7). We report that SYT7 mediates a hidden component of facilitation, which was unmasked by lowering initial release probability, or by low-frequency stimulation of nerve terminals. In Syt7 KO neurons, there was profound synaptic depression that significantly reduced release during stimulations that mimic in vivo firing patterns of SNc neurons. D2-mediated inhibitory postsynaptic currents in the SNc revealed a similar role for SYT7 in somatodendritic release. Our results indicate that SYT7 drives short-term facilitation of release from dopamine neurons, which likely underlies frequency-dependence of dopamine signaling in vivo.
]]></description>
<dc:creator>Jackman, S. L.</dc:creator>
<dc:creator>Kissiwaa, S. A.</dc:creator>
<dc:creator>Lebowitz, J. J.</dc:creator>
<dc:creator>Engeln, K. A.</dc:creator>
<dc:creator>Bowman, A. M.</dc:creator>
<dc:creator>Williams, J. T.</dc:creator>
<dc:date>2021-10-17</dc:date>
<dc:identifier>doi:10.1101/2021.10.17.464710</dc:identifier>
<dc:title><![CDATA[Synaptotagmin-7 enhances phasic dopamine release]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-10-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.10.18.464853v1?rss=1">
<title>
<![CDATA[
A peroxide-responding sRNA evolved from a peroxidase mRNA 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.10.18.464853v1?rss=1</link>
<description><![CDATA[
Small RNAs (sRNAs) are critical regulators of gene expression in bacteria, but we lack a clear understanding of how new sRNAs originate and get integrated into regulatory networks. A major obstacle to elucidating their evolution is the difficulty in tracing sRNAs across large phylogenetic distances. To overcome this roadblock, we investigated the prevalence of sRNAs in more than a thousand genomes across Enterobacterales, a bacterial order with a rare confluence of factors that allows robust genome-scale sRNA analyses: several well-studied organisms with fairly conserved genome structures, an established phylogeny, and substantial nucleotide diversity within a narrow evolutionary space. Using a covariance modeling-based approach, we analyzed the presence of hundreds of sRNAs and discovered that a majority of sRNAs arose recently, and uncovered protein-coding genes as a potential source for the generation of new sRNA genes. A detailed investigation of the emergence of OxyS, a peroxide-responding sRNA, demonstrated that it evolved from a 3' end fragment of a peroxidase mRNA. Collectively, our data show that the erosion of protein-coding genes can result in the formation of new sRNAs that continue to be part of the original proteins regulon. This novel insight provides a fresh framework for understanding how new sRNAs originate and get incorporated into preexisting regulatory networks.

AUTHOR SUMMARYSmall RNAs (sRNAs) are important gene regulators in bacteria, but it is unclear how new sRNAs originate and become part of regulatory networks that coordinate bacterial response to environmental stimuli. Here, we show that new sRNAs could arise from protein-coding genes and potentially be incorporated into the ancestral proteins regulatory networks. We illustrate this process by defining the origin of OxyS. This peroxide-responding sRNA evolved from and replaced a peroxidase gene, but continues to be part of the peroxide-response regulon. In sum, we describe the source from which OxyS, one of the most well-studied sRNAs, arose, identify protein-coding genes as a potential raw material from which new sRNAs could emerge, and suggest a novel evolutionary path through which new sRNAs could get incorporated into pre-existing regulatory networks.
]]></description>
<dc:creator>Krieger, M. C.</dc:creator>
<dc:creator>Dutcher, H. A.</dc:creator>
<dc:creator>Ashford, A. J.</dc:creator>
<dc:creator>Raghavan, R.</dc:creator>
<dc:date>2021-10-18</dc:date>
<dc:identifier>doi:10.1101/2021.10.18.464853</dc:identifier>
<dc:title><![CDATA[A peroxide-responding sRNA evolved from a peroxidase mRNA]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-10-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.10.18.464856v1?rss=1">
<title>
<![CDATA[
Preventing ovariectomy-induced weight gain decreases tumor burden in rodent models of obesity and postmenopausal breast cancer. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.10.18.464856v1?rss=1</link>
<description><![CDATA[
Obesity and adult weight gain are linked to increased breast cancer risk and poorer clinical outcomes in postmenopausal women, particularly for hormone-dependent tumors. Menopause is a time when significant weight gain occurs in many women, and clinical and preclinical studies have identified menopause (or ovariectomy) as a period of vulnerability for breast cancer development and promotion. We hypothesized that preventing weight gain after ovariectomy (OVX) may be sufficient to prevent the formation of new tumors and decrease growth of existing mammary tumors. Here, we tested this hypothesis in a rat model of obesity and carcinogen-induced postmenopausal mammary cancer and validated our findings in a murine xenograft model with implanted human tumors. In both models, preventing weight gain after OVX significantly decreased obesity-associated tumor development and growth. Importantly, we did not induce weight loss in these animals, but simply prevented weight gain. In both lean and obese rats, preventing weight gain reduced visceral fat accumulation and associated insulin resistance. Similarly, the intervention decreased circulating tumor-promoting growth factors and inflammatory cytokines (ie, BNDF, TNF, FGF2), with greater effects in obese compared to lean rats. In obese rats, preventing weight gain decreased adipocyte size, adipose tissue macrophage infiltration, reduced expression of the tumor-promoting growth factor FGF-1, and reduced phosphorylated FGFR in tumors. Together, these findings suggest that the underlying mechanisms associated with the anti-tumor effects of weight maintenance are multifactorial, and that weight maintenance during the peri-/post-menopausal period may be a viable strategy for reducing obesity-associated breast cancer risk and progression in women.
]]></description>
<dc:creator>Wellberg, E. A.</dc:creator>
<dc:creator>Corleto, K. A.</dc:creator>
<dc:creator>Checkley, L. A.</dc:creator>
<dc:creator>Jindal, S.</dc:creator>
<dc:creator>Johnson, G.</dc:creator>
<dc:creator>Higgins, J. A.</dc:creator>
<dc:creator>Obeid, S.</dc:creator>
<dc:creator>Anderson, S. M.</dc:creator>
<dc:creator>Thor, A. D.</dc:creator>
<dc:creator>Schedin, P. J.</dc:creator>
<dc:creator>MacLean, P. S.</dc:creator>
<dc:creator>Giles, E. D.</dc:creator>
<dc:date>2021-10-19</dc:date>
<dc:identifier>doi:10.1101/2021.10.18.464856</dc:identifier>
<dc:title><![CDATA[Preventing ovariectomy-induced weight gain decreases tumor burden in rodent models of obesity and postmenopausal breast cancer.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-10-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.11.01.465976v1?rss=1">
<title>
<![CDATA[
Transcriptional integration of distinct microbial and nutritional signals by the small intestinal epithelium 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.11.01.465976v1?rss=1</link>
<description><![CDATA[
To preserve its physiologic functions, the intestine must interpret and adapt to complex combinations of stimuli from dietary and microbial sources. However, the transcriptional strategies by which the intestinal epithelium integrates and adapts to dietary and microbial information remains unresolved. We compared adult mice reared germ free (GF) or conventionalized with a microbiota (CV) either fed normally or after a single high-fat meal (HFM). Jejunal epithelium preparations were queried using genomewide assays for RNA-seq, the activating histone mark H3K27ac ChIP-seq, and ChIP-seq of the microbially-responsive transcription factor HNF4A. We identified distinct nutritional and microbial responses at certain genes, but also apparent simultaneous influence of both stimuli at many other loci and regulatory regions. Increased expression levels and H3K27ac enrichment following HFM at a subset of these sites was dependent on microbial status. H3K27ac sites that were preferentially increased by HFM in the presence of microbes neighbor lipid anabolism and proliferation genes as well as intestinal stem cell (ISC) markers, were usually active only in ISCs, and were not HNF4A targets. In contrast, H3K27ac sites that were preferentially increased by HFM in the absence of microbes neighbored targets of the nuclear receptor and energy homeostasis regulator PPARA, were frequently accessible only in enterocytes, and were HNF4A bound. These results reveal that HNF4A supports a differentiated enterocyte and FAO program in GF, and that suppression of HNF4A by the combination of microbes and HFM may result in preferential activation of IEC proliferation programs. Microbial and nutritional responses are therefore integrated with some of the same transcriptional programs that regulate intestinal proliferation and differentiation.
]]></description>
<dc:creator>Lickwar, C. R.</dc:creator>
<dc:creator>Davison, J. M.</dc:creator>
<dc:creator>Kelly, C.</dc:creator>
<dc:creator>Padilla Mercado, G.</dc:creator>
<dc:creator>Wen, J.</dc:creator>
<dc:creator>Davis, B. R.</dc:creator>
<dc:creator>Tillman, M. C.</dc:creator>
<dc:creator>Semova, I.</dc:creator>
<dc:creator>Andres, S. F.</dc:creator>
<dc:creator>Vale, G.</dc:creator>
<dc:creator>McDonald, J. G.</dc:creator>
<dc:creator>Rawls, J. F.</dc:creator>
<dc:date>2021-11-03</dc:date>
<dc:identifier>doi:10.1101/2021.11.01.465976</dc:identifier>
<dc:title><![CDATA[Transcriptional integration of distinct microbial and nutritional signals by the small intestinal epithelium]]></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/10.1101/2021.11.01.466782v1?rss=1">
<title>
<![CDATA[
Btbd11 is an inhibitory interneuron specific synaptic scaffolding protein that supports excitatory synapse structure and function 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.11.01.466782v1?rss=1</link>
<description><![CDATA[
Synapses in the brain exhibit cell-type-specific differences in basal synaptic transmission and plasticity. Here, we evaluated cell-type-specific differences in the composition of glutamatergic synapses, identifying Btbd11, as an inhibitory interneuron-specific synapse-enriched protein. Btbd11 is highly conserved across species and binds to core postsynaptic proteins including Psd-95. Intriguingly, we show that Btbd11 can undergo liquid-liquid phase separation when expressed with Psd-95, supporting the idea that the glutamatergic post synaptic density in synapses in inhibitory and excitatory neurons exist in a phase separated state. Knockout of Btbd11 from inhibitory interneurons decreased glutamatergic signaling onto parvalbumin-positive interneurons. Further, both in vitro and in vivo, we find that Btbd11 knockout disrupts network activity. At the behavioral level, Btbd11 knockout from interneurons sensitizes mice to pharmacologically induced hyperactivity following NMDA receptor antagonist challenge. Our findings identify a cell-type-specific protein that supports glutamatergic synapse function in inhibitory interneurons--with implication for circuit function and animal behavior.
]]></description>
<dc:creator>Bygrave, A. M.</dc:creator>
<dc:creator>Sengupta, A.</dc:creator>
<dc:creator>Jackert, E. P.</dc:creator>
<dc:creator>Ahmed, M.</dc:creator>
<dc:creator>Adenuga, B.</dc:creator>
<dc:creator>Nelson, E.</dc:creator>
<dc:creator>Goldschmidt, H. L.</dc:creator>
<dc:creator>Johnson, R. C.</dc:creator>
<dc:creator>Zhong, H.</dc:creator>
<dc:creator>Yeh, F. L.</dc:creator>
<dc:creator>Sheng, M.</dc:creator>
<dc:creator>Huganir, R. L.</dc:creator>
<dc:date>2021-11-02</dc:date>
<dc:identifier>doi:10.1101/2021.11.01.466782</dc:identifier>
<dc:title><![CDATA[Btbd11 is an inhibitory interneuron specific synaptic scaffolding protein that supports excitatory synapse structure and function]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-11-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.11.03.467130v1?rss=1">
<title>
<![CDATA[
Transcription factor control of dendrite targeting via combinatorial cell-surface codes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.11.03.467130v1?rss=1</link>
<description><![CDATA[
Transcription factors specify the fate and connectivity of developing neurons. We investigate how a lineage-specific transcription factor, Acj6, controls the precise dendrite targeting of Drosophila olfactory projection neurons (PNs) by regulating the expression of cell-surface proteins. Quantitative cell-surface proteomic profiling of wild-type and acj6 mutant PNs in intact developing brains and a proteome-informed genetic screen identified PN surface proteins that execute Acj6-regulated wiring decisions. These include canonical cell adhesion molecules and proteins previously not associated with wiring, such as Piezo, whose mechanosensitive ion channel activity is dispensable for its function in PN dendrite targeting. Comprehensive genetic analyses revealed that Acj6 employs unique sets of cell-surface proteins in different PN types for dendrite targeting. Combinatorial expression of Acj6 wiring executors rescued acj6 mutant phenotypes with higher efficacy and breadth than expression of individual executors. Thus, Acj6 controls wiring specificity of different neuron types by specifying distinct combinatorial expression of cell- surface executors.
]]></description>
<dc:creator>Xie, Q.</dc:creator>
<dc:creator>Li, J.</dc:creator>
<dc:creator>Li, H.</dc:creator>
<dc:creator>Udeshi, N. D.</dc:creator>
<dc:creator>Svinkina, T.</dc:creator>
<dc:creator>Kohani, S.</dc:creator>
<dc:creator>Guajardo, R.</dc:creator>
<dc:creator>Mani, D. R.</dc:creator>
<dc:creator>Xu, C.</dc:creator>
<dc:creator>Li, T.</dc:creator>
<dc:creator>Han, S.</dc:creator>
<dc:creator>Wei, W.</dc:creator>
<dc:creator>Shuster, S. A.</dc:creator>
<dc:creator>Luginbuhl, D. J.</dc:creator>
<dc:creator>Ting, A. Y.</dc:creator>
<dc:creator>Carr, S. A.</dc:creator>
<dc:creator>Luo, L.</dc:creator>
<dc:date>2021-11-04</dc:date>
<dc:identifier>doi:10.1101/2021.11.03.467130</dc:identifier>
<dc:title><![CDATA[Transcription factor control of dendrite targeting via combinatorial cell-surface codes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-11-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.11.04.467367v1?rss=1">
<title>
<![CDATA[
Mutant-SETBP1 activates transcription of Myc programs to accelerate CSF3R-driven myeloproliferative neoplasms 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.11.04.467367v1?rss=1</link>
<description><![CDATA[
Colony stimulating factor 3 receptor (CSF3R) mutations lead to JAK pathway activation and are the molecular hallmark of chronic neutrophilic leukemia (CNL). Approximately half of CNL patients also have mutations in SET binding protein 1 (SETBP1). In this study, we developed models of SETBP1-mutant leukemia to understand the role that SETBP1 plays in CNL. SETBP1 mutations promote self-renewal of CSF3R-mutant hematopoietic progenitors in vitro and prevent cells from undergoing terminal differentiation. In vivo, SETBP1 mutations accelerate leukemia progression, leading to the rapid development of hepatosplenomegaly and granulocytosis. Through transcriptomic and epigenomic profiling, we found that SETBP1 enhances progenitor-associated programs--most strongly upregulating Myc and Myc target genes. This upregulation of Myc can be reversed by epigenetic modulatory drugs. In summary, we find that SETBP1 mutations promote aggressive hematopoietic cell expansion when expressed with mutant CSF3R through the upregulation of Myc-associated gene expression programs.

Statement of SignificanceSETBP1 is frequently mutated in chronic neutrophilic leukemia, but its role in the biology of this disease is unclear. We find that mutant SETBP1 enhances transcription of Myc and Myc target genes to promote aggressive disease biology, and that these oncogenic transcriptional programs can be reversed by epigenetic modulatory drugs.
]]></description>
<dc:creator>Carratt, S. A.</dc:creator>
<dc:creator>Kong, G. L.</dc:creator>
<dc:creator>Smith, B. M.</dc:creator>
<dc:creator>Schonrock, Z.</dc:creator>
<dc:creator>Maloney, L.</dc:creator>
<dc:creator>Baris, A.</dc:creator>
<dc:creator>Druker, B. J.</dc:creator>
<dc:creator>Braun, T. P.</dc:creator>
<dc:creator>Maxson, J. E.</dc:creator>
<dc:date>2021-11-05</dc:date>
<dc:identifier>doi:10.1101/2021.11.04.467367</dc:identifier>
<dc:title><![CDATA[Mutant-SETBP1 activates transcription of Myc programs to accelerate CSF3R-driven myeloproliferative neoplasms]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-11-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.11.16.468913v1?rss=1">
<title>
<![CDATA[
Genome-wide identification of novel sRNAs in Streptococcus mutans 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.11.16.468913v1?rss=1</link>
<description><![CDATA[
Streptococcus mutans is a major pathobiont involved in the development of dental caries. Its ability to utilize numerous sugars and to effectively respond to environmental stress promotes S. mutans proliferation in oral biofilms. Because of their quick action and low energetic cost, non-coding small RNAs (sRNAs) represent an ideal mode of gene regulation in stress response networks, yet their roles in oral pathogens have remained largely unexplored. We identified 15 novel sRNAs in S. mutans and show that they respond to four stress-inducing conditions commonly encountered by the pathogen in human mouth: sugar-phosphate stress, hydrogen peroxide exposure, high temperature, and low pH. To better understand the role of sRNAs in S. mutans, we further explored the function of the novel sRNA, SmsR4. Our data demonstrate that SmsR4 regulates the EIIA component of the sorbitol phosphotransferase system, which transports and phosphorylates the sugar alcohol sorbitol. The fine-tuning of EIIA availability by SmsR4 likely promotes S. mutans growth while using sorbitol as the main carbon source. Our work lays a foundation for understanding the role of sRNAs in regulating gene expression in stress response networks in S. mutans and highlights the importance of the underexplored phenomenon of posttranscriptional gene regulation in oral bacteria.

IMPORTANCESmall RNAs (sRNAs) are important gene regulators in bacteria, but the identities and functions of sRNAs in Streptococcus mutans, the principal bacterium involved in the formation of dental caries, are unknown. In this study, we identified 15 putative sRNAs in S. mutans and show that they respond to four common stress-inducing conditions present in human mouth: sugar-phosphate stress, hydrogen peroxide exposure, high temperature, and low pH. We further show that the novel sRNA SmsR4 likely modulates sorbitol transport into the cell by regulating SMU_313 mRNA, which encodes the EIIA subunit of the sorbitol phosphotransferase system. Gaining a better understanding of sRNA-based gene regulation may provide new opportunities to develop specific inhibitors of S. mutans growth, thereby improving oral health.
]]></description>
<dc:creator>Krieger, M.</dc:creator>
<dc:creator>Merritt, J.</dc:creator>
<dc:creator>Raghavan, R.</dc:creator>
<dc:date>2021-11-17</dc:date>
<dc:identifier>doi:10.1101/2021.11.16.468913</dc:identifier>
<dc:title><![CDATA[Genome-wide identification of novel sRNAs in Streptococcus mutans]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-11-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.11.22.469425v1?rss=1">
<title>
<![CDATA[
Investigation of Human Endogenous Retrovirus-K (ERVK) Expression and Function in Normal Placentation and Preterm Pregnancy 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.11.22.469425v1?rss=1</link>
<description><![CDATA[
BackgroundThere is a growing body of evidence indicating the importance of endogenous retrovirus (ERV) derived proteins during early development and reproduction in mammals. Recently, a protein derived from the youngest ERV in humans, ERVK (HML2), was shown to be expressed during human placentation. Since a number of highly similar ERVK proviral loci exist across the human genome, locus-specific analysis of ERVK transcription and identification of the coding sequence expressed in the human placenta is difficult. Thus, despite its activity in early human development, the native expression and function of ERVK in the human placenta remains largely uncharacterized.

ResultsIn this study, we comprehensively examined locus-specific ERVK transcription across several human placental tissues and cell types. Through a combination of RNA-seq and siRNA knock-down analyses, we identified the expression of a single ERVK locus, ERVK11q23.3, as (1) being significantly upregulated in preterm compared to term placenta, (2) predominantly expressed by mononuclear trophoblasts, (3) capable of encoding a truncated viral-like envelope protein, and (4) contributing to the expression cytokines involved in both antiviral and anti-inflammatory innate immune responses in human placental trophoblasts and BeWo choriocarcinoma cells, respectively.

ConclusionsCollectively, the results of this study highlight the utility of studying locus-specific ERVK expression, provide a thorough characterization of locus-specific ERVK transcription from human placental tissues, and indicate that altered expression of placental ERVK11q23.3 influences interferon antiviral response, which may contribute to preterm birth and other pregnancy complications.
]]></description>
<dc:creator>Rosenkrantz, J. L.</dc:creator>
<dc:creator>Martinez, M.</dc:creator>
<dc:creator>Mahankali, A.</dc:creator>
<dc:creator>Carbone, L.</dc:creator>
<dc:creator>Chavez, S. L.</dc:creator>
<dc:date>2021-11-22</dc:date>
<dc:identifier>doi:10.1101/2021.11.22.469425</dc:identifier>
<dc:title><![CDATA[Investigation of Human Endogenous Retrovirus-K (ERVK) Expression and Function in Normal Placentation and Preterm Pregnancy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-11-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.11.29.470281v1?rss=1">
<title>
<![CDATA[
A framework for multiplex imaging optimization and reproducible analysis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.11.29.470281v1?rss=1</link>
<description><![CDATA[
Multiplex imaging technologies are increasingly used for single-cell phenotyping and spatial characterization of tissues; however, transparent methods are needed for comparing the performance of platforms, protocols and analytical pipelines. We developed a python software, mplexable, for reproducible image processing and utilize Jupyter notebooks to share our optimization of signal removal, antibody specificity, background correction and batch normalization of the multiplex imaging with a focus on cyclic immunofluorescence (CyCIF). Our work both improves the CyCIF methodology and provides a framework for multiplexed image analytics that can be easily shared and reproduced.
]]></description>
<dc:creator>Eng, J. R.</dc:creator>
<dc:creator>Bucher, E.</dc:creator>
<dc:creator>Hu, Z.</dc:creator>
<dc:creator>Zheng, T.</dc:creator>
<dc:creator>Gibbs, S.</dc:creator>
<dc:creator>Chin, K.</dc:creator>
<dc:creator>Gray, J. W.</dc:creator>
<dc:date>2021-11-30</dc:date>
<dc:identifier>doi:10.1101/2021.11.29.470281</dc:identifier>
<dc:title><![CDATA[A framework for multiplex imaging optimization and reproducible analysis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-11-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.11.29.470416v1?rss=1">
<title>
<![CDATA[
Distinct roles of the Na+ binding sites in the allosteric coupling mechanism of the glutamate transporter homolog, GltPh. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.11.29.470416v1?rss=1</link>
<description><![CDATA[
Glutamate transporters carry out the concentrative uptake of glutamate by harnessing the ionic gradients present across cellular membranes. A central step in the transport mechanism is the coupled binding of Na+ and substrate. The sodium coupled Asp transporter, GltPh is an archaeal homolog of glutamate transporters that has been extensively used to probe the transport mechanism. Previous studies have shown that hairpin-2 (HP2) functions as the extracellular gate for the aspartate binding site and plays a key role in the coupled binding of sodium and aspartate to GltPh. The binding sites for three Na+ ions (Na1-3) have been identified in GltPh but the specific roles of the individual Na+ sites in the binding process has not been elucidated. In this study, we developed assays to probe Na+ binding to the Na1 and Na3 sites and to monitor the conformational switch in the NMDGT motif. We used these assays along with a fluorescence assay to monitor HP2 movement and EPR spectroscopy to show that Na+ binding to the Na3 site is required for the NMDGT conformational switch while Na+ binding to the Na1 site is responsible for the partial opening of HP2. Complete opening of HP2 requires the conformational switch of the NMDGT motif and therefore Na+ binding to both the Na1 and the Na3 sites. Based on our studies we also propose an alternate pathway for the coupled binding of Na+ and Asp.
]]></description>
<dc:creator>Valiyaveetil, F.</dc:creator>
<dc:creator>Riederer, E.</dc:creator>
<dc:creator>Moenne-Loccoz, P.</dc:creator>
<dc:date>2021-11-30</dc:date>
<dc:identifier>doi:10.1101/2021.11.29.470416</dc:identifier>
<dc:title><![CDATA[Distinct roles of the Na+ binding sites in the allosteric coupling mechanism of the glutamate transporter homolog, GltPh.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-11-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.11.30.470531v1?rss=1">
<title>
<![CDATA[
Potassium effects on NCC are attenuated during inhibition of Cullin E3-ubiquitin ligases 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.11.30.470531v1?rss=1</link>
<description><![CDATA[
The thiazide sensitive sodium-chloride co-transporter (NCC) plays a vital role in maintaining sodium (Na+) and potassium (K+) homeostasis. NCC activity is modulated by the with-no-lysine kinases 1 and 4 (WNK1 and WNK4), the abundance of which are controlled by the RING-type E3 ligase Cullin 3 (Cul3) and its substrate adapter Kelch-like protein 3. Dietary K+ intake has an inverse correlation with NCC activity, but the mechanism underlying this phenomenon remains to be fully elucidated. Here, we investigated the involvement of other members of the Cullin family in mediating K+ effects on NCC phosphorylation (active form) and abundance. In kidneys from mice fed diets varying in K+ content, there were negative correlations between NCC (phosphorylated and total) and active (neddylated) forms of Cullins (Cul1, 3, 4 and 5). High dietary K+ effects on phosphorylated NCC were attenuated in Cul3 mutant mice (CUL3-Het/{Delta}9). Short-term (30 min) and long-term (24 h) alterations in the extracellular K+ concentration did not affect Cullin neddylation levels in ex vivo renal tubules. Short-term, the ability of high extracellular K+ to decrease NCC phosphorylation was preserved in the presence of MLN4924 (pan Cullin inhibitor), but the response to low extracellular K+ was absent. Long-term, MLN4924 attenuated the effects of high extracellular K+ on NCC phosphorylation and responses to low extracellular K+ were absent. Our data suggest that in addition to Cul3, other Cullins are involved in mediating the effects of K+ on NCC phosphorylation and abundance.
]]></description>
<dc:creator>Murali, S. K.</dc:creator>
<dc:creator>Little, R.</dc:creator>
<dc:creator>Poulsen, S. B.</dc:creator>
<dc:creator>Ferdaus, M.</dc:creator>
<dc:creator>Ellison, D. H.</dc:creator>
<dc:creator>McCormick, J. A.</dc:creator>
<dc:creator>Fenton, R. A.</dc:creator>
<dc:date>2021-12-01</dc:date>
<dc:identifier>doi:10.1101/2021.11.30.470531</dc:identifier>
<dc:title><![CDATA[Potassium effects on NCC are attenuated during inhibition of Cullin E3-ubiquitin ligases]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-12-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.11.30.470628v1?rss=1">
<title>
<![CDATA[
The Long Noncoding RNA FEDORA is a Cell-Type- and Sex-Specific Regulator of Depression 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.11.30.470628v1?rss=1</link>
<description><![CDATA[
Women suffer from depression at twice the rate of men, but the underlying molecular mechanisms are poorly understood. Here, we identify dramatic baseline sex differences in expression of long noncoding RNAs (lncRNAs) in human postmortem brain tissue that are profoundly lost in depression. One such lncRNA, RP11-298D21.1 (which we termed FEDORA), is enriched in oligodendrocytes and neurons and upregulated in several cortical regions of depressed females but not males. We found that virally-expressing FEDORA selectively either in neurons or in oligodendrocytes of prefrontal cortex promoted depression-like behavioral abnormalities in female mice only, changes associated with cell-type-specific regulation of synaptic properties, myelin thickness, and gene expression. We also found that blood FEDORA levels have diagnostic significance for depressed women. These findings demonstrate the important role played by lncRNAs, and FEDORA in particular, in shaping the sex-specific landscape of the brain and contributing to sex differences in depression.
]]></description>
<dc:creator>Issler, O.</dc:creator>
<dc:creator>van der Zee, Y. Y.</dc:creator>
<dc:creator>Ramakrishnan, A.</dc:creator>
<dc:creator>Xia, S.</dc:creator>
<dc:creator>Zinsmaier, A. K.</dc:creator>
<dc:creator>Tan, C.</dc:creator>
<dc:creator>Li, W.</dc:creator>
<dc:creator>Browne, C. J.</dc:creator>
<dc:creator>Walker, D. M.</dc:creator>
<dc:creator>Salery, M.</dc:creator>
<dc:creator>Torres-Berrio, A.</dc:creator>
<dc:creator>Futamura, R.</dc:creator>
<dc:creator>Duffy, J. E.</dc:creator>
<dc:creator>Labonte, B.</dc:creator>
<dc:creator>Tamminga, C. A.</dc:creator>
<dc:creator>Dupree, J. L.</dc:creator>
<dc:creator>Dong, Y.</dc:creator>
<dc:creator>Murrough, J. W.</dc:creator>
<dc:creator>Shen, L.</dc:creator>
<dc:creator>Nestler, E. J.</dc:creator>
<dc:date>2021-12-01</dc:date>
<dc:identifier>doi:10.1101/2021.11.30.470628</dc:identifier>
<dc:title><![CDATA[The Long Noncoding RNA FEDORA is a Cell-Type- and Sex-Specific Regulator of Depression]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-12-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.12.01.470698v1?rss=1">
<title>
<![CDATA[
Babesia duncani as a model organism to study the development, virulence and drug susceptibility of intraerythrocytic parasites in vitro and in vivo 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.12.01.470698v1?rss=1</link>
<description><![CDATA[
Hematozoa are a subclass of protozoan parasites that invade and develop within vertebrate red blood cells to cause the pathological symptoms associated with diseases of both medical and veterinary importance such as malaria and babesiosis. A major limitation in the study of the most prominent hematozoa, Plasmodium spp, the causative agents of malaria, is the lack of a broadly accessible mouse model to evaluate parasite infection in vivo as is the case for P. falciparum or altogether the lack of an in vitro culture and mouse models as is the case for P. vivax, P. malariae and P. ovale. Similarly, no in vitro culture system exists for Babesia microti, the predominant agent of human babesiosis. In this study, we show that human red blood cells infected with the human pathogen Babesia duncani continuously propagated in culture, as well as merozoites purified from parasite cultures, can cause lethal infection in immunocompetent C3H/HeJ mice. Furthermore, highly reproducible parasitemia and survival outcomes were established using specific parasite loads and different mouse genetic backgrounds. Using the combined in culture-in mouse (ICIM) model of B. duncani infection, we demonstrate that current recommended combination therapies for the treatment of human babesiosis, while synergistic in cell culture, have weak potency in vitro and failed to clear infection or prevent death in mice. Interestingly, using the ICIM model, we identified two new endochin-like quinolone prodrugs, ELQ-331 and ELQ-468, that alone or in combination with atovaquone are highly efficacious against B. duncani and B. microti. The novelty, ease of use and scalability of the B. duncani ICIM dual model make it an ideal system to study intraerythrocytic parasitism by protozoa, unravel the molecular mechanisms underlying parasite virulence and pathogenesis, and accelerate the development of innovative therapeutic strategies that could be translated to unculturable parasites and important pathogens for which an animal model is lacking.

Author SummaryUse of model organisms is vital to the understanding of virulence and pathogenesis of a large number of human and animal pathogens. In case of hematozoan parasites that invade and develop within vertebrate erythrocytes, the studies are challenging because of the dearth of small animal model systems and the lack of continuous parasite growth in in vitro culture conditions. Here, we report a small animal model of lethal infection of Babesia duncani, one of the causative agents of human babesiosis. We show that in vitro cultured parasites and as well as parasites propagated in vivo can establish highly reproducible parasitemia which is dependent on the parasite load and also defined by different mouse genetic backgrounds. We further use this combined in culture-in mouse (ICIM) model of B. duncani infection to demonstrate the anti-babesial efficacy of two novel endochin like quinoline compounds. We propose that this ICIM dual model of B. duncani is an ideal system to get insights into protozoan intraerythrocytic parasitism, virulence, pathogenesis, and therapeutics and will open the vista to other important pathogens which are unculturable or lack an animal model.
]]></description>
<dc:creator>Mamoun, C.</dc:creator>
<dc:creator>Pal, A. C.</dc:creator>
<dc:creator>Renard, I.</dc:creator>
<dc:creator>Singh, P.</dc:creator>
<dc:creator>Vydyam, P.</dc:creator>
<dc:creator>Chiu, J. E.</dc:creator>
<dc:creator>Pou, S.</dc:creator>
<dc:creator>W. Winter, R.</dc:creator>
<dc:creator>Dodean, R.</dc:creator>
<dc:creator>Frueh, L.</dc:creator>
<dc:creator>Nilsen, A. C.</dc:creator>
<dc:creator>Riscoe, M. K.</dc:creator>
<dc:creator>Doggett, J. S.</dc:creator>
<dc:date>2021-12-01</dc:date>
<dc:identifier>doi:10.1101/2021.12.01.470698</dc:identifier>
<dc:title><![CDATA[Babesia duncani as a model organism to study the development, virulence and drug susceptibility of intraerythrocytic parasites in vitro and in vivo]]></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/10.1101/2021.12.05.471196v1?rss=1">
<title>
<![CDATA[
Sleep Dependent Changes of Lactate Concentration in Human Brain 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.12.05.471196v1?rss=1</link>
<description><![CDATA[
Lactate is an important cellular metabolite that is present at high concentrations in the brain, both within cells and in the extracellular space between cells. Small animal studies demonstrated high extracellular concentrations of lactate during wakefulness with reductions during sleep and/or anesthesia with a recent study suggesting the glymphatic activity as the mechanism for the reduction of lactate concentrations. We have recently developed a rigorous non-invasive imaging approach combining simultaneous magnetic resonance spectroscopy (MRS) and polysomnography (PSG) measurements, and here, we present the first in-vivo evaluation of brain lactate levels during sleep-wake cycles in young healthy humans. First, we collected single voxel proton MRS (1H-MRS) data at the posterior cingulate with high temporal resolution (every 7.5 sec), and simultaneously recorded PSG data while temporally registering with 1H-MRS time-series. Second, we evaluated PSG data in 30 s epochs, and classified into four stages Wake (W), Non-REM sleep stage 1 (N1), Non-REM sleep stage 2 (N2), and Non-REM sleep stage 3 (N3). Third, we determined lactate signal intensity from each 7.5-s spectrum, normalized to corresponding water signal, and averaged over 30-s for each PSG epoch. In examinations of nine healthy participants (four females, five males; mean age 24.2 ({+/-}2; SD) years; age range: 21-27 years) undergoing up to 3-hour simultaneous MRS/PSG recordings, we observed a group mean reduction of [4.9 {+/-} 4.9] % in N1, [10.4 {+/-} 5.2] % in N2, and [24.0 {+/-} 5.8] % in N3 when compared to W. Our finding is consistent with more than 70 years of invasive lactate measurements from small animal studies. In addition, reduced brain lactate was accompanied by a significant reduction the apparent diffusion coefficient of brain lactate. Taken together, these findings are consistent with the loss of lactate from the extracellular space during sleep while suggesting lactate metabolism is altered and/or lactate clearance via glymphatic exchange is increased during sleep.

Significance StatementThis study describes a non-invasive magnetic resonance spectroscopy/polysomnography technique that allows rigorous measurement of brain metabolite levels together with simultaneous characterization of brain arousal state as either wakeful or one of the several sleep states. The results provide the first in-vivo demonstration of reductions in brain lactate concentration and diffusivity during sleep versus wakefulness in young healthy human brain. These findings are consistent with invasive small-animal studies showing the loss of extracellular lactate during sleep, and support the notion of altered lactate metabolism and/or increased glymphatic activity in sleeping human brain.
]]></description>
<dc:creator>Yildiz, S.</dc:creator>
<dc:creator>Lim, M. M.</dc:creator>
<dc:creator>Sammi, M. K.</dc:creator>
<dc:creator>Powers, K.</dc:creator>
<dc:creator>Murchison, C. F.</dc:creator>
<dc:creator>Iliff, J. J.</dc:creator>
<dc:creator>Rooney, W. D.</dc:creator>
<dc:date>2021-12-07</dc:date>
<dc:identifier>doi:10.1101/2021.12.05.471196</dc:identifier>
<dc:title><![CDATA[Sleep Dependent Changes of Lactate Concentration in Human Brain]]></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/10.1101/2021.12.05.471280v1?rss=1">
<title>
<![CDATA[
WESTPA 2.0: High-performance upgrades for weighted ensemble simulations and analysis of longer-timescale applications 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.12.05.471280v1?rss=1</link>
<description><![CDATA[
The weighted ensemble (WE) family of methods is one of several statistical-mechanics based path sampling strategies that can provide estimates of key observables (rate constants, pathways) using a fraction of the time required by direct simulation methods such as molecular dynamics or discrete-state stochastic algorithms. WE methods oversee numerous parallel trajectories using intermittent overhead operations at fixed time intervals, enabling facile interoperability with any dynamics engine. Here, we report on major upgrades to the WESTPA software package, an open-source, high-performance framework that implements both basic and recently developed WE methods. These upgrades offer substantial improvements over traditional WE. Key features of the new WESTPA 2.0 software enhance efficiency and ease of use: an adaptive binning scheme for more efficient surmounting of large free energy barriers, streamlined handling of large simulation datasets, exponentially improved analysis of kinetics, and developer-friendly tools for creating new WE methods, including a Python API and resampler module for implementing both binned and "binless" WE strategies.

Table of Contents/Abstract ImageFor the manuscript "WESTPA 2.0: High-performance upgrades for weighted ensemble simulations and analysis of longer-timescale applications" by Russo et al.



O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=108 SRC="FIGDIR/small/471280v2_ufig1.gif" ALT="Figure 1">
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]]></description>
<dc:creator>Russo, J.</dc:creator>
<dc:creator>Zhang, S.</dc:creator>
<dc:creator>Leung, J.</dc:creator>
<dc:creator>Bogetti, A.</dc:creator>
<dc:creator>Thompson, J.</dc:creator>
<dc:creator>DeGrave, A.</dc:creator>
<dc:creator>Torrillo, P.</dc:creator>
<dc:creator>Pratt, A. J.</dc:creator>
<dc:creator>Wong, K.</dc:creator>
<dc:creator>Xia, J.</dc:creator>
<dc:creator>Copperman, J. T.</dc:creator>
<dc:creator>Adelman, J.</dc:creator>
<dc:creator>Zwier, M.</dc:creator>
<dc:creator>LeBard, D.</dc:creator>
<dc:creator>Zuckerman, D. M.</dc:creator>
<dc:creator>Chong, L.</dc:creator>
<dc:date>2021-12-06</dc:date>
<dc:identifier>doi:10.1101/2021.12.05.471280</dc:identifier>
<dc:title><![CDATA[WESTPA 2.0: High-performance upgrades for weighted ensemble simulations and analysis of longer-timescale applications]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-12-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.12.09.471904v1?rss=1">
<title>
<![CDATA[
Neurexins in serotonergic neurons regulate serotonin transmission and complex mouse behaviors 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.12.09.471904v1?rss=1</link>
<description><![CDATA[
Extensive serotonin (5-HT) innervation throughout the brain corroborates 5-HTs modulatory role in numerous cognitive activities. Volume transmission is the major mode for 5-HT transmission but mechanisms underlying 5-HT signaling are still largely unknown. Abnormal brain 5-HT levels and function have been implicated in autism spectrum disorder (ASD). Neurexin (Nrxn) genes encode presynaptic cell adhesion molecules important for the regulation of synaptic neurotransmitter release, notably glutamatergic and GABAergic transmission. Mutations in Nrxn genes are associated with neurodevelopmental disorders including ASD. However, the role of Nrxn genes in the 5-HT system is poorly understood. Here, we generated a mouse model with all three Nrxn genes disrupted specifically in 5-HT neurons to study how Nrxns affect 5-HT transmission. Loss of Nrxns in 5-HT neurons reduced the number of serotonin neurons in the early postnatal stage, impaired 5-HT release, and decreased 5-HT release sites and serotonin transporter expression. Furthermore, 5-HT neuron-specific Nrxn knockout reduced sociability and increased depressive-like behavior. Our results highlight functional roles for Nrxns in 5-HT neurotransmission, 5-HT neuron survival, and the execution of complex behaviors.
]]></description>
<dc:creator>Cheung, A.</dc:creator>
<dc:creator>Matsui, A.</dc:creator>
<dc:creator>Abe, M.</dc:creator>
<dc:creator>Sakimura, K.</dc:creator>
<dc:creator>Sasaoka, T.</dc:creator>
<dc:creator>Uemura, T.</dc:creator>
<dc:creator>Imamura Kawasawa, Y.</dc:creator>
<dc:creator>Futai, K.</dc:creator>
<dc:date>2021-12-09</dc:date>
<dc:identifier>doi:10.1101/2021.12.09.471904</dc:identifier>
<dc:title><![CDATA[Neurexins in serotonergic neurons regulate serotonin transmission and complex mouse behaviors]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-12-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.12.09.472040v1?rss=1">
<title>
<![CDATA[
Salmonella Typhimurium induces NAIP/NLRC4- and NLRP3/ASC-independent, caspase-1/4-dependent inflammasome activation in human intestinal epithelial cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.12.09.472040v1?rss=1</link>
<description><![CDATA[
Salmonella enterica serovar Typhimurium is a gram-negative pathogen that causes diseases ranging from gastroenteritis to systemic infection and sepsis. Salmonella uses type III secretion systems (T3SSs) to inject effectors into host cells. While these effectors are necessary for bacterial invasion and intracellular survival, intracellular delivery of T3SS products also enables detection of Salmonella by cytosolic immune sensors. Upon detecting translocated Salmonella ligands, these sensors form multimeric complexes called inflammasomes, which activate caspases that lead to proinflammatory cytokine release and pyroptosis. In particular, the Salmonella T3SS needle, inner rod, and flagellin proteins activate the NAIP/NLRC4 inflammasome in murine intestinal epithelial cells (IECs), which leads to restriction of bacterial replication and extrusion of infected IECs into the intestinal lumen, thereby preventing systemic dissemination of Salmonella. While these processes are studied quite well in mice, the role of the NAIP/NLRC4 inflammasome in human IECs remains unknown. Unexpectedly, we found the NAIP/NLRC4 inflammasome is dispensable for early inflammasome responses to Salmonella in both human intestinal epithelial cell lines and organoids. Additionally, the NLRP3 inflammasome and the adaptor protein ASC are not required for inflammasome activation in Caco-2 cells. Instead, we observed a partial requirement for caspase-1, and a necessity for caspase-4 and GSDMD pore-forming activity in mediating inflammasome responses to Salmonella in Caco-2 cells. These findings suggest that unlike murine IECs, human IECs do not rely on NAIP/NLRC4, and also do not use NLRP3/ASC. Instead, they primarily use caspases-1 and -4 to mediate early inflammasome responses to SPI-1-expressing Salmonella.
]]></description>
<dc:creator>Naseer, N.</dc:creator>
<dc:creator>Bauer, R.</dc:creator>
<dc:creator>Zhang, J.</dc:creator>
<dc:creator>Brodsky, I. E.</dc:creator>
<dc:creator>Rauch, I.</dc:creator>
<dc:creator>Shin, S.</dc:creator>
<dc:date>2021-12-10</dc:date>
<dc:identifier>doi:10.1101/2021.12.09.472040</dc:identifier>
<dc:title><![CDATA[Salmonella Typhimurium induces NAIP/NLRC4- and NLRP3/ASC-independent, caspase-1/4-dependent inflammasome activation in human intestinal epithelial cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-12-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.12.11.472223v1?rss=1">
<title>
<![CDATA[
Transcriptional and Cytotoxic Responses of Human Intestinal Organoids to Interferon Types I, II, and III 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.12.11.472223v1?rss=1</link>
<description><![CDATA[
The three types of interferon (IFN) have roles in antimicrobial immunity and inflammation that must be properly balanced to maintain tissue homeostasis. For example, IFNs are elevated in the context of inflammatory bowel disease and may synergize with inflammatory cytokines such as tumor necrosis factor alpha (TNF) to promote tissue damage. Prior studies suggest that in mouse intestinal epithelial cells (IECs), type III IFNs are preferentially produced during viral infections and are less cytotoxic than type I IFN. Here, we generated human IEC organoid lines from biopsies of ileum, ascending colon, and sigmoid colon of three healthy subjects to establish the baseline responses of normal human IECs to types I, II, and III IFN. We found that all IFN types elicited responses that were qualitatively consistent across intestinal biopsy sites. However, IFN types differed in magnitude of STAT1 phosphorylation and identity of genes in their downstream transcriptional programs. Specifically, there was a core transcriptional module shared by IFN types, but types I and II IFN stimulated unique transcriptional modules beyond this core gene signature. The transcriptional modules of type I and II IFN included pro-apoptotic genes, and expression of these genes correlated with potentiation of TNF cytotoxicity. These data define the response profiles of healthy human IEC organoids across IFN types, and suggest that cytotoxic effects mediated by TNF in inflamed tissues may be amplified by a simultaneous high-magnitude IFN response.
]]></description>
<dc:creator>Constant, D. A.</dc:creator>
<dc:creator>Van Winkle, J. A.</dc:creator>
<dc:creator>VanderHoek, E.</dc:creator>
<dc:creator>Dekker, S. E.</dc:creator>
<dc:creator>Sofia, M. A.</dc:creator>
<dc:creator>Regner, E.</dc:creator>
<dc:creator>Tsikitis, V. L.</dc:creator>
<dc:creator>Modiano, N.</dc:creator>
<dc:creator>Nice, T. J.</dc:creator>
<dc:date>2021-12-13</dc:date>
<dc:identifier>doi:10.1101/2021.12.11.472223</dc:identifier>
<dc:title><![CDATA[Transcriptional and Cytotoxic Responses of Human Intestinal Organoids to Interferon Types I, II, and III]]></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/10.1101/2021.12.15.472757v1?rss=1">
<title>
<![CDATA[
Plexins Promote Hedgehog Signaling Through Their Cytoplasmic GAP Activity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.12.15.472757v1?rss=1</link>
<description><![CDATA[
Hedgehog signaling controls tissue patterning during embryonic and postnatal development and continues to play important roles throughout life. Characterizing the full complement of Hedgehog pathway components is essential to understanding its wide- ranging functions. Previous work has identified Neuropilins, established Semaphorin receptors, as positive regulators of Hedgehog signaling. Neuropilins require Plexin co- receptors to mediate Semaphorin signaling, but a role for Plexins in Hedgehog signaling has not yet been explored. Here, we provide evidence that multiple Plexins promote Hedgehog signaling in NIH/3T3 fibroblasts and that Plexin loss-of-function in these cells results in significantly reduced Hedgehog pathway activity. Catalytic activity of the Plexin GTPase activating protein (GAP) domain is required for Hedgehog signal promotion, and constitutive activation of the GAP domain further amplifies Hedgehog signaling. Additionally, we demonstrate that Plexins promote Hedgehog signaling at the level of GLI transcription factors and that this promotion requires intact primary cilia.

Finally, we find that Plexin loss-of-function significantly reduces the response to Hedgehog pathway activation in the mouse dentate gyrus. Together, these data identify Plexins as novel components of the Hedgehog pathway and provide insight into their mechanism of action.
]]></description>
<dc:creator>Pinskey, J. M.</dc:creator>
<dc:creator>Hoard, T. M.</dc:creator>
<dc:creator>Zhao, X.-F.</dc:creator>
<dc:creator>Franks, N. E.</dc:creator>
<dc:creator>Frank, Z. C.</dc:creator>
<dc:creator>McMellen, A. N.</dc:creator>
<dc:creator>Giger, R. J.</dc:creator>
<dc:creator>Allen, B. L.</dc:creator>
<dc:date>2021-12-16</dc:date>
<dc:identifier>doi:10.1101/2021.12.15.472757</dc:identifier>
<dc:title><![CDATA[Plexins Promote Hedgehog Signaling Through Their Cytoplasmic GAP Activity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-12-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.12.20.473199v1?rss=1">
<title>
<![CDATA[
The origins and functional effects of postzygotic mutations throughout the human lifespan 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.12.20.473199v1?rss=1</link>
<description><![CDATA[
Postzygotic mutations (PZMs) begin to accrue in the human genome immediately after fertilization, but how and when PZMs affect development and lifetime health remains unclear. To study the origins and functional consequences of PZMs, we generated a multi-tissue atlas of PZMs from 948 donors using the final major release of the Genotype-Tissue Expression (GTEx) project. Nearly half the variation in mutation burden among tissue samples can be explained by measured technical and biological effects, while 9% can be attributed to donor-specific effects. Through phylogenetic reconstruction of PZMs, we find that their type and predicted functional impact varies during prenatal development, across tissues, and the germ cell lifecycle. Remarkably, a class of prenatal mutations was predicted to be more deleterious than any other category of genetic variation investigated and under positive selection as strong as somatic mutations in cancers. In total, the data indicate that PZMs can contribute to phenotypic variation throughout the human lifespan, and, to better understand the relationship between genotype and phenotype, we must broaden the long-held assumption of one genome per individual to multiple, dynamic genomes per individual.

One-Sentence SummaryThe predicted rates, functional effects and selection pressure of postzygotic mutations vary through the human lifecycle.
]]></description>
<dc:creator>Rockweiler, N. B.</dc:creator>
<dc:creator>Ramu, A.</dc:creator>
<dc:creator>Nagirnaja, L.</dc:creator>
<dc:creator>Wong, W. H.</dc:creator>
<dc:creator>Noordam, M. J.</dc:creator>
<dc:creator>Drubin, C. W.</dc:creator>
<dc:creator>Huang, N.</dc:creator>
<dc:creator>Miller, B.</dc:creator>
<dc:creator>Todres, E. Z.</dc:creator>
<dc:creator>Vigh-Conrad, K. A.</dc:creator>
<dc:creator>Zito, A.</dc:creator>
<dc:creator>Small, K. S.</dc:creator>
<dc:creator>Ardlie, K. G.</dc:creator>
<dc:creator>Cohen, B. A.</dc:creator>
<dc:creator>Conrad, D. F.</dc:creator>
<dc:date>2021-12-21</dc:date>
<dc:identifier>doi:10.1101/2021.12.20.473199</dc:identifier>
<dc:title><![CDATA[The origins and functional effects of postzygotic mutations throughout the human lifespan]]></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/10.1101/2021.12.22.473921v1?rss=1">
<title>
<![CDATA[
Effects of α-crystallin gene knockout on zebrafish lens development 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.12.22.473921v1?rss=1</link>
<description><![CDATA[
The -crystallin small heat shock proteins contribute to the transparency and refractive properties of the vertebrate eye lens and prevent the protein aggregation that would otherwise produce lens cataracts, the leading cause of human blindness. There are conflicting data in the literature as to what role the -crystallins may play in early lens development. In this study, we used CRISPR gene editing to produce zebrafish lines with null mutations for each of the three -crystallin genes (cryaa, cryaba and cryabb). The absence of normal protein was confirmed by mass spectrometry, and lens phenotypes were assessed with differential interference contrast microscopy and histology. Loss of A-crystallin produced a variety of lens defects with varying severity in larval lenses at 3 and 4 dpf but little substantial change in normal fiber cell denucleation. Loss of either Ba- or full-length Bb-crystallin produced no substantial lens defects. Mutation of each -crystallin gene did not alter the mRNA levels of the remaining two, suggesting a lack of genetic compensation. These data confirm a developmental role for A-crystallin in lens development, but the range of phenotype severity suggests that its loss simply increases the chance for defects and that the protein is not essential. Our finding that cryaba and cryabb mutants lack noticeable lens defects is congruent with insubstantial transcript levels in lens epithelial and fiber cells. Future experiments can explore the molecular consequences of cryaa mutation and causes of lens defects in this null mutant, as well as the roles of other genes in lens development and function.
]]></description>
<dc:creator>Posner, M.</dc:creator>
<dc:creator>Murray, K. L.</dc:creator>
<dc:creator>Andrew, B.</dc:creator>
<dc:creator>Brdicka, S.</dc:creator>
<dc:creator>Butterbaugh-Roberts, A.</dc:creator>
<dc:creator>Franklin, K.</dc:creator>
<dc:creator>Hussen, A.</dc:creator>
<dc:creator>Kaye, T.</dc:creator>
<dc:creator>Kepp, E.</dc:creator>
<dc:creator>McDonald, M.</dc:creator>
<dc:creator>Snodgrass, T.</dc:creator>
<dc:creator>Zientek, K.</dc:creator>
<dc:creator>David, L.</dc:creator>
<dc:date>2021-12-23</dc:date>
<dc:identifier>doi:10.1101/2021.12.22.473921</dc:identifier>
<dc:title><![CDATA[Effects of α-crystallin gene knockout on zebrafish lens development]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-12-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.12.27.474267v1?rss=1">
<title>
<![CDATA[
Mistimed feeding disrupts circadian rhythms of male mating behavior and female preovulatory LH surges in mice 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.12.27.474267v1?rss=1</link>
<description><![CDATA[
In rodents, eating at atypical circadian times, such as during the biological rest phase when feeding is normally minimal, reduces fertility. Prior findings suggest this fertility impairment is due, at least in part, to reduced mating success. However, the physiological and behavioral mechanisms underlying this reproductive suppression are not known. In the present study, we tested the hypothesis that mistimed feeding-induced infertility is due to a disruption in the normal circadian timing of mating behavior and/or the generation of pre-ovulatory luteinizing hormone (LH) surges (estrogen positive feedback). In the first experiment, male+female mouse pairs, acclimated to be food restricted to either the light (mistimed feeding) or dark (control feeding) phase, were scored for mounting frequency and ejaculations over 96 hours. Male mounting behavior and ejaculations were distributed much more widely across the day in light-fed mice than in dark-fed controls and fewer light-fed males ejaculated. In the second experiment, the timing of the LH surge, a well characterized circadian event driven by estradiol (E2) and the SCN, was analyzed from serial blood samples taken from ovariectomized and E2-primed female mice that were light-, dark-, or ad-lib-fed. LH concentrations peaked 2h after lights-off in both dark-fed and ad-lib control females, as expected, but not in light-fed females. Instead, the normally clustered LH surges were distributed widely with high inter-mouse variability in the light-fed group. These data indicate that mistimed feeding disrupts the temporal control of the neural processes underlying both ovulation and mating behavior, contributing to subfertility.
]]></description>
<dc:creator>Kukino, A.</dc:creator>
<dc:creator>Walbeek, T. J.</dc:creator>
<dc:creator>Sun, L. J.</dc:creator>
<dc:creator>Watt, A. T.</dc:creator>
<dc:creator>Park, J. H.</dc:creator>
<dc:creator>Kauffman, A. S.</dc:creator>
<dc:creator>Butler, M. P.</dc:creator>
<dc:date>2021-12-28</dc:date>
<dc:identifier>doi:10.1101/2021.12.27.474267</dc:identifier>
<dc:title><![CDATA[Mistimed feeding disrupts circadian rhythms of male mating behavior and female preovulatory LH surges in mice]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-12-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.12.29.473853v1?rss=1">
<title>
<![CDATA[
Diversity of microglial transcriptional responses during opioid exposure and neuropathic pain 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.12.29.473853v1?rss=1</link>
<description><![CDATA[
Microglia take on an altered morphology during chronic opioid treatment. This morphological change is broadly used to identify the activated microglial state associated with opioid side effects, including tolerance and opioid-induced hyperalgesia (OIH). Following chronic opioid treatment and peripheral nerve injury (PNI) microglia in the spinal cord display similar morphological responses. Consistent with this observation, functional studies have suggested that microglia activated by PNI or opioids engage common molecular mechanisms to induce hypersensitivity. Here we conducted deep RNA sequencing of acutely isolated spinal cord microglia from male mice to comprehensively interrogate transcriptional states and mechanistic commonality between multiple OIH and PNI models. Following PNI, we identify a common early proliferative transcriptional event across models that precedes the upregulation of histological markers of activation, followed by a delayed and injury-specific transcriptional response. Strikingly, we found no such transcriptional responses associated with opioid-induced microglial activation, consistent with histological data indicating that microglia number remain stable during morphine treatment. Collectively, these results reveal the diversity of pain-associated microglial transcriptomes and point towards the targeting of distinct insult-specific microglial responses to treat OIH, PNI, or other CNS pathologies.
]]></description>
<dc:creator>Sypek, E. I.</dc:creator>
<dc:creator>Collins, H. Y.</dc:creator>
<dc:creator>McCallum, W. M.</dc:creator>
<dc:creator>Bourdillon, A. T.</dc:creator>
<dc:creator>Bohlen, C. J.</dc:creator>
<dc:creator>Scherrer, G.</dc:creator>
<dc:date>2021-12-29</dc:date>
<dc:identifier>doi:10.1101/2021.12.29.473853</dc:identifier>
<dc:title><![CDATA[Diversity of microglial transcriptional responses during opioid exposure and neuropathic pain]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-12-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.01.07.475344v1?rss=1">
<title>
<![CDATA[
PD1+ T-cells correlate with Nerve Fiber Density as a prognostic biomarker in patients with resected perihilar cholangiocarcinoma. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.01.07.475344v1?rss=1</link>
<description><![CDATA[
Background & AimsPerihilar cholangiocarcinoma (pCCA) is a rare hepatobiliary malignancy. Nerve fiber invasion (NFI) shows cancer invading the nerve and is considered an aggressive feature. Nerve fiber density (NFD) consists of small nerve fibers without cancer invasion and is divided into high NFD (high numbers of small nerve fibers) or low NFD (low numbers of small nerve fibers). We aim to explore differences in immune cell populations and survival.

MethodsWe applied multiplex immunofluorescence (mIF) on 47 pCCA surgically resected patients and investigated the immune cell composition in the tumor microenvironment (TME) of different nerve fiber phenotypes (NFI, high and low NFD). Extensive group comparison was carried out and the association with overall survival (OS) was assessed.

ResultsThe NFI ROI was measured with highest CD68+ macrophage levels among 3 ROIs (NFI compared to tumor free p= 0.016 and to tumor p=0.034). Further, for NFI patients the density of co-inhibitory markers CD8+PD1+ and CD68+PD1+ were more abundant in the tumor rather than NFI ROI (p= 0.004 and p= 0.0029 respectively). Comparison between patients with NFD and NFI groups, the signals of co-expression of CD8+PD1+ as well as CD68+PD1+ were significantly higher in the high NFD group (p= 0.027 and p= 0.044, respectively). The OS for high NFD patients was 92 months median OS (95% CI:41-142), for low NFD patients 20 months ((95% CI: 4-36) and for NFI group of patients 19 months (95% CI 7-33). The OS for high NFD patients was significantly better compared to low NFD (p= 0.046) and NFI (p= 0.032).

ConclusionsPD1+ T-cells correlate with high NFD as a prognostic biomarker, the biological pathway behind this needs to be investigated.

Lay summaryNerve fibers play a dual role in the tumor microenvironment in pCCA. In our previous study, we showed that the presence of high numbers of small nerve fibers is associated with a better overall survival. In addition, we found that in high NFD patients PD1+ T-cells are significantly overexpressed. Therefore, we present high NFD as a promising prognostic biomarker.

Graphical abstract

O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=128 SRC="FIGDIR/small/475344v1_ufig1.gif" ALT="Figure 1">
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]]></description>
<dc:creator>Tan, X.</dc:creator>
<dc:creator>Rosin, M.</dc:creator>
<dc:creator>Appinger, S.</dc:creator>
<dc:creator>Bednarsch, J.</dc:creator>
<dc:creator>Liu, D.</dc:creator>
<dc:creator>Wiltberger, G.</dc:creator>
<dc:creator>Garcia Vallejo, J.</dc:creator>
<dc:creator>Jang, S.</dc:creator>
<dc:creator>Czigany, Z.</dc:creator>
<dc:creator>Boroojerdi, S.</dc:creator>
<dc:creator>Gaisa, N.</dc:creator>
<dc:creator>Boor, P.</dc:creator>
<dc:creator>Buelow, R.</dc:creator>
<dc:creator>de Vos-Geelen, J.</dc:creator>
<dc:creator>Valkenburg-van Iersel, L.</dc:creator>
<dc:creator>Clahsen-van Groningen, M.</dc:creator>
<dc:creator>de Jong, E.</dc:creator>
<dc:creator>Groot Koerkamp, B.</dc:creator>
<dc:creator>Doukas, M.</dc:creator>
<dc:creator>Rocha, F.</dc:creator>
<dc:creator>Luedde, T.</dc:creator>
<dc:creator>Klinge, U.</dc:creator>
<dc:creator>Sivakumar, S.</dc:creator>
<dc:creator>Neumann, U.</dc:creator>
<dc:creator>Heij, L.</dc:creator>
<dc:date>2022-01-11</dc:date>
<dc:identifier>doi:10.1101/2022.01.07.475344</dc:identifier>
<dc:title><![CDATA[PD1+ T-cells correlate with Nerve Fiber Density as a prognostic biomarker in patients with resected perihilar cholangiocarcinoma.]]></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/10.1101/2022.01.10.475735v1?rss=1">
<title>
<![CDATA[
GoPeaks: Histone Modification Peak Calling for CUT&Tag 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.01.10.475735v1?rss=1</link>
<description><![CDATA[
Genome-wide mapping of the histone modification landscape is critical to understanding tran-scriptional regulation. Cleavage Under Targets and Tagmentation (CUT&Tag) is a new method for profiling the localization of covalent histone modifications, offering improved sensitivity and decreased cost compared with Chromatin Immunoprecipitation Sequencing (ChIP-seq). Here, we present GoPeaks, a peak calling method specifically designed for histone modification CUT&Tag data. GoPeaks implements a Binomial distribution and stringent read count cut-off to nominate candidate genomic regions. We compared the performance of GoPeaks against commonly used peak calling algorithms to detect H3K4me3, H3K4me1, and H3K27Ac peaks from CUT&Tag data. These histone modifications display a range of peak profiles and are frequently used in epigenetic studies. We found GoPeaks robustly detects genome-wide histone modifications and, notably, identifies H3K27Ac with improved sensitivity compared to other standard peak calling algorithms.
]]></description>
<dc:creator>Yashar, W. M.</dc:creator>
<dc:creator>Kong, G.</dc:creator>
<dc:creator>VanCampen, J.</dc:creator>
<dc:creator>Smith, B. M.</dc:creator>
<dc:creator>Coleman, D. J.</dc:creator>
<dc:creator>Carbone, L.</dc:creator>
<dc:creator>Yardimci, G. G.</dc:creator>
<dc:creator>Maxson, J. E.</dc:creator>
<dc:creator>Braun, T. P.</dc:creator>
<dc:date>2022-01-12</dc:date>
<dc:identifier>doi:10.1101/2022.01.10.475735</dc:identifier>
<dc:title><![CDATA[GoPeaks: Histone Modification Peak Calling for CUT&Tag]]></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/10.1101/2022.01.11.475714v1?rss=1">
<title>
<![CDATA[
Responses to song playback differ in sleeping versus anesthetized songbirds 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.01.11.475714v1?rss=1</link>
<description><![CDATA[
Vocal learning in songbirds is mediated by a highly localized system of interconnected forebrain regions, including recurrent loops that traverse the cortex, basal ganglia, and thalamus. This brain-behavior system provides a powerful model for elucidating mechanisms of vocal learning, with implications for learning speech in human infants, as well as for advancing our understanding of skill learning in general. A long history of experiments in this area has tested neural responses to playback of different song stimuli in anesthetized birds at different stages of vocal development. These studies have demonstrated selectivity for different song types that provide neural signatures of learning. In contrast to the ease of obtaining responses to song playback in anesthetized birds, song-evoked responses in awake birds are greatly reduced or absent, indicating that behavioral state is an important determinant of neural responsivity. Song-evoked responses can be elicited in sleeping as well as anesthetized zebra finches, and the selectivity of responses to song playback in adult birds tends to be highly similar between anesthetized and sleeping states, encouraging the idea that anesthesia and sleep are highly similar. In contrast to that idea, we report evidence that cortical responses to song playback in juvenile zebra finches (Taeniopygia guttata) differ greatly between sleep and urethane anesthesia. This finding indicates that behavioral states differ in sleep versus anesthesia and raises questions about relationships between developmental changes in sleep activity, selectivity for different song types, and the neural substrate for vocal learning.

SignificancePatterns of spiking activity based on electrophysiological recordings in many different taxa are known to be heavily dependent on behavioral state. Neural activity patterns are frequently similar between sleep and anesthesia, which has encouraged the idea that similar states characterize sleep and anesthesia. Based on comparisons across studies, we report that activity patterns are highly dissimilar between sleep and urethane anesthesia in a cortical region of juvenile songbirds. These data argue against the idea that similar behavioral states are achieved in sleep versus anesthesia.
]]></description>
<dc:creator>Bottjer, S. W.</dc:creator>
<dc:creator>Le Moing, C. L.</dc:creator>
<dc:creator>Li, E. J.</dc:creator>
<dc:creator>Yuan, R. C.</dc:creator>
<dc:date>2022-01-12</dc:date>
<dc:identifier>doi:10.1101/2022.01.11.475714</dc:identifier>
<dc:title><![CDATA[Responses to song playback differ in sleeping versus anesthetized songbirds]]></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/10.1101/2022.01.12.475422v1?rss=1">
<title>
<![CDATA[
A Precision Functional Atlas of Network Probabilities and Individual-Specific Network Topography 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.01.12.475422v1?rss=1</link>
<description><![CDATA[
The brain is organized into a broad set of functional neural networks. These networks and their various characteristics have been described and scrutinized through in vivo resting state functional magnetic resonance imaging (rs-fMRI). While the basic properties of networks are generally similar between healthy individuals, there is vast variability in the precise topography across the population. These individual differences are often lost in population studies due to population averaging which assumes topographical uniformity. We leveraged precision brain mapping methods to establish a new open-source, method-flexible set of precision functional network atlases: the Masonic Institute for the Developing Brain (MIDB) Precision Brain Atlas. Using participants from the Adolescent Brain Cognitive Development (ABCD) study, single subject precision network maps were generated with two supervised network-matching procedures (template matching and non-negative matrix factorization), an overlapping template matching method for identifying integration zones, as well as an unsupervised community detection algorithm (Infomap). From these individualized maps we also generated probabilistic network maps and integration zones for two demographically-matched groups of n[~]3000 each. We demonstrate high reproducibility between groups (Pearsons r >0.999) and between methods (r=0.96), revealing both regions of high invariance and high variability. Compared to using parcellations based on groups averages, the MIDB Precision Brain Atlas allowed us to derive a set of brain regions that are largely invariant in network topography across populations, which provides more reproducible statistical maps of executive function in brain-wide associations. We also explore an example use case for probabilistic maps, highlighting their potential for use in targeted neuromodulation. The MIDB Precision Brain Atlas is expandable to alternative datasets and methods and is provided open-source with an online web interface to encourage the scientific community to experiment with probabilistic atlases and individual-specific topographies to more precisely relate network phenomenon to functional organization of the human brain.
]]></description>
<dc:creator>Hermosillo, R. J.</dc:creator>
<dc:creator>Moore, L. A.</dc:creator>
<dc:creator>Feczko, E. J.</dc:creator>
<dc:creator>Pines, A. R.</dc:creator>
<dc:creator>Dworetsky, A.</dc:creator>
<dc:creator>Conan, G.</dc:creator>
<dc:creator>Mooney, M. A.</dc:creator>
<dc:creator>Randolph, A.</dc:creator>
<dc:creator>Adeyemo, B.</dc:creator>
<dc:creator>Earl, E.</dc:creator>
<dc:creator>Perrone, A.</dc:creator>
<dc:creator>Carrasco, C. M.</dc:creator>
<dc:creator>Uriarte-Lopez, J.</dc:creator>
<dc:creator>Snider, K.</dc:creator>
<dc:creator>Doyle, O.</dc:creator>
<dc:creator>Cordova, M.</dc:creator>
<dc:creator>Nagel, B. J.</dc:creator>
<dc:creator>Ewing, S. W. F.</dc:creator>
<dc:creator>Satterthwaite, T. D.</dc:creator>
<dc:creator>Dosenbach, N.</dc:creator>
<dc:creator>Gratton, C.</dc:creator>
<dc:creator>Petersen, S.</dc:creator>
<dc:creator>Dominguez, O. M.</dc:creator>
<dc:creator>Fair, D. A.</dc:creator>
<dc:date>2022-01-13</dc:date>
<dc:identifier>doi:10.1101/2022.01.12.475422</dc:identifier>
<dc:title><![CDATA[A Precision Functional Atlas of Network Probabilities and Individual-Specific Network Topography]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-01-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.01.12.476110v1?rss=1">
<title>
<![CDATA[
Cryo-EM Structure of Adeno-associated virus-4 at 2.2 	A resolution 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.01.12.476110v1?rss=1</link>
<description><![CDATA[
Adeno-associated virus (AAV) is the vector of choice for several approved gene therapy treatments and is the basis for many ongoing clinical trials. Various strains of AAV exist (referred to as serotypes), each with their own transfection characteristics. Here, we present a high-resolution cryo-electron microscopy structure (2.2 [A]) for AAV serotype 4 (AAV4). The receptor responsible for transduction of the AAV4 clade of AAV viruses (including AAV11, 12 and rh32.33) is unknown. Other AAVs interact with the same cell receptor, Adeno-associated virus receptor (AAVR), in one of two different ways. AAV5-like viruses interact exclusively with the polycystic kidney disease-like [PKD]-1 domain of AAVR while most other AAVs interact primarily with the PKD2 domain. A comparison of the present AAV4 structure with prior corresponding structures of AAV5, AAV2 and AAV1 in complex with AAVR, provides a foundation for understanding why the AAV4-like clade is unable to interact with either PKD1 or PKD2. The conformation of the AAV4 capsid in variable regions I, III, IV and V on the viral surface appears to be sufficiently different from AAV2 to ablate binding with PKD2. Differences between AAV4 and AAV5 in variable region VII appear sufficient to exclude binding with PKD1.
]]></description>
<dc:creator>Zane, G. M.</dc:creator>
<dc:creator>Silveria, M. A.</dc:creator>
<dc:creator>Meyer, N. L.</dc:creator>
<dc:creator>White, T. A.</dc:creator>
<dc:creator>Chapman, M. S.</dc:creator>
<dc:date>2022-01-13</dc:date>
<dc:identifier>doi:10.1101/2022.01.12.476110</dc:identifier>
<dc:title><![CDATA[Cryo-EM Structure of Adeno-associated virus-4 at 2.2 	A resolution]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-01-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.01.17.476469v1?rss=1">
<title>
<![CDATA[
Dual Targeting of FLT3 and LSD1 Disrupts the MYC Super-Enhancer Complex in Acute Myeloid Leukemia 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.01.17.476469v1?rss=1</link>
<description><![CDATA[
Mutations in Fms-like tyrosine kinase 3 (FLT3) are common drivers in acute myeloid leukemia (AML) yet FLT3 inhibitors only provide modest clinical benefit. Prior work has shown that inhibitors of lysine-specific demethylase 1 (LSD1) enhance kinase inhibitor activity in AML. Here we show that combined LSD1 and FLT3 inhibition induces synergistic cell death in FLT3-mutant AML. Multi-omic profiling revealed that the drug combination disrupts STAT5, LSD1, and GFI1 binding at the MYC blood super-enhancer, suppressing super-enhancer activation as well as MYC expression and activity. The drug combination simultaneously results in the accumulation of repressive H3K9me1 methylation, an LSD1 substrate, at MYC target genes. We validated these findings in 72 primary AML samples with the nearly every sample demonstrating synergistic responses to the drug combination. Collectively, these studies provide preclinical rationale for the investigation of dual FLT3/LSD1 inhibition in a clinical trial.
]]></description>
<dc:creator>Yashar, W. M.</dc:creator>
<dc:creator>Smith, B. M.</dc:creator>
<dc:creator>Coleman, D. J.</dc:creator>
<dc:creator>VanCampen, J.</dc:creator>
<dc:creator>Kong, G.</dc:creator>
<dc:creator>Macaraeg, J.</dc:creator>
<dc:creator>Estabrook, J.</dc:creator>
<dc:creator>Demir, E.</dc:creator>
<dc:creator>Long, N.</dc:creator>
<dc:creator>Bottomly, D.</dc:creator>
<dc:creator>McWeeney, S. K.</dc:creator>
<dc:creator>Tyner, J. W.</dc:creator>
<dc:creator>Druker, B. J.</dc:creator>
<dc:creator>Maxson, J. E.</dc:creator>
<dc:creator>Braun, T. P.</dc:creator>
<dc:date>2022-01-18</dc:date>
<dc:identifier>doi:10.1101/2022.01.17.476469</dc:identifier>
<dc:title><![CDATA[Dual Targeting of FLT3 and LSD1 Disrupts the MYC Super-Enhancer Complex in Acute Myeloid Leukemia]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-01-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.01.18.476801v1?rss=1">
<title>
<![CDATA[
A potent alpaca-derived nanobody that neutralizes SARS-CoV-2 variants 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.01.18.476801v1?rss=1</link>
<description><![CDATA[
The spike glycoprotein of SARS-CoV-2 engages with human angiotensin-converting enzyme 2 (ACE2) to facilitate infection. Here, we describe an alpaca-derived heavy chain antibody fragment (VHH), saRBD-1, that disrupts this interaction by competitively binding to the spike protein receptor-binding domain. We further generated an engineered bivalent nanobody construct engineered by a flexible linker, and a dimeric Fc conjugated nanobody construct. Both multivalent nanobodies blocked infection at picomolar concentrations and demonstrated no loss of potency against emerging variants of concern including Alpha (B.1.1.7), Beta (B.1.351), Gamma (P.1), Epsilon (B.1.427/429), and Delta (B.1.617.2). saRBD-1 tolerates elevated temperature, freeze-drying, and nebulization, making it an excellent candidate for further development into a therapeutic approach for COVID-19.
]]></description>
<dc:creator>Weinstein, J. B.</dc:creator>
<dc:creator>Bates, T. A.</dc:creator>
<dc:creator>Leier, H. C.</dc:creator>
<dc:creator>McBride, S. K.</dc:creator>
<dc:creator>Barklis, E.</dc:creator>
<dc:creator>Tafesse, F. G.</dc:creator>
<dc:date>2022-01-20</dc:date>
<dc:identifier>doi:10.1101/2022.01.18.476801</dc:identifier>
<dc:title><![CDATA[A potent alpaca-derived nanobody that neutralizes SARS-CoV-2 variants]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-01-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.01.18.476818v1?rss=1">
<title>
<![CDATA[
Identification of differential hypothalamic DNA methylation and gene expression associated with sexual partner preferences in rams 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.01.18.476818v1?rss=1</link>
<description><![CDATA[
The sheep is a valuable model to test whether hormone mechanisms that sexually differentiate the brain underlie the expression of sexual partner preferences because as many as 8% of rams prefer same-sex partners. Epigenetic factors such as DNA methylation act as mediators in the interaction between steroid hormones and the genome. Variations in the epigenome could be important in determining morphological or behavior differences among individuals of the same species. In this study, we explored DNA methylation differences in the hypothalamus of male oriented rams (MORs) and female oriented rams (FORs). We employed reduced representation bisulfite sequencing (RRBS) to generate a genome-wide map of DNA methylation and RNA-Seq to profile the transcriptome. We found substantial DNA methylation and gene expression differences between FORs and MORs. Although none of the differentially methylated genes yielded significant functional terms directly associated with sex development, three differentially expressed genes were identified that have been associated previously with sexual behaviors. We hypothesize that these differences are involved in the phenotypic variation in ram sexual partner preferences, whereas future studies will have to find the specific mechanisms. Our results add an intriguing new dimension to sheep behavior that should be useful for further understanding epigenetic and transcriptomic involvement.
]]></description>
<dc:creator>Bhattacharya, S.</dc:creator>
<dc:creator>Amodei, R.</dc:creator>
<dc:creator>Vilain, E.</dc:creator>
<dc:creator>Roselli, C. E.</dc:creator>
<dc:date>2022-01-18</dc:date>
<dc:identifier>doi:10.1101/2022.01.18.476818</dc:identifier>
<dc:title><![CDATA[Identification of differential hypothalamic DNA methylation and gene expression associated with sexual partner preferences in rams]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-01-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.01.24.477547v1?rss=1">
<title>
<![CDATA[
A human DNA methylation atlas reveals principles of cell type-specific methylation and identifies thousands of cell type-specific regulatory elements 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.01.24.477547v1?rss=1</link>
<description><![CDATA[
DNA methylation is a fundamental epigenetic mark that governs chromatin organization, cell identity, and gene expression. Here we describe a human methylome atlas, based on deep whole-genome bisulfite sequencing allowing fragment-level analysis across thousands of unique markers for 39 cell types sorted from 207 healthy tissue samples.

Replicates of the same cell-type are >99.5% identical, demonstrating robustness of cell identity programs to genetic variation and environmental perturbation. Unsupervised clustering of the atlas recapitulates key elements of tissue ontogeny, and identifies methylation patterns retained since gastrulation. Loci uniquely unmethylated in an individual cell type often reside in transcriptional enhancers and contain DNA binding sites for tissue-specific transcriptional regulators. Uniquely hyper-methylated loci are rare and are enriched for CpG islands, polycomb targets, and CTCF binding sites, suggesting a novel role in shaping cell type-specific chromatin looping. The atlas provides an essential resource for interpretation of disease-associated genetic variants, and a wealth of potential tissue-specific biomarkers for use in liquid biopsies.

Summary paragraphDNA methylation, a fundamental epigenetic mark, governs chromatin organization and gene expression1, thus defining the molecular identity of cells and providing a window into developmental processes with wide-ranging physiologic and clinical ramifications. Current DNA methylation datasets have limitations, typically including only a fraction of methylation sites, many from cell lines that underwent massive changes in culture or from tissues containing unspecified mixtures of cells2-6.

We present a human methylome atlas based on deep whole-genome bisulfite sequencing of 39 sorted, primary cell types and use this dataset to address fundamental questions in developmental biology, physiology and pathology. Biological replicates are >99.5% identical, demonstrating unappreciated robustness to genetic variation and environmental perturbations. Clustering recapitulates key elements of tissue ontogeny, identifying methylation patterns retained since gastrulation. Loci uniquely unmethylated in individual cell types identify novel transcriptional enhancers and are enriched for tissue-specific transcription factors binding motifs. In contrast, loci uniquely hyper-methylated in specific cell types are rare, enriched for CpG islands and polycomb targets, and overlap CTCF binding sites, suggesting a novel role in shaping cell-type-specific chromatin looping. Finally, the atlas facilitates fragment-level deconvolution of tissue and plasma methylomes across thousands of cell-type specific regions to quantify their individual components at unprecedented resolution.

The human cell-type-specific methylation atlas provides an essential resource for studying gene regulation by defining cell-type-specific distal enhancers and regulators of 3D organization, for identifying pathological changes in DNA methylation, and for the interpretation of methylation-based liquid biopsies.

O_FIG_DISPLAY_L [Figure 1] M_FIG_DISPLAY C_FIG_DISPLAY

O_LIA deep methylation atlas of 39 human cell types, sorted from healthy samples
C_LIO_LIMethylomes record developmental history of cells
C_LIO_LIThousands of novel cell type-specific methylation markers
C_LIO_LIHypo-methylation uncovers cell type-specific regulatory map of distal enhancers
C_LIO_LIHyper-methylation across CTCF sites
C_LIO_LICell type-specific biomarkers facilitate fragment-level deconvolution of tissues and cfDNA
C_LI
]]></description>
<dc:creator>Loyfer, N.</dc:creator>
<dc:creator>Magenheim, J.</dc:creator>
<dc:creator>Peretz, A.</dc:creator>
<dc:creator>Cann, G.</dc:creator>
<dc:creator>Bredno, J.</dc:creator>
<dc:creator>Klochendler, A.</dc:creator>
<dc:creator>Fox-Fisher, I.</dc:creator>
<dc:creator>Shabi-Porat, S.</dc:creator>
<dc:creator>Hecht, M.</dc:creator>
<dc:creator>Pelet, T.</dc:creator>
<dc:creator>Moss, J.</dc:creator>
<dc:creator>Drawshy, Z.</dc:creator>
<dc:creator>Amini, H.</dc:creator>
<dc:creator>Moradi, P.</dc:creator>
<dc:creator>Nagaraju, S.</dc:creator>
<dc:creator>Bauman, D.</dc:creator>
<dc:creator>Shveiky, D.</dc:creator>
<dc:creator>Porat, S.</dc:creator>
<dc:creator>Rivkin, G.</dc:creator>
<dc:creator>Or, O.</dc:creator>
<dc:creator>Hirshoren, N.</dc:creator>
<dc:creator>Carmon, E.</dc:creator>
<dc:creator>Pikarsky, A.</dc:creator>
<dc:creator>Khalaileh, A.</dc:creator>
<dc:creator>Zamir, G.</dc:creator>
<dc:creator>Grinboim, R.</dc:creator>
<dc:creator>Abu Gazala, M.</dc:creator>
<dc:creator>Mizrahi, I.</dc:creator>
<dc:creator>Shussman, N.</dc:creator>
<dc:creator>Korach, A.</dc:creator>
<dc:creator>Wald, O.</dc:creator>
<dc:creator>Izhar, U.</dc:creator>
<dc:creator>Erez, E.</dc:creator>
<dc:creator>Yutkin, V.</dc:creator>
<dc:creator>Samet, Y.</dc:creator>
<dc:creator>Rotnemer Golinkin, D.</dc:creator>
<dc:creator>Spalding, K. L.</dc:creator>
<dc:creator>Druid, H.</dc:creator>
<dc:creator>Arner, P.</dc:creator>
<dc:creator>Shapiro, A. M. J.</dc:creator>
<dc:creator>Grompe, M.</dc:creator>
<dc:creator>Aravanis, A.</dc:creator>
<dc:creator>Venn, O.</dc:creator>
<dc:creator>Jamshidi, A.</dc:creator>
<dc:creator>Shemer, R.</dc:creator>
<dc:creator>Dor, Y.</dc:creator>
<dc:creator>Glaser, B</dc:creator>
<dc:date>2022-01-25</dc:date>
<dc:identifier>doi:10.1101/2022.01.24.477547</dc:identifier>
<dc:title><![CDATA[A human DNA methylation atlas reveals principles of cell type-specific methylation and identifies thousands of cell type-specific regulatory elements]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-01-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.01.25.477730v1?rss=1">
<title>
<![CDATA[
Resveratrol increases tear production and spontaneous ocular pain after corneal abrasion in male, but not female, rats using a preclinical model of photorefractive keratectomy (PRK) 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.01.25.477730v1?rss=1</link>
<description><![CDATA[
Photorefractive keratectomy (PRK) is an alternative to LASIK and can cause intense acute pain that is often not relieved by standard treatments. To assess potential therapeutics for this type of acute pain, appropriate preclinical models are needed. Herein we describe a rodent preclinical model of PRK and a multi-faceted approach to determine the therapeutic potential of resveratrol, a natural phytoestrogen, on pain, tear production, and the corneal epithelium. Studies were conducted in male and female Sprague-Dawley rats. Heptanol was applied to one eye and the superficial corneal epithelium was removed, mimicking the abrasion seen in PRK. Spontaneous pain was assessed with orbital tightening (OT) scores for 7 days. Corneal abrasion increased OT scores in both male and female rats with peak responses at 24 - 48 hours. Topical application of resveratrol had a sex-specific effect on OT scores and tear production. Resveratrol increased OT scores in abraded males, but not females, at 72 hours and 1 week after abrasion. Resveratrol dose-dependently increased tear production in abraded males, but had no effect in abraded females. While there was no correlation between OT score at 1 week and tear production, CGRP content of corneal nerves was positively correlated with 1 week OT score. There was also a significant increase in CD68-labeled macrophages in resveratrol-treated abraded corneas as compared to naive corneas. These findings demonstrate the usefulness of our preclinical PRK model for the assessment of ocular pain therapeutics and indicate that topical resveratrol may not be useful for managing PRK-induced pain.
]]></description>
<dc:creator>Hegarty, D. M.</dc:creator>
<dc:creator>Carroll, J. R.</dc:creator>
<dc:creator>Nguyen, D.</dc:creator>
<dc:creator>Halls, V. S.</dc:creator>
<dc:creator>Robbins, D. I.</dc:creator>
<dc:creator>Price, T. J.</dc:creator>
<dc:creator>Dussor, G.</dc:creator>
<dc:creator>Aicher, S. A.</dc:creator>
<dc:date>2022-01-28</dc:date>
<dc:identifier>doi:10.1101/2022.01.25.477730</dc:identifier>
<dc:title><![CDATA[Resveratrol increases tear production and spontaneous ocular pain after corneal abrasion in male, but not female, rats using a preclinical model of photorefractive keratectomy (PRK)]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-01-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.01.25.477760v1?rss=1">
<title>
<![CDATA[
Grandmaternal allergen exposure causes distinct epigenetic trajectories in offspring associated with airway hyperreactivity and inflammation. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.01.25.477760v1?rss=1</link>
<description><![CDATA[
Maternal asthma increases childhood asthma risk through multiple mechanisms including epigenetic regulation of asthma-associated genes. DNA methylation is one form of epigenetic regulation that is both inherited and modified by environmental exposures throughout life. In this study, we tested whether grandmaternal house dust mite (HDM) allergen exposure altered airway physiology and inflammation, as well as DNA methylation in both airway epithelium and airway sensory neurons of second-generation offspring. Grandmaternal allergen exposure induced a limited number of epigenetic changes in offspring at baseline that were not associated with increased airway reactivity or inflammation. In contrast, grandmaternal allergen exposure significantly altered offsprings response to HDM sensitization and challenge, inducing airway hyperreactivity to inhaled serotonin, increased airway inflammation, and potentiated DNA methylation. Gene sequences susceptible to methylation after allergen sensitization, and their corresponding biological processes and enriched pathways, were unique in offspring from HDM-exposed founders, indicating that grandmaternal allergen exposure established an epigenetic trajectory in offspring at birth that directed epigenetic and physiologic responses to subsequent allergen sensitization and challenge, contributing to inheritance of asthma risk.

SUMMARYGrandmaternal allergen exposure establishes an intergenerational, tissue-specific epigenetic trajectory in offspring at birth, which uniquely directs responses to allergen sensitization and challenge later in life and contributes to inheritance of asthma risk.
]]></description>
<dc:creator>Lebold, K. M.</dc:creator>
<dc:creator>Cook, M.</dc:creator>
<dc:creator>Pincus, A. B.</dc:creator>
<dc:creator>Nevonen, K. A.</dc:creator>
<dc:creator>Davis, B. A.</dc:creator>
<dc:creator>Carbone, L.</dc:creator>
<dc:creator>Calco, G. N.</dc:creator>
<dc:creator>Fryer, A. D.</dc:creator>
<dc:creator>Jacoby, D. B.</dc:creator>
<dc:creator>Drake, M. G.</dc:creator>
<dc:date>2022-01-27</dc:date>
<dc:identifier>doi:10.1101/2022.01.25.477760</dc:identifier>
<dc:title><![CDATA[Grandmaternal allergen exposure causes distinct epigenetic trajectories in offspring associated with airway hyperreactivity and inflammation.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-01-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.01.26.477791v1?rss=1">
<title>
<![CDATA[
Abnormal CCK/CB1R axon targeting in mouse models of dystroglycanopathy 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.01.26.477791v1?rss=1</link>
<description><![CDATA[
Dystroglycan (Dag1) is a transmembrane glycoprotein that links the extracellular matrix to the actin cytoskeleton. Mutations in Dag1 or the genes required for its glycosylation result in dystroglycanopathy, a type of congenital muscular dystrophy characterized by a wide range of phenotypes including muscle weakness, brain defects, and cognitive impairment. We investigated interneuron (IN) development, synaptic function, and associated seizure susceptibility in multiple mouse models that reflect the wide phenotypic range of dystroglycanopathy neuropathology. Mice that model severe dystroglycanopathy due to forebrain deletion of Dag1 or Pomt2, which is required for Dystroglycan glycosylation, show significant impairment of CCK+/CB1R+ IN development. CCK+/CB1R+ IN axons failed to properly target the somatodendritic compartment of pyramidal neurons in the hippocampus, resulting in synaptic defects and increased seizure susceptibility. Mice lacking the intracellular domain of Dystroglycan have milder defects in CCK+/CB1R+ IN axon targeting, but exhibit dramatic changes in inhibitory synaptic function, indicating a critical postsynaptic role of this domain. In contrast, CCK+/CB1R+ IN synaptic function and seizure susceptibility was normal in mice that model mild dystroglycanopathy due to partially reduced Dystroglycan glycosylation. Collectively, these data show that inhibitory synaptic defects and elevated seizure susceptibility are hallmarks of severe dystroglycanopathy, and show that Dystroglycan plays an important role in organizing functional inhibitory synapse assembly.
]]></description>
<dc:creator>Miller, D. S.</dc:creator>
<dc:creator>Jahncke, J. N.</dc:creator>
<dc:creator>Schnell, E.</dc:creator>
<dc:creator>Wright, K. M.</dc:creator>
<dc:date>2022-01-26</dc:date>
<dc:identifier>doi:10.1101/2022.01.26.477791</dc:identifier>
<dc:title><![CDATA[Abnormal CCK/CB1R axon targeting in mouse models of dystroglycanopathy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-01-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.01.27.477930v1?rss=1">
<title>
<![CDATA[
Development of the First Tractable Genetic System for Parvimonas micra, a Ubiquitous Pathobiont in Human Dysbiotic Disease 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.01.27.477930v1?rss=1</link>
<description><![CDATA[
Parvimonas micra is a Gram-positive obligate anaerobe and a typical member of the human microbiome. P. micra is among the most highly enriched species at numerous sites of mucosal dysbiotic disease and is closely associated with the development of multiple types of malignant tumors. Despite its strong association with disease, surprisingly little is known about P. micra pathobiology, which is directly attributable to its longstanding genetic intractability. To address this problem, we directly isolated a collection of P. micra strains from odontogenic abscess clinical specimens and then screened these isolates for natural competence. Amazingly, all of the P. micra clinical isolates exhibited various levels of natural competence, including the reference strain ATCC 33270. By exploiting this ability, we were able to employ cloning-independent methodologies to engineer and complement a variety of targeted chromosomal genetic mutations directly within low passage clinical isolates. To create the first P. micra genetic system, we employed renilla-based bioluminescence for highly sensitive reporter studies. This reporter system was then applied for the development of the novel Theo+ theophylline-inducible riboswitch for tunable gene expression studies over a broad dynamic range. Finally, we demonstrate the feasibility of generating Mariner-based Tn-seq libraries for forward genetic screening in P. micra. With the availability of a highly efficient transformation protocol and the current suite of genetic tools, P. micra should now be considered as a fully genetically tractable organism suitable for molecular genetic research. The methods presented here provide a clear path to investigate the understudied role of P. micra in polymicrobial infections and tumorigenesis.
]]></description>
<dc:creator>Higashi, D. L.</dc:creator>
<dc:creator>McGuire, S.</dc:creator>
<dc:creator>Abdelrahman, Y.</dc:creator>
<dc:creator>Zou, Z.</dc:creator>
<dc:creator>Qin, H.</dc:creator>
<dc:creator>Anderson, D.</dc:creator>
<dc:creator>Palmer, E. A.</dc:creator>
<dc:creator>Kreth, J.</dc:creator>
<dc:creator>Merritt, J.</dc:creator>
<dc:date>2022-01-28</dc:date>
<dc:identifier>doi:10.1101/2022.01.27.477930</dc:identifier>
<dc:title><![CDATA[Development of the First Tractable Genetic System for Parvimonas micra, a Ubiquitous Pathobiont in Human Dysbiotic Disease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-01-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.01.27.478064v1?rss=1">
<title>
<![CDATA[
Shared and distinct functional effects of patient-specific Tbr1 mutations on cortical development 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.01.27.478064v1?rss=1</link>
<description><![CDATA[
Predicted loss-of-function and missense heterozygous de novo mutations of TBR1 are strongly associated with intellectual disability and autism. The functional effects of these heterogeneous mutations on cortical development and genotype-phenotype relationships have yet to be explored. We characterized mouse models carrying patient mutations A136PfsX80 and K228E, finding convergent and discordant phenotypes. The A136PfsX80 mutation is loss-of-function and allelic to the Tbr1 knockout. In contrast, K228E causes significant upregulation of TBR1. Heterozygosity of either mutation produces axon defects, including reduction of the anterior commissure, and CTIP2 downregulation in adult cortex. While mice lacking TBR1 show extensive cortical apoptosis and inverted layering, K228E homozygotes show normal apoptosis levels and a complex layering phenotype--suggesting partial, yet abnormal, function of the allele. The construct and face validity of these Tbr1 patient mutation mice suggests they will be valuable translational models for studying the function of this essential brain transcription factor.
]]></description>
<dc:creator>Co, M.</dc:creator>
<dc:creator>Barnard, R. A.</dc:creator>
<dc:creator>Jahncke, J. N.</dc:creator>
<dc:creator>Grindstaff, S.</dc:creator>
<dc:creator>Fedorov, L. M.</dc:creator>
<dc:creator>Adey, A. C.</dc:creator>
<dc:creator>Wright, K. M.</dc:creator>
<dc:creator>O'Roak, B. J.</dc:creator>
<dc:date>2022-01-28</dc:date>
<dc:identifier>doi:10.1101/2022.01.27.478064</dc:identifier>
<dc:title><![CDATA[Shared and distinct functional effects of patient-specific Tbr1 mutations on cortical development]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-01-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.02.02.478920v1?rss=1">
<title>
<![CDATA[
A novel rhesus macaque model of Huntington's disease recapitulates key neuropathological changes along with progressive motor and cognitive decline 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.02.02.478920v1?rss=1</link>
<description><![CDATA[
We created a new nonhuman primate model of the genetic neurodegenerative disorder, Huntingtons disease (HD), by injecting a mixture of recombinant adeno-associated viral vectors, serotypes AAV2 and AAV2.retro, each expressing a fragment of human mutant HTT (mHTT) into the caudate and putamen of adult rhesus macaques. This novel modeling strategy results in robust expression of mutant huntingtin protein (mHTT) in the injected brain regions, as well as dozens of other cortical and subcortical brain regions that are also affected in human HD patients. We queried the disruption of cortico-basal ganglia circuitry for 20-months post-surgery using a variety of behavioral and imaging readouts. Compared to controls, mHTT-treated macaques developed progressive working memory decline and motor impairment. Multimodal imaging revealed circuit-wide white and gray matter degenerative processes in several key brain regions affected in HD. This novel model will aid in the development of disease biomarkers and therapeutic strategies for this devastating disorder.
]]></description>
<dc:creator>Weiss, A. R.</dc:creator>
<dc:creator>Liguore, W. A.</dc:creator>
<dc:creator>Brandon, K.</dc:creator>
<dc:creator>Wang, X.</dc:creator>
<dc:creator>Liu, Z.</dc:creator>
<dc:creator>Domire, J. S.</dc:creator>
<dc:creator>Button, D.</dc:creator>
<dc:creator>Kroenke, C. D.</dc:creator>
<dc:creator>McBride, J. L.</dc:creator>
<dc:date>2022-02-04</dc:date>
<dc:identifier>doi:10.1101/2022.02.02.478920</dc:identifier>
<dc:title><![CDATA[A novel rhesus macaque model of Huntington's disease recapitulates key neuropathological changes along with progressive motor and cognitive decline]]></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/10.1101/2022.02.10.479848v1?rss=1">
<title>
<![CDATA[
CD8 T Cell Memory Inflation is Driven by Latent CMV Infection of Lymphatic Endothelial Cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.02.10.479848v1?rss=1</link>
<description><![CDATA[
CMV, a ubiquitous herpesvirus, elicits an extraordinarily large T cell response that is sustained or increases over time, a phenomenon termed  memory inflation. Remarkably, even latent, non-productive infection can drive memory inflation. Despite intense research on this phenomenon, the infected cell type(s) involved are unknown. To identify the responsible cell type(s), we designed a Cre-lox murine CMV (MCMV) system, where a spread-deficient ({Delta}gL) virus expresses recombinant SIINFEKL only in Cre+ host cells. We found that latent infection of endothelial cells (ECs), but not dendritic cells (DCs) or hepatocytes, was sufficient to drive CD8 T cell memory inflation. Infection of Lyve-1-Cre and Prox1-CreERT2 mice revealed that amongst EC subsets, infection of lymphatic ECs was sufficient. Genetic ablation of {beta}2m on lymphatic ECs did not prevent inflation, suggesting another unidentified cell type can also present antigen to CD8 T cells during latency. This novel system definitively shows that antigen presentation by lymphatic ECs drives robust CD8 T cell memory inflation.

SUMMARYActive T cell immunosurveillance during latent CMV infection results in T cell  memory inflation. A novel Cre-lox genetic system for cell-specific antigen expression reveals that lymphatic ECs, but not DCs or hepatocytes, can drive CD8 T cell memory inflation.
]]></description>
<dc:creator>Hill, A. B.</dc:creator>
<dc:creator>Munks, M.</dc:creator>
<dc:creator>Rott, K.</dc:creator>
<dc:creator>Nesterenko, P.</dc:creator>
<dc:creator>Smart, S.</dc:creator>
<dc:creator>Smith, T.</dc:creator>
<dc:creator>Xu, G.</dc:creator>
<dc:creator>Williams, V.</dc:creator>
<dc:creator>Murray, S.</dc:creator>
<dc:creator>Tatum, A.</dc:creator>
<dc:date>2022-02-10</dc:date>
<dc:identifier>doi:10.1101/2022.02.10.479848</dc:identifier>
<dc:title><![CDATA[CD8 T Cell Memory Inflation is Driven by Latent CMV Infection of Lymphatic Endothelial Cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-02-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.02.14.480430v1?rss=1">
<title>
<![CDATA[
A global lipid map reveals host dependency factors conserved across SARS-CoV-2 variants 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.02.14.480430v1?rss=1</link>
<description><![CDATA[
A comprehensive understanding of host dependency factors for SARS-CoV-2 remains elusive. We mapped alterations in host lipids following SARS-CoV-2 infection using nontargeted lipidomics. We found that SARS-CoV-2 rewires host lipid metabolism, altering 409 lipid species up to 64-fold relative to controls. We correlated these changes with viral protein activity by transfecting human cells with each viral protein and performing lipidomics. We found that lipid droplet plasticity is a key feature of infection and that viral propagation can be blocked by small-molecule glycerolipid biosynthesis inhibitors. We found that this inhibition was effective against the main variants of concern (alpha, beta, gamma, and delta), indicating that glycerolipid biosynthesis is a conserved host dependency factor that supports this evolving virus.
]]></description>
<dc:creator>Farley, S. E.</dc:creator>
<dc:creator>Kyle, J.</dc:creator>
<dc:creator>Leier, H.</dc:creator>
<dc:creator>Bramer, L. M.</dc:creator>
<dc:creator>Weinstein, J.</dc:creator>
<dc:creator>Bates, T. A.</dc:creator>
<dc:creator>Lee, J.-Y.</dc:creator>
<dc:creator>Metz, T. O.</dc:creator>
<dc:creator>Schultz, C.</dc:creator>
<dc:creator>Tafesse, F.</dc:creator>
<dc:date>2022-02-15</dc:date>
<dc:identifier>doi:10.1101/2022.02.14.480430</dc:identifier>
<dc:title><![CDATA[A global lipid map reveals host dependency factors conserved across SARS-CoV-2 variants]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-02-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.02.15.480607v1?rss=1">
<title>
<![CDATA[
Contribution of individual excitatory synapses on dendritic spines to electrical signaling 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.02.15.480607v1?rss=1</link>
<description><![CDATA[
Dendritic spines, small ([~]1 {micro}m) membrane protrusions from neuronal dendrites which receive most of the excitatory synaptic inputs in the mammalian brain, are widely considered the elementary computational units of the brain. Our understanding of electrical signalling in spines is currently being debated, primarily for methodological reasons. We combined the standard techniques of whole-cell recording and voltage imaging methods to study excitatory postsynaptic potentials evoked by two-photon glutamate uncaging (uEPSPs) on individual dendritic spines on basal dendrites in rat cortical slices. We analyzed the initiation, temporal summation, and propagation of uEPSPs from the spine head to the parent dendrites in three principal neocortical pyramidal neuron classes. Our measurements show no significant attenuation of uEPSPs across the spine neck in most tested mushroom spines on basal dendrites. This result implies that spine synapses are not electrically isolated from parent dendrites and that these spines do not serve a meaningful electrical role. Using the same high-sensitivity voltage imaging techniques, we characterized the temporal summation of uEPSPs induced by repetitive glutamate uncaging mimicking burst activity of presynaptic neurons. We found that responses to high-frequency repetitive quantal EPSPs are strictly limited in amplitude and waveform. This finding reveals a biophysical mechanism for preventing synaptic saturation.

Significance StatementWe used an electrochromic voltage-sensitive dye, which acts as a transmembrane optical voltmeter, to define the electrical role of dendritic spines, small membrane protrusions that receive most of the excitatory synaptic inputs in the brain. The data argue that investigated spine synapses of principal neurons are not electrically isolated from the parent dendrites. We also found that the amplitude of temporal uEPSP summation during repetitive synaptic activation is restricted at the site of origin, preventing synaptic saturation. These results facilitate our understanding of how a complex assembly of receptors and ion channels in spines generates and processes electrical signals and mediate plasticity in response to the quantal release of chemical transmitters caused by patterned activity in presynaptic axons.
]]></description>
<dc:creator>Weng, J.-Y.</dc:creator>
<dc:creator>Ceballos, C.</dc:creator>
<dc:creator>Zecevic, D.</dc:creator>
<dc:date>2022-02-17</dc:date>
<dc:identifier>doi:10.1101/2022.02.15.480607</dc:identifier>
<dc:title><![CDATA[Contribution of individual excitatory synapses on dendritic spines to electrical signaling]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-02-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.02.16.480718v1?rss=1">
<title>
<![CDATA[
Oligodendrocytes form paranodal bridges that generate chains of myelin sheaths vulnerable to degeneration with age 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.02.16.480718v1?rss=1</link>
<description><![CDATA[
Myelin sheaths in the CNS are generated by the tips of oligodendrocyte processes, which wrap axons to accelerate action potential conduction, provide metabolic support and control excitability. Here we identify a distinct mode of myelination, conserved between zebrafish, mouse and human, in which oligodendrocytes extend myelin along individual axons by linking myelin sheaths across nodes of Ranvier (NoR). By forming thin extensions that cross NoR, which we term paranodal bridges, multiple sheaths can be connected to the soma by a single cytoplasmic process. Extensive in vivo live imaging-based analyses, complemented by serial electron microscopic reconstruction of paranodal bridges, revealed that many oligodendrocytes use this strategy to generate longer stretches of myelin along individual axons. In the mouse somatosensory cortex, paranodal bridges were particularly prevalent along the highly branched axons of parvalbumin expressing (PV) interneurons, which enabled oligodendrocytes to extend myelin sheaths around axon bifurcations. Sheaths at the distal ends of these chains of myelin degenerated more frequently in aged mice, suggesting that they may be more vulnerable to the aging brain environment. This previously undescribed and evolutionarily conserved feature of oligodendrocytes extends myelin coverage of individual axons without new oligodendrogenesis, which may reduce metabolic demand and preserve the fidelity of action potential propagation at axon branch points.
]]></description>
<dc:creator>Call, C. L.</dc:creator>
<dc:creator>Neely, S. A.</dc:creator>
<dc:creator>Early, J. J.</dc:creator>
<dc:creator>James, O. G.</dc:creator>
<dc:creator>Zoupi, L.</dc:creator>
<dc:creator>Williams, A. C.</dc:creator>
<dc:creator>Chandran, S.</dc:creator>
<dc:creator>Lyons, D. A.</dc:creator>
<dc:creator>Bergles, D. E.</dc:creator>
<dc:date>2022-02-17</dc:date>
<dc:identifier>doi:10.1101/2022.02.16.480718</dc:identifier>
<dc:title><![CDATA[Oligodendrocytes form paranodal bridges that generate chains of myelin sheaths vulnerable to degeneration with age]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-02-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.02.22.481478v1?rss=1">
<title>
<![CDATA[
Temporal continuity shapes visual responses of macaque face patch neurons 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.02.22.481478v1?rss=1</link>
<description><![CDATA[
Macaque inferior temporal cortex neurons respond selectively to complex visual images, with recent work showing that they are also entrained reliably by the evolving content of natural movies. To what extent does visual continuity itself shape the responses of high-level visual neurons? We addressed this question by measuring how cells in face-selective regions of the macaque temporal cortex were affected by the manipulation of a movies temporal structure. Sampling the movie at 1s intervals, we measured neural responses to randomized, brief stimuli of different lengths, ranging from 800 ms dynamic movie snippets to 100 ms static frames. We found that the disruption of temporal continuity strongly altered neural response profiles, particularly in the early onset response period of the randomized stimulus. The results suggest that models of visual system function based on discrete and randomized visual presentations may not translate well to the brains natural modes of operation.
]]></description>
<dc:creator>Russ, B. E.</dc:creator>
<dc:creator>Koyano, K. W.</dc:creator>
<dc:creator>Day-Cooney, J.</dc:creator>
<dc:creator>Perwez, N.</dc:creator>
<dc:creator>Leopold, D. A.</dc:creator>
<dc:date>2022-02-24</dc:date>
<dc:identifier>doi:10.1101/2022.02.22.481478</dc:identifier>
<dc:title><![CDATA[Temporal continuity shapes visual responses of macaque face patch neurons]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-02-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.02.25.482016v1?rss=1">
<title>
<![CDATA[
NEDD8 activity is important for direct antigen MHC class I antigen presentation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.02.25.482016v1?rss=1</link>
<description><![CDATA[
Successful direct MHC class I antigen presentation is dependent on the protein degradation machinery of the cell to generate antigenic peptides which can be loaded onto MHC class I molecules for surveillance by CD8+ T cells of the immune system. Most often this process involves the ubiquitin-proteasome system, however other ubiquitin-like (UBL) proteins have also been implicated in protein degradation and direct antigen presentation. Here, we examine the role of neuronal precursor cell-expressed developmentally down-regulated protein 8 (NEDD8) in direct antigen presentation. NEDD8 is the UBL with highest similarity to ubiquitin and fusion of NEDD8 to the amino-terminus of a target protein can lead to the target proteins degradation. We find that appending NEDD8 to the N-terminus of the model antigen ovalbumin resulted in degradation by both the proteasome and autophagy protein degradation pathways, but only proteasomal degradation, involving the proteasomal subunit NEDD8 ultimate buster 1 (NUB1), resulted in peptide presentation. When directly compare to ubiquitin, NEDD8-fusion was less efficient at generating peptides. However, inactivation of the NEDD8-conugation machinery by treating cells with MLN4924, inhibited the presentation of peptides from Defective Ribosomal Products (DRiPs) derived from a model antigen. These results demonstrate that NEDD8 activity in the cell is important for direct antigen presentation, but not by directly targeting proteins for degradation.
]]></description>
<dc:creator>Vijayasimha, K.</dc:creator>
<dc:creator>Leestemaker-Palmer, A. L.</dc:creator>
<dc:creator>Gibbs, J. S.</dc:creator>
<dc:creator>Yewdell, J. W.</dc:creator>
<dc:creator>Dolan, B. P.</dc:creator>
<dc:date>2022-02-27</dc:date>
<dc:identifier>doi:10.1101/2022.02.25.482016</dc:identifier>
<dc:title><![CDATA[NEDD8 activity is important for direct antigen MHC class I antigen presentation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-02-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.02.28.482383v1?rss=1">
<title>
<![CDATA[
Chronic obstructive pulmonary disease and cigarette smoke exposure lead to dysregulated MAIT cell activation by bronchial epithelial cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.02.28.482383v1?rss=1</link>
<description><![CDATA[
Chronic obstructive pulmonary disease (COPD) is associated with airway inflammation, increased infiltration by CD8+ T lymphocytes, and infection-driven exacerbations. COPD is most commonly caused by cigarette smoke (CS), however the mechanisms driving development of COPD in some smokers but not others are incompletely understood. Lung-resident mucosal-associated invariant T (MAIT) cells play a role in both microbial infections and inflammatory diseases. MAIT cell frequency is reduced in the peripheral blood of individuals with COPD, however the role of MAIT cells in COPD pathology is unknown. Here, we examined MAIT cell activation in response to CS-exposed primary human bronchial epithelial cells (BEC) from healthy, COPD, or smoker donors. We observed significantly higher MAIT cell responses to COPD BEC than healthy BEC. However, COPD BEC stimulated a smaller fold-increase in MAIT cell response despite increased microbial infection. For all donor groups, CS-exposed BEC elicited reduced MAIT cell responses; conversely, CS exposure increased ligand-mediated MR1 surface translocation in healthy and COPD BEC. Our data demonstrate MAIT cell activation is dysregulated in the context of CS and COPD. MAIT cells could contribute to CS- and COPD-associated inflammation through both inappropriate activation and reduced early recognition of bacterial infection, contributing to microbial persistence and COPD exacerbations.
]]></description>
<dc:creator>Huber, M. E.</dc:creator>
<dc:creator>Larson, E.</dc:creator>
<dc:creator>Lust, T. N.</dc:creator>
<dc:creator>Heisler, C. M.</dc:creator>
<dc:creator>Harriff, M. J.</dc:creator>
<dc:date>2022-03-02</dc:date>
<dc:identifier>doi:10.1101/2022.02.28.482383</dc:identifier>
<dc:title><![CDATA[Chronic obstructive pulmonary disease and cigarette smoke exposure lead to dysregulated MAIT cell activation by bronchial epithelial cells]]></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/10.1101/2022.03.02.482686v1?rss=1">
<title>
<![CDATA[
Taxonomic and functional shifts in the perinatal gut microbiome of rhesus macaques 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.03.02.482686v1?rss=1</link>
<description><![CDATA[
Pregnancy and the postpartum period result in some of the most dramatic metabolic, hormonal, and physiological changes that can be experienced by an otherwise healthy adult. The timing and magnitude of these changes is key for both maternal and fetal health. One of the factors believed to critically modulate these physiological changes is the maternal gut microbiome. However, the dynamic changes in this community during the perinatal period remain understudied. Clinical studies can be complicated by confounding variables such as diet and other drivers of heterogeneity in the human microbiome. Therefore, in this study, we conducted a longitudinal analysis of the fecal microbiome obtained during the perinatal and post-partum periods in 25 rhesus macaques using 16S rRNA gene amplicon sequencing and shotgun metagenomics. Shifts at both the taxonomic and functional potential level were detected when comparing pregnancy to postpartum samples. Taxonomically, Alloprevotella, Actinobacillus, and Anaerovibrio were enriched during pregnancy while Treponema, Lachnospiraceae, and Methanosphaera were more abundant post-partum. Functionally, pregnancy was associated with increased abundance in the pathway to produce the beneficial short chain fatty acid (SCFA), butyrate, while pathways associated with starch degradation and folate transformation were more abundant postpartum. These data demonstrate dramatic changes in the maternal gut even in the absence of dietary changes and suggest that rhesus macaques could provide a valuable model to determine how changes in the microbiome correlate to other physiological changes in pregnancy.
]]></description>
<dc:creator>Rhoades, N. S.</dc:creator>
<dc:creator>Cinco, I. R.</dc:creator>
<dc:creator>Hendrickson, S. M.</dc:creator>
<dc:creator>Slifka, M. K.</dc:creator>
<dc:creator>Messaoudi, I.</dc:creator>
<dc:date>2022-03-03</dc:date>
<dc:identifier>doi:10.1101/2022.03.02.482686</dc:identifier>
<dc:title><![CDATA[Taxonomic and functional shifts in the perinatal gut microbiome of rhesus macaques]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-03-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.03.02.482692v1?rss=1">
<title>
<![CDATA[
Ligand-mediated structural dynamics of a mammalian pancreatic KATP channel 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.03.02.482692v1?rss=1</link>
<description><![CDATA[
Regulation of pancreatic KATP channels involves orchestrated interactions of channel subunits, Kir6.2 and SUR1, and their ligands. How ligand interactions affect channel conformations and activity is not well understood. To elucidate the structural correlates pertinent to ligand interactions and channel gating, we compared cryo-EM structures of channels in the presence and absence of pharmacological inhibitors and ATP, focusing on channel conformational dynamics. We found pharmacological inhibitors and ATP enrich a channel conformation in which the Kir6.2 cytoplasmic domain is closely associated with the transmembrane domain relative to one where the Kir6.2 cytoplasmic domain is extended away into the cytoplasm. This conformation change remodels a network of intra and inter-subunit interactions as well as both the ATP and PIP2 binding pockets. The structures resolved key contacts between the distal N-terminus of Kir6.2 and SUR1s ABC module involving residues implicated in channel function. A SUR1 residue, K134, is identified to directly contribute to the PIP2 binding pocket. Molecular dynamics simulations revealed two Kir6.2 residues, K39 and R54, that mediate both ATP and PIP2 binding, suggesting a mechanism for competitive gating by ATP and PIP2.
]]></description>
<dc:creator>Sung, M. W.</dc:creator>
<dc:creator>Driggers, C. M.</dc:creator>
<dc:creator>Mostofian, B.</dc:creator>
<dc:creator>Russo, J. D.</dc:creator>
<dc:creator>Patton, B. L.</dc:creator>
<dc:creator>Zuckerman, D. M.</dc:creator>
<dc:creator>Shyng, S.-L.</dc:creator>
<dc:date>2022-03-02</dc:date>
<dc:identifier>doi:10.1101/2022.03.02.482692</dc:identifier>
<dc:title><![CDATA[Ligand-mediated structural dynamics of a mammalian pancreatic KATP channel]]></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/10.1101/2022.03.02.482705v1?rss=1">
<title>
<![CDATA[
Kv7 channel antagonists block glycine receptors 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.03.02.482705v1?rss=1</link>
<description><![CDATA[
XE991 (10,10-bis(4-pyridinylmethyl)-9(10H)-anthracenone) is currently the most widely used and specific antagonist of the Kv7 (KCNQ) family of K+ channels. We report an unexpected antagonistic effect of this drug on ionotropic glycine receptors. In recordings of synaptic transmission in two brainstem nuclei (the medial nucleus of the trapezoid body and the dorsal cochlear nucleus), 10 M XE991, a concentration typical for Kv7 studies in brain tissue, inhibited evoked glycinergic inhibitory postsynaptic currents (IPSCs) without altering paired-pulse ratio, and also reduced the amplitude of glycinergic miniature IPSCs. These results are indicative of a direct effect of the drug on postsynaptic glycine receptors. XE991 also produced dose-dependent block of the response to exogenously applied glycine, to a degree comparable to the block of synaptic transmission. Moreover, the drug inhibited homomeric glycine receptors expressed on presynaptic membrane of the calyx of Held. The degree of block was independent of glycine concentration, suggesting an allosteric interaction. The effects of XE991 on glycine responses are not likely to reflect block of the glycine-activated Cl- channels themselves, because block was voltage independent, and because GABA-activated Cl- currents were resistant to XE991 at concentrations up to 100 {micro}M. Linopirdine, but not retigabine, also antagonized glycine receptor currents. Given the prevalence of glycine receptor signaling in the brain, these observations should be taken into account in studies of the roles of Kv7 channels in neural circuit function and disease.
]]></description>
<dc:creator>Lu, H.-W.</dc:creator>
<dc:creator>Romero, G. E.</dc:creator>
<dc:creator>Apostolides, P. F.</dc:creator>
<dc:creator>Huang, H.</dc:creator>
<dc:creator>Trussell, L. O.</dc:creator>
<dc:date>2022-03-02</dc:date>
<dc:identifier>doi:10.1101/2022.03.02.482705</dc:identifier>
<dc:title><![CDATA[Kv7 channel antagonists block glycine receptors]]></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/10.1101/2022.03.04.483011v1?rss=1">
<title>
<![CDATA[
Rapid in vivo multiplexed editing (RIME) of the adult mouse liver 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.03.04.483011v1?rss=1</link>
<description><![CDATA[
Background & AimsAssessing mammalian gene function in vivo has traditionally relied on manipulation of the mouse genome in embryonic stem cells or peri-zygotic embryos. These approaches are time consuming and require extensive breeding when simultaneous mutations in multiple genes is desired. The aim of this study is to introduce a Rapid In vivo Multiplexed Editing (RIME), and to provide a proof-of-concept of this system.

Approach & ResultsRIME, a system wherein CRISPR/Cas9 technology, paired with adeno-associated viruses (AAVs), permits the inactivation of one or more genes in the adult mouse liver. The method is quick, requiring as little as 1 month from conceptualization to knockout (KO), and highly efficient, enabling editing in >95% of target cells. To highlight its utility, we used this system to inactivate, alone or in combination, genes with functions spanning metabolism, mitosis, mitochondrial maintenance, and cell proliferation.

ConclusionRIME enables the rapid, efficient, and inexpensive analysis of multiple genes in the mouse liver in vivo.
]]></description>
<dc:creator>Katsuda, T.</dc:creator>
<dc:creator>Cure, H.</dc:creator>
<dc:creator>Simeonov, K. P.</dc:creator>
<dc:creator>Arany, Z.</dc:creator>
<dc:creator>Grompe, M.</dc:creator>
<dc:creator>Stanger, B.</dc:creator>
<dc:date>2022-03-05</dc:date>
<dc:identifier>doi:10.1101/2022.03.04.483011</dc:identifier>
<dc:title><![CDATA[Rapid in vivo multiplexed editing (RIME) of the adult mouse liver]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-03-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.03.07.483324v1?rss=1">
<title>
<![CDATA[
In silico screening and testing of FDA approved small molecules to block SARS-CoV-2 entry to the host cell by inhibiting Spike protein cleavage 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.03.07.483324v1?rss=1</link>
<description><![CDATA[
The COVID-19 pandemic began in 2019, but it is still active. The development of an effective vaccine reduced the number of deaths; however, a treatment is still needed. Here, we aimed to inhibit viral entry to the host cell by inhibiting Spike (S) protein cleavage by several proteases. We develop a computational pipeline to repurpose FDA-approved drugs to inhibit protease activity and thus prevent S protein cleavage. We tested some of our drug candidates and demonstrated a decrease in protease activity. We believe our pipeline will be beneficial in identifying a drug regimen for COVID-19 patients.
]]></description>
<dc:creator>Ozdemir, E. S.</dc:creator>
<dc:creator>Le, H. H.</dc:creator>
<dc:creator>Yildirim, A.</dc:creator>
<dc:creator>Ranganathan, S. V.</dc:creator>
<dc:date>2022-03-07</dc:date>
<dc:identifier>doi:10.1101/2022.03.07.483324</dc:identifier>
<dc:title><![CDATA[In silico screening and testing of FDA approved small molecules to block SARS-CoV-2 entry to the host cell by inhibiting Spike protein cleavage]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-03-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.03.10.483746v1?rss=1">
<title>
<![CDATA[
Chondroitin Sulfate Proteoglycan 4,6 sulfation regulates sympathetic nerve regeneration after myocardial infarction 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.03.10.483746v1?rss=1</link>
<description><![CDATA[
Sympathetic denervation of the heart following myocardial infarction (MI) is sustained by chondroitin sulfate proteoglycans (CSPGs) in the cardiac scar, and denervation predicts risk of sudden cardiac death. Blocking CSPG signaling restores sympathetic axon outgrowth into the cardiac scar, decreasing arrhythmia susceptibility. Axon growth inhibition by CSPGs is thought to depend on the sulfation status of the glycosaminoglycans (CS-GAGs) attached to the core protein. Tandem sulfation of CS-GAGs at the 4th (4S) and 6th (6S) positions of n-acetyl-galactosamine inhibits outgrowth in several types of neurons within the central nervous system, but it is not known if sulfation is similarly critical during peripheral nerve regeneration. We asked if CSPG sulfation prevented sympathetic axon outgrowth. Sympathetic neurite outgrowth across purified CSPGs is restored in vitro by reducing 4S with the 4-sulfatase enzyme Arylsulfatase-B (ARSB). Additionally, we co-cultured cardiac scar tissue with sympathetic ganglia ex vivo and found that reducing 4S with ARSB restored axon outgrowth to control levels. We examined levels of the enzymes responsible for adding and removing sulfation to CS-GAGs by western blot to determine if they were altered in the left ventricle after MI. We found that CHST15 (4S dependent 6-sulfotransferase) was upregulated, and ARSB was downregulated after MI. Increased CHST15 combined with decreased ARSB suggests a mechanism for production and maintenance of sulfated CSPGs in the cardiac scar. We altered tandem sulfated 4S,6S CS-GAGs in vivo by transient siRNA knockdown of Chst15 and found that reducing 4S,6S restored Tyrosine Hydroxylase (TH) positive sympathetic nerve fibers in the cardiac scar after MI and reduced arrhythmias. Overall, our results suggest that modulating CSPG-sulfation after MI may be a therapeutic target to promote sympathetic nerve regeneration in the cardiac scar and reduce post-MI cardiac arrhythmias.
]]></description>
<dc:creator>Blake, M. R.</dc:creator>
<dc:creator>Parrish, D. C.</dc:creator>
<dc:creator>Staffenson, M. A.</dc:creator>
<dc:creator>Sueda, S.</dc:creator>
<dc:creator>Woodward, W. R.</dc:creator>
<dc:creator>Beth Habecker, B. A.</dc:creator>
<dc:date>2022-03-13</dc:date>
<dc:identifier>doi:10.1101/2022.03.10.483746</dc:identifier>
<dc:title><![CDATA[Chondroitin Sulfate Proteoglycan 4,6 sulfation regulates sympathetic nerve regeneration after myocardial infarction]]></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/10.1101/2022.03.11.484016v1?rss=1">
<title>
<![CDATA[
Retained introns in long RNA-seq reads are not reliably detected in sample-matched short reads 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.03.11.484016v1?rss=1</link>
<description><![CDATA[
There is growing interest in retained introns in a variety of disease contexts including cancer and aging. Many software tools have been developed to detect retained introns from short RNA-seq reads, but reliable detection is complicated by overlapping genes and transcripts as well as the presence of unprocessed or partially processed RNAs. We compared introns detected by 5 tools using short RNA-seq reads with introns observed in long RNA-seq reads from the same biological specimens and found: (1) significant disagreement among tools (Fleiss{kappa} = 0.231) such that 52.4% of all detected intron retentions were not called by more than one tool; (2) that no tool achieved greater than 20% precision or 35% recall under generous conditions; and (3) that retained intron detectability was adversely affected by greater intron length and overlap with annotated exons.
]]></description>
<dc:creator>David, J. K.</dc:creator>
<dc:creator>Maden, S. K.</dc:creator>
<dc:creator>Wood, M. A.</dc:creator>
<dc:creator>Thompson, R. F.</dc:creator>
<dc:creator>Nellore, A.</dc:creator>
<dc:date>2022-03-12</dc:date>
<dc:identifier>doi:10.1101/2022.03.11.484016</dc:identifier>
<dc:title><![CDATA[Retained introns in long RNA-seq reads are not reliably detected in sample-matched short reads]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-03-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.03.18.484955v1?rss=1">
<title>
<![CDATA[
Synapses in oligodendrocyte precursor cells are dynamic and contribute to Ca2+ activity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.03.18.484955v1?rss=1</link>
<description><![CDATA[
In the nervous system, only one type of neuron-glial synapse is known to exist: that between neurons and oligodendrocyte precursor cells (OPCs). Neuron-OPC synapses are thought to bridge neuronal activity to OPCs. However, their composition, assembly, downstream signaling, and in vivo functions remain largely unclear. Here, we use zebrafish to address these questions and identify postsynaptic molecules PSD-95 and Gephyrin in OPCs. They increase during early development and decrease upon OPC differentiation. PSD-95 and Gephyrin in OPCs are highly dynamic and frequently assemble at "hotspots." Gephyrin hotspots and synapse-associated Ca2+ activity in OPCs predict where a subset of myelin sheaths form in oligodendrocytes. Further analyses reveal that spontaneous synaptic release is integral to OPC Ca2+ activity, while evoked synaptic release contributes only in early development. Finally, disruption of the synaptic genes dlg4a&b, gephyrinb, and nlgn3b impairs OPC differentiation and myelination. Together, we propose that neuron-OPC synapses are dynamically assembled and can predetermine myelination patterns through Ca2+ signaling.
]]></description>
<dc:creator>Li, J.</dc:creator>
<dc:creator>Miramontes, T.</dc:creator>
<dc:creator>Czopka, T.</dc:creator>
<dc:creator>Monk, K. R.</dc:creator>
<dc:date>2022-03-19</dc:date>
<dc:identifier>doi:10.1101/2022.03.18.484955</dc:identifier>
<dc:title><![CDATA[Synapses in oligodendrocyte precursor cells are dynamic and contribute to Ca2+ activity]]></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/10.1101/2022.03.21.484586v1?rss=1">
<title>
<![CDATA[
Three-dimensional structure of the single domain cupredoxin AcoP 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.03.21.484586v1?rss=1</link>
<description><![CDATA[
Cupredoxins are widely occurring copper-binding proteins with a typical Greek-key beta barrel fold. They are generally described as electron carriers that rely on a T1 copper center coordinated by four ligands provided by the folded polypeptide. The discovery of novel cupredoxins demonstrates the high diversity of this family, with variations in term of copper-binding ligands, copper center geometry, redox potential, as well as biological function. AcoP is a periplasmic protein belonging to the iron respiratory chain of the acidophilic bacterium Acidithiobacillus ferrooxidans. AcoP presents original features: highly resistant to acidic pH, it possesses a constrained green-type copper center of high redox potential. To understand the unique properties of AcoP, we undertook structural and biophysical characterization of wild-type AcoP and of two Cu-ligand mutants (H166A and M171A). The crystallographic structure of AcoP at 1.65 [A] resolution unveils a typical cupredoxin fold with extended loops, never observed in previously characterized cupredoxins, that might be involved in the interaction of AcoP with its physiological partners. Moreover, the structure shows that the green color of AcoP cannot be attributed to nonclassical copper ligands, its green-colored copper center raising from a long Cu-S (Cys) bond, determined by both X-ray diffraction and EXAFS. The crystal structures of two AcoP mutants confirm that the active center of AcoP is highly constrained. Comparative analysis with other cupredoxins of known structures, suggests that in AcoP the second coordination sphere might be an important determinant of active center rigidity due to the presence of an extensive hydrogen bond network.
]]></description>
<dc:creator>Roger, M.</dc:creator>
<dc:creator>Leone, P.</dc:creator>
<dc:creator>Blackburn, N.</dc:creator>
<dc:creator>Horrel, S.</dc:creator>
<dc:creator>Chicano, T. M.</dc:creator>
<dc:creator>Giudici-Orticoni, M.-T.</dc:creator>
<dc:creator>Abriata, L. A.</dc:creator>
<dc:creator>Hura, G. L.</dc:creator>
<dc:creator>Hough, M. A.</dc:creator>
<dc:creator>Sciara, G.</dc:creator>
<dc:creator>Ilbert, M.</dc:creator>
<dc:date>2022-03-21</dc:date>
<dc:identifier>doi:10.1101/2022.03.21.484586</dc:identifier>
<dc:title><![CDATA[Three-dimensional structure of the single domain cupredoxin AcoP]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-03-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.03.21.485088v1?rss=1">
<title>
<![CDATA[
Fetal loss in pregnant rhesus macaques infected with high-dose African-lineage Zika virus 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.03.21.485088v1?rss=1</link>
<description><![CDATA[
Countermeasures against Zika virus (ZIKV), including vaccines, are frequently tested in nonhuman primates (NHP). Macaque models are important for understanding how ZIKV infections impact human pregnancy due to similarities in placental development. The lack of consistent adverse pregnancy outcomes in ZIKV-affected pregnancies poses a challenge in macaque studies where group sizes are often small (4-8 animals). Studies in small animal models suggest that African-lineage Zika viruses can cause more frequent and severe fetal outcomes. No adverse outcomes were observed in macaques inoculated with a low dose of African-lineage ZIKV at gestational day (GD) 45. Here, we inoculate eight pregnant rhesus macaques with a higher dose of African-lineage ZIKV at GD 45 to test the hypothesis that adverse pregnancy outcomes are dose-dependent. Three of eight pregnancies ended prematurely with fetal death. ZIKV was detected in both fetal and placental tissues from all cases of early fetal loss. Further refinements of this challenge system (e.g., varying the dose and timing of infection) could lead to an even more consistent, unambiguous fetal loss phenotype for assessing ZIKV countermeasures in pregnancy. These data demonstrate that high-dose inoculation with African-lineage ZIKV causes pregnancy loss in macaques and also suggest that ZIKV-induced first trimester pregnancy could be strain-specific.

Author summaryAlthough pregnant rhesus macaques are susceptible to infection with Zika virus (ZIKV), fetal phenotypes can be subtle and variable. Most macaque studies of ZIKV have involved infection with Asian-lineage viruses because these viruses caused Western Hemisphere outbreaks beginning in 2015. African-lineage ZIKV yields more severe adverse fetal outcomes in small animal models. Here, we provide evidence that pregnant macaques infected late in the first trimester using a high dose of African-lineage ZIKV have frequent ZIKV-associated pregnancy loss. This severe phenotype establishes a new model for evaluating countermeasures and reinforces the idea that African-lineage ZIKV infection may be a frequent cause of pregnancy loss in areas where it is endemic depending on the amount of transmitted virus during infection.
]]></description>
<dc:creator>Raasch, L.</dc:creator>
<dc:creator>Yamamoto, K.</dc:creator>
<dc:creator>Newman, C. M.</dc:creator>
<dc:creator>Rosinski, J. R.</dc:creator>
<dc:creator>Shepherd, P. M.</dc:creator>
<dc:creator>Razo, E.</dc:creator>
<dc:creator>Crooks, C. M.</dc:creator>
<dc:creator>Bliss, M. I.</dc:creator>
<dc:creator>Breitbach, M. E.</dc:creator>
<dc:creator>Sneed, E. L.</dc:creator>
<dc:creator>Weiler, A. M.</dc:creator>
<dc:creator>Zeng, X.</dc:creator>
<dc:creator>Noguchi, K. K.</dc:creator>
<dc:creator>Morgan, T. K.</dc:creator>
<dc:creator>Fuhler, N. A.</dc:creator>
<dc:creator>Bohm, E. K.</dc:creator>
<dc:creator>Alberts, A. J.</dc:creator>
<dc:creator>Havlicek, S. J.</dc:creator>
<dc:creator>Kabakov, S.</dc:creator>
<dc:creator>Mitzey, A. M.</dc:creator>
<dc:creator>Antony, K. M.</dc:creator>
<dc:creator>Ausderau, K. K.</dc:creator>
<dc:creator>Mejia, A.</dc:creator>
<dc:creator>Basu, P.</dc:creator>
<dc:creator>Simmons, H. A.</dc:creator>
<dc:creator>Eickhoff, J. C.</dc:creator>
<dc:creator>Aliota, M. T.</dc:creator>
<dc:creator>Mohr, E. L.</dc:creator>
<dc:creator>Friedrich, T. C.</dc:creator>
<dc:creator>Golos, T. G.</dc:creator>
<dc:creator>O'Connor, D. H.</dc:creator>
<dc:creator>Dudley, D. M.</dc:creator>
<dc:date>2022-03-22</dc:date>
<dc:identifier>doi:10.1101/2022.03.21.485088</dc:identifier>
<dc:title><![CDATA[Fetal loss in pregnant rhesus macaques infected with high-dose African-lineage Zika virus]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-03-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.03.23.485409v1?rss=1">
<title>
<![CDATA[
PRMT5 as a Novel Druggable Vulnerability for EWSR1-ATF1-driven Clear Cell Sarcoma 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.03.23.485409v1?rss=1</link>
<description><![CDATA[
Clear cell sarcoma of soft tissue (CCSST) is an ultra-rare sarcoma with poor prognosis presently with no cure. It is characterized by a balanced t(12;22) (q13;q12) chromosomal translocation, resulting in a fusion of the Ewings sarcoma gene EWSR1 with activating transcription factor 1 (ATF1) to give an oncogene EWSR1-ATF1. Unlike normal ATF1, whose transcription activity is dependent on phosphorylation, EWSR1-ATF1 is constitutively active to drive ATF1-dependent gene transcription to cause tumorigenesis. No EWSR1-ATF1-targeted therapies have been identified due to the challenges in targeting intracellular transcription factors. To identify potential druggable targets for CCSST, we show that protein arginine methyltransferase 5 (PRMT5) is a novel enzyme in enhancing EWSR1-ATF1-mediated gene transcription to sustain CCSST cell proliferation. Genetic silencing of PRMT5 in CCSST cells resulted in severely impaired cell proliferation and EWSR1-ATF1-driven transcription. Furthermore, the clinical-stage PRMT5 inhibitor JNJ-64619178 potently and efficaciously inhibited CCSST cell growth in vitro and in vivo. These results provide new insights into PRMT5 as a transcription regulator and warrant JNJ-64619178 for further clinical development to treat CCSST patients.
]]></description>
<dc:creator>Li, B. X.</dc:creator>
<dc:creator>Xiao, X.</dc:creator>
<dc:creator>David, L. L.</dc:creator>
<dc:creator>Davis, L. E.</dc:creator>
<dc:date>2022-03-23</dc:date>
<dc:identifier>doi:10.1101/2022.03.23.485409</dc:identifier>
<dc:title><![CDATA[PRMT5 as a Novel Druggable Vulnerability for EWSR1-ATF1-driven Clear Cell Sarcoma]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-03-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.03.28.486094v1?rss=1">
<title>
<![CDATA[
Classification of Movement-Related Oscillations in sEEG Recordings with Machine Learning 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.03.28.486094v1?rss=1</link>
<description><![CDATA[
Previous electrophysiological research has characterized canonical oscillatory patterns associated with movement mostly from recordings of primary sensorimotor cortex. Less work has attempted to decode movement based on electrophysiological recordings from a broader array of brain areas such as those sampled by stereoelectroencephalography (sEEG). Here we decoded movement using a linear support vector machine (SVM). We were able to accurately classify sEEG spectrograms during a keypress movement in a task versus those during the inter-trial interval. Furthermore, the important time-frequency patterns for this classification recapitulated findings from previous studies that used non-invasive electroencephalography (EEG) and electrocorticography (ECoG) and identified brain regions that were not associated with movement in previous studies. Specifically, we found these previously described patterns: beta (13 - 30 Hz) desynchronization, beta synchronization (rebound), pre-movement alpha (8 - 15 Hz) modulation, a post-movement broadband gamma (60 - 90 Hz) increase and an event-related potential. These oscillatory patterns were newly observed in a wide range of brain areas accessible with sEEG that are not accessible with other electrophysiology recording methods. For example, the presence of beta desynchronization in the frontal lobe was more widespread than previously described, extending outside primary and secondary motor cortices. We provide evidence for a system of putative motor networks that exhibit unique oscillatory patterns by describing the anatomical extent of the movement-related oscillations that were observed most frequently across all sEEG contacts.

Significance StatementSeveral major motor networks have been previously delineated in humans, however, much less is known about the population-level oscillations that coordinate this neural circuitry, especially in cortex. Therapies that modulate brain circuits to treat movement disorders, such as deep brain stimulation (DBS), or use brain signals to control movement, such as brain-computer interfaces (BCIs), rely on our basic scientific understanding of this movement neural circuitry. In order to bridge this gap, we used stereoelectroencephalography (sEEG) collected in human patients being monitored for epilepsy to assess oscillatory patterns during movement.
]]></description>
<dc:creator>Rockhill, A. P.</dc:creator>
<dc:creator>Mantovani, A.</dc:creator>
<dc:creator>Stedelin, B.</dc:creator>
<dc:creator>Raslan, A. M.</dc:creator>
<dc:creator>Swann, N. C.</dc:creator>
<dc:date>2022-03-29</dc:date>
<dc:identifier>doi:10.1101/2022.03.28.486094</dc:identifier>
<dc:title><![CDATA[Classification of Movement-Related Oscillations in sEEG Recordings with Machine Learning]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-03-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.03.28.486115v1?rss=1">
<title>
<![CDATA[
Profiling the diversity of agonist-selective effects on the proximal proteome environment of G protein-coupled receptors 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.03.28.486115v1?rss=1</link>
<description><![CDATA[
The mu opioid receptor (OR), a prototypic member of the large G protein-coupled receptor (GPCR) family, represents an important target of therapeutic and abused drugs. To date, most of our understanding of OR activity has focused on signal transducers and regulatory molecules including G proteins, GPCR kinases, and beta-arrestins. Yet it is clear that signaling through the OR is coordinated by additional proteins recruited into the proximal interaction network of the activated receptor, which have largely remained invisible given the lack of technologies to interrogate these networks systematically.

Here, we implement a quantitative proteomics pipeline leveraging the chemical diversity of OR agonists and APEX-based proximity labeling to investigate the protein networks that underlie OR signaling. We leverage a novel computational framework to extract subcellular location, trafficking, and functional partners of GPCR activity from the proximity labeling datasets. Applying this unbiased, systematic approach to the OR, we demonstrate that opioid agonists exert differences in the OR proximal proteome mediated by endocytosis and subsequent endosomal sorting, exemplified by VPS35 and COMMD3. Moreover, we identify two novel OR network components, EYA4 and KCTD12, that are recruited into the receptor proximal network irrespective of the activating ligand and independent of receptor trafficking but based on receptor-triggered G protein activation. We provide functional evidence that these network components form a previously unrecognized buffering system for G protein activity which broadly modulates cellular GPCR signaling.
]]></description>
<dc:creator>Polacco, B. J.</dc:creator>
<dc:creator>Lobingier, B. T.</dc:creator>
<dc:creator>Blythe, E. E.</dc:creator>
<dc:creator>Abreu, N.</dc:creator>
<dc:creator>Xu, J.</dc:creator>
<dc:creator>Li, Q.</dc:creator>
<dc:creator>Naing, Z. Z. C.</dc:creator>
<dc:creator>Shoichet, B. K.</dc:creator>
<dc:creator>Levitz, J.</dc:creator>
<dc:creator>Krogan, N. J.</dc:creator>
<dc:creator>Von Zastrow, M.</dc:creator>
<dc:creator>Huttenhain, R.</dc:creator>
<dc:date>2022-03-29</dc:date>
<dc:identifier>doi:10.1101/2022.03.28.486115</dc:identifier>
<dc:title><![CDATA[Profiling the diversity of agonist-selective effects on the proximal proteome environment of G protein-coupled receptors]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-03-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.03.29.486217v1?rss=1">
<title>
<![CDATA[
A Distinct Chromatin State Drives Therapeutic Resistance in Invasive Lobular Breast Cancer 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.03.29.486217v1?rss=1</link>
<description><![CDATA[
Most invasive lobular breast cancers (ILC) are of the luminal A subtype and strongly hormone receptor positive. Yet, they are relatively resistant to tamoxifen and are associated with inferior long-term outcomes compared to invasive ductal cancers (IDC). In this study, we sought to gain mechanistic insights into these clinical findings that are not explained by the genetic landscape of ILC and to identify strategies to improve patient outcomes. Through a comprehensive analysis of the epigenome of ILC in pre-clinical models and clinical samples we found that compared to IDC, ILC has a distinct chromatin state that is linked to gained recruitment of FOXA1, a lineage-defining pioneer transcription factor. This results in an ILC-unique FOXA1-estrogen receptor (ER) axis that promotes the transcription of genes associated with tumor progression and poor outcomes. The ILC-unique FOXA1-ER axis leads to retained ER chromatin binding after tamoxifen treatment thereby facilitating tamoxifen resistance while remaining strongly dependent on ER signaling. Mechanistically, gained FOXA1 binding was associated with the auto-induction of FOXA1 in ILC through an ILC-unique FOXA1 binding site. Targeted silencing of this regulatory site resulted in the disruption of the feed-forward loop and growth inhibition in ILC.

In summary, we show that ILC is characterized by a unique cell state and FOXA1-ER axis that dictate tumor progression and offer a novel mechanism of tamoxifen resistance. These results underscore the importance of conducting clinical trials dedicated to patients with ILC to optimize endocrine treatments in this breast cancer subtype.
]]></description>
<dc:creator>Nardone, A.</dc:creator>
<dc:creator>Qui, X.</dc:creator>
<dc:creator>Feiglin, A.</dc:creator>
<dc:creator>Fu, X.</dc:creator>
<dc:creator>Spisak, S.</dc:creator>
<dc:creator>Feit, A.</dc:creator>
<dc:creator>Cohen Feit, G.</dc:creator>
<dc:creator>Xie, Y.</dc:creator>
<dc:creator>Font-Tello, A.</dc:creator>
<dc:creator>Guarducci, C.</dc:creator>
<dc:creator>Hermida Prado, F.</dc:creator>
<dc:creator>Syamala, S.</dc:creator>
<dc:creator>Lim, K.</dc:creator>
<dc:creator>Pun, M.</dc:creator>
<dc:creator>Cornwell, M.</dc:creator>
<dc:creator>Liu, W.</dc:creator>
<dc:creator>Ors, A.</dc:creator>
<dc:creator>Mohammed, H.</dc:creator>
<dc:creator>Brock, J. E.</dc:creator>
<dc:creator>Freedman, M. L.</dc:creator>
<dc:creator>Schiff, R.</dc:creator>
<dc:creator>Winer, E.</dc:creator>
<dc:creator>Long, H. W.</dc:creator>
<dc:creator>Metzger Filho, O.</dc:creator>
<dc:creator>Jeselsohn, R.</dc:creator>
<dc:date>2022-03-30</dc:date>
<dc:identifier>doi:10.1101/2022.03.29.486217</dc:identifier>
<dc:title><![CDATA[A Distinct Chromatin State Drives Therapeutic Resistance in Invasive Lobular Breast Cancer]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-03-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.03.29.486234v1?rss=1">
<title>
<![CDATA[
Learning of probabilistic punishment as a model of anxiety produces changes in action but not punishment encoding in the dmPFC and VTA 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.03.29.486234v1?rss=1</link>
<description><![CDATA[
Previously, we developed a novel model for anxiety during motivated behavior by training rats to perform a task where actions executed to obtain a reward were probabilistically punished and observed that after learning, neuronal activity in the ventral tegmental area (VTA) and dorsomedial prefrontal cortex (dmPFC) encode the relationship between action and punishment risk (Park & Moghaddam, 2017). Here we used male and female rats to expand on the previous work by focusing on neural changes in the dmPFC and VTA that were associated with the learning of probabilistic punishment, and with anxiolytic treatment with diazepam after learning. We find that adaptive neural responses of dmPFC and VTA during the learning of anxiogenic contingencies are independent from the punishment experience and occur primarily during the peri-action period. Our results further identify peri-action ramping of VTA neural activity, and VTA-dmPFC correlated activity, as potential markers for the anxiolytic properties of diazepam.
]]></description>
<dc:creator>Jacobs, D. S.</dc:creator>
<dc:creator>Allen, M. C.</dc:creator>
<dc:creator>Park, J.</dc:creator>
<dc:creator>Moghaddam, B.</dc:creator>
<dc:date>2022-03-29</dc:date>
<dc:identifier>doi:10.1101/2022.03.29.486234</dc:identifier>
<dc:title><![CDATA[Learning of probabilistic punishment as a model of anxiety produces changes in action but not punishment encoding in the dmPFC and VTA]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-03-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.03.30.486449v1?rss=1">
<title>
<![CDATA[
An open protocol for modeling T Cell Clonotype repertoires using TCRβ CDR3 sequences 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.03.30.486449v1?rss=1</link>
<description><![CDATA[
T cell receptor (TCR) repertoires can be profiled using next generation sequencing (NGS) to monitor dynamical changes in response to disease and other perturbations. Several strategies for profiling TCRs have been recently developed with different benefits and drawbacks. Genomic DNA-based bulk sequencing, however, remains the most cost-effective method to profile TCRs. The major disadvantage of this method is the need for multiplex target amplification with a large set of primer pairs with potentially very different amplification efficiencies. One approach addressing this problem is by iteratively adjusting the concentrations of the primers based on their efficiencies, and then computationally correcting any remaining bias. Yet there are no standard, publicly available protocols to process and analyze raw sequencing data generated by this method. Here, we utilize an equimolar primer mixture and propose a single statistical normalization step that efficiently corrects for amplification bias post sequencing. Using samples analyzed by both approaches, we show that the concordance between bulk clonality metrics obtained from using the commercial kits and that developed herein is high. Therefore, we suggest the method presented here as an inexpensive and non-commercial alternative for measuring and monitoring adaptive dynamics in TCR clonotype repertoire.
]]></description>
<dc:creator>Gurun, B.</dc:creator>
<dc:creator>Horton, W.</dc:creator>
<dc:creator>Murugan, D.</dc:creator>
<dc:creator>Zhu, B.</dc:creator>
<dc:creator>Leyshock, P.</dc:creator>
<dc:creator>Kumar, S.</dc:creator>
<dc:creator>Byrne, K. T.</dc:creator>
<dc:creator>Vonderheide, R. H.</dc:creator>
<dc:creator>Margolin, A. A.</dc:creator>
<dc:creator>Mori, M.</dc:creator>
<dc:creator>Spellman, P. T.</dc:creator>
<dc:creator>Coussens, L. M.</dc:creator>
<dc:creator>Speed, T. P.</dc:creator>
<dc:date>2022-04-01</dc:date>
<dc:identifier>doi:10.1101/2022.03.30.486449</dc:identifier>
<dc:title><![CDATA[An open protocol for modeling T Cell Clonotype repertoires using TCRβ CDR3 sequences]]></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/10.1101/2022.04.01.486784v1?rss=1">
<title>
<![CDATA[
Appia: a web interface for chromatography 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.04.01.486784v1?rss=1</link>
<description><![CDATA[
Chromatography is an essential family of assays for molecular biology and chemistry. Typically, only a qualitative assessment of peak height, position, and shape are sufficient to proceed. However, there are more advanced forms of chromatography that require integration of peak width, baseline subtraction, and precise peak position calling. To handle these precise workflows, instrument manufacturer software is complicated and arcane; this makes simpler day-to-day analyses inefficient. Additionally, each manufacturer reports the data in their own proprietary format, requiring that analysis is performed at instrument computers and making data sharing between labs difficult. Here we present Appia, a free, open-source chromatography processing and visualization package focused on making analysis, collaboration, and publication quick and easy.
]]></description>
<dc:creator>Posert, R.</dc:creator>
<dc:creator>Hartfield, K.</dc:creator>
<dc:creator>Baconguis, I.</dc:creator>
<dc:date>2022-04-02</dc:date>
<dc:identifier>doi:10.1101/2022.04.01.486784</dc:identifier>
<dc:title><![CDATA[Appia: a web interface for chromatography]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-04-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.04.05.487240v1?rss=1">
<title>
<![CDATA[
Vascular inflammation exposes perivascular cells to SARS-CoV-2 infection 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.04.05.487240v1?rss=1</link>
<description><![CDATA[
Pericytes stabilize blood vessels and promote vascular barrier function. However, vessels subjected to pro-inflammatory conditions have impaired barrier function, which has been suggested to potentially expose perivascular cells to SARS-CoV-2. To test this hypothesis, we engineered pericyte-supported vascular capillaries on-a-chip, and determined that the extravasation and binding of spike protein (S1) on perivascular cells of inflamed vessels to be significantly higher that in healthy controls, indicating a potential target to understand COVID-19 vascular complications.
]]></description>
<dc:creator>Franca, C. M.</dc:creator>
<dc:creator>Mansoorifar, A.</dc:creator>
<dc:creator>Athirasala, A.</dc:creator>
<dc:creator>Subbiah, R.</dc:creator>
<dc:creator>Tahayeri, A.</dc:creator>
<dc:creator>Bertassoni, L. E.</dc:creator>
<dc:date>2022-04-06</dc:date>
<dc:identifier>doi:10.1101/2022.04.05.487240</dc:identifier>
<dc:title><![CDATA[Vascular inflammation exposes perivascular cells to SARS-CoV-2 infection]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-04-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.04.06.485949v1?rss=1">
<title>
<![CDATA[
A subtype of melanopsin ganglion cells encodes ground luminance 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.04.06.485949v1?rss=1</link>
<description><![CDATA[
Visual input to the hypothalamus from intrinsically photosensitive retinal ganglion cells (ipRGCs) influences several functions including circadian entrainment, body temperature, and sleep. ipRGCs also project to nuclei such as the supraoptic nucleus (SON), which is involved in systemic fluid homeostasis, maternal behavior, and appetite. However, little is known about the SON-projecting ipRGCs or their relationship to well-characterized ipRGC subtypes. Using a GlyT2Cre mouse line, we identify a subtype of ipRGCs restricted to the dorsal retina that selectively project to the SON. These ipRGCs form a non-overlapping tiled mosaic that is limited to a dorsal region of the retina, forming a substrate for encoding ground luminance. Optogenetic activation of their axons demonstrates they release the neurotransmitter glutamate and that the SON is retinorecipient, receiving synaptic input from dorsal ipRGCs. Our results challenge the idea that ipRGC dendrites overlap to optimize photon capture and suggests non-image forming vision operates to sample local regions of the visual field.
]]></description>
<dc:creator>Berry, M. H.</dc:creator>
<dc:creator>Moldavan, M.</dc:creator>
<dc:creator>Garrett, T.</dc:creator>
<dc:creator>Meadows, M.</dc:creator>
<dc:creator>Cravetchi, O.</dc:creator>
<dc:creator>White, E.</dc:creator>
<dc:creator>von Gersdorff, H.</dc:creator>
<dc:creator>Wright, K. M.</dc:creator>
<dc:creator>Allen, C.</dc:creator>
<dc:creator>Sivyer, B. M.</dc:creator>
<dc:date>2022-04-10</dc:date>
<dc:identifier>doi:10.1101/2022.04.06.485949</dc:identifier>
<dc:title><![CDATA[A subtype of melanopsin ganglion cells encodes ground luminance]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-04-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.04.11.487493v1?rss=1">
<title>
<![CDATA[
Reduced affinity of calcium sensing-receptor heterodimers and reduced mutant homodimer trafficking combine to impair function in a mouse model of Familial Hypocalciuric Hypercalcemia Type 1 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.04.11.487493v1?rss=1</link>
<description><![CDATA[
Heterozygous loss-of-function mutation of the calcium sensing-receptor (CaSR), causes familial hypocalciuric hypercalcemia type 1 (FHH1), a typically benign condition characterized by mild hypercalcemia. In contrast, homozygous mutation of this dimer-forming G-protein coupled receptor manifests as the lethal neonatal severe hyperparathyroidism (NSHPT). To investigate the mechanisms by which CaSR mutations lead to these distinct disease states, we engineered wild-type (WT) and an exon 5-deficient disease-causing mutation, and transfected expression constructs into human embryonic kidney (HEK) cells. WT protein was mainly membrane-expressed whereas the mutant CaSR protein (mCaSR) was confined to the cytoplasm. Co-expression of WT CaSR directed mCaSR to the cell membrane. In assays of CaSR function, increases in extracellular [Ca2+] ([Ca2+]o) increased intracellular [Ca2+] ([Ca2+]i) in cells expressing WT CaSR while the response was reduced in cells coexpressingmutant and WT receptor. Untransfected cells or those expressing mCaSR alone, showed minimal, equivalent responses to increased [Ca2+]o. Immunoprecipitation experiments confirmed an association between mutant and wild-type CaSR. The affinity of the WT CaSR for calcium was three times greater than that of the heterodimer. The maximal functional response to [Ca]o was dependent on localization of CaSR to the membrane level and independent of homo- or heterodimerizations. In summary, these results suggest that heterodimerization of WT and mCaSR receptors, rescues the trafficking defect of the mutant receptors and also reduces the affinity of the WT-mutant heterodimer for [Ca]o. In contrast, the homozygous mutants do not produce functional receptors on cell membrane. These data indicate how substantial differences between signaling of hetero- and homodimeric mutants lead to profound differences in the severity of disease in heterozygous and homozygous carriers of these mutations.
]]></description>
<dc:creator>Wang, X.</dc:creator>
<dc:creator>Lundblad, J.</dc:creator>
<dc:creator>Smith, S. M.</dc:creator>
<dc:date>2022-04-11</dc:date>
<dc:identifier>doi:10.1101/2022.04.11.487493</dc:identifier>
<dc:title><![CDATA[Reduced affinity of calcium sensing-receptor heterodimers and reduced mutant homodimer trafficking combine to impair function in a mouse model of Familial Hypocalciuric Hypercalcemia Type 1]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-04-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.04.13.488220v1?rss=1">
<title>
<![CDATA[
Effect of Positive Allosteric Modulation and Orthosteric Agonism of Dopamine D2 Receptors on Respiration in Mouse Models of Rett Syndrome 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.04.13.488220v1?rss=1</link>
<description><![CDATA[
Rett syndrome (RTT) is an autism spectrum disorder caused by loss-of-function mutations in the methyl-CPG-binding protein 2 (Mecp2) gene. Frequent apneas and irregular breathing are prevalent in RTT, and also occur in rodent models of the disorder, including Mecp2Bird and Mecp2R168X mice. Sarizotan, a serotonin 5-HT1a and dopamine D2-like receptor agonist, reduces the incidence of apneas and irregular breathing in mouse models of RTT (Abdala et al., 2014). Targeting the 5HT1a receptor alone also improves respiration in RTT mice (Levitt et al., 2013). However, the contribution of D2 receptors in correcting these respiratory disturbances remains untested. PAOPA, a dopamine D2 receptor positive allosteric modulator, and quinpirole, a dopamine D2 receptor orthosteric agonist, were used in conjunction with whole-body plethysmography to evaluate whether activation of D2 receptors is sufficient to improve breathing disturbances in female heterozygous Mecp2Bird/+ and Mecp2R168X/+ mice. PAOPA did not significantly change apnea incidence or irregularity score in RTT mice. PAOPA also had no effect on the ventilatory response to hypercapnia (7% CO2). In contrast, quinpirole reduced apnea incidence and irregularity scores and improved the hypercapnic ventilatory response in Mecp2R168X/+ and Mecp2Bird/+ mice, while also reducing respiratory rate. These results suggest that D2 receptors do contribute to the positive effects of sarizotan in the correction of respiratory abnormalities in Rett syndrome. However, positive allosteric modulation of the D2 receptor alone is not sufficient to evoke these effects.
]]></description>
<dc:creator>Maletz, S. N.</dc:creator>
<dc:creator>Reid, B. T.</dc:creator>
<dc:creator>Baekey, D. M.</dc:creator>
<dc:creator>Whitaker-Fornek, J. R.</dc:creator>
<dc:creator>Bateman, J. T.</dc:creator>
<dc:creator>Bissonnette, J. M.</dc:creator>
<dc:creator>Levitt, E. S.</dc:creator>
<dc:date>2022-04-14</dc:date>
<dc:identifier>doi:10.1101/2022.04.13.488220</dc:identifier>
<dc:title><![CDATA[Effect of Positive Allosteric Modulation and Orthosteric Agonism of Dopamine D2 Receptors on Respiration in Mouse Models of Rett Syndrome]]></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/10.1101/2022.04.16.488549v1?rss=1">
<title>
<![CDATA[
SATINN: An automated neural network-based classification of testicular sections allows for high-throughput histopathology of mouse mutants 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.04.16.488549v1?rss=1</link>
<description><![CDATA[
MotivationThe mammalian testis is a complex organ with a hierarchical organization that changes smoothly and stereotypically over time in normal adults. While testis histology is already an invaluable tool for identifying and describing developmental differences in evolution and disease, methods for standardized, digital image analysis of testis are needed to expand the utility of this approach.

ResultsWe developed SATINN (Software for Analysis of Testis Images with Neural Networks), a multi-level framework for automated analysis of multiplexed immunofluorescence images from mouse testis. This approach uses a convolutional neural network (CNN) to classify nuclei from seminiferous tubules into 7 distinct cell types with an accuracy of 94.2%. These cell classifications are then used in a second-level tubule CNN, which places seminiferous tubules into one of 7 distinct tubule stages with 90.4% accuracy. We further describe numerous cell- and tubule-level statistics that can be derived from wildtype testis. Finally, we demonstrate how the classifiers and derived statistics can be used to rapidly and precisely describe pathology by applying our methods to image data from two mutant mouse lines. Our results demonstrate the feasibility and potential of using computer-assisted analysis for testis histology, an area poised to evolve rapidly on the back of emerging, spatially-resolved genomic and proteomic technologies.

Availability and implementationScripts to apply the methods described here are available from http://github.com/conradlab/SATINN.
]]></description>
<dc:creator>Yang, R.</dc:creator>
<dc:creator>Stendahl, A.</dc:creator>
<dc:creator>Vigh-Conrad, K.</dc:creator>
<dc:creator>Held, M.</dc:creator>
<dc:creator>Lima, A.</dc:creator>
<dc:creator>Conrad, D.</dc:creator>
<dc:date>2022-04-16</dc:date>
<dc:identifier>doi:10.1101/2022.04.16.488549</dc:identifier>
<dc:title><![CDATA[SATINN: An automated neural network-based classification of testicular sections allows for high-throughput histopathology of mouse mutants]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-04-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.04.21.489073v1?rss=1">
<title>
<![CDATA[
Dissection of a sensorimotor circuit that regulates aversion to odors and pathogenic bacteria in C. elegans by whole-brain simulation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.04.21.489073v1?rss=1</link>
<description><![CDATA[
Altering behavior to reduce pathogen exposure is a key line of defense against pathogen attack for nearly all animals. The use of Caenorhabditis elegans bacterial infection models have allowed for many insights into the molecular mechanisms of behavioral immunity. However, the neural circuitry between chemosensory neurons that sense pathogenic bacterial cues and the motor neurons responsible for avoidance-associated locomotion remains unknown. We found that backward locomotion was a component of learned pathogen avoidance, as animals pre-exposed to Pseudomonas aeruginosa or Enterococcus faecalis showed reflexive aversion to drops of the bacteria, requiring ASI, AWB, and AWC neurons and ASE, AWB, and AWC neurons, respectively. This response also involved intestinal distention and, for E. faecalis, required expression of TRPM channels in the intestine and excretory system. Using whole-brain simulation and functional assays, we uncovered a sensorimotor circuit governing learned reflexive aversion. This behavior is controlled by a four-layer neural circuit composed of olfactory neurons, interneurons, and motor neurons that control backward locomotion crucial for learned reflexive aversion to pathogenic bacteria, learned avoidance, and a repulsive odor. The discovery of a complete sensorimotor circuit for reflexive aversion demonstrates the utility of using the C. elegans connectome and computational modeling in uncovering new neuronal regulators of behavior.
]]></description>
<dc:creator>Filipowicz, A.</dc:creator>
<dc:creator>Lalsiamthara, J.</dc:creator>
<dc:creator>Aballay, A.</dc:creator>
<dc:date>2022-04-22</dc:date>
<dc:identifier>doi:10.1101/2022.04.21.489073</dc:identifier>
<dc:title><![CDATA[Dissection of a sensorimotor circuit that regulates aversion to odors and pathogenic bacteria in C. elegans by whole-brain simulation]]></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/10.1101/2022.04.25.489405v1?rss=1">
<title>
<![CDATA[
Epigenetic Control of Hundreds of Chromosome-Associated lncRNA Genes Essential for Replication and Stability 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.04.25.489405v1?rss=1</link>
<description><![CDATA[
ASARs are long noncoding RNA genes that control replication timing of entire human chromosomes in cis. The three known ASAR genes are located on human chromosomes 6 and 15, and are essential for chromosome integrity. To identify ASARs on all human chromosomes we utilized a set of distinctive ASAR characteristics that allowed for the identification of hundreds of autosomal loci with epigenetically controlled, allele-restricted behavior in expression and replication timing of coding and noncoding genes, and is distinct from genomic imprinting. Disruption of noncoding RNA genes at five of five tested loci resulted in chromosome-wide delayed replication and chromosomal instability, validating their ASAR activity. In addition to the three known essential cis-acting chromosomal loci, origins, centromeres, and telomeres, we propose that all mammalian chromosomes also contain "Inactivation/Stability Centers" that display allele-restricted epigenetic regulation of protein coding and noncoding ASAR genes that are essential for replication and stability of each chromosome.
]]></description>
<dc:creator>Heskett, M. B.</dc:creator>
<dc:creator>Vouzas, A. E.</dc:creator>
<dc:creator>Smith, L. G.</dc:creator>
<dc:creator>Yates, P. A.</dc:creator>
<dc:creator>Boniface, C.</dc:creator>
<dc:creator>Bouhassira, E. E.</dc:creator>
<dc:creator>Spellman, P. T.</dc:creator>
<dc:creator>Gilbert, D. M.</dc:creator>
<dc:creator>Thayer, M. J.</dc:creator>
<dc:date>2022-04-25</dc:date>
<dc:identifier>doi:10.1101/2022.04.25.489405</dc:identifier>
<dc:title><![CDATA[Epigenetic Control of Hundreds of Chromosome-Associated lncRNA Genes Essential for Replication and Stability]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-04-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.04.26.489507v1?rss=1">
<title>
<![CDATA[
Chemotherapy Signatures Map Evolution of Therapy-Related Myeloid Neoplasms 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.04.26.489507v1?rss=1</link>
<description><![CDATA[
Patients treated with cytotoxic therapies, including autologous stem cell transplantation, are at risk for developing therapy-related myeloid neoplasms1, 2. Pre-leukemic clones (i.e., clonal hematopoiesis) are detectable years before the development of these aggressive malignancies3-5, though the genomic events leading to transformation and expansion are not well-defined. Here, leveraging distinctive chemotherapy-associated mutational signatures6-12 from whole-genome sequencing data and targeted sequencing of pre-chemotherapy samples, we reconstruct the evolutionary life-history of 39 therapy-related myeloid malignancies. A dichotomy is revealed, in which neoplasms with evidence of chemotherapy-induced mutagenesis from platinum and melphalan are relatively hypermutated and enriched for complex structural variants (i.e., chromothripsis), while neoplasms with alternative exposures bear a similar profile to de novo acute myeloid leukemia. Using chemotherapy-associated mutational signatures as a temporal barcode in each patients life, we estimate that several complex events and genomic drivers are acquired after chemotherapy exposure. In the case of treatment with high-dose melphalan and autologous stem cell transplantation, we demonstrate that the procedure allows clonal hematopoiesis to escape chemotherapy exposure entirely, and to be reinfused to expand to malignancy. This information reveals a novel mode of malignant progression for therapy-related malignancies that is not reliant on direct mutagenesis or even exposure to chemotherapy, itself, and prompts further investigation into leukemia-permissive effects of cytotoxic drugs.
]]></description>
<dc:creator>Diamond, B. T.</dc:creator>
<dc:creator>Ziccheddu, B.</dc:creator>
<dc:creator>Maclachlan, K. H.</dc:creator>
<dc:creator>Taylor, J.</dc:creator>
<dc:creator>Boyle, E.</dc:creator>
<dc:creator>Arrango Ossa, J.</dc:creator>
<dc:creator>Jahn, J. T.</dc:creator>
<dc:creator>Affer, M.</dc:creator>
<dc:creator>Totiger, T. M.</dc:creator>
<dc:creator>Coffey, D. G.</dc:creator>
<dc:creator>Watts, J.</dc:creator>
<dc:creator>Lu, S. X.</dc:creator>
<dc:creator>Bolli, N.</dc:creator>
<dc:creator>Bolton, K.</dc:creator>
<dc:creator>Park, J. H.</dc:creator>
<dc:creator>Landau, H.</dc:creator>
<dc:creator>Ganesh, K.</dc:creator>
<dc:creator>McPherson, A.</dc:creator>
<dc:creator>Sekeres, M.</dc:creator>
<dc:creator>Lesokhin, A.</dc:creator>
<dc:creator>Chung, D.</dc:creator>
<dc:creator>Zhang, Y.</dc:creator>
<dc:creator>Ho, C.</dc:creator>
<dc:creator>Roshal, M.</dc:creator>
<dc:creator>Tyner, J.</dc:creator>
<dc:creator>Nimer, S.</dc:creator>
<dc:creator>Papaemmanuil, E.</dc:creator>
<dc:creator>Usmani, S.</dc:creator>
<dc:creator>Morgan, G.</dc:creator>
<dc:creator>Landgren, O.</dc:creator>
<dc:creator>Maura, F.</dc:creator>
<dc:date>2022-04-27</dc:date>
<dc:identifier>doi:10.1101/2022.04.26.489507</dc:identifier>
<dc:title><![CDATA[Chemotherapy Signatures Map Evolution of Therapy-Related Myeloid Neoplasms]]></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/10.1101/2022.04.26.489579v1?rss=1">
<title>
<![CDATA[
Cancer stem cell-derived extracellular vesicles preferentially target MHCII- macrophages and PD1+ T cells in the tumor microenvironment 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.04.26.489579v1?rss=1</link>
<description><![CDATA[
Immunotherapy is an approved treatment option for head and neck squamous cell carcinoma (HNSCC). However, the response rate to immune checkpoint blockade is only 13% for recurrent HNSCC, highlighting the urgent need to better understand tumor-immune interplay, with the ultimate goal of improving patient outcomes. HNSCC present high local recurrence rates and therapy resistance that can be attributed to the presence of cancer stem cells (CSC) within tumors. CSC exhibit singular properties that enable them to avoid immune detection and eradication. The immune cell types that directly engage with CSC to allow immune escape and cancer recurrence are still unknown. Here, we genetically engineered CSC-derived extracellular vesicles (EVs) to perform sortase-mediated in vivo proximity labeling. We identified specific immune cell subsets recruited into the CSC niche. We demonstrated that unmanipulated CSC-EVs preferentially target MHC-II- macrophages and PD1+ T cells, and that such EV-mediated intercellular communication between CSC and these immune cells contributed to the observed spatial interactions and niche sharing. These results suggest that combination therapies targeting CSC, tumor macrophages and PD1 may synergize and lower local recurrence rates in HNSCC patients.
]]></description>
<dc:creator>Gonzalez-Callejo, P.</dc:creator>
<dc:creator>Guo, Z.</dc:creator>
<dc:creator>Ziglari, T.</dc:creator>
<dc:creator>Claudio, N. M.</dc:creator>
<dc:creator>Oshimori, N.</dc:creator>
<dc:creator>Seras-Franzoso, J.</dc:creator>
<dc:creator>Pucci, F.</dc:creator>
<dc:date>2022-04-26</dc:date>
<dc:identifier>doi:10.1101/2022.04.26.489579</dc:identifier>
<dc:title><![CDATA[Cancer stem cell-derived extracellular vesicles preferentially target MHCII- macrophages and PD1+ T cells in the tumor microenvironment]]></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/10.1101/2022.04.26.489617v1?rss=1">
<title>
<![CDATA[
Systematic Replication Enables Normalization of High-throughput Imaging Assays 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.04.26.489617v1?rss=1</link>
<description><![CDATA[
MotivationHigh-throughput fluorescent microscopy is a popular class of techniques for studying tissues and cells through automated imaging and feature extraction of hundreds to thousands of samples. Like other high-throughput assays, these approaches can suffer from unwanted noise and technical artifacts that obscure the biological signal. In this work we consider how an experimental design incorporating multiple levels of replication enables removal of technical artifacts from such image-based platforms.

ResultsWe develop a general approach to remove technical artifacts from high-throughput image data that leverages an experimental design with multiple levels of replication. To illustrate the methods we consider microenvironment microarrays (MEMAs), a high-throughput platform designed to study cellular responses to microenvironmental perturbations. In application on MEMAs, our approach removes unwanted spatial artifacts and thereby enhances the biological signal. This approach has broad applicability to diverse biological assays.

AvailabilityRaw data is on synapse (syn2862345), analysis code is on github (gjhunt/mema norm), a Docker image is available on dockerhub (gjhunt/memanorm). online.
]]></description>
<dc:creator>Hunt, G. J.</dc:creator>
<dc:creator>Dane, M. A.</dc:creator>
<dc:creator>Korkola, J. E.</dc:creator>
<dc:creator>Heiser, L. M.</dc:creator>
<dc:creator>Gagnon-Bartsch, J. A.</dc:creator>
<dc:date>2022-04-28</dc:date>
<dc:identifier>doi:10.1101/2022.04.26.489617</dc:identifier>
<dc:title><![CDATA[Systematic Replication Enables Normalization of High-throughput Imaging Assays]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-04-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.04.28.489781v1?rss=1">
<title>
<![CDATA[
Human Lung-resident Mucosal-Associated Invariant T cells are Abundant, Express Antimicrobial Proteins, and are Cytokine Responsive 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.04.28.489781v1?rss=1</link>
<description><![CDATA[
Mucosal-associated Invariant T (MAIT) cells are an innate-like T cell subset that recognize a broad array of microbial pathogens, including respiratory pathogens. Here we investigate the transcriptional profile of MAIT cells localized to the human lung, and postulate that MAIT cells may play a role in maintaining homeostasis at this mucosal barrier. Using the MR1/5-OP-RU tetramer, we identified MAIT cells and non-MAIT CD8+ T cells in lung tissue not suitable for transplant from human donors. We used RNA-sequencing of MAIT cells compared to non-MAIT CD8+ T cells to define the transcriptome of MAIT cells in the human lung. We show that, as a population, lung MAIT cells are polycytotoxic, secrete the directly antimicrobial molecule IL-26, express genes associated with persistence, and selectively express cytokine and chemokine-related molecules distinct from other lung-resident CD8+ T cells, such as interferon-{gamma}- and IL-12-receptors. These data highlight MAIT cells predisposition to rapid pro-inflammatory cytokine responsiveness and antimicrobial mechanisms in human lung tissue, concordant with findings of blood-derived counterparts, and support a function for MAIT cells as early sensors in the defense of respiratory barrier function.
]]></description>
<dc:creator>Meermeier, E.</dc:creator>
<dc:creator>Zheng, C.</dc:creator>
<dc:creator>Soma, S.</dc:creator>
<dc:creator>Worley, A.</dc:creator>
<dc:creator>Weiss, D.</dc:creator>
<dc:creator>Modlin, R.</dc:creator>
<dc:creator>Swarbrick, G.</dc:creator>
<dc:creator>Karamooz, E.</dc:creator>
<dc:creator>Khuzwayo, S.</dc:creator>
<dc:creator>Wong, E.</dc:creator>
<dc:creator>Gold, M.</dc:creator>
<dc:creator>Lewinsohn, D.</dc:creator>
<dc:date>2022-05-02</dc:date>
<dc:identifier>doi:10.1101/2022.04.28.489781</dc:identifier>
<dc:title><![CDATA[Human Lung-resident Mucosal-Associated Invariant T cells are Abundant, Express Antimicrobial Proteins, and are Cytokine Responsive]]></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/10.1101/2022.04.29.490090v1?rss=1">
<title>
<![CDATA[
Increased glucose availability sensitizes pancreatic cancer to chemotherapy 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.04.29.490090v1?rss=1</link>
<description><![CDATA[
Pancreatic cancer (Pancreatic Ductal Adenocarcinoma; PDAC) is highly resistant to chemotherapy. Effective alternative therapies have yet to emerge, leaving chemotherapy as the best available systematic treatment. The discovery of safe and available adjuncts that improve chemotherapeutic efficacy would potentially improve survival outcomes. We show that a hyperglycemic state enhances the efficacy of conventional single- and multi-agent chemotherapies against PDAC. Molecular analyses of tumors exposed to relatively high glucose levels revealed that a key metabolic pathway, glutathione biosynthesis, is diminished and underlies chemo-sensitization by enhancing oxidative injury to cancer cells. Inhibition of this pathway under normal conditions phenocopied a hyperglycemic state by enhancing chemotherapeutic efficacy in mouse PDAC, while rescuing the pathway under high glucose abrogated the anti-tumor effects observed with chemotherapy.
]]></description>
<dc:creator>Vaziri-Gohar, A.</dc:creator>
<dc:creator>Hue, J. J.</dc:creator>
<dc:creator>Graor, H. J.</dc:creator>
<dc:creator>Abbas, A.</dc:creator>
<dc:creator>Zarei, M.</dc:creator>
<dc:creator>Hajihassani, O.</dc:creator>
<dc:creator>Titomihelakis, G.</dc:creator>
<dc:creator>Feczko, J.</dc:creator>
<dc:creator>Rathore, M.</dc:creator>
<dc:creator>Wang, R.</dc:creator>
<dc:creator>Zarei, M.</dc:creator>
<dc:creator>Goudarzi, M.</dc:creator>
<dc:creator>Zhang, R.</dc:creator>
<dc:creator>Willard, B.</dc:creator>
<dc:creator>Zhang, L.</dc:creator>
<dc:creator>Wang, G.-M.</dc:creator>
<dc:creator>Tatsuoka, C.</dc:creator>
<dc:creator>Salvino, J. M.</dc:creator>
<dc:creator>Bederman, I.</dc:creator>
<dc:creator>Brunengraber, H.</dc:creator>
<dc:creator>Lyssiotis, C. A.</dc:creator>
<dc:creator>Brody, J. R.</dc:creator>
<dc:creator>Winter, J. M.</dc:creator>
<dc:date>2022-05-01</dc:date>
<dc:identifier>doi:10.1101/2022.04.29.490090</dc:identifier>
<dc:title><![CDATA[Increased glucose availability sensitizes pancreatic cancer to chemotherapy]]></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/10.1101/2022.04.30.490175v1?rss=1">
<title>
<![CDATA[
An in situ quantitative map of initial human colorectal HIV transmission 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.04.30.490175v1?rss=1</link>
<description><![CDATA[
The initial immune response to HIV is critical in determining transmission. However, due to technical limitations we still do not have a comparative map of early mucosal transmission events. We combined RNAscope, cyclic-immunofluorescence and novel image analysis tools to quantify HIV transmission dynamics in intact human colorectal tissue. We mapped HIV enrichment to mucosal dendritic cells (DC) and submucosal macrophages, but not CD4+ T-cells, the primary targets of downstream infection. DCs appeared to funnel virus to lymphoid aggregates which acted as early sanctuaries of high viral titres whilst facilitating HIV passage to the submucosa. Finally, HIV entry induced rapid recruitment and clustering of target cells, facilitating DC and macrophage mediated HIV transfer and enhanced infection of CD4+ T-cells. These data demonstrate a rapid response to HIV structured to maximise the likelihood of mucosal infection, and provide a framework for in situ studies of host pathogen interactions and immune mediated pathologies.

Highlights- in situ quantification of host cellular microenvironment response to pathogen invasion in human colorectal tissue.
- HIV first localises to mucosal DCs and submucosal macrophages, but not CD4+ T cells.
- Viral enrichment first occurs in lymphoid aggregates which is associated with passage into the submucosa.
- Early localisation of HIV to CD4+ T cells is associated with interactions with DCs and macrophages.


Graphical Abstract

O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=200 SRC="FIGDIR/small/490175v1_ufig1.gif" ALT="Figure 1">
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]]></description>
<dc:creator>Baharlou, H.</dc:creator>
<dc:creator>Canete, N. P.</dc:creator>
<dc:creator>Vine, E. E.</dc:creator>
<dc:creator>hu, k.</dc:creator>
<dc:creator>Yuan, D.</dc:creator>
<dc:creator>Sandgren, K. J.</dc:creator>
<dc:creator>Bertram, K. M.</dc:creator>
<dc:creator>Nasr, `Najla</dc:creator>
<dc:creator>Rhodes, J. W.</dc:creator>
<dc:creator>Gosselink, M.</dc:creator>
<dc:creator>Di Re, A.</dc:creator>
<dc:creator>Reza, F.</dc:creator>
<dc:creator>Ctercteko, G.</dc:creator>
<dc:creator>Pathma-Nathan, N. `</dc:creator>
<dc:creator>Collins, G.</dc:creator>
<dc:creator>Toh, J.</dc:creator>
<dc:creator>Patrick, E.</dc:creator>
<dc:creator>Haniffa, M. A.</dc:creator>
<dc:creator>Estes, J. D.</dc:creator>
<dc:creator>Byrne, S. N.</dc:creator>
<dc:creator>Cunningham, T. L.</dc:creator>
<dc:creator>Harman, A. N.</dc:creator>
<dc:date>2022-05-01</dc:date>
<dc:identifier>doi:10.1101/2022.04.30.490175</dc:identifier>
<dc:title><![CDATA[An in situ quantitative map of initial human colorectal HIV transmission]]></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/10.1101/2022.05.03.490487v1?rss=1">
<title>
<![CDATA[
Aminomethanesulfonic acid illuminates the boundary between full and partial agonists of the pentameric glycine receptor 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.05.03.490487v1?rss=1</link>
<description><![CDATA[
To clarify the determinants of agonist efficacy in pentameric ligand-gated ion channels we examined a new compound, aminomethanesulfonic acid (AMS), a molecule intermediate in structure between glycine and taurine. Despite wide availability, to date there are no reports of AMS action on glycine receptors, perhaps because AMS is unstable at physiological pH. Here we show that at pH 5, AMS is an efficacious agonist, eliciting in zebrafish 1 glycine receptors a maximum single channel open probability of 0.85, much greater than that of {beta}-alanine (0.54) or taurine (0.12), and second only to that of glycine itself (0.96). Thermodynamic cycle analysis of the efficacy of these closely related agonists shows supra-additive interaction between changes in the length of the agonist molecule and the size of the anionic moiety. Single particle cryo-EM structures of AMS-bound glycine receptors show that the AMS-bound agonist pocket is as compact as with glycine, and three-dimensional classification demonstrates that the channel populates the open and the desensitized states, like glycine, but not the closed intermediate state associated with the weaker partial agonists, {beta}-alanine and taurine. Because AMS is on the cusp between full and partial agonists, it provides a new tool to help us understand agonist action in the pentameric superfamily of ligand-gated ion channels.
]]></description>
<dc:creator>Sivilotti, L.</dc:creator>
<dc:creator>Ivica, J.</dc:creator>
<dc:creator>Zhu, H.</dc:creator>
<dc:creator>Lape, R.</dc:creator>
<dc:creator>Gouaux, E.</dc:creator>
<dc:date>2022-05-04</dc:date>
<dc:identifier>doi:10.1101/2022.05.03.490487</dc:identifier>
<dc:title><![CDATA[Aminomethanesulfonic acid illuminates the boundary between full and partial agonists of the pentameric glycine receptor]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-05-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.05.04.490552v1?rss=1">
<title>
<![CDATA[
Ongoing Replication Stress Response and New Clonal T Cell Development Discriminate Between Liver and Lung Recurrence Sites and Patient Outcomes in Pancreatic Ductal Adenocarcinoma 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.05.04.490552v1?rss=1</link>
<description><![CDATA[
Background and AimsMetastatic pancreatic adenocarcinoma (mPDAC) is lethal, yet a subset of patients who have metastatic disease that spreads only to the lung have better outcomes. We identified unique transcriptomic and immune features that distinguish patients who develop metastases in the liver (liver cohort) versus those with lung-avid but liver-averse mPDAC (lung cohort).

MethodsWe used clinical data from the Oregon Pancreas Tissue Registry to identify PDAC patients with liver and/or lung metastases. Gene expression and genomic alteration data from 290 PDAC tumors were used to identify features unique to patients from the liver and lung cohorts. In parallel, T cell receptor sequencing data from 289 patients were used to identify immune features unique to patients in the lung cohort.

ResultsLung cohort patients had better survival outcomes than liver cohort patients. Primary tumors from patients in the liver cohort expressed a novel gene signature associated with ongoing replication stress (RS) response predictive of poor patient outcome independent from known subtypes. In contrast, patients with tumors lacking the RS response signature survived longer, especially if their tumors had alterations in DNA damage repair genes. A subset of patients in the lung cohort demonstrated new T cell clonal development in their primary and metastatic tumors leading to diverse peripheral blood TCR repertoires.

ConclusionLiver-avid metastatic PDAC is associated with an ongoing RS response, whereas tumors lacking the RS response with ongoing T cell clonal responses may have unique vulnerabilities allowing long-term survival in patients with lung-avid, liver-averse metastatic PDAC.
]]></description>
<dc:creator>Link, J. M.</dc:creator>
<dc:creator>Pelz, C.</dc:creator>
<dc:creator>Worth, P. J.</dc:creator>
<dc:creator>Owen, S.</dc:creator>
<dc:creator>Keith, D.</dc:creator>
<dc:creator>Langer, E. M.</dc:creator>
<dc:creator>Grossblatt-Wait, A.</dc:creator>
<dc:creator>Creason, A. L.</dc:creator>
<dc:creator>Egger, J.</dc:creator>
<dc:creator>Holly, H.</dc:creator>
<dc:creator>English, I.</dc:creator>
<dc:creator>MacPherson, K.</dc:creator>
<dc:creator>Tsuda, M.</dc:creator>
<dc:creator>Goecks, J.</dc:creator>
<dc:creator>Demir, E.</dc:creator>
<dc:creator>Kardosh, A.</dc:creator>
<dc:creator>Lopez, C. D.</dc:creator>
<dc:creator>Sheppard, B. C.</dc:creator>
<dc:creator>Guimaraes, A.</dc:creator>
<dc:creator>Brinkerhoff, B.</dc:creator>
<dc:creator>Morgan, T. K. C.</dc:creator>
<dc:creator>Mills, G. B.</dc:creator>
<dc:creator>Brody, J.</dc:creator>
<dc:creator>Sears, R. C.</dc:creator>
<dc:date>2022-05-06</dc:date>
<dc:identifier>doi:10.1101/2022.05.04.490552</dc:identifier>
<dc:title><![CDATA[Ongoing Replication Stress Response and New Clonal T Cell Development Discriminate Between Liver and Lung Recurrence Sites and Patient Outcomes in Pancreatic Ductal Adenocarcinoma]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-05-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.05.06.490478v1?rss=1">
<title>
<![CDATA[
Structure of C. elegans TMC-1 complex illuminates auditory mechanosensory transduction 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.05.06.490478v1?rss=1</link>
<description><![CDATA[
The initial step in the sensory transduction pathway underpinning hearing and balance in mammals involves the conversion of force into the gating of a mechanosensory transduction (MT) channel. Despite the profound socioeconomic impacts of hearing disorders and the fundamental biological significance of understanding MT, the composition, structure and mechanism of the MT complex has remained elusive. Here we report the single particle cryo-EM structure of the native MT TMC-1 complex isolated from C. elegans. The 2-fold symmetric complex is composed of 2 copies each of the pore-forming TMC-1 subunit, the calcium-binding protein CALM-1 and the transmembrane inner ear protein TMIE. CALM-1 makes extensive contacts with the cytoplasmic face of the TMC-1 subunits while the single-pass TMIE subunits reside on the periphery of the complex, poised like the handles of an accordion. A subset of particles in addition harbors a single arrestin-like protein, ARRD-6, bound to a CALM-1 domain. Single- particle reconstructions and molecular dynamics simulations show how the MT complex deforms the membrane bilayer and suggest crucial roles for lipid-protein interactions in the mechanism by which mechanical force is transduced to ion channel gating.
]]></description>
<dc:creator>Jeong, H.</dc:creator>
<dc:creator>Clark, S.</dc:creator>
<dc:creator>Goehring, A.</dc:creator>
<dc:creator>Dehghani-Ghahnaviyeh, S.</dc:creator>
<dc:creator>Rasouli, A.</dc:creator>
<dc:creator>Tajkhorshid, E.</dc:creator>
<dc:creator>Gouaux, E.</dc:creator>
<dc:date>2022-05-06</dc:date>
<dc:identifier>doi:10.1101/2022.05.06.490478</dc:identifier>
<dc:title><![CDATA[Structure of C. elegans TMC-1 complex illuminates auditory mechanosensory transduction]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-05-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.05.09.490804v1?rss=1">
<title>
<![CDATA[
Evolution avoids a pathological stabilizing interaction in the immune protein S100A9 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.05.09.490804v1?rss=1</link>
<description><![CDATA[
Stability constrains evolution. While much is known about constraints on destabilizing mutations, less is known about the constraints on stabilizing mutations. We recently identified a mutation in the innate immune protein S100A9 that provides insight into such constraints. When introduced into human S100A9, M63F simultaneously increases the stability of the protein and disrupts its natural ability to activate Toll-like receptor 4. Using chemical denaturation, we found that M63F stabilizes a calcium-bound conformation of hS100A9. We then used NMR to solve the structure of the mutant protein, revealing that the mutation distorts the hydrophobic binding surface of hS100A9, explaining its deleterious effect on function. Hydrogen deuterium exchange (HDX) experiments revealed stabilization of the region around M63F in the structure, notably Phe37. In the structure of the M63F mutant, the Phe37 and Phe63 sidechains are in contact, plausibly forming an edge-face {nu}-stack. Mutating Phe37 to Leu abolished the stabilizing effect of M63F as probed by both chemical denaturation and HDX. It also restored the biological activity of S100A9 disrupted by M63F. These findings reveal that Phe63 creates a "molecular staple" with Phe37 that stabilizes a non-functional conformation of the protein, thus disrupting function. Using a bioinformatic analysis, we found that S100A9 proteins from different organisms rarely have Phe at both positions 37 and 63, suggesting that avoiding a pathological stabilizing interaction indeed constrains S100A9 evolution. This work highlights an important evolutionary constraint on stabilizing mutations: they must avoid inappropriately stabilizing non-functional protein conformations.

SIGNIFICANCE STATEMENTProtein stability constrains protein evolution. While much is known about evolutionary constraints on destabilizing mutations, less is known about the constraints on stabilizing mutations. We recently found a mutation to an innate immune protein that increases its stability but disrupts its function. Here we show, through careful biophysical and functional studies, that this mutation stabilizes a nonfunctional form of the protein through a direct interaction with another amino acid. We find that specific amino acids can be tolerated at each of the interacting positions individually, but not at both simultaneously. This pattern has been conserved over millions of years of evolution. Our work highlights an underappreciated evolutionary constraint on stabilizing mutations: they must avoid inappropriately stabilizing non-functional protein conformations.
]]></description>
<dc:creator>Harman, J. L.</dc:creator>
<dc:creator>Reardon, P. N.</dc:creator>
<dc:creator>Costello, S. M.</dc:creator>
<dc:creator>Warren, G. D.</dc:creator>
<dc:creator>Phillips, S. R.</dc:creator>
<dc:creator>Connor, P. J.</dc:creator>
<dc:creator>Marqusee, S.</dc:creator>
<dc:creator>Harms, M. J.</dc:creator>
<dc:date>2022-05-09</dc:date>
<dc:identifier>doi:10.1101/2022.05.09.490804</dc:identifier>
<dc:title><![CDATA[Evolution avoids a pathological stabilizing interaction in the immune protein S100A9]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-05-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.05.11.491531v1?rss=1">
<title>
<![CDATA[
Deaza-modification of MR1 ligands modulates recognition by MR1-restricted T cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.05.11.491531v1?rss=1</link>
<description><![CDATA[
MR1-restricted T (MR1T) cells recognize microbial small molecule metabolites presented on the MHC Class I-like molecule MR1 and have been implicated in early effector responses to microbial infection. As a result, there is considerable interest in identifying chemical properties of metabolite ligands that permit recognition by MR1T cells, for consideration in therapeutic or vaccine applications. Here, we made chemical modifications to known MR1 ligands to evaluate the effect on MR1T cell activation. Specifically, we modified 6,7-dimethyl-8-D-ribityllumazine (DMRL) to generate 6,7-dimethyl-8-D-ribityldeazalumazine (DZ), and then further derivatized DZ to determine the requirements for retaining MR1 surface stabilization and agonistic properties. Interestingly, the IFN-{gamma} response toward DZ varied widely across a panel of T cell receptor (TCR)-diverse MR1T cell clones; while one clone was agnostic toward the modification, most displayed either an enhancement or depletion of IFN-{gamma} production when compared with its response to DMRL. To gain insight into a putative mechanism behind this phenomenon, we used in silico molecular docking techniques for DMRL and its derivatives and performed molecular dynamics simulations of the complexes. In assessing the dynamics of each ligand in the MR1 pocket, we found that DMRL and DZ exhibit differential dynamics of both the ribityl moiety and the aromatic backbone, which may contribute to ligand recognition. Together, our results support an emerging hypothesis for flexibility in MR1:ligand-MR1T TCR interactions and enable further exploration of the relationship between MR1:ligand structures and MR1T cell recognition for downstream applications targeting MR1T cells.
]]></description>
<dc:creator>Jin, H.</dc:creator>
<dc:creator>Ladd, N. A.</dc:creator>
<dc:creator>Peev, A. M.</dc:creator>
<dc:creator>Swarbrick, G. M.</dc:creator>
<dc:creator>Cansler, M.</dc:creator>
<dc:creator>Null, M.</dc:creator>
<dc:creator>Boughter, C. T.</dc:creator>
<dc:creator>McMurtrey, C.</dc:creator>
<dc:creator>Nilsen, A.</dc:creator>
<dc:creator>Dobos, K. M.</dc:creator>
<dc:creator>Hildebrand, W. H.</dc:creator>
<dc:creator>Lewinsohn, D. A.</dc:creator>
<dc:creator>Adams, E. J.</dc:creator>
<dc:creator>Lewinsohn, D. M.</dc:creator>
<dc:creator>Harriff, M.</dc:creator>
<dc:date>2022-05-11</dc:date>
<dc:identifier>doi:10.1101/2022.05.11.491531</dc:identifier>
<dc:title><![CDATA[Deaza-modification of MR1 ligands modulates recognition by MR1-restricted T cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-05-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.05.11.491541v1?rss=1">
<title>
<![CDATA[
Mechanism of Lys6 poly-ubiquitin specificity by the L. pneumophila deubiquitinase LotA 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.05.11.491541v1?rss=1</link>
<description><![CDATA[
The versatility of ubiquitination to impose control over vast domains of eukaryotic biology is due, in part, to diversification through differently-linked poly-ubiquitin chains. Deciphering the signaling roles for some poly-ubiquitin chain types, including those linked via K6, has been stymied by a lack of stringent linkage specificity among the implicated regulatory proteins. Forged through strong evolutionary pressures, pathogenic bacteria have evolved intricate mechanisms to regulate host ubiquitin, and in some cases even with exquisite specificity for distinct poly-ubiquitin signals. Herein, we identify and characterize a deubiquitinase domain of the secreted effector protein LotA from Legionella pneumophila that specifically regulates K6-linked poly-ubiquitin during infection. We demonstrate the utility of LotA as a tool for studying K6 poly-ubiquitin. By determining apo and diUb-bound structures, we identify the mechanism of LotA activation and K6 poly-ubiquitin specificity, and identify a novel ubiquitin-binding domain utilized among bacterial deubiquitinases.
]]></description>
<dc:creator>Warren, G. D.</dc:creator>
<dc:creator>Kitao, T.</dc:creator>
<dc:creator>Franklin, T. G.</dc:creator>
<dc:creator>Nguyen, J. V.</dc:creator>
<dc:creator>Geurink, P. P.</dc:creator>
<dc:creator>Kubori, T.</dc:creator>
<dc:creator>Nagai, H.</dc:creator>
<dc:creator>Pruneda, J. N.</dc:creator>
<dc:date>2022-05-11</dc:date>
<dc:identifier>doi:10.1101/2022.05.11.491541</dc:identifier>
<dc:title><![CDATA[Mechanism of Lys6 poly-ubiquitin specificity by the L. pneumophila deubiquitinase LotA]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-05-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.05.15.491411v1?rss=1">
<title>
<![CDATA[
CITE-Viz: Replicating the Interactive Flow Cytometry Workflow in CITE-Seq 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.05.15.491411v1?rss=1</link>
<description><![CDATA[
SummaryThe rapid advancement of new genomic sequencing technology has enabled the development of multi-omic single-cell sequencing assays. These assays profile multiple modalities in the same cell and can often yield new insights not revealed with a single modality. For example, CITE-Seq (Cellular Indexing of Transcriptomes and Epitopes by Sequencing) simultaneously profiles the single-cell RNA transcriptome and the surface protein expression. The extra dimension of surface protein markers can be used to further identify cell clusters - an essential step for downstream analyses and interpretation. Additionally, multi-dimensional datasets like CITE-Seq require nuanced visualization methods to accurately assess the data. To facilitate cell cluster classification and visualization in CITE-Seq, we developed CITE-Viz. CITE-Viz is a single-cell visualization platform with a custom module that replicates the interactive flow-cytometry gating workflow. With CITE-Viz, users can investigate CITE-Seq specific quality control (QC) metrics, view multi-omic co-expression feature plots, and classify cell clusters by iteratively gating on the abundance of cell surface markers. CITE-Viz was developed to make multi-modal single-cell analysis accessible to a wide variety of biologists, with the aim to discover new insights into their data and to facilitate novel hypothesis generation.

Availability and ImplementationCITE-Viz installation and usage instructions can be found in the GitHub repository https://github.com/maxsonBraunLab/CITE-Viz

Contactmaxsonj@ohsu.edu

Supplementary InformationDown-sampled peripheral blood mononuclear dataset (Hao et al. 2021): https://bit.ly/3vxbhfW
]]></description>
<dc:creator>Kong, G. L.</dc:creator>
<dc:creator>Nguyen, T. T.</dc:creator>
<dc:creator>Rosales, W. K.</dc:creator>
<dc:creator>Panikar, A. D.</dc:creator>
<dc:creator>Cheney, J. H. W.</dc:creator>
<dc:creator>Curtiss, B. M.</dc:creator>
<dc:creator>Carratt, S. A.</dc:creator>
<dc:creator>Braun, T. P.</dc:creator>
<dc:creator>Maxson, J. E.</dc:creator>
<dc:date>2022-05-16</dc:date>
<dc:identifier>doi:10.1101/2022.05.15.491411</dc:identifier>
<dc:title><![CDATA[CITE-Viz: Replicating the Interactive Flow Cytometry Workflow in CITE-Seq]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-05-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.05.16.491863v1?rss=1">
<title>
<![CDATA[
Therapeutic neutralizing monoclonal antibody administration protects against lethal viscerotropic Yellow Fever infection 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.05.16.491863v1?rss=1</link>
<description><![CDATA[
Few countermeasures to treat Yellow Fever virus (YFV) infection are under development, because vaccines have helped to limit new infections. Unfortunately, vaccine hesitancy, supply deficits, and a paucity of therapeutic options have left individuals at risk. Here, we tested potent YFV-specific neutralizing monoclonal antibodies in rodents and non-human primates. We administered antibodies during acute pathogenic YFV infection and demonstrate that we can prevent severe disease and death. Given the severity of YFV-induced disease, our results show that these antibodies could be effective in saving lives and fill a much-needed void in managing Yellow Fever cases during outbreaks around the world.

One Sentence SummaryTherapeutic monoclonal antibodies prevent death from YFV infection.
]]></description>
<dc:creator>Ricciardi, M. J.</dc:creator>
<dc:creator>Rust, L. N.</dc:creator>
<dc:creator>Pedreno-Lopez, N.</dc:creator>
<dc:creator>Yusova, S.</dc:creator>
<dc:creator>Biswas, S.</dc:creator>
<dc:creator>Webb, G. M.</dc:creator>
<dc:creator>Fischer, M.</dc:creator>
<dc:creator>Gonsales-Nieto, L.</dc:creator>
<dc:creator>Voigt, T. B.</dc:creator>
<dc:creator>Louw, J. J.</dc:creator>
<dc:creator>Laurino, F. D.</dc:creator>
<dc:creator>DiBello, J. R.</dc:creator>
<dc:creator>Raue, H.-P.</dc:creator>
<dc:creator>Raphael, L. M. S.</dc:creator>
<dc:creator>Yrizarry-Medina, A.</dc:creator>
<dc:creator>Rosen, B. C.</dc:creator>
<dc:creator>Agnor, R.</dc:creator>
<dc:creator>Gao, L.</dc:creator>
<dc:creator>Labriola, C.</dc:creator>
<dc:creator>Axthelm, M.</dc:creator>
<dc:creator>Smedley, J.</dc:creator>
<dc:creator>Julander, J. G.</dc:creator>
<dc:creator>Bonaldo, M. C.</dc:creator>
<dc:creator>Walker, L. M.</dc:creator>
<dc:creator>Slifka, M. K.</dc:creator>
<dc:creator>Messaoudi, I.</dc:creator>
<dc:creator>Burton, D. R.</dc:creator>
<dc:creator>Kallas, E. G.</dc:creator>
<dc:creator>Sacha, J. B.</dc:creator>
<dc:creator>Watkins, D. I.</dc:creator>
<dc:creator>Burwitz, B. J.</dc:creator>
<dc:date>2022-05-18</dc:date>
<dc:identifier>doi:10.1101/2022.05.16.491863</dc:identifier>
<dc:title><![CDATA[Therapeutic neutralizing monoclonal antibody administration protects against lethal viscerotropic Yellow Fever infection]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-05-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.05.16.492053v1?rss=1">
<title>
<![CDATA[
Infection of the maternal-fetal interface and vertical transmission following low-dose inoculation of pregnant rhesus macaques (Macaca mulatta) with an African-lineage Zika virus 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.05.16.492053v1?rss=1</link>
<description><![CDATA[
BackgroundCongenital Zika virus (ZIKV) infection can result in birth defects, including malformations in the fetal brain and visual system. There are two distinct genetic lineages of ZIKV: African and Asian. Asian lineage ZIKVs have been associated with adverse pregnancy outcomes in humans; however, recent evidence from experimental models suggests that African-lineage viruses can also be vertically transmitted and cause fetal harm.

Methodology/Principal FindingsTo evaluate the potential for vertical transmission of African-lineage ZIKV, we inoculated nine pregnant rhesus macaques (Macaca mulatta) subcutaneously with 44 plaque- forming units of a ZIKV strain from Senegal, (ZIKV-DAK). Dams were inoculated either at gestational day 30 or 45. Following maternal inoculation, pregnancies were surgically terminated seven or 14 days later and fetal and maternal-fetal interface tissues were collected and evaluated. Infection in the dams was evaluated via plasma viremia and neutralizing antibody titers pre- and post- ZIKV inoculation. All dams became productively infected and developed strong neutralizing antibody responses. ZIKV RNA was detected in maternal-fetal interface tissues (placenta, decidua, and fetal membranes) by RT-qPCR and in situ hybridization. In situ hybridization detected ZIKV predominantly in the decidua and revealed that the fetal membranes may play a role in ZIKV vertical transmission. Infectious ZIKV was detected in the amniotic fluid of three pregnancies and one fetus had ZIKV RNA detected in multiple tissues. No significant pathology was observed in any fetus; however, we did find an increase in the occurrence of decidual vasculitis and necrosis in ZIKV-exposed pregnancies compared to gestational-age-matched controls.

Conclusions/SignificanceThis study demonstrates that African-lineage ZIKV, like Asian-lineage ZIKV, can be vertically transmitted to the macaque fetus during pregnancy. The low inoculating dose used in this study suggests a low minimal infectious dose for rhesus macaques. Vertical transmission with a low dose in macaques further supports the high epidemic potential of African ZIKV strains.

Author SummaryZika virus infection during pregnancy can result in adverse pregnancy outcomes including birth defects and miscarriage. There are two distinct genetic backgrounds of Zika virus: Asian-lineage and African-lineage. Currently, only Asian-lineage Zika virus is causally associated with adverse pregnancy outcomes in people. However, experimental studies have shown that African-lineage Zika virus can infect the fetus during pregnancy and cause adverse outcomes. Adverse pregnancy outcomes may not be associated with infection in people until there is an outbreak in a naive population. Thus, as African-lineage Zika virus continues to spread globally, the risk that it may pose to pregnant people remains a public health concern. In this study, we demonstrate that African-lineage Zika virus can be transmitted from the mother to the fetus during pregnancy. This study is significant because we used rhesus macaques, an animal that shares many key elements of Zika virus infection in pregnant people. This study is also significant because we inoculated pregnant macaques with a very small amount of virus, suggesting that fetal infection reported in previously published macaque studies is not limited to high-dose inoculation.
]]></description>
<dc:creator>Koenig, M. R.</dc:creator>
<dc:creator>Mitzey, A. M.</dc:creator>
<dc:creator>Morgan, T. K.</dc:creator>
<dc:creator>Zeng, X.</dc:creator>
<dc:creator>Simmons, H. A.</dc:creator>
<dc:creator>Mejia, A.</dc:creator>
<dc:creator>Leyva Jaimes, F.</dc:creator>
<dc:creator>Keding, L. T.</dc:creator>
<dc:creator>Crooks, C. M.</dc:creator>
<dc:creator>Weiler, A. M.</dc:creator>
<dc:creator>Bohm, E. K.</dc:creator>
<dc:creator>Aliota, M. T.</dc:creator>
<dc:creator>Friedrich, T. C.</dc:creator>
<dc:creator>Mohr, E. L.</dc:creator>
<dc:creator>Golos, T. G.</dc:creator>
<dc:date>2022-05-16</dc:date>
<dc:identifier>doi:10.1101/2022.05.16.492053</dc:identifier>
<dc:title><![CDATA[Infection of the maternal-fetal interface and vertical transmission following low-dose inoculation of pregnant rhesus macaques (Macaca mulatta) with an African-lineage Zika virus]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-05-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.05.19.492679v1?rss=1">
<title>
<![CDATA[
Timed Notch Inhibition drives Photoreceptor fate specification in Human Retinal Organoids 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.05.19.492679v1?rss=1</link>
<description><![CDATA[
PurposeTransplanting photoreceptors from human pluripotent stem cell derived retinal organoids have the potential to reverse vision loss in affected individuals. However, transplantable photoreceptors are only a subset of all cells in the organoids. Hence the goal of our current study was to accelerate and synchronize photoreceptor differentiation in retinal organoids by inhibiting the Notch signaling pathway at different developmental time-points using a small molecule, PF-03084014 (PF).

MethodsHuman induced pluripotent stem cell (hiPSC)- and embryonic stem cells (hESC)-derived retinal organoids were treated with 10M PF for three days at day 45 (D45), D60, D90 and D120 of differentiation. Organoids collected at 14-, 28-, and 42-days post-PF treatment were analyzed for progenitor and photoreceptor markers and Notch pathway inhibition by immunohistochemistry (IHC), quantitative PCR (qPCR) and bulk RNA-seq (n=3-5 organoids from 3 independent experiments).

ResultsRetinal organoids collected at 14-days post-PF treatment showed a decrease in progenitor markers (KI67, VSX2, PAX6, and LHX2) and an increase in differentiated pan-photoreceptor markers (OTX2, CRX, and RCVRN) at all organoid stages except D120. PF-treated organoids at D45 and D60 exhibited an increase in cone photoreceptor markers (RXRG and ARR3). PF-treatment at D90 revealed an increase in cone and rod photoreceptors markers (ARR3, NRL, and NR2E3). Bulk RNA-seq analysis mirrored the IHC data and qPCR confirmed Notch effector inhibition.

ConclusionsTiming the Notch pathway inhibition in human retinal organoids to align with progenitor competency stages can yield an enriched population of early cone or rod photoreceptors.
]]></description>
<dc:creator>Chew, S.</dc:creator>
<dc:creator>Martinez, C.</dc:creator>
<dc:creator>Chirco, K.</dc:creator>
<dc:creator>Kandoi, S.</dc:creator>
<dc:creator>Lamba, D. A.</dc:creator>
<dc:date>2022-05-20</dc:date>
<dc:identifier>doi:10.1101/2022.05.19.492679</dc:identifier>
<dc:title><![CDATA[Timed Notch Inhibition drives Photoreceptor fate specification in Human Retinal Organoids]]></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/10.1101/2022.05.19.492680v1?rss=1">
<title>
<![CDATA[
recountmethylation enables flexible analysis ofpublic blood DNA methylation array data 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.05.19.492680v1?rss=1</link>
<description><![CDATA[
Thousands of DNA methylation (DNAm) array samples from human blood are publicly available on the Gene Expression Omnibus (GEO), but they remain underutilized for experiment planning, replication, and cross-study and cross-platform analyses. To facilitate these tasks, we augmented our recountmethylation R/Bioconductor package with 12,537 uniformly processed EPIC and HM450K blood samples on GEO as well as several new features. We subsequently used our updated package in several illustrative analyses, finding (1) study ID bias adjustment increased variation explained by biological and demographic variables, (2) most variation in autosomal DNAm was explained by genetic ancestry and CD4+ T-cell fractions, and (3) the dependence of power to detect differential methylation on sample size was similar for each of peripheral blood mononuclear cells (PBMC), whole blood, and umbilical cord blood. Finally, we used PBMC and whole blood to perform independent validations, and we recovered 40-46% of differentially methylated probes (DMPs) between sexes from two previously published epigenome-wide association studies (EWAS).
]]></description>
<dc:creator>Maden, S. K.</dc:creator>
<dc:creator>Walsh, B.</dc:creator>
<dc:creator>Ellrott, K.</dc:creator>
<dc:creator>Hansen, K. D.</dc:creator>
<dc:creator>Thompson, R.</dc:creator>
<dc:creator>Nellore, A.</dc:creator>
<dc:date>2022-05-20</dc:date>
<dc:identifier>doi:10.1101/2022.05.19.492680</dc:identifier>
<dc:title><![CDATA[recountmethylation enables flexible analysis ofpublic blood DNA methylation array data]]></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/10.1101/2022.05.24.493132v1?rss=1">
<title>
<![CDATA[
Single-Cell RNA Sequencing Reveals the Effects of Chemotherapy on Human Pancreatic Adenocarcinoma and its Tumor Microenvironment 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.05.24.493132v1?rss=1</link>
<description><![CDATA[
The tumor microenvironment (TME) in pancreatic ductal adenocarcinoma (PDAC) is a complex ecosystem that drives tumor progression; however, in-depth single cell characterization of the PDAC TME and its role in response to therapy is lacking. We performed single-cell RNA sequencing on freshly collected human PDAC samples either before or after chemotherapy. Overall, we found a heterogeneous mixture of basal and classical cancer cell subtypes, along with distinct cancer-associated fibroblast and macrophage subpopulations. Strikingly, classical and basal-like cancer cells exhibited similar transcriptional responses to chemotherapy, and did not demonstrate a shift towards a basal-like transcriptional program among treated samples. We observed decreased ligand-receptor interactions in treated samples, particularly TIGIT on CD8+ T cells and its receptor on cancer cells, and identified TIGIT as the major inhibitory checkpoint molecule of CD8+ T cells. Our results suggest that chemotherapy profoundly impacts the PDAC TME and may promote resistance to immunotherapy.
]]></description>
<dc:creator>Werba, G.</dc:creator>
<dc:creator>Weissinger, D.</dc:creator>
<dc:creator>Kawaler, E. A.</dc:creator>
<dc:creator>Zhao, E.</dc:creator>
<dc:creator>Kalfakakou, D.</dc:creator>
<dc:creator>Dhara, S.</dc:creator>
<dc:creator>Oh, G.</dc:creator>
<dc:creator>Jing, X.</dc:creator>
<dc:creator>Beri, N.</dc:creator>
<dc:creator>Khanna, L.</dc:creator>
<dc:creator>Gonda, T.</dc:creator>
<dc:creator>Oberstein, P.</dc:creator>
<dc:creator>Hajdu, C.</dc:creator>
<dc:creator>Loomis, C. A.</dc:creator>
<dc:creator>Heguy, A.</dc:creator>
<dc:creator>Sherman, M. H.</dc:creator>
<dc:creator>Lund, A. W.</dc:creator>
<dc:creator>Welling, T. H.</dc:creator>
<dc:creator>Dolgalev, I.</dc:creator>
<dc:creator>Tsirigos, A.</dc:creator>
<dc:creator>Simeone, D. M.</dc:creator>
<dc:date>2022-05-25</dc:date>
<dc:identifier>doi:10.1101/2022.05.24.493132</dc:identifier>
<dc:title><![CDATA[Single-Cell RNA Sequencing Reveals the Effects of Chemotherapy on Human Pancreatic Adenocarcinoma and its Tumor Microenvironment]]></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/10.1101/2022.05.24.493187v1?rss=1">
<title>
<![CDATA[
Hemin shows antiviral activity in vitro, possibly through suppression of viral entry mediators. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.05.24.493187v1?rss=1</link>
<description><![CDATA[
Heme oxygenase-1 (HO-1) is a stress-induced enzyme that catalyzes the breakdown of heme into biliverdin, carbon monoxide, and iron. Targeting HO-1 to treat severe COVID-19 has been suggested by several groups, yet the role of HO-1 in SARS-CoV-2 infection remains unclear. Based on this, we aimed to investigate the antiviral activity of Hemin, an activator of HO-1. Infectivity of SARS-CoV-2 was decreased in Vero E6 cells treated with Hemin. Hemin also decreased TMPRSS2 and ACE2 mRNA levels in non-infected cells, possibly explaining the observed decrease in infectivity. TMPRSS2 protein expression and proteolytic activity were decreased in Vero E6 cells treated with Hemin. Besides that, experimental studies supported with in silico calculations. Overall, our study supports further exploration of Hemin as a potential antiviral and inflammatory drug for the treatment of COVID-19.
]]></description>
<dc:creator>UNAL, M. A.</dc:creator>
<dc:creator>BITIRIM, C. V.</dc:creator>
<dc:creator>SOMERS, J.</dc:creator>
<dc:creator>SUMMAK, G. Y.</dc:creator>
<dc:creator>BUL BESBINAR, O.</dc:creator>
<dc:creator>KOCAKAYA, E.</dc:creator>
<dc:creator>GURCAN, C.</dc:creator>
<dc:creator>NAZIR, H.</dc:creator>
<dc:creator>Aksoy Ozer, Z. B.</dc:creator>
<dc:creator>OZKAN, S. A.</dc:creator>
<dc:creator>BEREKETOGLU, S.</dc:creator>
<dc:creator>OZKUL, A.</dc:creator>
<dc:creator>DEMIR, E.</dc:creator>
<dc:creator>AKCALI, K. C.</dc:creator>
<dc:creator>YILMAZER, A.</dc:creator>
<dc:date>2022-05-25</dc:date>
<dc:identifier>doi:10.1101/2022.05.24.493187</dc:identifier>
<dc:title><![CDATA[Hemin shows antiviral activity in vitro, possibly through suppression of viral entry mediators.]]></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/10.1101/2022.05.26.493614v1?rss=1">
<title>
<![CDATA[
Leucine-Rich Repeat Kinase 2 limits dopamine D1 receptor signaling in the striatum and biases against heavy persistent alcohol drinking 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.05.26.493614v1?rss=1</link>
<description><![CDATA[
The transition from hedonic alcohol drinking to problematic drinking is a hallmark of alcohol use disorder that occurs only in a subset of drinkers. This transition is known to require long-lasting changes in the synaptic drive and the activity of striatal neurons expressing dopamine D1 receptor (D1R). The molecular mechanisms that generate vulnerability in some individuals to undergo the transition are less understood. Here, we report that the Parkinsons-related protein leucine-rich repeat kinase 2 (LRRK2) modulates striatal D1R function to affect the behavioral response to alcohol and the likelihood that mice transition to heavy, persistent alcohol drinking. Deletion of the Lrrk2 gene specifically from D1R-expressing neurons potentiates D1R signaling at the cellular and synaptic level, enhancing alcohol-related behaviors and drinking. Mice with cell-specific deletion of Lrrk2 are more prone to heavy alcohol drinking and consumption is insensitive to punishment. These findings identify a novel role for LRRK2 function in the striatum in promoting resilience against heavy and persistent alcohol drinking.
]]></description>
<dc:creator>da Silva e Silva, D.</dc:creator>
<dc:creator>Matsui, A.</dc:creator>
<dc:creator>Murray, E. M.</dc:creator>
<dc:creator>Mamais, A.</dc:creator>
<dc:creator>Shaw, M.</dc:creator>
<dc:creator>Ron, D.</dc:creator>
<dc:creator>Cookson, M. R.</dc:creator>
<dc:creator>Alvarez, V. A.</dc:creator>
<dc:date>2022-05-29</dc:date>
<dc:identifier>doi:10.1101/2022.05.26.493614</dc:identifier>
<dc:title><![CDATA[Leucine-Rich Repeat Kinase 2 limits dopamine D1 receptor signaling in the striatum and biases against heavy persistent alcohol drinking]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-05-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.05.30.494080v1?rss=1">
<title>
<![CDATA[
T cell Egress via Lymphatic Vessels Limits the Intratumoral T cell Repertoire in Melanoma 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.05.30.494080v1?rss=1</link>
<description><![CDATA[
Antigen-specific CD8+ T cell accumulation in tumors is a prerequisite for effective immunotherapy, and yet, the mechanisms of lymphocyte transit remain poorly defined. We find that tumor-associated lymphatic vessels control T cell exit from tumors via the chemokine CXCL12, and intratumoral antigen encounter tunes CXCR4 expression on effector CD8+ T cells. Only high affinity antigen downregulates CXCR4 and upregulates the CXCL12 decoy receptor, ACKR3, thereby reducing CXCL12 sensitivity and promoting T cell retention. A diverse repertoire of functional tumor-specific CD8+ T cells exit the tumor, thereby limiting tumor control. CXCR4 inhibition and loss of lymphatic-specific CXCL12 boosts T cell retention and enhances response to therapeutic immune checkpoint blockade. Strategies that limit T cell egress, therefore, provide a new tool to boost immunotherapy response.

One-Sentence SummaryLymphatic vessel-mediated, antigen-dependent CD8+ T cell egress limits T cell accumulation in melanomas and impairs anti-tumor immunity.
]]></description>
<dc:creator>Steele, M. M.</dc:creator>
<dc:creator>Dryg, I. D.</dc:creator>
<dc:creator>Murugan, D.</dc:creator>
<dc:creator>Femel, J.</dc:creator>
<dc:creator>du Bois, H.</dc:creator>
<dc:creator>Hill, C.</dc:creator>
<dc:creator>Leachman, S. A.</dc:creator>
<dc:creator>Chang, Y. H.</dc:creator>
<dc:creator>Coussens, L. M.</dc:creator>
<dc:creator>Lund, A. W.</dc:creator>
<dc:date>2022-05-30</dc:date>
<dc:identifier>doi:10.1101/2022.05.30.494080</dc:identifier>
<dc:title><![CDATA[T cell Egress via Lymphatic Vessels Limits the Intratumoral T cell Repertoire in Melanoma]]></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/10.1101/2022.06.01.494442v1?rss=1">
<title>
<![CDATA[
Recording large-scale, cellular-resolution neuronal activity from freely-moving mice 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.06.01.494442v1?rss=1</link>
<description><![CDATA[
Current methods for recording large-scale neuronal activity from behaving mice with single-cell resolution require either fixing the mouse head under a microscope or attachment of a recording device to the animals skull. Both of these options significantly affect the animal behavior and hence also the recorded brain activity patterns. Here, we introduce a new method to acquire snapshots of single-cell cortical activity maps from freely-moving mice using a calcium sensor called CaMPARI. CaMPARI has a unique property of irreversibly changing its color from green to red inside active neurons when illuminated with 400nm light. We capitalize on this property to demonstrate cortex-wide activity recording without any head fixation or attachment of a miniaturized device to the mouses head. Multiple cortical regions were recorded while the mouse was performing a battery of behavioral and cognitive tests. We identified task-dependent activity patterns across motor and somatosensory cortices, with significant differences across sub-regions of the motor cortex. This new CaMPARI-based recording method expands the capabilities of recording neuronal activity from freely-moving and behaving mice under minimally-restrictive experimental conditions and provides large-scale volumetric data that are not accessible otherwise.
]]></description>
<dc:creator>Das, A.</dc:creator>
<dc:creator>Holden, S.</dc:creator>
<dc:creator>Borovicka, J.</dc:creator>
<dc:creator>Icardi, J.</dc:creator>
<dc:creator>Patel, D.</dc:creator>
<dc:creator>Patel, R.</dc:creator>
<dc:creator>Raber, J.</dc:creator>
<dc:creator>Dana, H.</dc:creator>
<dc:date>2022-06-03</dc:date>
<dc:identifier>doi:10.1101/2022.06.01.494442</dc:identifier>
<dc:title><![CDATA[Recording large-scale, cellular-resolution neuronal activity from freely-moving mice]]></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/10.1101/2022.06.03.494221v1?rss=1">
<title>
<![CDATA[
Community assessment of methods to deconvolve cellular composition from bulk gene expression 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.06.03.494221v1?rss=1</link>
<description><![CDATA[
We evaluate deconvolution methods, which infer levels of immune infiltration from bulk expression of tumor samples, through a community-wide DREAM Challenge. We assess six published and 22 community-contributed methods using in vitro and in silico transcriptional profiles of admixed cancer and healthy immune cells. Several published methods predict most cell types well, though they either were not trained to evaluate all functional CD8+ T cell states or do so with low accuracy. Several community-contributed methods address this gap, including a deep learning-based approach, whose strong performance establishes the applicability of this paradigm to deconvolution. Despite being developed largely using immune cells from healthy tissues, deconvolution methods predict levels of tumor-derived immune cells well. Our admixed and purified transcriptional profiles will be a valuable resource for developing deconvolution methods, including in response to common challenges we observe across methods, such as sensitive identification of functional CD4+ T cell states.
]]></description>
<dc:creator>White, B. S.</dc:creator>
<dc:creator>de Reynies, A.</dc:creator>
<dc:creator>Newman, A. M.</dc:creator>
<dc:creator>Waterfall, J. J.</dc:creator>
<dc:creator>Lamb, A.</dc:creator>
<dc:creator>Petitprez, F.</dc:creator>
<dc:creator>Yu, T.</dc:creator>
<dc:creator>Guo, X.</dc:creator>
<dc:creator>Valdeolivas, A.</dc:creator>
<dc:creator>Lin, Y.</dc:creator>
<dc:creator>Li, H.</dc:creator>
<dc:creator>Xiao, X.</dc:creator>
<dc:creator>Wang, S.</dc:creator>
<dc:creator>Zheng, F.</dc:creator>
<dc:creator>Yang, W.</dc:creator>
<dc:creator>Yu, R.</dc:creator>
<dc:creator>Guerrero-Gimenez, M. E.</dc:creator>
<dc:creator>Catania, C. A.</dc:creator>
<dc:creator>Lang, B. J.</dc:creator>
<dc:creator>Domanskyi, S.</dc:creator>
<dc:creator>Bertus, T. J.</dc:creator>
<dc:creator>Piermarocchi, C.</dc:creator>
<dc:creator>Monaco, G.</dc:creator>
<dc:creator>Caruso, F. P.</dc:creator>
<dc:creator>Ceccarelli, M.</dc:creator>
<dc:creator>Coller, J.</dc:creator>
<dc:creator>Maecker, H.</dc:creator>
<dc:creator>Duault, C.</dc:creator>
<dc:creator>Sokoohi, V.</dc:creator>
<dc:creator>Patel, S.</dc:creator>
<dc:creator>Liliental, J. E.</dc:creator>
<dc:creator>Simon, S.</dc:creator>
<dc:creator>Tumor Deconvolution DREAM Challenge consortium,</dc:creator>
<dc:creator>Saez-Rodriguez, J.</dc:creator>
<dc:creator>Heiser, L. M.</dc:creator>
<dc:creator>Guinney, J.</dc:creator>
<dc:creator>Gentles, A. J.</dc:creator>
<dc:date>2022-06-05</dc:date>
<dc:identifier>doi:10.1101/2022.06.03.494221</dc:identifier>
<dc:title><![CDATA[Community assessment of methods to deconvolve cellular composition from bulk gene expression]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-06-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.06.09.495548v1?rss=1">
<title>
<![CDATA[
Gait Abnormalities and Aberrant D2 Receptor Expression and Signaling in a Mouse Model of the Human Pathogenic Mutation DRD2I212F 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.06.09.495548v1?rss=1</link>
<description><![CDATA[
A dopamine D2 receptor mutation was recently identified in a family with a novel hyperkinetic movement disorder (Mov Disord 36: 729-739, 2021). That allelic variant D2-I212F is a constitutively active and G protein-biased receptor. We now describe mice engineered to carry the D2-I212F variant, Drd2I212F. The mice exhibited gait abnormalities resembling those in other mouse models of chorea and/or dystonia, and had decreased striatal D2 receptor expression. Electrically evoked IPSCs in midbrain dopamine neurons and striatum from Drd2I212F mice exhibited slow onset and decay compared to wild type mice. In the presence of dopamine, current decay initiated by photolytic release of sulpiride from CyHQ-sulpiride was slower in midbrain slices from Drd2I212F mice than Drd2+/+ mice. Furthermore, in contrast to wild type mice in which dopamine is more potent at neurons in the nucleus accumbens than in the dorsal striatum, reflecting activation of Go vs. Gi1, dopamine had similar potencies in those two brain regions of Drd2I212F mice. Repeated cocaine treatment, which decreases dopamine potency in the nucleus accumbens of wild type mice, had no effect on dopamine potency in Drd2I212F mice. The results demonstrate the utility of this mouse model for investigating the role of pathogenic DRD2 variants in early-onset hyperkinetic movement disorders.
]]></description>
<dc:creator>Rodriguez-Contreras, D.</dc:creator>
<dc:creator>Gong, S.</dc:creator>
<dc:creator>Lebowitz, J. J.</dc:creator>
<dc:creator>Fedorov, L. M.</dc:creator>
<dc:creator>Asad, N.</dc:creator>
<dc:creator>Dore, T. M.</dc:creator>
<dc:creator>Ford, C. P.</dc:creator>
<dc:creator>Williams, J. T.</dc:creator>
<dc:creator>Neve, K. A.</dc:creator>
<dc:date>2022-06-11</dc:date>
<dc:identifier>doi:10.1101/2022.06.09.495548</dc:identifier>
<dc:title><![CDATA[Gait Abnormalities and Aberrant D2 Receptor Expression and Signaling in a Mouse Model of the Human Pathogenic Mutation DRD2I212F]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-06-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.06.10.495698v1?rss=1">
<title>
<![CDATA[
Can Deep Learning Provide a Generalizable Model for Dynamic Sound Encoding in Auditory Cortex? 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.06.10.495698v1?rss=1</link>
<description><![CDATA[
Convolutional neural networks (CNNs) can provide powerful and flexible models of neural sensory processing. However, the utility of CNNs in studying the auditory system has been limited by their requirement for large datasets and the complex response properties of single auditory neurons. To address these limitations, we developed a population encoding model: a CNN that simultaneously predicts activity of several hundred neurons recorded during presentation of a large set of natural sounds. This approach defines a shared spectro-temporal space and pools statistical power across neurons. Population models of varying architecture performed consistently better than traditional linear-nonlinear models on data from primary and non-primary auditory cortex. Moreover, population models were highly generalizable. The output layer of a model pre-trained on one population of neurons could be fit to novel single units, achieving performance equivalent to that of neurons in the original fit data. This ability to generalize suggests that population encoding models capture a general set of computations performed by auditory cortex.
]]></description>
<dc:creator>Pennington, J. R.</dc:creator>
<dc:creator>David, S. V.</dc:creator>
<dc:date>2022-06-13</dc:date>
<dc:identifier>doi:10.1101/2022.06.10.495698</dc:identifier>
<dc:title><![CDATA[Can Deep Learning Provide a Generalizable Model for Dynamic Sound Encoding in Auditory Cortex?]]></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/10.1101/2022.06.14.495937v1?rss=1">
<title>
<![CDATA[
Crowd-sourced benchmarking of single-sample tumour subclonal reconstruction 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.06.14.495937v1?rss=1</link>
<description><![CDATA[
Tumours are dynamically evolving populations of cells. Subclonal reconstruction algorithms use bulk DNA sequencing data to quantify parameters of tumour evolution, allowing assessment of how cancers initiate, progress and respond to selective pressures. A plethora of subclonal reconstruction algorithms have been created, but their relative performance across the varying biological and technical features of real-world cancer genomic data is unclear. We therefore launched the ICGC-TCGA DREAM Somatic Mutation Calling -- Tumour Heterogeneity and Evolution Challenge. This seven-year community effort used cloud-computing to benchmark 31 containerized subclonal reconstruction algorithms on 51 simulated tumours. Each algorithm was scored for accuracy on seven independent tasks, leading to 12,061 total runs. Algorithm choice influenced performance significantly more than tumour features, but purity-adjusted read-depth, copy number state and read mappability were associated with performance of most algorithms on most tasks. No single algorithm was a top performer for all seven tasks and existing ensemble strategies were surprisingly unable to outperform the best individual methods, highlighting a key research need. All containerized methods, evaluation code and datasets are available to support further assessment of the determinants of subclonal reconstruction accuracy and development of improved methods to understand tumour evolution.
]]></description>
<dc:creator>Salcedo, A.</dc:creator>
<dc:creator>Tarabichi, M.</dc:creator>
<dc:creator>Buchanan, A.</dc:creator>
<dc:creator>Espiritu, S. M. G.</dc:creator>
<dc:creator>Zhang, H.</dc:creator>
<dc:creator>Zhu, K.</dc:creator>
<dc:creator>Yang, T.-H. O.</dc:creator>
<dc:creator>Leshchiner, I.</dc:creator>
<dc:creator>Anastassiou, D.</dc:creator>
<dc:creator>Guan, Y.</dc:creator>
<dc:creator>Jang, G. H.</dc:creator>
<dc:creator>Haase, K.</dc:creator>
<dc:creator>Deshwar, A. G.</dc:creator>
<dc:creator>Zou, W.</dc:creator>
<dc:creator>Umar, I.</dc:creator>
<dc:creator>Dentro, S.</dc:creator>
<dc:creator>Wintersinger, J. A.</dc:creator>
<dc:creator>Chiotti, K.</dc:creator>
<dc:creator>Demeulemeester, J.</dc:creator>
<dc:creator>Jolly, C.</dc:creator>
<dc:creator>Scyza, L.</dc:creator>
<dc:creator>Ko, M.</dc:creator>
<dc:creator>PCAWG-11 Working Group,</dc:creator>
<dc:creator>SMC-Het Participants,</dc:creator>
<dc:creator>Wedge, D. C.</dc:creator>
<dc:creator>Morris, Q. D.</dc:creator>
<dc:creator>Ellrot, K.</dc:creator>
<dc:creator>Van Loo, P.</dc:creator>
<dc:creator>Boutros, P. C.</dc:creator>
<dc:date>2022-06-15</dc:date>
<dc:identifier>doi:10.1101/2022.06.14.495937</dc:identifier>
<dc:title><![CDATA[Crowd-sourced benchmarking of single-sample tumour subclonal reconstruction]]></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/10.1101/2022.06.15.492526v1?rss=1">
<title>
<![CDATA[
Daily administration of low-dose daunorubicin or doxorubicin inhibits hypoxia-inducible factor 1 and tumor vascularization 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.06.15.492526v1?rss=1</link>
<description><![CDATA[
Using a hypoxia-inducible factor 1 (HIF-1)-dependent luciferase reporter in Hep3B human hepatocellular carcinoma cells, we screened over 3,000 drugs that have been used in clinical trials and identified multiple anthracyclines as inhibitors of HIF-1 activity. Anthracyclines interfered with the ability of HIF-1 to bind to DNA. Daily injection of tumor-bearing mice with anthracyclines at low dose inhibited expression of the luciferase reporter and HIF-1 target genes that encode vascular endothelial growth factor A (VEGFA; ligand of VEGFR2), stromal-derived factor 1 (SDF-1; ligand of CXCR4), and stem cell factor (SCF; ligand of CD117) in tumor tissue. Increased numbers of circulating CXCR4+/Sca1+, VEGFR2+/CD34+, and VEGFR2+/CD117+ cells were demonstrated in immunodeficient mice bearing prostate cancer xenografts but not in tumor-bearing mice treated with anthracyclines, which also significantly inhibited angiogenesis in tumor tissue. Our findings indicate that HIF-1 inhibition underlies the anti-angiogenic effect associated with daunorubicin or doxorubicin metronomic therapy and suggest that these drugs may be particularly effective in patients with high levels of HIF-1 in their diagnostic tumor biopsy.
]]></description>
<dc:creator>Yang, Y.</dc:creator>
<dc:creator>Chen, D. Z.</dc:creator>
<dc:creator>Rey, S.</dc:creator>
<dc:creator>Liu, J. O.</dc:creator>
<dc:creator>Semenza, G. L.</dc:creator>
<dc:date>2022-06-17</dc:date>
<dc:identifier>doi:10.1101/2022.06.15.492526</dc:identifier>
<dc:title><![CDATA[Daily administration of low-dose daunorubicin or doxorubicin inhibits hypoxia-inducible factor 1 and tumor vascularization]]></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/10.1101/2022.06.20.496891v1?rss=1">
<title>
<![CDATA[
Persistent inflammation promotes endocannabinoid release and presynaptic cannabinoid 1 receptor desensitization 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.06.20.496891v1?rss=1</link>
<description><![CDATA[
Pain therapies targeting the cannabinoid system are increasing with expansion of cannabis legalization but adaptations in the endogenous cannabinoid system during inflammatory pain could limit their efficacy. Presynaptic inhibition of GABA release mediated by cannabinoid 1 receptor (CB1R) agonists in the ventrolateral periaqueductal gray (vlPAG) is markedly reduced in male and female Sprague Dawley rats after persistent inflammation induced by Complete Freunds Adjuvant (CFA). Inflammation results in increased endocannabinoid (eCB) synthesis and desensitization of presynaptic CB1Rs that is reversed by a GRK2/3 inhibitor, Compound 101. Despite CB1R desensitization, eCB activation of CB1Rs is maintained after inflammation. Depolarization-induced suppression of inhibition (DSI) in naive animals is rapid and transient, but is prolonged in recordings after inflammation. Prolonged DSI is mediated by 2-arachidonoylglycerol (2-AG) indicating reduced monoacylglycerol lipase (MAGL) activity. These adaptations within the endogenous cannabinoid system have important implications for the development of future pain therapies targeting CB1Rs or MAGL.
]]></description>
<dc:creator>Bouchet, C. A.</dc:creator>
<dc:creator>Janowsky, A.</dc:creator>
<dc:creator>Ingram, S. L.</dc:creator>
<dc:date>2022-06-22</dc:date>
<dc:identifier>doi:10.1101/2022.06.20.496891</dc:identifier>
<dc:title><![CDATA[Persistent inflammation promotes endocannabinoid release and presynaptic cannabinoid 1 receptor desensitization]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-06-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.06.24.497507v1?rss=1">
<title>
<![CDATA[
Liver Endothelium Microenvironment Promotes HER3-mediated Cell Growth in Pancreatic ductal adenocarcinoma 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.06.24.497507v1?rss=1</link>
<description><![CDATA[
[~]90% metastatic pancreatic ductal adenocarcinoma (mPDAC) occurs in the liver, and the 5-year survival rate for patients with mPDAC is only at 3%. We previously reported that liver endothelial cells (ECs) secreted soluble factors to promote colorectal cancer cell survival in a paracrine fashion. However, the effects of liver ECs on mPDAC have not been elucidated. In this study, we used primary ECs from non-neoplastic liver tissues. We treated PDAC cells with conditioned medium (CM) from liver ECs, with CM from PDAC as controls, and determined that liver EC-secreted factors increased PDAC cell growth. Using an unbiased receptor tyrosine kinase array, we identified human epidermal growth factor receptor 3 (HER3, also known as ErbB3) as a key mediator in EC-induced growth in PDAC cells that have HER3 expression (HER3 +ve). We found that EC-secreted neuregulins activated the HER3-AKT signaling axis, and that depleting neuregulins from EC CM or blocking HER3 with an antibody, seribantumab, attenuated EC-induced proliferation in HER3 +ve PDAC cells, but not in cells without HER3 expression. Furthermore, we determined that EC CM increased PDAC xenograft growth in vivo, and that seribantumab blocked EC-induced growth in xenografts with HER3 expression. These findings elucidated a paracrine role of liver ECs in promoting PDAC cell growth, and identified the HER3-AKT axis as a key mediator in EC-induced functions in PDAC cells with HER3 expression.

Implicationsover 70% mPDAC express HER3. This study suggests the potential of using HER3-targeted therapies for treating patients with HER3 +ve mPDAC.
]]></description>
<dc:creator>Zhang, W.</dc:creator>
<dc:creator>Wright, M.</dc:creator>
<dc:creator>Rathore, M.</dc:creator>
<dc:creator>Zarei, M.</dc:creator>
<dc:creator>Vaziri-Gohar, A.</dc:creator>
<dc:creator>Winter, J.</dc:creator>
<dc:creator>Brody, J.</dc:creator>
<dc:creator>Wang, R.</dc:creator>
<dc:date>2022-06-28</dc:date>
<dc:identifier>doi:10.1101/2022.06.24.497507</dc:identifier>
<dc:title><![CDATA[Liver Endothelium Microenvironment Promotes HER3-mediated Cell Growth in Pancreatic ductal adenocarcinoma]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-06-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.06.27.497817v1?rss=1">
<title>
<![CDATA[
Electrical Signaling in Cochlear Efferents is Driven by an Intrinsic Neuronal Oscillator 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.06.27.497817v1?rss=1</link>
<description><![CDATA[
Efferent neurons are believed to play essential roles in maintaining auditory function. The lateral olivocochlear (LOC) neurons, which project from the brainstem to the inner ear where they release multiple transmitters including peptides, catecholamines and acetylcholine, are the most numerous yet least understood elements of efferent control of the cochlea. Using in vitro calcium imaging and patch-clamp recordings, we found that LOC neurons in juvenile and young adult mice exhibited extremely slow waves of activity (~0.1 Hz). These seconds-long bursts of Na+ spikes were driven by an intrinsic oscillator dependent on L-type Ca2+ channels, and were not observed in prehearing mice, suggesting an age-dependent mechanism underlying the intrinsic oscillator. Using optogenetic approaches, we identified both ascending (cochlear nucleus) and descending (auditory cortex) sources of synaptic excitation, as well as the synaptic receptors used for such excitation. Additionally, we identified potent inhibition originating in the glycinergic medial nucleus of trapezoid body (MNTB). Conductance-clamp experiments revealed an unusual mechanism of electrical signaling in LOC neurons, in which synaptic excitation and inhibition served to switch on and off the intrinsically generated spike burst mechanism, allowing for prolonged periods of activity or silence controlled by brief synaptic events. Protracted bursts of action potentials may be essential for effective exocytosis of the diverse transmitters released by LOC fibers in the cochlea.

Significance StatementThe lateral olivocochlear (LOC) neurons, being the most abundant auditory efferent control of the ear, remained largely unexplored. Here we reported that LOC neurons displayed patterned electrical activity at an unusually slow pace (~0.1 Hz), mediated by a calcium-dependent intrinsic oscillator. This is surprising given the speed and precision were believed to be the currency of signaling in the lower auditory system. Optogenetic experiments determined the glutamatergic and glycinergic sources of synaptic inputs to these neurons, while conductance-clamp experiments revealed that synaptic activity acts like switches for turning on or off prolonged spike activity driven by the intrinsic oscillator. This extended spike activity may be essential for effective exocytosis of the diverse transmitters released by LOC fibers in the cochlea.
]]></description>
<dc:creator>Hong, H.</dc:creator>
<dc:creator>Trussell, L. O.</dc:creator>
<dc:date>2022-06-30</dc:date>
<dc:identifier>doi:10.1101/2022.06.27.497817</dc:identifier>
<dc:title><![CDATA[Electrical Signaling in Cochlear Efferents is Driven by an Intrinsic Neuronal Oscillator]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-06-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.06.29.498022v1?rss=1">
<title>
<![CDATA[
Quantifying cooperative multisite binding through Bayesian inference 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.06.29.498022v1?rss=1</link>
<description><![CDATA[
Multistep protein-protein interactions underlie most biological processes, but their characterization through methods such as isothermal titration calorimetry (ITC) is largely confined to simple models that provide little information on the intermediate, individual steps. In this study, we primarily examine the essential hub protein LC8, a small dimer that binds disordered regions of 100+ client proteins in two symmetrical grooves at the dimer interface. Mechanistic details of LC8 binding have remained elusive, hampered in part by ITC data analyses employing simple models that treat bivalent binding as a single event with a single binding affinity. We build on existing Bayesian ITC approaches to quantify thermodynamic parameters for multi-site binding interactions impacted by significant uncertainty in protein concentration. Using a two-site binding model, we model LC8 binding and identify positive cooperativity with high confidence for multiple client peptides. Application of an identical model to two-site binding between the coiled-coil dimer NudE and the intermediate chain of dynein reveals little evidence of cooperativity, in contrast to LC8. We propose that cooperativity in the LC8 system drives the formation of saturated 2:2 bound states, which play a functional role in many LC8 complexes. In addition to these system-specific findings, our work advances general ITC analysis in two ways. First, we describe a previously unrecognized mathematical ambiguity in concentrations in standard binding models and clarify how it impacts the precision with which binding parameters can be determined in cases of high uncertainty in analyte concentrations. Second, building on observations in the LC8 system, we develop a system-agnostic heat map of practical parameter identifiability calculated from synthetic data which demonstrates that certain binding parameters intrinsically inflate parameter uncertainty in ITC analysis, independent of experimental uncertainties.

Author SummaryMulti-site protein-protein interactions govern many protein functions throughout the cell. Precise determination of thermodynamic constants of multi-site binding is a significant biophysical challenge, however. The application of complex models to multi-step interactions is difficult and hampered further by complications arising from uncertainty in analyte concentrations. To address these issues, we utilize Bayesian statistical techniques which calculate the  likelihood of parameters giving rise to experimental observations to build probability density distributions for thermodynamic parameters of binding. To demonstrate the method and improve our understanding how the hub protein LC8 promotes dimerization of its 100+ binding partners, we test the pipeline on several of these partners and demonstrate that LC8 can bind clients cooperatively, driving interactions towards a  fully bound functional state. We additionally examine an interaction between the dimer NudE and the intermediate chain of dynein, which does not appear to bind with cooperativity. Our work provides a solid foundation for future analysis of more complicated binding interactions, including oligomeric complexes formed between LC8 and clients with multiple LC8-binding sites.
]]></description>
<dc:creator>Estelle, A. B.</dc:creator>
<dc:creator>George, A.</dc:creator>
<dc:creator>Barbar, E.</dc:creator>
<dc:creator>Zuckerman, D. M.</dc:creator>
<dc:date>2022-07-01</dc:date>
<dc:identifier>doi:10.1101/2022.06.29.498022</dc:identifier>
<dc:title><![CDATA[Quantifying cooperative multisite binding through Bayesian inference]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-07-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.06.30.498366v1?rss=1">
<title>
<![CDATA[
Senescent cell-derived extracellular vesicles recruit antigen presenting cells and limit squamous carcinoma recurrence 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.06.30.498366v1?rss=1</link>
<description><![CDATA[
Extracellular vesicles (EVs) are key signaling mediators. To explore the role of senescent cell-derived extracellular vesicles (senEVs) in inflammatory responses to senescence, we developed an engraftment-based senescence model in wild-type mice and genetically blocked senEV release in vivo, without significantly affecting soluble mediators. Our results demonstrate that senEVs are both necessary and sufficient to trigger immune-mediated clearance of senescent cells, thereby suppressing tumor growth. In the absence of senEVs, the recruitment of MHC-II+ antigen-presenting cells to the senescence microenvironment was markedly impaired. Blocking senEV release redirected the primary target of senescent cell signaling from antigen-presenting cells to neutrophils.

Through comprehensive transcriptional and proteomic analyses, we identified six ligands specific to senEVs, highlighting their role in promoting antigen-presenting cell-T cell adhesion and synapse formation. Antigen-presenting cells activated CCR2+CD4+ TH17 cells, which appeared to inhibit B cell activation. CD4 T cells were essential for preventing tumor recurrence, indicating that CCR2+ TH17 cells function downstream of senEVs during senescence surveillance.

Our findings suggest that senEVs complement the activity of secreted inflammatory mediators by recruiting and activating distinct immune cell subsets, thereby enhancing the efficient clearance of senescent cells. These conclusions may have implications not only for tumor recurrence but also for understanding senescence during de novo carcinogenesis. Consequently, this work could inform the development of novel cancer early detection strategies based on the biology of cellular senescence.
]]></description>
<dc:creator>Ziglari, T.</dc:creator>
<dc:creator>Claudio, N. M.</dc:creator>
<dc:creator>Kyle, J. E.</dc:creator>
<dc:creator>Nakayasu, E. S.</dc:creator>
<dc:creator>Guo, Z.</dc:creator>
<dc:creator>Pucci, F.</dc:creator>
<dc:date>2022-07-02</dc:date>
<dc:identifier>doi:10.1101/2022.06.30.498366</dc:identifier>
<dc:title><![CDATA[Senescent cell-derived extracellular vesicles recruit antigen presenting cells and limit squamous carcinoma recurrence]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-07-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.07.05.498896v1?rss=1">
<title>
<![CDATA[
Epigenetic biomarkers of autoimmune risk and protective antioxidant signaling in methylmercury-exposed adults 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.07.05.498896v1?rss=1</link>
<description><![CDATA[
I.BackgroundEpigenome-wide association studies (EWAS) are a highly promising approach that can inform precision environmental health. However, current EWAS are underpowered for biomarker detection and increasing sample sizes will require substantial resources. Therefore, alternative approaches for identifying candidate biomarkers through EWAS are critical for moving the field forward.

ObjectivesTo provide proof-of-principle that maximizing exposure variance in EWAS by selecting participants from disproportionately exposed global populations enables effective candidate biomarker detection, even in small sample sizes.

MethodsWe profiled genome-wide DNA methylation using Illumina Infinium MethylationEPIC BeadChip in whole blood from N=32 individuals from Madre de Dios, Peru with high methylmercury (MeHg) exposure due to artisanal and small-scale gold mining. We compared DNA methylation in N=16 individuals with high (>10 g/g) vs. N=16 individuals with low (<1 g/g) total hair mercury (a proxy for methylmercury exposure), matched on age and sex.

ResultsWe identified nine differentially methylated CpG sites (FDR<0.05), including several with known links to MeHg toxicity. The most significantly different CpG site was in an intronic enhancer of the SLC5A7 gene, which encodes the L-type amino acid transporter 1 (LAT1) that facilitates MeHg transport into protein-rich tissue, including muscle and brain. Our Gene Ontology and transcription factor motif enrichment analyses identified differential methylation of genes involved in several outcomes with established links to MeHg, including immune response, neurotoxicity, and type 2 diabetes (T2D) risk. Last, we identified candidate epigenetic biomarkers of PUFA-mediated protection against MeHg toxicity.

DiscussionHere, we show that a small EWAS on samples with high MeHg exposure variance can detect candidate differentially methylated CpGs and pathways of interest relevant to MeHg biology. Similar EWAS in global populations with known high exposure variance can be leveraged to develop targeted, custom sequencing panels and microarrays limited to replicated, validated biomarkers of a given exposure.
]]></description>
<dc:creator>Weinhouse, C.</dc:creator>
<dc:creator>Perez, L.</dc:creator>
<dc:creator>Ryde, I.</dc:creator>
<dc:creator>Goodrich, J. M.</dc:creator>
<dc:creator>Miranda, J. J.</dc:creator>
<dc:creator>Hsu-Kim, H.</dc:creator>
<dc:creator>Murphy, S. K.</dc:creator>
<dc:creator>Meyer, J. N.</dc:creator>
<dc:creator>Pan, W. K.</dc:creator>
<dc:date>2022-07-06</dc:date>
<dc:identifier>doi:10.1101/2022.07.05.498896</dc:identifier>
<dc:title><![CDATA[Epigenetic biomarkers of autoimmune risk and protective antioxidant signaling in methylmercury-exposed adults]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-07-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.07.11.499573v1?rss=1">
<title>
<![CDATA[
Delivery of loaded MR1 Monomer Results in Efficient Ligand Exchange to Host MR1 and Subsequent MR1T cell activation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.07.11.499573v1?rss=1</link>
<description><![CDATA[
MR1-restricted T cells have been implicated in microbial infections, sterile inflammation, wound healing and cancer. Similar to other antigen presentation molecules, evidence supports multiple, complementary MR1 antigen presentation pathways. To investigate ligand exchange pathways for MR1, we used MR1 monomers and tetramers loaded with 5-(2-oxopropylideneamino)-6-d-ribitylaminouracil (5-OP-RU) to deliver the antigen. Using MR1-deficient cells reconstituted with wild-type MR1 or MR1 molecules that cannot bind 5-OP-RU, we show that presentation of monomer-delivered 5-OP-RU is dependent on cellular MR1 and requires the transfer of ligand from the soluble molecule onto MR1 expressed by the antigen presenting cell. This mode of antigen delivery strengthens the evidence for post-ER ligand exchange pathways for MR1, which could represent an important avenue by which MR1 acquires antigens derived from endocytosed pathogens.
]]></description>
<dc:creator>Kulicke, C. A.</dc:creator>
<dc:creator>Swarbrick, G. M.</dc:creator>
<dc:creator>Ladd, N. A.</dc:creator>
<dc:creator>Cansler, M.</dc:creator>
<dc:creator>Null, M.</dc:creator>
<dc:creator>Worley, A.</dc:creator>
<dc:creator>Lemon, C.</dc:creator>
<dc:creator>Ahmed, T.</dc:creator>
<dc:creator>Bennett, J.</dc:creator>
<dc:creator>Lewinsohn, D. A.</dc:creator>
<dc:creator>Adams, E. J.</dc:creator>
<dc:creator>Lewinsohn, D. M.</dc:creator>
<dc:creator>Harriff, M. J.</dc:creator>
<dc:date>2022-07-12</dc:date>
<dc:identifier>doi:10.1101/2022.07.11.499573</dc:identifier>
<dc:title><![CDATA[Delivery of loaded MR1 Monomer Results in Efficient Ligand Exchange to Host MR1 and Subsequent MR1T cell activation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-07-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.07.11.499612v1?rss=1">
<title>
<![CDATA[
REM sleep has minute-scale rhythms in mice and humans: A non-binary continuum between phasic and tonic microstates 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.07.11.499612v1?rss=1</link>
<description><![CDATA[
Rapid eye movement sleep (REM) is believed to have a binary temporal structure with "phasic" and "tonic" microstates, defined by motoric activity versus quiescence, respectively. However, we observed in mice that the frequency of theta activity (a marker of rodent REM) fluctuates in a non-binary fashion, with the extremes of that fluctuation correlating with phasic-type and tonic-type facial motricity. This demonstrates that phasic and tonic REM rather represent ends of a continuum. These cycles of brain physiology and facial movement occurred at 0.01-0.06 Hz, or infraslow frequencies, and affected cross-frequency coupling and neuronal assembly activity in the neocortex, suggesting network functional impact. We then confirmed that humans also demonstrate non-binary phasic/tonic microstates, with continuous 0.01-0.04 Hz respiratory rate cycles matching the incidence of eye movements. Thus, we discovered a fundamental property of REM, which can yield new insights into our understanding of sleep health.
]]></description>
<dc:creator>Bueno-Junior, L. S.</dc:creator>
<dc:creator>Ruckstuhl, M. S.</dc:creator>
<dc:creator>Lim, M. M.</dc:creator>
<dc:creator>Watson, B. O.</dc:creator>
<dc:date>2022-07-13</dc:date>
<dc:identifier>doi:10.1101/2022.07.11.499612</dc:identifier>
<dc:title><![CDATA[REM sleep has minute-scale rhythms in mice and humans: A non-binary continuum between phasic and tonic microstates]]></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/10.1101/2022.07.12.499810v1?rss=1">
<title>
<![CDATA[
NudC regulated Lis1 stability is essential for maintenance of dynamic microtubule ends in the axon terminal 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.07.12.499810v1?rss=1</link>
<description><![CDATA[
Axon terminal structure is critical for neuronal function. This cellular compartment houses synaptic terminals and is a site of high metabolic and functional demand. Axon terminals are also the site of a change in microtubule structure within the neuron. Microtubule stability is decreased relative to the axon shaft due to an enrichment of microtubule plus ends and increase in microtubule dynamics. These dynamic microtubule plus ends have many functions including serving as a docking site for the microtubule motor protein complex Cytoplasmic dynein. Here, we report an unexplored function of the dynein motor in axon terminals: regulation of microtubule stability. Using a forward genetic screen, we identified a mutant with abnormal axon terminal structure due to a loss of function mutation in the dynein interacting protein NudC. We show that the primary function of NudC in the axon terminal is as a chaperone for the protein Lis1. Loss of NudC results in decreased Lis1 protein in this neuronal compartment. Decreased Lis1 in nudc mutants causes dynein/dynactin accumulation and increased microtubule stability in axon terminals. Microtubules in the proximal axon are unaffected. Abnormal microtubule stability and structure can be suppressed by pharmacologically inhibiting dynein, implicating excess dynein motor activity as causal in the enhanced axon terminal microtubule stability. Together, our data support a model in which local NudC-Lis1 modulation of dynein motor activity is critical for regulation of microtubule stability in the axon terminal.
]]></description>
<dc:creator>Kawano, D.</dc:creator>
<dc:creator>Pinter, K.</dc:creator>
<dc:creator>Chlebowski, M.</dc:creator>
<dc:creator>Petralia, R. S.</dc:creator>
<dc:creator>Wang, Y.-X.</dc:creator>
<dc:creator>Nechiporuk, A.</dc:creator>
<dc:creator>Drerup, C.</dc:creator>
<dc:date>2022-07-13</dc:date>
<dc:identifier>doi:10.1101/2022.07.12.499810</dc:identifier>
<dc:title><![CDATA[NudC regulated Lis1 stability is essential for maintenance of dynamic microtubule ends in the axon terminal]]></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/10.1101/2022.07.19.500665v1?rss=1">
<title>
<![CDATA[
Cas9+ conditionally immortalized neutrophil progenitors as a tool for genome wide CRISPR screening for neutrophil differentiation and function 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.07.19.500665v1?rss=1</link>
<description><![CDATA[
Neutrophils are short-lived cells of the innate immune system that play numerous roles in defense against infection, regulation of immune responses, tissue damage and repair, autoimmunity, and other non-communicable diseases. Understanding neutrophil function at a mechanistic level has been hampered by the difficulty of working with primary neutrophils, which die rapidly upon isolation, and the relative paucity of neutrophil cell lines. Murine neutrophil progenitors that are immortalized with estrogen-regulated expression of Hoxb8 differentiate into neutrophils upon withdrawal of estrogen and facilitate the quantitative production of neutrophils in vitro. Here we report the creation of a Cas9+ER-Hoxb8 neutrophil progenitor cell line that enables both forward and reverse genetic analysis of neutrophils for the first time. By editing progenitors via transduction with sgRNAs, and then withdrawing estrogen, Cas9 edited neutrophils are produced with high efficiency. Importantly, neutrophil differentiation of edited progenitors occurs both in vitro in cell culture and when transferred into murine recipients. To demonstrate the utility of Cas9+ER-Hoxb8 progenitors for forward genetics, we performed a pooled CRISPR screen to identify factors required for survival during neutrophil differentiation. This screen identified hundreds of genes both negatively and positively selected under differentiation conditions. One of the top hits from this screen was Cebpe, a transcription factor known to be required for neutrophil differentiation from pre-neutrophils to immature neutrophils. Using the progenitor cell line, we also confirmed that Cepbe is required for neutrophil differentiation in vivo, validating the utility of this cell line both for screening and for studying in vivo phenotypes. The genome-wide screen also identified all components of the WASH complex as being required for neutrophil differentiation, a finding that extends the known role of WASH in hematopoietic stem cell differentiation to later stages of neutrophil development. Taken together, we demonstrate that Cas9+ER-Hoxb8 immortalized neutrophils can be used to study neutrophil function both in vitro and in vivo. This new resource will enable the analysis of the role of neutrophils in numerous disease states using genetics for the first time.
]]></description>
<dc:creator>Jong, R. M.</dc:creator>
<dc:creator>Ching, K.</dc:creator>
<dc:creator>Garelis, N. E.</dc:creator>
<dc:creator>Zilinskas, A.</dc:creator>
<dc:creator>Nguyenla, X.</dc:creator>
<dc:creator>Rawal, S.</dc:creator>
<dc:creator>Hill, B.</dc:creator>
<dc:creator>Luckie, B.</dc:creator>
<dc:creator>Shallow, L.</dc:creator>
<dc:creator>Cox, J. S.</dc:creator>
<dc:creator>Barton, G.</dc:creator>
<dc:creator>Stanley, S.</dc:creator>
<dc:date>2022-07-20</dc:date>
<dc:identifier>doi:10.1101/2022.07.19.500665</dc:identifier>
<dc:title><![CDATA[Cas9+ conditionally immortalized neutrophil progenitors as a tool for genome wide CRISPR screening for neutrophil differentiation and function]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-07-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.07.20.500827v1?rss=1">
<title>
<![CDATA[
Gabapentin Disrupts Binding of Perlecan to the α2δ1 Voltage Sensitive Calcium Channel Subunit and Impairs Skeletal Mechanosensation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.07.20.500827v1?rss=1</link>
<description><![CDATA[
Our understanding of how osteocytes, the principal mechanosensors within bone, sense and perceive force remains unclear. Previous work identified "tethering elements" (TEs) spanning the pericellular space of osteocytes and transmitting mechanical information into biochemical signals. While we identified the heparan sulfate proteoglycan perlecan (PLN) as a component of these TEs, PLN must attach to the cell surface to induce biochemical responses. As voltage-sensitive calcium channels (VSCCs) are critical for bone mechanotransduction, we hypothesized that PLN binds the extracellular 2{delta}1 subunit of VSCCs to couple the bone matrix to the osteocyte membrane. Here, we showed co-localization of PLN and 2{delta}1 along osteocyte dendritic processes. Additionally, we quantified the molecular interactions between 2{delta}1 and PLN domains and demonstrated for the first time that 2{delta}1 strongly associates with PLN via its domain III. Furthermore, 2{delta}1 is the binding site for the commonly used pain drug, gabapentin (GBP), which is associated with adverse skeletal effects when used chronically. We found that GBP disrupts PLN::2{delta}1 binding in vitro, and GBP treatment in vivo results in impaired bone mechanosensation. Our work identified a novel mechanosensory complex within osteocytes composed of PLN and 2{delta}1, necessary for bone force transmission and sensitive to the drug GBP. This work provides insights into the mechanisms underlying mechanotransduction and will inform future studies to understand the mechanisms responsible for the negative effects of GBP on bone.
]]></description>
<dc:creator>Fernandez, P. C. R.</dc:creator>
<dc:creator>Wright, C. S.</dc:creator>
<dc:creator>Masterson, A. N.</dc:creator>
<dc:creator>Yi, X.</dc:creator>
<dc:creator>Tellman, T. V.</dc:creator>
<dc:creator>Bonteanu, A.</dc:creator>
<dc:creator>Rust, K.</dc:creator>
<dc:creator>Noonan, M. L.</dc:creator>
<dc:creator>White, K. E.</dc:creator>
<dc:creator>Lewis, K. J.</dc:creator>
<dc:creator>Sankar, U.</dc:creator>
<dc:creator>Hum, J. M.</dc:creator>
<dc:creator>Bix, G.</dc:creator>
<dc:creator>Wu, D.</dc:creator>
<dc:creator>Robling, A. G.</dc:creator>
<dc:creator>Sardar, R.</dc:creator>
<dc:creator>Farach-Carson, M. C.</dc:creator>
<dc:creator>Thompson, W. R.</dc:creator>
<dc:date>2022-07-21</dc:date>
<dc:identifier>doi:10.1101/2022.07.20.500827</dc:identifier>
<dc:title><![CDATA[Gabapentin Disrupts Binding of Perlecan to the α2δ1 Voltage Sensitive Calcium Channel Subunit and Impairs Skeletal Mechanosensation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-07-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.07.24.501297v1?rss=1">
<title>
<![CDATA[
Multi-Omics Binary Integration via Lasso Ensembles (MOBILE) for identification of context-specific networks and new regulatory mechanisms 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.07.24.501297v1?rss=1</link>
<description><![CDATA[
Cell phenotypes are dictated by both extra- and intra-cellular contexts, and robust identification of context-specific network features that control phenotypes remains challenging. Here, we developed a multi-omics data integration strategy called MOBILE (Multi-Omics Binary Integration via Lasso Ensembles) to nominate molecular features associated with specific cellular phenotypes. We applied this method to chromatin accessibility, mRNA, protein, and phospho-protein time course datasets and focus on two illustrative use cases after we show MOBILE could recover known biology. First, MOBILE nominated new mechanisms of interferon-{gamma} (IFN{gamma}) regulated PD-L1 expression, where analyses suggested, and literature supported that IFN{gamma}-controlled PD-L1 expression involves BST2, CLIC2, FAM83D, ACSL5, and HIST2H2AA3 genes. Second, we explored differences between the highly similar transforming growth factor-beta 1 (TGF{beta}1) and bone morphogenetic protein 2 (BMP2) and showed that differential cell size and clustering properties induced by TGF{beta}1, but not BMP2, were related to the laminin/collagen pathway activity. Given the ever-growing availability of multi-omics datasets, we envision that MOBILE will be broadly applicable to identify context-specific molecular features associated with cellular phenotypes.

Graphical Summary

O_FIG O_LINKSMALLFIG WIDTH=162 HEIGHT=200 SRC="FIGDIR/small/501297v1_ufig1.gif" ALT="Figure 1">
View larger version (76K):
org.highwire.dtl.DTLVardef@12fa3a5org.highwire.dtl.DTLVardef@a0b6a8org.highwire.dtl.DTLVardef@137efb7org.highwire.dtl.DTLVardef@15aec85_HPS_FORMAT_FIGEXP  M_FIG C_FIG Multi-Omics Binary Integration via Lasso Ensembles (MOBILE) pipeline yields statistically robust, context-specific association networksThe MOBILE pipeline integrates omics datasets in a data-driven, biologically-structured manner.

The pipeline outputs are gene-level, contextspecific association networks.

These association networks nominate differentially enriched pathways, subnetworks, and new connections.

Broadly applicable to find condition specific networks using multi-omics datasets.
]]></description>
<dc:creator>Erdem, C.</dc:creator>
<dc:creator>Gross, S. M.</dc:creator>
<dc:creator>Heiser, L.</dc:creator>
<dc:creator>Birtwistle, M. R.</dc:creator>
<dc:date>2022-07-24</dc:date>
<dc:identifier>doi:10.1101/2022.07.24.501297</dc:identifier>
<dc:title><![CDATA[Multi-Omics Binary Integration via Lasso Ensembles (MOBILE) for identification of context-specific networks and new regulatory mechanisms]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-07-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.07.25.501380v1?rss=1">
<title>
<![CDATA[
Trophoblast glycoprotein is required for efficient synaptic vesicle exocytosis from retinal rod bipolar cells. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.07.25.501380v1?rss=1</link>
<description><![CDATA[
Rod bipolar cells (RBCs) faithfully transmit light-driven signals from rod photoreceptors in the outer retina to third order neurons in the inner retina. Recently, significant work has focused on the role of leucine-rich repeat (LRR) proteins in synaptic development and signal transduction at RBC synapses. We previously identified trophoblast glycoprotein (TPBG) as a novel transmembrane LRR protein localized to the dendrites and axon terminals of RBCs. We now examine the effects on RBC physiology and retinal processing of TPBG genetic knockout in mice using immunofluorescence and electrophysiological techniques. The scotopic electroretinogram showed a modest increase in the b-wave and a marked attenuation in oscillatory potentials in the TPBG knockout. No effect of TPBG knockout was observed on the RBC dendritic morphology, TRPM1 currents, or RBC excitability. Because scotopic oscillatory potentials primarily reflect RBC-driven rhythmic activity of the inner retina, we investigated the contribution of TPBG to downstream transmission from RBCs to third-order neurons. Using electron microscopy, we found shorter synaptic ribbons in TPBG knockout axon terminals in RBCs. Time-resolved capacitance measurements indicated that TPBG knockout reduces synaptic vesicle exocytosis and subsequent GABAergic reciprocal feedback without altering voltage-gated Ca2+ currents. Thus, TPBG is required for normal synaptic ribbon development and efficient neurotransmitter release from RBCs to downstream cells. Our results highlight a novel synaptic role for TPBG at RBC ribbon synapses and support further examination into the mechanisms by which TPBG regulates RBC physiology.

Graphical abstract

O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=113 SRC="FIGDIR/small/501380v2_ufig1.gif" ALT="Figure 1">
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org.highwire.dtl.DTLVardef@13d5916org.highwire.dtl.DTLVardef@a3c259org.highwire.dtl.DTLVardef@12c74e0org.highwire.dtl.DTLVardef@1ad43a8_HPS_FORMAT_FIGEXP  M_FIG C_FIG
]]></description>
<dc:creator>Wakeham, C. M.</dc:creator>
<dc:creator>Shi, Q.</dc:creator>
<dc:creator>Ren, G.</dc:creator>
<dc:creator>Haley, T. L.</dc:creator>
<dc:creator>Duvoisin, R.</dc:creator>
<dc:creator>von Gersdorff, H.</dc:creator>
<dc:creator>Morgans, C. W.</dc:creator>
<dc:date>2022-07-26</dc:date>
<dc:identifier>doi:10.1101/2022.07.25.501380</dc:identifier>
<dc:title><![CDATA[Trophoblast glycoprotein is required for efficient synaptic vesicle exocytosis from retinal rod bipolar cells.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-07-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.07.28.501447v1?rss=1">
<title>
<![CDATA[
JBrowse 2: A modular genome browser with views of synteny and structural variation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.07.28.501447v1?rss=1</link>
<description><![CDATA[
We present JBrowse 2, a general-purpose genome annotation browser offering enhanced visualization of complex structural variation and evolutionary relationships. JBrowse 2 retains the core features of the open-source JavaScript genome browser JBrowse while adding new views for synteny, dotplots, breakpoints, gene fusions, and whole-genome overviews. The software readily allows users to share sessions, open multiple genomes or views, and navigate quickly between these views. It can be embedded in a web page, used as a standalone desktop application, or run from Jupyter notebooks or R sessions. Using a plugin framework, developers can create new data adapters, track types, and visualizations. These improvements are enabled by a ground-up redesign of the JBrowse architecture using modern web technology. We describe application functionality, use cases, performance benchmarks, and implementation notes for web administrators and developers.
]]></description>
<dc:creator>Diesh, C.</dc:creator>
<dc:creator>Stevens, G. J.</dc:creator>
<dc:creator>Xie, P.</dc:creator>
<dc:creator>Martinez, T. D. J.</dc:creator>
<dc:creator>Hershberg, E. A.</dc:creator>
<dc:creator>Leung, A.</dc:creator>
<dc:creator>Guo, E.</dc:creator>
<dc:creator>Dider, S.</dc:creator>
<dc:creator>Zhang, J.</dc:creator>
<dc:creator>Bridge, C.</dc:creator>
<dc:creator>Hogue, G.</dc:creator>
<dc:creator>Duncan, A.</dc:creator>
<dc:creator>Morgan, M.</dc:creator>
<dc:creator>Flores, T.</dc:creator>
<dc:creator>Bimber, B. N.</dc:creator>
<dc:creator>Haw, R.</dc:creator>
<dc:creator>Cain, S.</dc:creator>
<dc:creator>Buels, R. M.</dc:creator>
<dc:creator>Stein, L. D.</dc:creator>
<dc:creator>Holmes, I.</dc:creator>
<dc:date>2022-07-31</dc:date>
<dc:identifier>doi:10.1101/2022.07.28.501447</dc:identifier>
<dc:title><![CDATA[JBrowse 2: A modular genome browser with views of synteny and structural variation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-07-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.07.29.502041v1?rss=1">
<title>
<![CDATA[
Optimal drug treatment for reducing long-term drug resistance 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.07.29.502041v1?rss=1</link>
<description><![CDATA[
The maximum-tolerated dose principle, the highest possible drug dose in the shortest possible time period, has been the standard care for cancer treatment. Although it is appealing in a homogeneous tumor settings, tumor heterogeneity and adaptation play a significant role in driving treatment failure. They are still major obstacles in cancer treatments despite great advances in modeling and cancer therapy using optimal control theory. To address this, we first generalize two population models and examine the long-term effects of differential selective treatment strategies. Second, we take into account different drug-imposed selective pressure into designing optimal treatment strategies. Numerical examples demonstrate that the proposed treatment strategy decreases long-term tumor burden by decreasing the rate of tumor adaptation.
]]></description>
<dc:creator>Ghodsi Asnaashari, T.</dc:creator>
<dc:creator>Chang, Y. H.</dc:creator>
<dc:date>2022-08-02</dc:date>
<dc:identifier>doi:10.1101/2022.07.29.502041</dc:identifier>
<dc:title><![CDATA[Optimal drug treatment for reducing long-term drug resistance]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-08-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.07.29.502048v1?rss=1">
<title>
<![CDATA[
De novo assembly and annotation of the singing mouse genome 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.07.29.502048v1?rss=1</link>
<description><![CDATA[
BackgroundDeveloping genomic resources for a diverse range of species is an important step towards understanding the mechanisms underlying complex traits.Specifically, organisms that exhibit unique, accessible phenotypes-of-interests, allow researchers to address questions that may be ill-suited to traditional model organisms. We sequenced the genome and transcriptome of Alstons singing mouse (Scotinomys teguina), an emerging model for social cognition and vocal communication. In addition to producing advertisement songs used for mate attraction and male-male competition, these rodents are diurnal, live at high-altitudes, and are obligate insectivores, providing opportunities to explore diverse physiological, ecological, and evolutionary questions.

ResultsUsing PromethION, Illumina, and PacBio sequencing, we produced an annotated genome and transcriptome, which were validated using gene expression and functional enrichment analyses. To assess the usefulness of our assemblies, we performed single nuclei sequencing on cells of the orofacial motor cortex, a brain region implicated in song coordination, identifying 12 cell types.

ConclusionsThese resources will provide the opportunity to identify the molecular basis of complex traits in singing mice as well as to contribute data that can be used for large-scale comparative analyses.
]]></description>
<dc:creator>Smith, S. K.</dc:creator>
<dc:creator>Frazel, P. W.</dc:creator>
<dc:creator>Khodadadi-Jamayran, A.</dc:creator>
<dc:creator>Zappile, P.</dc:creator>
<dc:creator>Marier, C.</dc:creator>
<dc:creator>Okhovat, M.</dc:creator>
<dc:creator>Brown, S. M.</dc:creator>
<dc:creator>Long, M. A.</dc:creator>
<dc:creator>Heguy, A.</dc:creator>
<dc:creator>Phelps, S. M.</dc:creator>
<dc:date>2022-07-29</dc:date>
<dc:identifier>doi:10.1101/2022.07.29.502048</dc:identifier>
<dc:title><![CDATA[De novo assembly and annotation of the singing mouse genome]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-07-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.08.01.502019v1?rss=1">
<title>
<![CDATA[
A tumour-promoting senescent secretome triggered by platinum chemotherapy exploits a targetable TGFβR1/Akt-mTOR axis in lung cancer 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.08.01.502019v1?rss=1</link>
<description><![CDATA[
Platinum-based chemotherapy is commonly used for non-small cell lung cancer (NSCLC) treatment, yet clinical outcomes remain poor. Cellular senescence and its associated secretory phenotype (SASP) can have multiple tumour-promoting activities, although these are largely unexplored in lung cancer. Here we show that cisplatin-derived SASP enhances the malignant phenotype of lung cancer cells. Using xenograft, orthotopic and KrasG12V-driven murine NSCLC models, we demonstrate that cisplatin-induced senescent cells strongly promote tumour progression. Mechanistically, we find that a TGF-{beta}-enriched SASP drives pro-proliferative effects through TGF{beta}R1 and Akt/mTOR pathway activation. We validate the translational relevance of chemotherapy-induced SASP using clinical NSCLC samples from patients who received neoadjuvant platinum-based chemotherapy. Importantly, TGF{beta}R1 inhibition with galunisertib or senolytic treatment significantly reduces tumour promotion driven by cisplatin-induced senescence. Finally, we demonstrate, using distinct murine NSCLC models, that addition of TGFBR1 inhibitors to platinum-based chemotherapy reduces tumour burden and improves survival, providing pre-clinical proof-of-concept for future trial designs.
]]></description>
<dc:creator>Gonzalez-Gualda, E.</dc:creator>
<dc:creator>Macias, D.</dc:creator>
<dc:creator>Morsli, S.</dc:creator>
<dc:creator>Martin, E.</dc:creator>
<dc:creator>Ou, H.-L.</dc:creator>
<dc:creator>Denholm, M.</dc:creator>
<dc:creator>Olan, I.</dc:creator>
<dc:creator>Hoffmann, R.</dc:creator>
<dc:creator>Dane, M.</dc:creator>
<dc:creator>Veroutis, D.</dc:creator>
<dc:creator>Medrano, G.</dc:creator>
<dc:creator>Mulero, F.</dc:creator>
<dc:creator>Martins, C.</dc:creator>
<dc:creator>Barbacid, M.</dc:creator>
<dc:creator>Gorgoulis, V.</dc:creator>
<dc:creator>Korkola, J.</dc:creator>
<dc:creator>Rassl, D. M.</dc:creator>
<dc:creator>Doherty, G. J.</dc:creator>
<dc:creator>Rintoul, R. C.</dc:creator>
<dc:creator>Narita, M.</dc:creator>
<dc:creator>Munoz-Espin, D.</dc:creator>
<dc:date>2022-08-03</dc:date>
<dc:identifier>doi:10.1101/2022.08.01.502019</dc:identifier>
<dc:title><![CDATA[A tumour-promoting senescent secretome triggered by platinum chemotherapy exploits a targetable TGFβR1/Akt-mTOR axis in lung cancer]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-08-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.08.01.502088v1?rss=1">
<title>
<![CDATA[
Mutational screens highlight glycosylation as a modulator of CSF3R activity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.08.01.502088v1?rss=1</link>
<description><![CDATA[
The colony-stimulating factor 3 receptor (CSF3R) controls the growth of neutrophils, the most abundant type of white blood cell. In healthy neutrophils, signaling is dependent on CSF3R binding to its ligand CSF3. A single amino acid mutation in CSF3R, T618I, instead allows for constitutive, ligand-independent cell growth and leads to a rare type of cancer called chronic neutrophilic leukemia (CNL). We investigated why this threonine to isoleucine substitution is the predominant mutation in CNL and how it leads to uncontrolled neutrophil growth. Using protein domain mapping, we demonstrated that the single CSF3R domain containing residue 618 is sufficient for ligand-independent activity. We then applied an unbiased mutational screening strategy focused on this domain and found that activating mutations are enriched at sites normally occupied by asparagine, threonine, and serine residues - the three amino acids which are commonly glycosylated. We confirmed glycosylation at multiple CSF3R residues by mass spectrometry, including the presence of GalNAc and Gal-GalNAc glycans at wild-type threonine 618. Using the same approach applied to other cell surface receptors, we identified an activating mutation, S489F, in the interleukin-31 receptor alpha chain (IL-31R). Combined, these results suggest a role for glycosylated hotspot residues in regulating receptor signaling, mutation of which can lead to ligand-independent, uncontrolled activity.
]]></description>
<dc:creator>Hollander, M. J.</dc:creator>
<dc:creator>Malaker, S. A.</dc:creator>
<dc:creator>Perez, I.</dc:creator>
<dc:creator>Maxson, J. E.</dc:creator>
<dc:creator>Cochran, J. R.</dc:creator>
<dc:creator>Bertozzi, C. R.</dc:creator>
<dc:date>2022-08-01</dc:date>
<dc:identifier>doi:10.1101/2022.08.01.502088</dc:identifier>
<dc:title><![CDATA[Mutational screens highlight glycosylation as a modulator of CSF3R activity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-08-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.08.05.502959v1?rss=1">
<title>
<![CDATA[
Persistent effects of acute trauma on Pavlovian-to-instrumental transfer 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.08.05.502959v1?rss=1</link>
<description><![CDATA[
In humans, an acutely traumatic experience can lead to post-traumatic stress disorder (PTSD), which is often characterized by changes in anxiety and motivation months after trauma. There are few demonstrations of the persistent motivational effects of an acute stressor in rodent approaches to PTSD. In two experiments, we evaluated the persistent effects of a battery of footshocks in one context on appetitive Pavlovian conditioning, instrumental learning, and Pavlovian-to-instrumental transfer (PIT) in a different context. A battery of footshocks before appetitive training caused deficits in single-outcome PIT in male Long Evans rats. The same battery of footshocks after appetitive training, but before testing had little effect on single-outcome PIT, but there were some qualitative deficits. Although males showed more generalized fear from the aversive to the appetitive context compared to females, there were no effects of shock on sensory-specific PIT in male or female rats. In general females showed less evidence for sensory-specific PIT compared to males, who showed robust sensory-specific PIT, with clear extinction and spontaneous recovery of the sensory-specific PIT effect across test sessions. These findings show that (a) an acute trauma can have persistent effects on general motivational processes and (b) sensory-specific PIT is a useful approach for exploring sex differences in strategies for instrumental learning. We discuss implications for current approaches to stress and motivation in preclinical and clinical studies.
]]></description>
<dc:creator>Derman, R. C.</dc:creator>
<dc:creator>Lattal, M.</dc:creator>
<dc:date>2022-08-06</dc:date>
<dc:identifier>doi:10.1101/2022.08.05.502959</dc:identifier>
<dc:title><![CDATA[Persistent effects of acute trauma on Pavlovian-to-instrumental transfer]]></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/10.1101/2022.08.07.503091v1?rss=1">
<title>
<![CDATA[
Multiplexed and millimeter-scale superresolution imaging of cells and tissue sections via prism-illumination and microfluidics-enhanced DNA-PAINT 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.08.07.503091v1?rss=1</link>
<description><![CDATA[
Fluorescence nanoscopy has become increasingly powerful for biomedical research, but it has historically afforded a small field-of-view (FOV) around 50 {micro}m x 50 {micro}m at once and more recently up to ~200 {micro}m x 200 {micro}m. Efforts to further increase the FOV in fluorescence nanoscopy have thus far relied on the use of fabricated waveguide substrates, adding cost and sample constraints on the applications. Here we report PRism-Illumination and Microfluidics-Enhanced DNA-PAINT (PRIME-PAINT) for multiplexed fluorescence nanoscopy across millimeter-scale FOVs. Built upon the well-established prism-type total internal reflection microscopy, PRIME-PAINT achieves robust single-molecule localization with up to ~520 {micro}m x 520 {micro}m single FOVs and 25-40 nm lateral resolutions. Through stitching, nanoscopic imaging over mm2 sample areas can be completed in as little as 40 minutes per target. An on-stage microfluidics chamber facilitates probe exchange for multiplexing and enhances image quality particularly for formalin-fixed paraffin-embedded (FFPE) tissue sections. We demonstrate the utility of PRIME-PAINT by analyzing ~106 caveolae structures in ~1,000 cells and imaging entire pancreatic cancer lesions from patient tissue biopsies. By imaging from nanometers to millimeters with multiplexity and broad sample compatibility, PRIME-PAINT will be useful for building multiscale, Google-Earth-like views of biological systems.
]]></description>
<dc:creator>Rames, M. J.</dc:creator>
<dc:creator>Kenison, J.</dc:creator>
<dc:creator>Heineck, D.</dc:creator>
<dc:creator>Civitci, F.</dc:creator>
<dc:creator>Szczepaniak, M.</dc:creator>
<dc:creator>Tao, K.</dc:creator>
<dc:creator>Zheng, T.</dc:creator>
<dc:creator>Shangguan, J.</dc:creator>
<dc:creator>Esener, S.</dc:creator>
<dc:creator>Nan, X.</dc:creator>
<dc:date>2022-08-07</dc:date>
<dc:identifier>doi:10.1101/2022.08.07.503091</dc:identifier>
<dc:title><![CDATA[Multiplexed and millimeter-scale superresolution imaging of cells and tissue sections via prism-illumination and microfluidics-enhanced DNA-PAINT]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-08-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.08.09.503379v1?rss=1">
<title>
<![CDATA[
Follicle-innervating Aδ-low threshold mechanoreceptors organize through a population-dependent mechanism 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.08.09.503379v1?rss=1</link>
<description><![CDATA[
The mammalian somatosensory system is comprised of multiple neuronal populations that form specialized, highly organized sensory endings in the skin. The organization of somatosensory endings is essential to their functions, yet the mechanisms which regulate this organization remain unclear. Using a combination of genetic and molecular labeling approaches, we examined the development of mouse hair follicle-innervating low-threshold mechanoreceptors (LTMRs) and explored competition for innervation targets as a mechanism involved in the patterning of their receptive fields. We show that follicle innervating neurons are present in the skin at birth and that LTMR receptive fields gradually add follicle-innervating endings during the first two postnatal weeks. Using a constitutive Bax knockout to increase the number of neurons in adult animals, we show that two LTMR subtypes have differential responses to an increase in neuronal population size: A{delta}-LTMR neurons shrink their receptive fields to accommodate the increased number of neurons innervating the skin, while C-LTMR neurons do not. Our findings suggest that competition for hair follicles to innervate plays a role in the patterning and organization of follicle-innervating LTMR neurons.

Summary StatementA{delta} follicle-innervating low-threshold mechanoreceptor neurons form tiled receptive fields through competition for hair follicles during the early postnatal period.
]]></description>
<dc:creator>Pomaville, M. B.</dc:creator>
<dc:creator>Wright, K. M.</dc:creator>
<dc:date>2022-08-10</dc:date>
<dc:identifier>doi:10.1101/2022.08.09.503379</dc:identifier>
<dc:title><![CDATA[Follicle-innervating Aδ-low threshold mechanoreceptors organize through a population-dependent mechanism]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-08-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.08.12.503665v1?rss=1">
<title>
<![CDATA[
Anandamide reduces excitability by preferentially targeting somatic VGSCs via CB1 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.08.12.503665v1?rss=1</link>
<description><![CDATA[
Endogenous cannabinoid signaling is vital for important brain functions and can be modified pharmacologically to treat pain, epilepsy, and posttraumatic stress disorder. Endocannabinoid mediated changes to excitability are predominantly attributed to 2-arachidonoylglycerol at synapses. Here we identify a pathway in the neocortex by which anandamide, the other major endocannabinoid, powerfully inhibits sodium conductances in the soma resulting in a loss of neuronal excitability. This pathway is mediated by the cannabinoid receptor, and its activation results in a decrease of recurrent action potential generation. The synthetic cannabinoid, WIN 55,212-2, also inhibits VGSC currents indicating this pathway is positioned to mediate the actions of exogenous cannabinoids.

HighlightsAnandamide (AEA), a major endocannabinoid, indirectly inhibits VGSC currents in neocortical neurons.

This prevalent signaling pathway involves AEA activation of CB1 and other G-protein-coupled receptors localized to the intracellular compartment of neurons.

CB1 activation by AEA reduces VGSC availability at the soma but not at the axonal compartment suggesting tighter functional coupling between VGSCs and CB1 at the cell body.

Cannabinoid action on somatic CB1 inhibits VGSCs with high efficacy, providing a parallel pathway outside of nerve terminals, by which these ligands reduce neuronal excitability in the neocortex.
]]></description>
<dc:creator>Steiger, L. J.</dc:creator>
<dc:creator>Tsintsadze, T.</dc:creator>
<dc:creator>Mattheisen, G. B.</dc:creator>
<dc:creator>Smith, S. M.</dc:creator>
<dc:date>2022-08-14</dc:date>
<dc:identifier>doi:10.1101/2022.08.12.503665</dc:identifier>
<dc:title><![CDATA[Anandamide reduces excitability by preferentially targeting somatic VGSCs via CB1]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-08-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.08.15.504002v1?rss=1">
<title>
<![CDATA[
Inactivated voltage-gated sodium channels are preferentially targeted in cinacalcet block of mouse neocortical action potentials 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.08.15.504002v1?rss=1</link>
<description><![CDATA[
Voltage-gated sodium channel (VGSC) activation is essential for action potential generation in the brain. Allosteric calcium-sensing receptor (CaSR) agonist, cinacalcet, strongly and ubiquitously inhibits VGSC currents in neocortical neurons via an unidentified, G-protein-dependent blocking molecule. The mechanisms by which VGSC characteristics influence cinacalcet-mediated inhibition are not well understood. Here, using whole-cell patch clamp methods, we investigated the voltage-dependence of cinacalcet-mediated inhibition of VGSCs and the channel state preference of cinacalcet. The rate of inhibition of VGSC currents was accelerated at more depolarized holding potentials. Cinacalcet shifted the voltage-dependence of both fast and slow inactivation of VGSCs in the hyperpolarizing direction. Utilizing a simple model, the voltage-dependence of VGSC current inhibition may be explained if the affinity of the blocking molecule to the channel states follows the sequence: fast-inactivated > slow-inactivated > resting. The state dependence of block contributes to the non-linearity of action potential block by cinacalcet. This dynamic and abundant signaling pathway by which G-proteins regulate VGSC currents provides an important voltage-dependent mechanism for modulating central neuronal excitability.

Key points summaryVoltage-gated sodium channels are essential for the action potential generation and propagation that is central to physiological function in excitable cells. VGSC inhibitors are useful therapies to treat epilepsy, pain, and cardiac arrhythmias.

Cinacalcet inhibits VGSC currents strongly and the underlying G-protein dependent signaling pathway occurs in the vast majority of neocortical and hippocampal neurons.

Here we demonstrate that cinacalcet inhibits the VGSC current by activating a downstream blocking molecule that preferentially binds to the fast-inactivated state, that the blocking molecule stabilizes the inactivated states, and that cinacalcet impacts neuronal excitability in a non-linear manner.

Characterization of the mechanism by which cinacalcet operates will facilitate the determination of its role in regulating neocortical excitability and the identification of new therapeutic targets for epilepsy, pain, and arrythmias.
]]></description>
<dc:creator>Lindner, J. S.</dc:creator>
<dc:creator>Rajayer, S. R.</dc:creator>
<dc:creator>Smith, S. M.</dc:creator>
<dc:date>2022-08-15</dc:date>
<dc:identifier>doi:10.1101/2022.08.15.504002</dc:identifier>
<dc:title><![CDATA[Inactivated voltage-gated sodium channels are preferentially targeted in cinacalcet block of mouse neocortical action potentials]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-08-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.08.18.504417v1?rss=1">
<title>
<![CDATA[
The pharmacokinetics and pharmacodynamics of 4-methylumbelliferone and its glucuronide metabolite in mice 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.08.18.504417v1?rss=1</link>
<description><![CDATA[
Hyaluronan (HA) is an extracellular matrix glycosaminoglycan, with important roles in chronic inflammation, cancer and autoimmunity. 4-methylumbelliferone (4-MU), a small molecule inhibitor of HA synthases, is widely used to study HAs interactions with the surrounding tissues and the immune cells. There is substantial experimental and therapeutic interest in using oral 4-MU to inhibit HA synthesis, but pharmacokinetic and pharmacodynamic data on treatment routes have been lacking. Moreover, it recently became clear that the main metabolite of 4-MU, 4-methlyumbelliferyl glucuronide (4-MUG), is bioactive. We therefore sought to define the pharmacokinetics and pharmacodynamics of 4-MU and its active metabolite 4-MUG in mice. Single dose mouse studies showed that 4-MU administered intravenously (i.v.) resulted in 100-fold higher 4-MU exposure compared to oral (p.o.) administration. The 4-MU ratio AUC i.v./AUC p.o. was 96/1. 4-MUG exposures were much higher than 4-MU exposures after both 4-MU i.v. and p.o. administration, but only small differences in 4-MUG exposure were seen after 4-MU i.v. versus p.o. administration. The 4-MUG metabolite was also administered as a single dose both i.v. and p.o. and showed a 25.9% bioavailability. Compared to 4-MUG p.o. dosing, 1.14 higher 4-MUG exposures were seen after 4-MU p.o. dosing. 4-MU exposure after 4-MUG p.o. administration was minimal but similar to 4-MU exposure after 4-MU p.o. administration. In mice treated for several weeks with 4-MU in chow, the 4-MU concentration immediately drops after treatment was stopped, whereas the 4-MUG concentration showed a peak 1 hour after treatment stop. In a build-up study, 4-MU and 4-MUG treatment in mice lead to a plateau of 4-MU concentration starting at 4 days post treatment start. These 4-MU and 4-MUG concentration findings in vivo will inform future clinical studies and experimental work with 4-MU.
]]></description>
<dc:creator>Nagy, N.</dc:creator>
<dc:creator>Kaber, G.</dc:creator>
<dc:creator>Haddock, N.</dc:creator>
<dc:creator>Hargil, A.</dc:creator>
<dc:creator>Rajadas, J.</dc:creator>
<dc:creator>Malhotra, S.</dc:creator>
<dc:creator>Unger, M.</dc:creator>
<dc:creator>Frymoyer, A.</dc:creator>
<dc:creator>Bollyky, P.</dc:creator>
<dc:date>2022-08-18</dc:date>
<dc:identifier>doi:10.1101/2022.08.18.504417</dc:identifier>
<dc:title><![CDATA[The pharmacokinetics and pharmacodynamics of 4-methylumbelliferone and its glucuronide metabolite in mice]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-08-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.08.18.504436v1?rss=1">
<title>
<![CDATA[
A Web-based Software Resource for Interactive Analysis of Multiplex Tissue Imaging Datasets 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.08.18.504436v1?rss=1</link>
<description><![CDATA[
Highly multiplexed tissue imaging (MTI) are powerful spatial proteomics technologies that enable in situ single-cell characterization of tissues. However, analysis and visualization of MTI datasets remains challenging, and we developed the Galaxy-ME software hub to address this challenge.Galaxy-ME is a web-based, interactive software hub that enables end-to-end analysis and visualization of MTI datasets and is accessible to everyone. To demonstrate its utility, Galaxy-ME was used to analyze datasets obtained from multiple MTI assays and evaluate assay concordance in both normal and cancerous tissues. Galaxy-ME is a publicly available web resource.
]]></description>
<dc:creator>Creason, A. L.</dc:creator>
<dc:creator>Watson, C.</dc:creator>
<dc:creator>Gu, Q.</dc:creator>
<dc:creator>Persson, D.</dc:creator>
<dc:creator>Sargent, L. L.</dc:creator>
<dc:creator>Chen, Y.-A.</dc:creator>
<dc:creator>Lin, J.-R.</dc:creator>
<dc:creator>Sivagnanam, S.</dc:creator>
<dc:creator>Wünnemann, F.</dc:creator>
<dc:creator>Nirmal, A. J.</dc:creator>
<dc:creator>Chin, K.</dc:creator>
<dc:creator>Feiler, H. S.</dc:creator>
<dc:creator>Coussens, L. M.</dc:creator>
<dc:creator>Schapiro, D.</dc:creator>
<dc:creator>Grüning, B. A.</dc:creator>
<dc:creator>Sorger, P. K.</dc:creator>
<dc:creator>Sokolov, A.</dc:creator>
<dc:creator>Goecks, J.</dc:creator>
<dc:date>2022-08-19</dc:date>
<dc:identifier>doi:10.1101/2022.08.18.504436</dc:identifier>
<dc:title><![CDATA[A Web-based Software Resource for Interactive Analysis of Multiplex Tissue Imaging Datasets]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-08-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.08.22.504741v1?rss=1">
<title>
<![CDATA[
Cortical Betz cells analogue in songbirds utilizes Kv3.1 to generate ultranarrow spikes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.08.22.504741v1?rss=1</link>
<description><![CDATA[
Complex motor skills in vertebrates require specialized upper motor neurons with precise action potential (AP) firing. To examine how diverse populations of upper motor neurons subserve distinct functions and the specific repertoire of ion channels involved, we conducted a thorough study of the excitability of upper motor neurons controlling somatic motor function in the zebra finch. We found that robustus arcopallialis projection neurons (RAPNs), key command neurons for song production, exhibit ultranarrow spikes and higher firing rates compared to neurons controlling non-vocal somatic motor functions (AId neurons). This striking difference was primarily due to the expression of a high threshold, fast-activating voltage-gated K+ channel, Kv3.1 (KCNC1). RAPN properties thus mirror those of the sparse, specialized Betz cells in the motor cortex of humans and other primates, which also fire ultranarrow spikes enabled by Kv3.1 expression. These large layer 5 pyramidal neurons are involved in fine digit control and are notably absent in rodents. Our study thus provides evidence that songbird RAPNs and primate Betz cells have convergently evolved the use of Kv3.1 to ensure precise, rapid AP firing required for fast and complex motor skills.
]]></description>
<dc:creator>Zemel, B.</dc:creator>
<dc:creator>Nevue, A.</dc:creator>
<dc:creator>Tavares, L. E. S.</dc:creator>
<dc:creator>Dadostin, A.</dc:creator>
<dc:creator>Lovell, P.</dc:creator>
<dc:creator>Jin, D. Z.</dc:creator>
<dc:creator>Mello, C.</dc:creator>
<dc:creator>von Gersdorff, H.</dc:creator>
<dc:date>2022-08-22</dc:date>
<dc:identifier>doi:10.1101/2022.08.22.504741</dc:identifier>
<dc:title><![CDATA[Cortical Betz cells analogue in songbirds utilizes Kv3.1 to generate ultranarrow spikes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-08-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.08.24.505142v1?rss=1">
<title>
<![CDATA[
Computational Multiplex Panel Reduction to Maximize Information Retention in Breast Cancer Tissue Microarrays 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.08.24.505142v1?rss=1</link>
<description><![CDATA[
Recent state-of-the-art multiplex imaging techniques have expanded the depth of information that can be captured within a single tissue sample by allowing for panels with dozens of markers. Despite this increase in capacity, space on the panel is still limited due to technical artifacts, tissue loss, and long imaging acquisition time. As such, selecting which markers to include on a panel is important, since removing important markers will result in a loss of biologically relevant information, but identifying redundant markers will provide a room for other markers. To address this, we propose computational approaches to determine the amount of shared information between markers and select an optimally reduced panel that captures maximum amount of information with the fewest markers. Here we examine several panel selection approaches and evaluate them based on their ability to reconstruct the full panel images and information within breast cancer tissue microarray datasets using cyclic immunofluorescence as a proof of concept. We show that all methods perform adequately and can re-capture cell types using only 18 of 25 markers (72% of the original panel size). The correlation-based selection methods achieved the best single-cell marker mean intensity predictions with a Spearman correlation of 0.90 with the reduced panel. Using the proposed methods shown here, it is possible for researchers to design more efficient multiplex imaging panels that maximize the amount of information retained with the limited number of markers with respect to certain evaluation metrics and architecture biases.

Author SummaryMultiplex tissue imaging techniques utilize large panels of markers that attempt to gather as much information as possible, but increasing the number of stains does come with the downsides of increased autofluorescence and tissue degradation. There exists a theoretical subsampling of markers that is able to recreate the same information as a full panel; therefore, removing the self-correlating information with such a subset would increase the efficiency of the imaging process and maximize the information collected. By selecting an idealized subsample of markers, a deep learning model can be trained to predict the same information as a full dataset with fewer rounds of staining. Here we evaluate several methods of subsample marker selection and demonstrate their ability to reconstruct the full panels information.
]]></description>
<dc:creator>Ternes, L.</dc:creator>
<dc:creator>Gray, J. W.</dc:creator>
<dc:creator>Chang, Y. H.</dc:creator>
<dc:date>2022-08-26</dc:date>
<dc:identifier>doi:10.1101/2022.08.24.505142</dc:identifier>
<dc:title><![CDATA[Computational Multiplex Panel Reduction to Maximize Information Retention in Breast Cancer Tissue Microarrays]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-08-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.08.26.505400v1?rss=1">
<title>
<![CDATA[
Alcohol dependence modifies brain networks activated during abstinence and reaccess: a c-fos-based analysis in mice 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.08.26.505400v1?rss=1</link>
<description><![CDATA[
High-level alcohol consumption causes neuroplastic changes in the brain that lead to negative affective and somatic symptoms when alcohol is withdrawn, promoting relapse drinking. We have some understanding of these plastic changes in defined brain circuits and cell types, but unbiased approaches are needed to explore broader patterns of adaptations. Here, we employed whole-brain c-fos mapping and network analysis to assess how brain-wide patterns of neuronal activity are altered during acute alcohol abstinence and reaccess in a well-characterized model of alcohol dependence. Mice underwent four cycles of chronic intermittent ethanol vapor exposure (CIE) with alternating weeks of voluntary alcohol drinking, and a subset of mice underwent forced swim stress (FSS) prior to drinking sessions to further escalate alcohol consumption. After four CIE cycles, brains were collected from mice in each group either 24 hours (abstinence) or immediately following a one-hour period of alcohol reaccess. Brains from CIE mice during acute abstinence displayed widespread neuronal activation relative to those from AIR mice, independent of FSS, and this increase in c-fos was reversed by reaccess drinking. For network analysis, mice were then classified as high or low drinkers (HD or LD). We computed Pearson correlations for all pairs of brain regions and used graph theoretical methods to identify changes in network properties associated with high-drinking behavior. Network modularity, a measure of network segregation into communities, was increased in HD mice after alcohol reaccess relative to abstinence. Within-community strength and diversity measures were computed for each region and condition, and highly coactive regions were identified. One high-diversity region, the cortical amygdala (COA), was further interrogated using a chemogenetic approach. COA silencing in CIE mice reduced voluntary drinking, validating our network analysis and indicating that this region may play an important but underappreciated role in alcohol dependence.
]]></description>
<dc:creator>Roland, A.</dc:creator>
<dc:creator>Coelho, C.</dc:creator>
<dc:creator>Haun, H.</dc:creator>
<dc:creator>Gianessi, C.</dc:creator>
<dc:creator>Lopez, M.</dc:creator>
<dc:creator>D'Ambrosio, S.</dc:creator>
<dc:creator>Machinski, S.</dc:creator>
<dc:creator>Kroenke, C.</dc:creator>
<dc:creator>Frankland, P.</dc:creator>
<dc:creator>Becker, H.</dc:creator>
<dc:creator>Kash, T.</dc:creator>
<dc:date>2022-08-26</dc:date>
<dc:identifier>doi:10.1101/2022.08.26.505400</dc:identifier>
<dc:title><![CDATA[Alcohol dependence modifies brain networks activated during abstinence and reaccess: a c-fos-based analysis in mice]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-08-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.08.29.504975v1?rss=1">
<title>
<![CDATA[
Alignment, Segmentation and Neighborhood Analysis in Cyclic Immunohistochemistry Data Using CASSATT 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.08.29.504975v1?rss=1</link>
<description><![CDATA[
Cyclic immunohistochemistry (cycIHC) uses sequential rounds of colorimetric immunostaining and imaging for quantitative mapping of location and number of cells of interest. In addition, cycIHC benefits from the speed and simplicity of brightfield microscopy for data collection, making the collection of entire tissue sections and slides possible at a trivial cost compared to other high dimensional imaging modalities. However, large cycIHC datasets (greater than 50 GB) currently require an expert data scientist to concatenate separate open-source tools for each step of image pre-processing, registration, and segmentation, or the use of proprietary software. Here, we present a unified and user-friendly pipeline for processing, aligning, and analyzing cycIHC data - Cyclic Analysis of Single-Cell Subsets and Tissue Territories (CASSATT). CASSATT registers scanned slide images across all rounds of staining, segments individual nuclei, and measures marker expression on each detected cell. Beyond straightforward single cell data analysis outputs, CASSATT explores the spatial relationships between cell populations. By calculating the logodds of interaction frequencies between cell populations within tissues and tissue regions, this pipeline helps users identify populations of cells that interact - or do not interact - at frequencies that are greater than those occurring by chance. It also identifies specific neighborhoods of cells based on the assortment of neighboring cell types that surround each cell in the sample. The presence and location of these neighborhoods can be compared across slides or within distinct regions within a tissue. CASSATT was first tested using a newly generated cycIHC dataset consisting of six GBM tissue sections processed through eight cycles of AEC based IHC staining. Further validation was completed on a previously published lung cancer tissue microarray dataset consisting of 107 cores processed through eighteen cycles of staining and imaging. CASSATT is a fully open-source workflow tool developed to process cycIHC data and will allow greater utilization of this powerful staining technique.
]]></description>
<dc:creator>Brockman, A.</dc:creator>
<dc:creator>Khurana, R.</dc:creator>
<dc:creator>Bartkowiak, T.</dc:creator>
<dc:creator>Thomas, P. L.</dc:creator>
<dc:creator>Sivagnanam, S.</dc:creator>
<dc:creator>Betts, C. B.</dc:creator>
<dc:creator>Coussens, L. M.</dc:creator>
<dc:creator>Lovly, C.</dc:creator>
<dc:creator>Irish, J. M.</dc:creator>
<dc:creator>Ihrie, R. A.</dc:creator>
<dc:date>2022-08-29</dc:date>
<dc:identifier>doi:10.1101/2022.08.29.504975</dc:identifier>
<dc:title><![CDATA[Alignment, Segmentation and Neighborhood Analysis in Cyclic Immunohistochemistry Data Using CASSATT]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-08-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.09.01.505886v1?rss=1">
<title>
<![CDATA[
Nuclear-enriched protein phosphatase 4 ensures outer kinetochore assemblyprior to nuclear dissolution 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.09.01.505886v1?rss=1</link>
<description><![CDATA[
A landmark event in the transition from interphase to mitosis in metazoans is nuclear envelope breakdown (NEBD). Many events important for mitosis occur prior to NEBD, including condensation of replicated chromosomes and assembly of kinetochores to rapidly engage spindle microtubules. Here we show that nuclear-enriched protein phosphatase 4 (PP4) ensures robust assembly of the microtubule-coupling outer kinetochore prior to NEBD. In the absence of PP4, chromosomes exhibit extended monopolar orientation after NEBD and subsequently mis-segregate. A secondary consequence of diminished outer kinetochore assembly is defective sister chromatid resolution. After NEBD, a cytoplasmic activity compensates for PP4 loss, leading to outer kinetochore assembly and recovery of chromosomes from monopolar orientation to significant biorientation. The Ndc80-Ska microtubule-binding module of the outer kinetochore is required for this recovery. PP4 associates with the inner kinetochore protein CENP-C; however, disrupting the PP4-CENP-C interaction does not perturb chromosome segregation. These results establish that PP4-dependent outer kinetochore assembly prior to NEBD is critical for timely and proper engagement of chromosomes with spindle microtubules.
]]></description>
<dc:creator>Rocha, H.</dc:creator>
<dc:creator>Simoes, P. A.</dc:creator>
<dc:creator>Budrewicz, J.</dc:creator>
<dc:creator>Lara-Gonzalez, P.</dc:creator>
<dc:creator>Xavier de Carvalho, A.</dc:creator>
<dc:creator>Dumont, J.</dc:creator>
<dc:creator>Desai, A.</dc:creator>
<dc:creator>Gassmann, R.</dc:creator>
<dc:date>2022-09-02</dc:date>
<dc:identifier>doi:10.1101/2022.09.01.505886</dc:identifier>
<dc:title><![CDATA[Nuclear-enriched protein phosphatase 4 ensures outer kinetochore assemblyprior to nuclear dissolution]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-09-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.09.03.506477v1?rss=1">
<title>
<![CDATA[
Structures and membrane interactions of native serotonin transporter in complexes with psychostimulants 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.09.03.506477v1?rss=1</link>
<description><![CDATA[
The serotonin transporter (SERT) is a member of the SLC6 neurotransmitter transporter family that mediates serotonin reuptake at presynaptic nerve terminals. SERT is the target of both therapeutic antidepressant drugs and illicit psychostimulant substances such as cocaine and methamphetamines, which are small molecules that perturb normal serotonergic transmission by interfering with serotonin transport. Despite decades of studies, important functional aspects of SERT such as the oligomerization state of native SERT and its interactions with potential proteins remain unresolved. Here we develop methods to isolate SERT from porcine brain (pSERT) using a mild, non-ionic detergent, utilize fluorescence- detection size-exclusion chromatography to investigate its oligomerization state and interactions with other proteins, and employ single-particle cryo-electron microscopy to elucidate the structures of pSERT in complexes with methamphetamine or cocaine, providing structural insights into psychostimulant recognition and accompanying pSERT conformations. Methamphetamine and cocaine both bind to SERT central site, stabilizing the transporter in an outward open conformation. We also identify densities attributable to multiple cholesterol or cholesteryl hemisuccinate (CHS) molecules, as well as to a detergent molecule bound to SERT allosteric site. Under our conditions of isolation, we find that pSERT is best described as a monomeric entity, isolated without interacting proteins, and is ensconced by multiple cholesterol or CHS molecules.

SignificanceThe serotonin transporter (SERT) is the target of antidepressants and illicit psychostimulants. Despite its importance in the nervous, cardiovascular and gastrointestinal systems, there is no direct knowledge of SERTs oligomerization state(s) and interactions with other proteins. Here, we develop methods to isolate porcine SERT (pSERT) from native brain tissue in the presence of a mild, non-ionic detergent, and investigated its properties by biochemical, structural and computational methods. We show how cocaine and methamphetamine exert their pharmacological effect on SERT, binding to a site halfway across the membrane-spanning region of the transporter, stabilizing pSERT in an outward open conformation. pSERT is best described as a monomeric entity, requiring neither oligomerization or additional proteins for its structure or function.
]]></description>
<dc:creator>Yang, D.</dc:creator>
<dc:creator>Zhao, Z.</dc:creator>
<dc:creator>Tajkhorshid, E.</dc:creator>
<dc:creator>Gouaux, E.</dc:creator>
<dc:date>2022-09-05</dc:date>
<dc:identifier>doi:10.1101/2022.09.03.506477</dc:identifier>
<dc:title><![CDATA[Structures and membrane interactions of native serotonin transporter in complexes with psychostimulants]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-09-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.09.05.506590v1?rss=1">
<title>
<![CDATA[
Rad51-mediated interhomolog recombination during budding yeast meiosis is promoted by the meiotic recombination checkpoint and the conserved Pif1 helicase 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.09.05.506590v1?rss=1</link>
<description><![CDATA[
During meiosis, recombination between homologous chromosomes (homologs) generates crossovers that promote proper segregation at the first meiotic division. Recombination is initiated by Spo11-catalyzed double strand breaks (DSBs). 5 end resection of the DSBs creates 3 single strand tails that two recombinases, Rad51 and Dmc1, bind to form presynaptic filaments that search for homology, mediate strand invasion and generate displacement loops (D-loops). D-loop processing then forms crossover and non-crossover recombinants. Meiotic recombination occurs in two temporally distinct phases. During Phase 1, Rad51 is inhibited and Dmc1 mediates the interhomolog recombination that promotes homolog synapsis. In Phase 2, Rad51 becomes active and functions with Rad54 to repair residual DSBs, making increasing use of sister chromatids. The transition from Phase 1 to Phase 2 is controlled by the meiotic recombination checkpoint through the meiosis-specific effector kinase Mek1. This work shows that constitutive activation of Rad51 in Phase 1 results in a subset of DSBs being repaired by a Rad51-mediated interhomolog recombination pathway that is distinct from that of Dmc1. Strand invasion intermediates generated by Rad51 require more time to be processed into recombinants, resulting in a meiotic recombination checkpoint delay in prophase I. Without the checkpoint, Rad51-generated intermediates are more likely to be repaired using a sister chromatid, thereby increasing Meiosis I chromosome nondisjunction. This Rad51 interhomolog recombination pathway is specifically promoted by the conserved 5-3 helicase PIF1 and its paralog, RRM3 and requires Pif1 helicase activity and its interaction with PCNA. This work demonstrates that (1) inhibition of Rad51 during Phase 1 is important to prevent competition with Dmc1 for DSB repair, (2) Rad51-mediated meiotic recombination intermediates are initially processed differently than those made by Dmc1, (3) the meiotic recombination checkpoint provides time during prophase 1 for processing of Rad51-generated recombination intermediates.

AUTHOR SUMMARYTo sexually reproduce, cells containing two copies of each chromosome must undergo the specialized cell division of meiosis to sort the chromosomes into gametes containing a single copy of each chromosome. But how do homologous chromosomes know who is who? The answer is by recombination, a process in which double strand breaks on one chromosome are converted to single stranded ends that can search for the complementary sequence on the homolog. In yeast and mammals, this homology search involves binding of single strand ends by two highly conserved recombinases, Rad51 and the meiosis specific Dmc1. Rad51 is used in mitotic cells to repair breaks, primarily using sister chromatids as templates, while Dmc1 functions in meiosis to generate interhomolog crossovers. In budding yeast, Rad51 strand exchange activity is normally inhibited while Dmc1 is active. We show here that when Rad51 and Dmc1 are active at the same time, Rad51 competes with Dmc1 to mediate interhomolog recombination of a subset of double strand breaks. However, because Rad51- generated recombination intermediates take longer to process, there is a need to keep Rad51 inactive while interhomolog recombination is occurring.
]]></description>
<dc:creator>Ziesel, A.</dc:creator>
<dc:creator>Weng, Q.</dc:creator>
<dc:creator>Ahuja, J. S.</dc:creator>
<dc:creator>Bhattacharya, A.</dc:creator>
<dc:creator>Dutta, R.</dc:creator>
<dc:creator>Cheng, E.</dc:creator>
<dc:creator>Börner, G. V.</dc:creator>
<dc:creator>Lichten, M.</dc:creator>
<dc:creator>Hollingsworth, N. M.</dc:creator>
<dc:date>2022-09-06</dc:date>
<dc:identifier>doi:10.1101/2022.09.05.506590</dc:identifier>
<dc:title><![CDATA[Rad51-mediated interhomolog recombination during budding yeast meiosis is promoted by the meiotic recombination checkpoint and the conserved Pif1 helicase]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-09-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.09.06.506760v1?rss=1">
<title>
<![CDATA[
Low HER2 enables dedifferentiation and transformation of normal breast epithelial cells via chromatin opening 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.09.06.506760v1?rss=1</link>
<description><![CDATA[
Overexpression of the human epidermal growth factor 2 (HER2) protein in breast cancer patients is a predictor of poor prognosis and resistance to therapies. Despite significant advances in the development of targeted therapies and improvements in the 5-year survival rate of metastatic HER2-positive breast cancer patients, a better understanding of the disease at an early stage is needed to prevent its progression. Here, we used an inducible breast cancer transformation system that allows investigation of early molecular changes at high temporal resolution. HER2 overexpression to similar levels as those observed in a subtype of HER2 positive breast cancer patients induced transformation of MCF10A cells and resulted in gross morphological changes, increased anchorage-independent growth of cells, and altered transcriptional programme of genes associated with oncogenic transformation. Global phosphoproteomic analysis during the first few hours of HER2 induction predominantly detected an increase in protein phosphorylation. Intriguingly, this correlated with a wave of chromatin opening, as measured by ATAC-seq on acini isolated from 3D cell culture. We observed that HER2 overexpression leads to reprogramming of many distal regulatory regions and promotes reprogramming-associated heterogeneity. We found that a subset of cells acquired a dedifferentiated breast stem-like phenotype, making them likely candidates for malignant transformation. Our data show that this population of cells, which counterintuitively enriches for relatively low HER2 protein abundance and increased chromatin accessibility, possesses transformational drive, resulting in increased anchorage-independent growth in vitro compared to cells not displaying a stem-like phenotype. Our data provide a discovery platform for signalling to chromatin pathways in HER2-driven cancers, offering an opportunity for biomarker discovery and identification of novel drug targets.
]]></description>
<dc:creator>Hayat, A.</dc:creator>
<dc:creator>Carter, E.</dc:creator>
<dc:creator>King, H. W.</dc:creator>
<dc:creator>Ors, A.</dc:creator>
<dc:creator>Doe, A.</dc:creator>
<dc:creator>Teijeiro, S. A.</dc:creator>
<dc:creator>Charrot, S.</dc:creator>
<dc:creator>Godinho, S.</dc:creator>
<dc:creator>Cutillas, P. R.</dc:creator>
<dc:creator>Mohammed, H.</dc:creator>
<dc:creator>Grose, R. P.</dc:creator>
<dc:creator>Ficz, G.</dc:creator>
<dc:date>2022-09-06</dc:date>
<dc:identifier>doi:10.1101/2022.09.06.506760</dc:identifier>
<dc:title><![CDATA[Low HER2 enables dedifferentiation and transformation of normal breast epithelial cells via chromatin opening]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-09-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.09.11.507477v1?rss=1">
<title>
<![CDATA[
High Resolution Imaging of the Osteogenic and Angiogenic Interface at the Site of Cranial Bone Defect Repair via Multiphoton Microscopy 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.09.11.507477v1?rss=1</link>
<description><![CDATA[
The spatiotemporal blood vessel formation and specification at the osteogenic and angiogenic interface of cranial bone defect repair were examined utilizing a high-resolution multiphoton-based imaging platform in conjunction with advanced optical techniques that allow interrogation of the oxygen microenvironment and cellular energy metabolism in living animals. Our study demonstrates the dynamic changes of vessel types, i.e. arterial, venous and capillary vessel networks at the superior and dura periosteum of cranial bone defect, suggesting a differential coupling of the vessel type with osteoblast expansion and bone tissue deposition/remodeling during repair. Employing transgenic reporter mouse models that label distinct types of vessels at the site of repair, we further show that oxygen distributions in capillary vessels at the healing site are heterogeneous as well as time and location-dependent. The endothelial cells coupling to osteoblasts prefer glycolysis and are less sensitive to microenvironmental oxygen changes than osteoblasts. In comparison, osteoblasts utilize relatively more OxPhos and potentially consume more oxygen at the site of repair. Taken together, our study highlights the dynamics and functional significance of blood vessel types at the site of defect repair, opening up opportunities for further delineating the oxygen and metabolic microenvironment at the interface of bone tissue regeneration.
]]></description>
<dc:creator>Schilling, K.</dc:creator>
<dc:creator>Zhai, Y.</dc:creator>
<dc:creator>Zhou, Z.</dc:creator>
<dc:creator>Zhou, B.</dc:creator>
<dc:creator>Brown, E.</dc:creator>
<dc:creator>Zhang, X.</dc:creator>
<dc:date>2022-09-13</dc:date>
<dc:identifier>doi:10.1101/2022.09.11.507477</dc:identifier>
<dc:title><![CDATA[High Resolution Imaging of the Osteogenic and Angiogenic Interface at the Site of Cranial Bone Defect Repair via Multiphoton Microscopy]]></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/10.1101/2022.09.11.507488v1?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/10.1101/2022.09.14.507917v1?rss=1">
<title>
<![CDATA[
Capillary electrophoresis mass spectrometry identifies new isomers of inositol pyrophosphates in mammalian tissues 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.09.14.507917v1?rss=1</link>
<description><![CDATA[
Technical challenges have to date prevented a complete profiling of the levels of myo-inositol phosphates (InsPs) and pyrophosphates (PP-InsPs) in mammalian tissues. Here, we have deployed capillary electrophoresis mass spectrometry to identify and record the levels of InsPs and PP-InsPs in several tissues obtained from wild type mice and a newly-created PPIP5K2 knockout strain. We observe that the mouse colon harbours unusually high levels of InsPs and PP-InsPs. Additionally, the PP-InsP profile is considerably more complex than previously reported for animal cells: using chemically synthesized internal stable isotope references, and high-resolution mass spectra, we characterize two new PP-InsP isomers as 4/6-PP-InsP5 and 2-PP-InsP5. The latter has not previously been described in Nature. Analysis of feces and the commercial mouse diet suggest the latter is one potential source of noncanonical isomers in the colon. However, we also identify both molecules in the heart, indicating unknown synthesis pathways in mammals. We also demonstrate that the CE-MS method is sensitive enough to measure PP-InsPs from patient samples such as colon biopsies and peripheral blood mononuclear cells (PBMCs). Strikingly, PBMCs also contain 4/6-PP-InsP5 and 2-PP-InsP5. In summary, our study substantially expands PP-InsP biology in mammals.
]]></description>
<dc:creator>Qiu, D.</dc:creator>
<dc:creator>Gu, C.</dc:creator>
<dc:creator>Liu, G.</dc:creator>
<dc:creator>Ritter, K.</dc:creator>
<dc:creator>Eisenbeis, V. B.</dc:creator>
<dc:creator>Bittner, T.</dc:creator>
<dc:creator>Gruzdev, A.</dc:creator>
<dc:creator>Seidel, L.</dc:creator>
<dc:creator>Bengsch, B.</dc:creator>
<dc:creator>Shears, S. B.</dc:creator>
<dc:creator>Jessen, H.</dc:creator>
<dc:date>2022-09-14</dc:date>
<dc:identifier>doi:10.1101/2022.09.14.507917</dc:identifier>
<dc:title><![CDATA[Capillary electrophoresis mass spectrometry identifies new isomers of inositol pyrophosphates in mammalian tissues]]></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/10.1101/2022.09.15.507688v1?rss=1">
<title>
<![CDATA[
The transcriptional and epigenetic reprogramming mediated by chronic IL1β exposure drives self-renewal ability and myeloid priming in TET2 deficient stem and progenitor cells. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.09.15.507688v1?rss=1</link>
<description><![CDATA[
Clonal hematopoiesis (CH) increases the risk for the development of hematological malignancy and cardiovascular disease. IL1{beta} is elevated in patients with CH and its inhibition mitigates cardiovascular risk in murine models with Tet2 loss-of-function. How IL1{beta} alters population dynamics of hematopoietic cells upon Tet2 deletion (Tet2-KO) is not well understood. We demonstrated IL1{beta} expands Tet2-KO monocytes/macrophages, and long-term hematopoietic stem cells. IL1{beta} promoted myeloid bias over other lineages of Tet2-KO HSPCs coinciding with the failure to demethylate lineage-associated enhancer and transcription factor binding sites. IL1{beta} enhanced the self-renewal ability of Tet2-KO HSPCs by upregulating genes associated with self-renewal and by resisting the demethylation of binding sites of transcription factors promoting terminal differentiation. The IL1{beta}-mediated premalignant phenotype is suppressed by the IL1{beta} antagonist or deletion of the IL1 receptor-1, in vivo in aged mice. Our results demonstrate that targeting IL1 signaling could be an efficient early intervention strategy in preleukemic disorders.

STATEMENT OF SIGNIFICANCEIL1{beta} promoted myeloid expansion and self-renewal capacity of TET2-null pre-leukemic cells. Lineage bias occurred early within progenitors towards pro-inflammatory macrophages. Genes specific to aging and with roles in promoting self-renewal capacity, and myeloid bias were upregulated. Hypermethylation occurred within lymphoid and erythroid lineage-specific regulatory elements. Targeting IL1R1 reduced aberrant myeloid bias and premalignant phenotype.
]]></description>
<dc:creator>McClatchy, J.</dc:creator>
<dc:creator>Strogantsev, R.</dc:creator>
<dc:creator>Wolfe, E.</dc:creator>
<dc:creator>Estabrook, J.</dc:creator>
<dc:creator>Lin, H.-Y.</dc:creator>
<dc:creator>Mohammadhosseini, M.</dc:creator>
<dc:creator>Davis, B.</dc:creator>
<dc:creator>Eden, C.</dc:creator>
<dc:creator>Goldman, D.</dc:creator>
<dc:creator>Fleming, W.</dc:creator>
<dc:creator>Cimmino, L.</dc:creator>
<dc:creator>Mohammed, H.</dc:creator>
<dc:creator>Agarwal, A.</dc:creator>
<dc:date>2022-09-16</dc:date>
<dc:identifier>doi:10.1101/2022.09.15.507688</dc:identifier>
<dc:title><![CDATA[The transcriptional and epigenetic reprogramming mediated by chronic IL1β exposure drives self-renewal ability and myeloid priming in TET2 deficient stem and progenitor cells.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-09-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.09.15.508188v1?rss=1">
<title>
<![CDATA[
The Tre1/S1pr1 phospholipid-binding G protein-coupled receptor signaling pathway is required for astrocyte morphogenesis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.09.15.508188v1?rss=1</link>
<description><![CDATA[
Astrocytes play crucial roles in regulating neural circuit function by forming a dense network of synapse-associated membrane specializations, but signaling pathways regulating astrocyte morphogenesis remain poorly defined. Here we show the Drosophila lipid-binding G protein-coupled receptor (GPCR) Tre1, likely acting through Rac1, is required for astrocytes to elaborate their complex morphology in vivo. The lipid phosphate phosphatases Wunen/Wunen2, which process phospholipid ligands, also regulate astrocyte morphology, and, via Tre1, mediate astrocyte-astrocyte competition for growth promoting lipids. Loss of s1pr1, the functional analog of Tre1 in zebrafish disrupts astrocyte process elaboration. Live-imaging and pharmacology demonstrate that S1pr1 balances proper astrocyte process extension/retraction dynamics during morphogenesis, and that S1pr1 signaling is required throughout astrocyte development. Tre1 and S1pr1 are thus potent evolutionarily conserved regulators of astrocyte growth and elaboration of morphological complexity. O_LIThe GPCR Tre1 and LPPs Wun/Wun2 promote astrocyte process outgrowth in Drosophila
C_LIO_LIAstrocytes compete for a growth{-}promoting phospholipid in the CNS
C_LIO_LIWun/Wun2 act locally to regulate process outgrowth through Tre1
C_LIO_LIVertebrate S1pr1 regulates astrocyte growth early, through modulation of process dynamics
C_LI

O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=162 SRC="FIGDIR/small/508188v2_ufig1.gif" ALT="Figure 1">
View larger version (24K):
org.highwire.dtl.DTLVardef@1e86db5org.highwire.dtl.DTLVardef@7abd6borg.highwire.dtl.DTLVardef@29f163org.highwire.dtl.DTLVardef@1a6c4b9_HPS_FORMAT_FIGEXP  M_FIG C_FIG
]]></description>
<dc:creator>Chen, J.</dc:creator>
<dc:creator>Stork, T.</dc:creator>
<dc:creator>Kang, Y.</dc:creator>
<dc:creator>Sheehan, A.</dc:creator>
<dc:creator>Paton, C.</dc:creator>
<dc:creator>Monk, K.</dc:creator>
<dc:creator>Freeman, M.</dc:creator>
<dc:date>2022-09-17</dc:date>
<dc:identifier>doi:10.1101/2022.09.15.508188</dc:identifier>
<dc:title><![CDATA[The Tre1/S1pr1 phospholipid-binding G protein-coupled receptor signaling pathway is required for astrocyte morphogenesis]]></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/10.1101/2022.09.16.508249v1?rss=1">
<title>
<![CDATA[
Purification and biochemical analysis of native AMPA receptors from three different mammalian species 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.09.16.508249v1?rss=1</link>
<description><![CDATA[
The majority of fast, excitatory synaptic transmission in the central nervous system (CNS) is mediated by -amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptors (AMPARs), which are glutamate-activated ion channels integral to synaptic plasticity, motor coordination, learning, and memory. AMPARs are multiprotein assemblies comprised of a tetrameric receptor core that co-assembles with a broad range of peripheral auxiliary proteins which shape subcellular localization and signaling properties of the resulting complexes. Structure determination of AMPARs has traditionally relied on recombinant expression systems; however, these methods are not well suited to elucidate the diverse array of AMPAR assemblies that are differentially expressed in mammalian brains. While recent studies of native receptor complexes have advanced our understanding of endogenous assemblies, receptors thus far have only been isolated from rodent brain tissue. Here, we employed an immunoaffinity purification strategy to isolate native AMPARs from the brains of three different mammals - pigs, sheep, and cows. Compared to rodents; pigs, sheep, and cows are from a unique clade of mammals - ungulates, that share a closer genomic identity with humans. We determined the molecular size, overall yield, and purity of native AMPARs isolated from these three mammals, thereby demonstrating that structural determination and biochemical analysis is possible from a clade of mammals evolutionarily distinct from rodents.
]]></description>
<dc:creator>Rao, P.</dc:creator>
<dc:creator>Gouaux, E.</dc:creator>
<dc:date>2022-09-16</dc:date>
<dc:identifier>doi:10.1101/2022.09.16.508249</dc:identifier>
<dc:title><![CDATA[Purification and biochemical analysis of native AMPA receptors from three different mammalian species]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-09-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.09.18.508448v1?rss=1">
<title>
<![CDATA[
Comparing acute IOP-induced lamina cribrosa deformations pre-mortem and post-mortem 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.09.18.508448v1?rss=1</link>
<description><![CDATA[
PurposeLamina cribrosa (LC) deformations caused by elevated intraocular pressure (IOP) are believed to contribute to glaucomatous neuropathy and have therefore been extensively studied, in many conditions from in-vivo to ex-vivo. We compare acute IOP-induced global and local LC deformations immediately before (pre-mortem) and after (post-mortem) sacrifice by exsanguination.

MethodsThe optic nerve heads of three healthy monkeys 12-15 years old were imaged with spectral-domain optical coherence tomography under controlled IOP pre-mortem and post-mortem. Volume scans were acquired at baseline IOP (8-10 mmHg) and at 15, 30, and 40 mmHg IOP. A digital volume correlation technique was used to determine the IOP-induced 3D LC deformations (strains) in regions visible pre-mortem and post-mortem.

ResultsBoth conditions exhibited similar nonlinear relationships between IOP increases and LC deformations. Median effective and shear strains were, on average over all eyes and pressures, smaller post-mortem than pre-mortem, by 14% and 11%, respectively (Ps < 0.001). Locally, however, the differences in LC deformation between conditions were variable. Some regions were subjected pre-mortem to triple the strains observed post-mortem, and others suffered smaller deformations pre-mortem than post-mortem.

ConclusionsIncreasing IOP acutely caused nonlinear LC deformations with an overall smaller effect post-mortem than pre-mortem. Locally, deformations pre-mortem and post-mortem were sometimes substantially different. We suggest that the differences may be due to weakened mechanical support from the unpressurized central retinal vessels post-mortem.

Translational RelevanceAdditional to the important pre-mortem information, comparison with post-mortem provides a unique context essential to understand the translational relevance of all post-mortem biomechanics literature.

PrecisThe authors compared in monkeys acute IOP-induced deformations of the lamina cribrosa pre-mortem and post-mortem. Deformation trends were similar pre-mortem and post-mortem, but deformations pre-mortem were generally smaller than those post-mortem, with substantial local variations. The differences are likely due to loss of vessel support post-mortem.
]]></description>
<dc:creator>Wei, J.</dc:creator>
<dc:creator>Hua, Y.</dc:creator>
<dc:creator>Yang, B.</dc:creator>
<dc:creator>Wang, B.</dc:creator>
<dc:creator>Schmitt, S. E.</dc:creator>
<dc:creator>Wang, B.</dc:creator>
<dc:creator>Lucy, K. A.</dc:creator>
<dc:creator>Ishikawa, H.</dc:creator>
<dc:creator>Schuman, J. S.</dc:creator>
<dc:creator>Smith, M. A.</dc:creator>
<dc:creator>Wollstein, G.</dc:creator>
<dc:creator>Sigal, I. A.</dc:creator>
<dc:date>2022-09-19</dc:date>
<dc:identifier>doi:10.1101/2022.09.18.508448</dc:identifier>
<dc:title><![CDATA[Comparing acute IOP-induced lamina cribrosa deformations pre-mortem and post-mortem]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-09-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.09.20.508779v1?rss=1">
<title>
<![CDATA[
A small molecule RIG-I agonist serves to adjuvant broad multifaceted influenza virus vaccine immunity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.09.20.508779v1?rss=1</link>
<description><![CDATA[
Retinoic acid-inducible gene I (RIG-I) is essential for activating host cell innate immunity to regulates the immune response against many RNA viruses. We identified a small molecule compound, KIN1148, that directly binds RIG-I to drive IRF3activation to impart the expression of IRF3-target genes, including specific immunomodulatory cytokines and chemokines. KIN1148 does not lead to ATPase activity or compete with ATP for binding but activates RIG-I to induce antiviral gene expression programs distinct from type I interferon (IFN) treatment. When administered in combination with a vaccine against influenza A virus (IAV), KIN1148 induces both neutralizing antibody and broadly cross-protective IAV-specific T cell responses compared to vaccination alone, which induces comparatively poor responses. This robust KIN1148-adjuvanted immune response protects mice from lethal H1N1 and H5N1 IAV challenge. Importantly, KIN1148 also augments human CD8+ T cell activation. Thus, we have identified a small molecule RIG-I agonist that serves as an effective adjuvant in inducing non-canonical RIG-I activation for induction of innate immune programs that enhance adaptive immune protection of antiviral vaccination.

SummaryHemann, et. al. identify a small-molecule RIG-I agonist (KIN1148) that directly binds RIG-I for non-canonical activation and adjuvants pandemic and avian influenza virus vaccination. KIN1148 augments broadly neutralizing antibody and T cell responses in mice and enhances human DC maturation and CD8+ T cell activation.
]]></description>
<dc:creator>Hemann, E. A.</dc:creator>
<dc:creator>Knoll, M. L.</dc:creator>
<dc:creator>Wilkins, C. R.</dc:creator>
<dc:creator>Subra, C.</dc:creator>
<dc:creator>Green, R.</dc:creator>
<dc:creator>Garcia-Sastre, A.</dc:creator>
<dc:creator>Thomas, P.</dc:creator>
<dc:creator>Trautmann, L.</dc:creator>
<dc:creator>Ireton, R.</dc:creator>
<dc:creator>Loo, Y.-M.</dc:creator>
<dc:creator>Gale, M.</dc:creator>
<dc:date>2022-09-20</dc:date>
<dc:identifier>doi:10.1101/2022.09.20.508779</dc:identifier>
<dc:title><![CDATA[A small molecule RIG-I agonist serves to adjuvant broad multifaceted influenza virus vaccine immunity]]></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/10.1101/2022.09.27.509541v1?rss=1">
<title>
<![CDATA[
A Genome Scale Transcriptional Regulatory Model of the Human Placenta 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.09.27.509541v1?rss=1</link>
<description><![CDATA[
Gene regulation is essential to placental function and fetal development. We report a genome-scale transcriptional regulatory network (TRN) of the human placenta built using digital genomic footprinting and transcriptomic data. We integrated 475 transcriptomes and 12 DNase hypersensitivity datasets from placental samples to globally and quantitatively map transcription factor (TF)-target gene interactions. In an independent dataset, the TRN model predicted target gene expression with an out of sample R2 value greater than 0.25 for 74% of target genes. We performed siRNA knockdowns of 4 TFs and achieved concordance between the predicted gene targets in our TRN and differences in expression of knockdowns with an accuracy of >0.7 for 3 of the 4 TFs. Our final model contained 113,158 interactions across 391 TFs and 7,712 target genes and is publicly available. We identified six TFs which were significantly enriched as regulators for genes previously associated with preterm birth.
]]></description>
<dc:creator>Paquette, A. G.</dc:creator>
<dc:creator>Ahuna, K.</dc:creator>
<dc:creator>Hwang, Y. M.</dc:creator>
<dc:creator>Pearl, J.</dc:creator>
<dc:creator>Liao, H.</dc:creator>
<dc:creator>Shannon, P.</dc:creator>
<dc:creator>Kadam, L.</dc:creator>
<dc:creator>Lapehn, S.</dc:creator>
<dc:creator>Bucher, M.</dc:creator>
<dc:creator>Roper, R.</dc:creator>
<dc:creator>Funk, C. C.</dc:creator>
<dc:creator>MacDonald, J.</dc:creator>
<dc:creator>Bammler, T.</dc:creator>
<dc:creator>Baloni, P.</dc:creator>
<dc:creator>Brockway, H.</dc:creator>
<dc:creator>Mason, W. A.</dc:creator>
<dc:creator>Bush, N.</dc:creator>
<dc:creator>LeWinn, K. Z.</dc:creator>
<dc:creator>Carr, C. J.</dc:creator>
<dc:creator>Stamatoyannopoulos, J.</dc:creator>
<dc:creator>Muglia, L. J.</dc:creator>
<dc:creator>Jones, H. M.</dc:creator>
<dc:creator>Sadovsky, Y.</dc:creator>
<dc:creator>Myatt, L.</dc:creator>
<dc:creator>Sathyanarayana, S.</dc:creator>
<dc:creator>Price, N.</dc:creator>
<dc:creator>Environmental influences on Child Health Outcomes,</dc:creator>
<dc:date>2022-09-28</dc:date>
<dc:identifier>doi:10.1101/2022.09.27.509541</dc:identifier>
<dc:title><![CDATA[A Genome Scale Transcriptional Regulatory Model of the Human Placenta]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-09-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.09.27.509669v1?rss=1">
<title>
<![CDATA[
MHC class I and MHC class II reporter mice enable analysis of immune oligodendroglia in mouse models of multiple sclerosis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.09.27.509669v1?rss=1</link>
<description><![CDATA[
Oligodendrocytes and their progenitors upregulate MHC pathways in response to inflammation, but the frequency of this phenotypic change is unknown and the features of these immune oligodendroglia are poorly defined. We generated MHC class I and II transgenic reporter mice to define their dynamics in response to inflammatory demyelination, providing a means to monitor MHC activation in diverse cell types in living mice and define their roles in aging, injury and disease.
]]></description>
<dc:creator>Harrington, E. P.</dc:creator>
<dc:creator>Catenacci, R. B.</dc:creator>
<dc:creator>Smith, M. D.</dc:creator>
<dc:creator>Heo, D.</dc:creator>
<dc:creator>Miller, C. E.</dc:creator>
<dc:creator>Meyers, K. R.</dc:creator>
<dc:creator>Glatzer, J.</dc:creator>
<dc:creator>Bergles, D. E.</dc:creator>
<dc:creator>Calabresi, P. A.</dc:creator>
<dc:date>2022-09-28</dc:date>
<dc:identifier>doi:10.1101/2022.09.27.509669</dc:identifier>
<dc:title><![CDATA[MHC class I and MHC class II reporter mice enable analysis of immune oligodendroglia in mouse models of multiple sclerosis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-09-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.09.27.509698v1?rss=1">
<title>
<![CDATA[
Pericytes control vascular stability and auditory spiral ganglion neuron survival 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.09.27.509698v1?rss=1</link>
<description><![CDATA[
The inner ear has a rich population of pericytes, a multi-functional mural cell essential for sensory hair cell heath and normal hearing. However, the mechanics of how pericytes contribute to the homeostasis of the auditory vascular-neuronal complex in the spiral ganglion is not yet known. In this study, using an inducible and conditional pericyte depletion mouse (PdgfrbCreERT2+/-; ROSA26iDTR+/-) model, we demonstrate, for the first time, that pericyte depletion causes loss of vascular volume and spiral ganglion neurons (SGNs) and adversely affects hearing sensitivity. Using an in vitro trans-well co-culture system, we show pericytes markedly promote neurite and vascular branch growth in neonatal SGN explants and adult SGNs. The pericyte-controlled neural growth is strongly mediated by pericyte-released exosomes containing vascular endothelial growth factor-A (VEGF-A). Treatment of neonatal SGN explants or adult SGNs with pericyte-derived exosomes significantly enhances angiogenesis, SGN survival, and neurite growth, all of which were inhibited by a selective blocker of the VEGF receptor 2 (Flk1). Our study demonstrates that pericytes in the adult ear are critical for vascular stability and SGN health. Cross-talk between pericytes and SGNs via exosomes is essential for neuronal and vascular health and normal hearing.
]]></description>
<dc:creator>Zhang, Y.</dc:creator>
<dc:creator>Neng, L.</dc:creator>
<dc:creator>Sharma, K.</dc:creator>
<dc:creator>Hou, Z.</dc:creator>
<dc:creator>Johnson, A.</dc:creator>
<dc:creator>Song, J.</dc:creator>
<dc:creator>Dabdoub, A.</dc:creator>
<dc:creator>Shi, X.</dc:creator>
<dc:date>2022-09-28</dc:date>
<dc:identifier>doi:10.1101/2022.09.27.509698</dc:identifier>
<dc:title><![CDATA[Pericytes control vascular stability and auditory spiral ganglion neuron survival]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-09-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.09.30.509798v1?rss=1">
<title>
<![CDATA[
Dual states of Bmi1-expressing intestinal stem cells drive epithelial development and tissue regeneration 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.09.30.509798v1?rss=1</link>
<description><![CDATA[
Intestinal development, response to injury and disease states rely upon balanced stem cell proliferation. Historically, two subtypes of intestinal epithelial stem cells (ISCs)--slow-cycling/label-retaining, and actively-cycling/canonical Wnt-dependent--coordinate to drive proliferation and regulate epithelial renewal during adult tissue homeostasis and injury response. Recent studies focused on Bmi1-expressing cells revealed that differentiated Bmi1+ enteroendocrine cells could dedifferentiate towards a canonical Wnt-dependent stem cell state, calling into question the dogma that a dual stem cell axis regulates epithelial proliferation. Herein, we identify stem cell function in a Bmi1+ cell population in early murine intestinal development prior to the establishment of canonical Wnt-dependent, Lgr5-expressing ISCs. In-depth analyses of developmental Bmi1+ ISCs using lineage-tracing and single cell RNA-sequencing reveal their distinct identity and capacity to differentiate into Lgr5+ ISCs and other differentiated lineages. Further, during in utero development, the Bmi1+ ISCs initially exists in a highly proliferative state then transitions to a slow-cycling state, with the emergence of actively-cycling Lgr5+ ISCs. In adult tissue, Bmi1+ ISCs are a distinct population that re-express developmental gene and protein profiles, and a non-canonical Wnt signaling signature in response to injury and in human colorectal tumors. Further, developmental Bmi1+ ISCs are distinct from Lgr5+ ISCs and the previously identified differentiated Bmi1+ progenitor cells. Re-evaluation of an under-appreciated Bmi1+ ISC population with fundamental importance in intestinal development re-establishes the importance of the dynamic interplay between discrete ISC populations that are regulated by opposing Wnt signaling pathways. This finding opens opportunities and targetable pathways to augment regeneration or inhibit tumorigenesis.
]]></description>
<dc:creator>Smith, N. R.</dc:creator>
<dc:creator>Sengupta, S. K.</dc:creator>
<dc:creator>Conley, P.</dc:creator>
<dc:creator>Giske, N. R.</dc:creator>
<dc:creator>Klocke, C.</dc:creator>
<dc:creator>Walker, B. S.</dc:creator>
<dc:creator>McPhail, N.</dc:creator>
<dc:creator>Swain, J. R.</dc:creator>
<dc:creator>Yoo, Y. J.</dc:creator>
<dc:creator>Anderson, A. N.</dc:creator>
<dc:creator>Davies, P. S.</dc:creator>
<dc:creator>Sanati, N.</dc:creator>
<dc:creator>Nguyen, T. N.</dc:creator>
<dc:creator>Torkenczy, K.</dc:creator>
<dc:creator>Adey, A. C.</dc:creator>
<dc:creator>Fischer, J. M.</dc:creator>
<dc:creator>Wu, G.</dc:creator>
<dc:creator>Wong, M. H.</dc:creator>
<dc:date>2022-10-01</dc:date>
<dc:identifier>doi:10.1101/2022.09.30.509798</dc:identifier>
<dc:title><![CDATA[Dual states of Bmi1-expressing intestinal stem cells drive epithelial development and tissue regeneration]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-10-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.10.01.510403v1?rss=1">
<title>
<![CDATA[
Three-dimensional structure of inner ear hair cell mitochondrial networks and ribbon synapses in a zebrafish model of Usher syndrome type 1B 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.10.01.510403v1?rss=1</link>
<description><![CDATA[
Inner ear hair cells are the fundamental unit of sound and vibration detection in inner ear and lateral line structures. Our understanding of the structure and ultrastructure of hair cells has heretofore relied upon two-dimensional imaging. The development of serial block-face scanning electron microscopy (SBFSEM) changes this paradigm and allows for ultrastructural evaluation of hair cells in three-dimensions. This is the first report of SBFSEM analysis in the myo7aa-/- mutant where we evaluated several attributes of zebrafish hair cells from the inner ear cristae in both wildtype and myo7aa-/- mutant zebrafish through three-dimensional reconstruction. We describe ribbon synapse number, location, and volume, mitochondrial localization, and innervation for individual hair cells. We determined that myo7aa-/- mutant ribbon synapses have a smaller volume and surface area; however, all other hair cell attributes investigated were not significantly different between the mutant and wildtype zebrafish. These findings are critical for the development of therapies for deafness caused by mutations in myo7aa, as this study supports that the necessary hair cell machinery for hearing is largely intact. In addition, the methodology and measurements developed in this study provide a guide for the evaluation of zebrafish hair cells using SBFSEM.
]]></description>
<dc:creator>Riley, K. C.</dc:creator>
<dc:creator>Koleilat, A.</dc:creator>
<dc:creator>Dugdale, J. A.</dc:creator>
<dc:creator>Cooper, S. A.</dc:creator>
<dc:creator>Christensen, T. A.</dc:creator>
<dc:creator>Schimmenti, L. A.</dc:creator>
<dc:date>2022-10-02</dc:date>
<dc:identifier>doi:10.1101/2022.10.01.510403</dc:identifier>
<dc:title><![CDATA[Three-dimensional structure of inner ear hair cell mitochondrial networks and ribbon synapses in a zebrafish model of Usher syndrome type 1B]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-10-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.10.03.510650v1?rss=1">
<title>
<![CDATA[
Rapid Cell Type-Specific Nascent Proteome Labeling in Drosophila 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.10.03.510650v1?rss=1</link>
<description><![CDATA[
Controlled protein synthesis is required to regulate gene expression and is often carried out in a cell type-specific manner. Protein synthesis is commonly measured by labeling the nascent proteome with amino acid analogs or isotope-containing amino acids. These methods have been difficult to implement in vivo as they require lengthy amino acid replacement procedures. O-propargyl-puromycin (OPP) is a puromycin analog that incorporates into nascent polypeptide chains. Through its terminal alkyne, OPP can be conjugated to a fluorophore-azide for directly visualizing nascent protein synthesis, or to a biotin-azide for capture and identification of newly-synthesized proteins. To achieve cell type-specific OPP incorporation, we developed phenylacetyl-OPP (PhAc-OPP), a puromycin analog harboring an enzyme-labile blocking group that can be removed by Penicillin G acylase (PGA). Here, we show that cell type-specific PGA expression in Drosophila can be used to achieve OPP labeling of newly-synthesized proteins in targeted cell populations within the brain. Following a brief 2-hour incubation of intact brains with PhAc-OPP, we observe robust imaging and affinity purification of OPP-labeled nascent proteins in PGA-targeted cell populations. We apply this method to show a pronounced age-related decline in neuronal protein synthesis in the fly brain, demonstrating the capability to quantitatively capture in vivo protein synthesis states using PhAc-OPP. This method, which we call POPPi (PGA-dependent OPP incorporation), should be applicable for rapidly visualizing protein synthesis and identifying nascent proteins synthesized under diverse physiological and pathological conditions with cellular specificity in vivo.
]]></description>
<dc:creator>Villalobos-Cantor, S.</dc:creator>
<dc:creator>Barrett, R. M.</dc:creator>
<dc:creator>Condon, A. F.</dc:creator>
<dc:creator>Arreola-Bustos, A.</dc:creator>
<dc:creator>Rodriguez, K. M.</dc:creator>
<dc:creator>Cohen, M. S.</dc:creator>
<dc:creator>Martin, I.</dc:creator>
<dc:date>2022-10-05</dc:date>
<dc:identifier>doi:10.1101/2022.10.03.510650</dc:identifier>
<dc:title><![CDATA[Rapid Cell Type-Specific Nascent Proteome Labeling in Drosophila]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-10-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.10.04.510803v1?rss=1">
<title>
<![CDATA[
A Suite of Advanced Tutorials for the WESTPA 2.0 Rare-Events Sampling Software 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.10.04.510803v1?rss=1</link>
<description><![CDATA[
We present six advanced tutorials instructing users in the best practices of using key new features and plugins/extensions of the WESTPA 2.0 software package, which consists of major upgrades for enabling applications of the weighted ensemble (WE) path sampling strategy to even larger systems and/or slower processes. The tutorials demonstrate the use of the following key features: (i) a generalized resampler module for the creation of "binless" schemes, (ii) a minimal adaptive binning scheme for more efficient surmounting of free energy barriers, (iii) streamlined handling of large simulation datasets using an HDF5 framework, (iv) two different schemes for more efficient rate-constant estimation, (v) a Python API for simplified analysis of WE simulations, and (vi) plugins/extensions for Markovian Weighted Ensemble Milestoning and WE rule-based modeling at the system biology level. Applications of the tutorials range from atomistic to residue-level to non-spatial models, and include complex processes such as protein folding and the membrane permeability of a drug-like molecule. Users are expected to already have significant experience with running conventional molecular dynamics simulations and completed the previous suite of WESTPA tutorials.
]]></description>
<dc:creator>Bogetti, A.</dc:creator>
<dc:creator>Leung, J.</dc:creator>
<dc:creator>Russo, J.</dc:creator>
<dc:creator>Zhang, S.</dc:creator>
<dc:creator>Thompson, J.</dc:creator>
<dc:creator>Saglam, A.</dc:creator>
<dc:creator>Ray, D.</dc:creator>
<dc:creator>Abraham, R.</dc:creator>
<dc:creator>Faeder, J.</dc:creator>
<dc:creator>Andricioaei, I.</dc:creator>
<dc:creator>Adelman, J.</dc:creator>
<dc:creator>Zwier, M.</dc:creator>
<dc:creator>LeBard, D.</dc:creator>
<dc:creator>Zuckerman, D. M.</dc:creator>
<dc:creator>Chong, L.</dc:creator>
<dc:date>2022-10-07</dc:date>
<dc:identifier>doi:10.1101/2022.10.04.510803</dc:identifier>
<dc:title><![CDATA[A Suite of Advanced Tutorials for the WESTPA 2.0 Rare-Events Sampling Software]]></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/10.1101/2022.10.07.511231v1?rss=1">
<title>
<![CDATA[
A Novel Mouse Model that Recapitulates the Heterogeneity of Human Triple Negative Breast Cancer 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.10.07.511231v1?rss=1</link>
<description><![CDATA[
Triple-negative breast cancer (TNBC) patients have a poor prognosis and few treatment options. Mouse models of TNBC are important for development of new targeted therapies, but few TNBC mouse models exist. Here, we developed a novel TNBC murine model by mimicking two common TNBC mutations with high co-occurrence: amplification of the oncogene MYC and deletion of the tumor suppressor PTEN. This Myc;Ptenfl murine model develops TN mammary tumors that display histological and molecular features commonly found in human TNBC. We performed deep omic analyses on Myc;Ptenfl tumors including machine learning for morphologic features, bulk and single-cell RNA-sequencing, multiplex immunohistochemistry and single-cell phenotyping. Through comparison with human TNBC, we demonstrated that this new genetic mouse model develops mammary tumors with differential survival that closely resemble the inter- and intra-tumoral and microenvironmental heterogeneity of human TNBC; providing a unique pre-clinical tool for assessing the spectrum of patient TNBC biology and drug response.

Statement of significanceThe development of cancer models that mimic triple-negative breast cancer (TNBC) microenvironment complexities is critical to develop effective drugs and enhance disease understanding. This study addresses a critical need in the field by identifying a murine model that faithfully mimics human TNBC heterogeneity and establishing a foundation for translating preclinical findings into effective human clinical trials.
]]></description>
<dc:creator>Doha, Z. O.</dc:creator>
<dc:creator>Wang, X.</dc:creator>
<dc:creator>Calistri, N.</dc:creator>
<dc:creator>Eng, J.</dc:creator>
<dc:creator>Daniel, C. J.</dc:creator>
<dc:creator>Ternes, L.</dc:creator>
<dc:creator>Kim, E. N.</dc:creator>
<dc:creator>Pelz, C.</dc:creator>
<dc:creator>Munks, M.</dc:creator>
<dc:creator>Betts, C.</dc:creator>
<dc:creator>Kirchberger, N.</dc:creator>
<dc:creator>Kwon, S.</dc:creator>
<dc:creator>Bucher, E.</dc:creator>
<dc:creator>Li, X.</dc:creator>
<dc:creator>Waugh, T.</dc:creator>
<dc:creator>Pietenpol, J. A.</dc:creator>
<dc:creator>Sanders, M. E.</dc:creator>
<dc:creator>Mills, G. B.</dc:creator>
<dc:creator>Chin, K.</dc:creator>
<dc:creator>Chang, Y. H.</dc:creator>
<dc:creator>Coussens, L. M.</dc:creator>
<dc:creator>Gray, J. W.</dc:creator>
<dc:creator>Heiser, L.</dc:creator>
<dc:creator>Sears, R. C.</dc:creator>
<dc:date>2022-10-07</dc:date>
<dc:identifier>doi:10.1101/2022.10.07.511231</dc:identifier>
<dc:title><![CDATA[A Novel Mouse Model that Recapitulates the Heterogeneity of Human Triple Negative Breast Cancer]]></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/10.1101/2022.10.11.511646v1?rss=1">
<title>
<![CDATA[
A genome-wide CRISPR interference screen using an engineered trafficking biosensor reveals a role for RME-8 in opioid receptor regulation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.10.11.511646v1?rss=1</link>
<description><![CDATA[
G protein-coupled receptors (GPCRs) are the largest family of membrane-bound signaling molecules. Activity of these receptors is critically regulated by their trafficking through the endo-lysosomal pathway. Identifying the genes involved in GPCR trafficking is challenging due the complexity of sorting operations and low affinity protein-protein interactions. Here we present a chemical biology fluorescence-based technique to interrogate GPCR trafficking. We show that the engineered enzyme APEX2 is a highly sensitive biosensor for GPCR trafficking to the lysosome, and this trafficking can be monitored through APEX-based activation of fluorogenic substrates such as Amplex UltraRed (AUR). We used this approach to perform a genome-wide CRISPR interference screen focused on the delta type opioid receptor (DOR), a GPCR which modulates anxiety, depression, and pain. The screen identified 492 genes including known- and novel-regulators of DOR expression and trafficking. We demonstrate that one of the novel genes, RME-8, localizes to early endosomes and plays a critical role in regulating DOR trafficking to the lysosome. Together, our data demonstrate that GPCR-APEX2/AUR is a flexible and highly sensitive chemical biology platform for genetic interrogation of receptor trafficking.
]]></description>
<dc:creator>Novy, B.</dc:creator>
<dc:creator>Adoff, H.</dc:creator>
<dc:creator>De Maria, M.</dc:creator>
<dc:creator>Kampmann, M.</dc:creator>
<dc:creator>Tsvetanova, N.</dc:creator>
<dc:creator>Von Zastrow, M.</dc:creator>
<dc:creator>Lobingier, B.</dc:creator>
<dc:date>2022-10-11</dc:date>
<dc:identifier>doi:10.1101/2022.10.11.511646</dc:identifier>
<dc:title><![CDATA[A genome-wide CRISPR interference screen using an engineered trafficking biosensor reveals a role for RME-8 in opioid receptor regulation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-10-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.10.12.511931v1?rss=1">
<title>
<![CDATA[
Early-life sleep disruption impairs subtle social behaviors during pair bond formation in prairie voles: A computer vision study 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.10.12.511931v1?rss=1</link>
<description><![CDATA[
Early life sleep disruption (ELSD) has been shown to have long lasting effects on social behavior in adult prairie voles (Microtus ochrogaster), including impaired expression of pair bond behavior during partner preference testing (e.g., reduced huddling with a pair bonded partner). However, due to the limitations of manual behavior tracking, the behavioral effects of ELSD across the entire time course of pair bond formation have not yet been described, hindering our ability to trace mechanisms. Here, we used computer vision to precisely track multiple behaviors during opposite-sex cohabitation of prairie voles. Male-female pairs were allowed to interact through a mesh divider in the home cage for 72 h, providing variables of body direction, distance-to-divider, and locomotion speed, with temporal resolution as high as the video frame rate (20-25 Hz). We found that control males displayed periodic, stereotyped patterns of body orientation towards females during pair bond formation. In contrast, ELSD males showed reduced duration and ultradian periodicity of these body orientation behaviors towards females. Furthermore, in both sexes, ELSD altered stereotypical spatial and temporal patterns of locomotion seen in control animals that typically varied across days of cohabitation and light/dark periods. This study highlights the utility of computer vision in deep characterization of subtle behaviors and allows a more comprehensive behavioral assessment of the profound and persistent effects of ELSD on later life social behavior. Our findings may shed light on causal mechanisms underlying human neurodevelopmental disorders featuring sleep disruption and social deficits, such as autism spectrum disorder.
]]></description>
<dc:creator>Bueno-Junior, L. S.</dc:creator>
<dc:creator>Jones-Tinsley, C. E.</dc:creator>
<dc:creator>Wickham, P. T.</dc:creator>
<dc:creator>Watson, B. O.</dc:creator>
<dc:creator>Lim, M. M.</dc:creator>
<dc:date>2022-10-17</dc:date>
<dc:identifier>doi:10.1101/2022.10.12.511931</dc:identifier>
<dc:title><![CDATA[Early-life sleep disruption impairs subtle social behaviors during pair bond formation in prairie voles: A computer vision study]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-10-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.10.17.512537v1?rss=1">
<title>
<![CDATA[
Agonistic anti-CD40 converts Tregs into Type 1 effectors within the tumor micro-environment 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.10.17.512537v1?rss=1</link>
<description><![CDATA[
Multiple cell types, molecules, and processes contribute to inhibition of anti-tumor effector responses, often frustrating effective immunotherapy. Among these, Foxp3+ CD4+ cells (Tregs) are well-recognized to play an immunosuppressive role in the tumor microenvironment. The first clinically successful checkpoint inhibitor, anti-CTLA-4 antibody, may deplete Tregs at least in part by antibody-dependent cellular cytotoxicity (ADCC), but this effect is unreliable in mice, including in a genetically engineered mouse model of pancreatic ductal adenocarcinoma (PDAC). In contrast, agonistic CD40 antibody, which serves as an effective therapy, is associated with notable Treg disappearance in the PDAC model. The mechanism of CD40-mediated Treg loss is poorly understood, as Tregs are CD40-negative. Here we have explored the mechanistic basis for the loss of Foxp3 T cells upon anti-CD40 treatment and find, using tissue-level multiplex immunostaining and orthogonal dissociated cell analyses, that Tregs are not depleted but converted into interferon-{gamma} (IFN-{gamma}) producing, Type I CD4+ T effector cells. This process depends on IL-12 and IFN-{gamma} signaling evoked by action of the anti-CD40 antibody on dendritic cells (DCs), especially BATF3-dependent cDC1s. These findings provide insight into a previously unappreciated mechanism of CD40 agonism as a potent anti-tumor intervention that promotes the re-programming of Tregs into tumor-reactive CD4+ effector T cells, markedly augmenting the anti-tumor response.
]]></description>
<dc:creator>Maltez, V.</dc:creator>
<dc:creator>Arora, C.</dc:creator>
<dc:creator>Sor, R.</dc:creator>
<dc:creator>Lian, Q.</dc:creator>
<dc:creator>Vonderheide, R. H.</dc:creator>
<dc:creator>Germain, R. N.</dc:creator>
<dc:creator>Byrne, K. T.</dc:creator>
<dc:date>2022-10-20</dc:date>
<dc:identifier>doi:10.1101/2022.10.17.512537</dc:identifier>
<dc:title><![CDATA[Agonistic anti-CD40 converts Tregs into Type 1 effectors within the tumor micro-environment]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-10-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.10.18.512732v1?rss=1">
<title>
<![CDATA[
Early life sleep disruption has long lasting, sex specific effects on later development of sleep in prairie voles 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.10.18.512732v1?rss=1</link>
<description><![CDATA[
In mammals, sleep duration is highest in the early postnatal period of life and is critical for shaping neural circuits that control the development of complex behaviors. The prairie vole is a wild, highly social rodent that serves as a unique model for the study of complex, species-typical social behaviors. Previous work in our laboratory has found that early life sleep disruption (ELSD) in prairie voles during a sensitive window of postnatal development leads to long lasting changes in social and cognitive behaviors as well as structural changes in excitatory and inhibitory neural circuits in the brain. However, it is currently unknown how later sleep is impacted by ELSD, both shortly after ELSD and over the long term. Therefore, the aim of this study was to describe the effects of ELSD on later life sleep, compared to sleep in normally developing prairie voles. First, we conducted tethered electroencephalogram/electromyogram (EEG/EMG) recordings in juvenile prairie voles undergoing ELSD, compared to Control conditions. Second, we conducted 24 hours of home cage tethered EEG/EMG recordings in either adolescent or adult male and female prairie voles that had previously undergone ELSD or Control conditions as juveniles. We found that, as adults, male ELSD prairie voles showed persistently lower REM sleep duration and female ELSD prairie voles showed persistently higher NREM sleep duration compared to Controls, but no other sleep parameters differed. We concluded that 1) persistent effects of ELSD on sleep into adulthood may contribute to the social and cognitive deficits observed in adult voles, and 2) sleep disruption early in life can influence later sleep patterns in adulthood.
]]></description>
<dc:creator>Jones-Tinsley, C. E.</dc:creator>
<dc:creator>Olson, R. J.</dc:creator>
<dc:creator>Mader, M.</dc:creator>
<dc:creator>Wickham, P. T.</dc:creator>
<dc:creator>Gutowsky, K.</dc:creator>
<dc:creator>Wong, C.</dc:creator>
<dc:creator>Chu, S. S.</dc:creator>
<dc:creator>Milman, N. E. P.</dc:creator>
<dc:creator>Cao, H.</dc:creator>
<dc:creator>Lim, M. M.</dc:creator>
<dc:date>2022-10-21</dc:date>
<dc:identifier>doi:10.1101/2022.10.18.512732</dc:identifier>
<dc:title><![CDATA[Early life sleep disruption has long lasting, sex specific effects on later development of sleep in prairie voles]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-10-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.10.18.512778v1?rss=1">
<title>
<![CDATA[
IL-6/STAT3 signaling drives early-stage pancreatic cancer cachexia via suppressed ketogenesis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.10.18.512778v1?rss=1</link>
<description><![CDATA[
Cancer cachexia is highly prevalent in patients with pancreatic ductal adenocarcinoma (PDAC). Although advanced cachexia is associated with inflammatory signaling, the early events driving wasting are poorly defined. Using an orthotopic mouse model of PDAC, we find that early cachexia is defined by a pronounced vulnerability to undernutrition, characterized by increased skeletal muscle wasting. PDAC suppresses lipid beta oxidation and impairs ketogenesis in the liver, which coordinates the adaptive response to nutritional scarcity. When PDAC mice are fed ketogenic diet, this effect is reversed, and muscle mass is preserved. Furthermore, physiologic levels of ketones are sufficient to protect myotubes against PDAC-associated wasting. Interleukin-6 (IL-6) drives liver metabolic reprogramming, and hepatocyte-specific loss of Signal Transducer and Activator of Transcription 3 (STAT3) is sufficient to prevent PDAC-associated muscle loss. Together, these studies define a key role for the liver in cachexia development and directly link skeletal muscle homeostasis to hepatic lipid oxidation.
]]></description>
<dc:creator>Arneson-Wissink, P. C.</dc:creator>
<dc:creator>Mendez, H.</dc:creator>
<dc:creator>Pelz, K.</dc:creator>
<dc:creator>Dickie, J.</dc:creator>
<dc:creator>Bartlett, A. Q.</dc:creator>
<dc:creator>Worley, B. L.</dc:creator>
<dc:creator>Krasnow, S. M.</dc:creator>
<dc:creator>Eil, R.</dc:creator>
<dc:creator>Grossberg, A. J.</dc:creator>
<dc:date>2022-10-21</dc:date>
<dc:identifier>doi:10.1101/2022.10.18.512778</dc:identifier>
<dc:title><![CDATA[IL-6/STAT3 signaling drives early-stage pancreatic cancer cachexia via suppressed ketogenesis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-10-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.10.21.512606v1?rss=1">
<title>
<![CDATA[
Modulation of type I interferon responses potently inhibits SARS-CoV-2 replication and inflammation in rhesus macaques 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.10.21.512606v1?rss=1</link>
<description><![CDATA[
Type-I interferons (IFN-I) are critical mediators of innate control of viral infections, but also drive recruitment of inflammatory cells to sites of infection, a key feature of severe COVID-19. Here, and for the first time, IFN-I signaling was modulated in rhesus macaques (RMs) prior to and during acute SARS-CoV-2 infection using a mutated IFN2 (IFN-modulator; IFNmod), which has previously been shown to reduce the binding and signaling of endogenous IFN-I. In SARS-CoV-2-infected RMs, IFNmod reduced both antiviral and inflammatory ISGs. Notably, IFNmod treatment resulted in a potent reduction in (i) SARS-CoV-2 viral load in Bronchoalveolar lavage (BAL), upper airways, lung, and hilar lymph nodes; (ii) inflammatory cytokines, chemokines, and CD163+MRC1-inflammatory macrophages in BAL; and (iii) expression of Siglec-1, which enhances SARS-CoV-2 infection and predicts disease severity, on circulating monocytes. In the lung, IFNmod also reduced pathogenesis and attenuated pathways of inflammasome activation and stress response during acute SARS-CoV-2 infection. This study, using an intervention targeting both IFN- and IFN-{beta} pathways, shows that excessive inflammation driven by type 1 IFN critically contributes to SARS-CoV-2 pathogenesis in RMs, and demonstrates the potential of IFNmod to limit viral replication, SARS-CoV-2 induced inflammation, and COVID-19 severity.
]]></description>
<dc:creator>Hoang, T. N.</dc:creator>
<dc:creator>Viox, E. G.</dc:creator>
<dc:creator>Upadhyay, A. A.</dc:creator>
<dc:creator>Strongin, Z.</dc:creator>
<dc:creator>Tharp, G. K.</dc:creator>
<dc:creator>Pino, M.</dc:creator>
<dc:creator>Nchioua, R.</dc:creator>
<dc:creator>Hirschenberger, M.</dc:creator>
<dc:creator>Gagne, M.</dc:creator>
<dc:creator>Nguyen, K.</dc:creator>
<dc:creator>Harper, J. L.</dc:creator>
<dc:creator>Marciano, S.</dc:creator>
<dc:creator>Boddapati, A. K.</dc:creator>
<dc:creator>Pellegrini, K. L.</dc:creator>
<dc:creator>Tisoncik-Go, J.</dc:creator>
<dc:creator>Whitmore, L. S.</dc:creator>
<dc:creator>Karunakaran, K. A.</dc:creator>
<dc:creator>Roy, M.</dc:creator>
<dc:creator>Kirejczyk, S.</dc:creator>
<dc:creator>Curran, E. H.</dc:creator>
<dc:creator>Wallace, C.</dc:creator>
<dc:creator>Wood, J. S.</dc:creator>
<dc:creator>Connor-Stroud, F.</dc:creator>
<dc:creator>Kasturi, S. P.</dc:creator>
<dc:creator>Levit, R. D.</dc:creator>
<dc:creator>Gale, M.</dc:creator>
<dc:creator>Vanderford, T. H.</dc:creator>
<dc:creator>Silvestri, G.</dc:creator>
<dc:creator>Busman-Sahay, K.</dc:creator>
<dc:creator>Estes, J. D.</dc:creator>
<dc:creator>Vaccari, M.</dc:creator>
<dc:creator>Douek, D. C.</dc:creator>
<dc:creator>Sparrer, K. M. J.</dc:creator>
<dc:creator>Kirchhoff, F.</dc:creator>
<dc:creator>Johnson, R. P.</dc:creator>
<dc:creator>Schreiber, G.</dc:creator>
<dc:creator>Bosinger, S. E.</dc:creator>
<dc:creator>Paiardini, M.</dc:creator>
<dc:date>2022-10-24</dc:date>
<dc:identifier>doi:10.1101/2022.10.21.512606</dc:identifier>
<dc:title><![CDATA[Modulation of type I interferon responses potently inhibits SARS-CoV-2 replication and inflammation in rhesus macaques]]></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/10.1101/2022.10.22.513357v1?rss=1">
<title>
<![CDATA[
Metformin use history and genome-wide DNA methylation profile: potential molecular mechanism for aging and longevity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.10.22.513357v1?rss=1</link>
<description><![CDATA[
BackgroundWhile there are medications that treat and manage age-related diseases, a compound that prolongs lifespan is yet to be discovered. Nonetheless, metformin, a commonly prescribed anti-diabetic medication, has repeatedly been shown to hinder aging in pre-clinical models and to be associated with lower mortality for humans, even among cancer patients. It is, however, not well understood how metformin can potentially prolong lifespan from a biological standpoint. We hypothesized that metformins potential mechanism of action for longevity is through its epigenetic modifications.

MethodsTo test our hypothesis, we conducted a post-hoc analysis of available genome-wide DNA methylation (DNAm) data obtained from whole blood collected from inpatients with and without a history of metformin use. We assessed the methylation profile of 171 patients (first run) and only among 63 diabetic patients (second run) and compared the DNAm rates between metformin users and nonusers.

ResultsEnrichment analysis from the Kyoto Encyclopedia of Genes and Genome (KEGG) showed pathways relevant to metformins mechanism of action, such as longevity, AMPK, and inflammatory pathways. We also identified several pathways related to delirium whose risk factor is aging. Moreover, top hits from the Gene Ontology (GO) included HIF-1 pathways. However, no individual CpG site showed genome-wide statistical significance (p<5E-08).

ConclusionThis study may elucidate metformins potential role in longevity through epigenetic modifications and other possible mechanisms of action.
]]></description>
<dc:creator>Marra, P. S.</dc:creator>
<dc:creator>Yamanashi, T.</dc:creator>
<dc:creator>Crutchley, K. J.</dc:creator>
<dc:creator>Wahba, N. E.</dc:creator>
<dc:creator>Anderson, Z.-E. M.</dc:creator>
<dc:creator>Modukuri, M.</dc:creator>
<dc:creator>Chang, G.</dc:creator>
<dc:creator>Tran, T.</dc:creator>
<dc:creator>Iwata, M.</dc:creator>
<dc:creator>Cho, H. R.</dc:creator>
<dc:creator>Shinozaki, G.</dc:creator>
<dc:date>2022-10-24</dc:date>
<dc:identifier>doi:10.1101/2022.10.22.513357</dc:identifier>
<dc:title><![CDATA[Metformin use history and genome-wide DNA methylation profile: potential molecular mechanism for aging and longevity]]></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/10.1101/2022.10.28.514267v1?rss=1">
<title>
<![CDATA[
Reversing gut microbiome-driven adipose tissue inflammation alleviates metabolic syndrome 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.10.28.514267v1?rss=1</link>
<description><![CDATA[
The gut microbiota contributes to macrophage-mediated inflammation in adipose tissue with consumption of an obesogenic diet, thus driving the development of metabolic syndrome. There is a need to identify and develop interventions that abrogate this condition. The hops-derived prenylated flavonoid xanthohumol (XN) and its semi-synthetic derivative tetrahydroxanthohumol (TXN) attenuate high-fat diet-induced obesity, hepatosteatosis and metabolic syndrome in C57Bl/6J mice. This coincides with a decrease in pro-inflammatory gene expression in the gut and adipose tissue, together with alterations in the gut microbiota and bile acid composition. In this study, we integrated and interrogated multi-omics data from different organs with fecal 16S sequences and systemic metabolic phenotypic data using a transkingdom network analysis. By incorporating cell type information from single cell RNA-seq data, we discovered TXN attenuates macrophage inflammatory processes in adipose tissue. TXN treatment also reversed levels of inflammation-inducing microbes, such as Oscillibacter valericigenes, that lead to adverse metabolic phenotypes. Furthermore, in vitro validation in macrophage cell lines and in vivo mouse supplementation showed addition of O. valericigenes supernatant induced the expression of metabolic macrophage signature genes that are downregulated by TXN in vivo. Our findings establish an important mechanism by which TXN mitigates adverse phenotypic outcomes from diet-induced obesity and metabolic syndrome. It primarily reduces the abundance of pro-inflammatory gut microbes that can otherwise promote macrophage-associated inflammation in adipose tissue.
]]></description>
<dc:creator>Newman, N. K.</dc:creator>
<dc:creator>Zhang, Y.</dc:creator>
<dc:creator>Padiadpu, J.</dc:creator>
<dc:creator>Miranda, C.</dc:creator>
<dc:creator>Magana, A. A.</dc:creator>
<dc:creator>Hioki, K.</dc:creator>
<dc:creator>Pederson, J. W.</dc:creator>
<dc:creator>Li, Z.</dc:creator>
<dc:creator>Gurung, M.</dc:creator>
<dc:creator>Bruce, A. M.</dc:creator>
<dc:creator>Brown, K.</dc:creator>
<dc:creator>Bobe, G.</dc:creator>
<dc:creator>Sharpton, T. J.</dc:creator>
<dc:creator>Shulzhenko, N.</dc:creator>
<dc:creator>Maier, C.</dc:creator>
<dc:creator>Stevens, J. F.</dc:creator>
<dc:creator>Gombart, A. F.</dc:creator>
<dc:creator>Morgun, A.</dc:creator>
<dc:date>2022-10-31</dc:date>
<dc:identifier>doi:10.1101/2022.10.28.514267</dc:identifier>
<dc:title><![CDATA[Reversing gut microbiome-driven adipose tissue inflammation alleviates metabolic syndrome]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-10-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.11.01.514598v1?rss=1">
<title>
<![CDATA[
Multimodal profiling of term human decidua reveals tissue-specific immune adaptations with maternal obesity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.11.01.514598v1?rss=1</link>
<description><![CDATA[
The leukocyte diversity of early maternal-fetal interface has been recently described, however, characterization of decidua at term is still pending. We, therefore, profiled the CD45+ compartment within human term decidua collected via cesarean section. Relative to the first trimester, our analyses revealed a shift from NK cells and macrophages to T-cells and enhanced activation of all immune cells. T-cells in the decidua were phenotypically distinct from their blood counterparts and demonstrated significant clonotype sharing. Using pseudo-temporal analyses of macrophages, we characterize previously undescribed diversity of bonafide tissue resident decidual macrophages. The frequency of this population correlated with high pregravid maternal body mass index but responded poorly to bacteria ligands. Collectively, these findings provide evidence of skewing of decidual myeloid cells towards immunoregulation representing a possible mechanism to safeguard the fetus. This immune cell atlas of term decidua is an asset for future studies investigating pathological conditions that compromise reproductive success.

One sentence summaryHuman immune cell atlas of term decidua and observed alterations with pre-gravid obesity.
]]></description>
<dc:creator>Sureshchandra, S.</dc:creator>
<dc:creator>Doratt, B. M.</dc:creator>
<dc:creator>Mendoza, N.</dc:creator>
<dc:creator>Rincon, M.</dc:creator>
<dc:creator>Marshall, N. E.</dc:creator>
<dc:creator>Messaoudi, I.</dc:creator>
<dc:date>2022-11-02</dc:date>
<dc:identifier>doi:10.1101/2022.11.01.514598</dc:identifier>
<dc:title><![CDATA[Multimodal profiling of term human decidua reveals tissue-specific immune adaptations with maternal obesity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-11-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.11.01.514719v1?rss=1">
<title>
<![CDATA[
A generalizable epigenetic clock captures aging in two nonhuman primates. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.11.01.514719v1?rss=1</link>
<description><![CDATA[
Epigenetic clocks generated from DNA methylation array data provide important insights into biological aging, disease susceptibility, and mortality risk. However, these clocks cannot be applied to high-throughput, sequence-based datasets more commonly used to study nonhuman animals. Here, we built a generalizable epigenetic clock using genome-wide DNA methylation data from 493 free-ranging rhesus macaques. Using a sliding-window approach that maximizes generalizability across datasets and species, this model predicted age with high accuracy ({+/-} 1.42 years) in held-out test samples, as well as in two independent test sets: rhesus macaques from a captive population (n=43) and wild baboons in Kenya (n=271). Our model can also be used to generate insight into the factors hypothesized to alter epigenetic aging, including social status and exposure to traumatic events. Our results thus provide a flexible tool for predicting age in other populations and species and illustrate how connecting behavioral data with the epigenetic clock can uncover social influences on biological age.
]]></description>
<dc:creator>Goldman, E. A.</dc:creator>
<dc:creator>Watowich, M. M.</dc:creator>
<dc:creator>Chiou, K. L.</dc:creator>
<dc:creator>Mercer, A.</dc:creator>
<dc:creator>Sams, S. N.</dc:creator>
<dc:creator>Horvath, J. E.</dc:creator>
<dc:creator>Anderson, J.</dc:creator>
<dc:creator>Cayo Biobank Research Unit,</dc:creator>
<dc:creator>Tung, J.</dc:creator>
<dc:creator>Higham, J. P.</dc:creator>
<dc:creator>Brent, L. J.</dc:creator>
<dc:creator>Martinez, M. I.</dc:creator>
<dc:creator>Montague, M. J.</dc:creator>
<dc:creator>Platt, M. L.</dc:creator>
<dc:creator>Sterner, K. N.</dc:creator>
<dc:creator>Snyder-Mackler, N.</dc:creator>
<dc:date>2022-11-02</dc:date>
<dc:identifier>doi:10.1101/2022.11.01.514719</dc:identifier>
<dc:title><![CDATA[A generalizable epigenetic clock captures aging in two nonhuman primates.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-11-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.11.03.515127v1?rss=1">
<title>
<![CDATA[
Magnetic Levitation and Sorting of Neoplastic Circulating Cell Hybrids 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.11.03.515127v1?rss=1</link>
<description><![CDATA[
Circulating hybrid cells (CHCs) are a novel, rare cell population that harbor tumor and immune cell phenotypes and genotypes and are detectible in peripheral blood. Several recent reports implicated CHCs in the metastatic cascade and found their enumeration to provide better prognostic value than conventionally-defined circulating tumor cells (CTCs). However, methods for isolation and enrichment of CHCs are not well-studied or established. Here, we developed an ultrasensitive, antigen-independent platform leveraging the principles of magnetic levitation for the detection and isolation of disseminated neoplastic CHCs. For the first time, we demonstrate that CHCs can be magnetically focused to different levitation heights, under various paramagnetic conditions using a static levitation system, and we quantified the biophysical properties of CHCs (i.e., levitation heights). In addition, we investigated whether magnetic levitation approach can be combined with the affinity-based strategies to enrich CHCs under the magnetic field. Using clinical samples from breast and colorectal cancer patients, we demonstrated that neoplastic cells can be sorted with a magnetic levitation-based sorting device, without relying on any surface markers. Overall, we demonstrated the feasibility of the magnetic levitation method for unbiased enrichment of rare neoplastic-immune hybrid cells from peripheral blood specimens from cancer patients. This approach can be expanded to more clinical samples and cancer types to unprecedentedly explore the biology of rare neoplastic cells and develop metastasis-tailored therapies broadly impacting personalized and precision clinical treatments.
]]></description>
<dc:creator>Liang, K.</dc:creator>
<dc:creator>Yaman, S.</dc:creator>
<dc:creator>Patel, R. K.</dc:creator>
<dc:creator>Parappilly, M. S.</dc:creator>
<dc:creator>Walker, B. S.</dc:creator>
<dc:creator>Wong, M. H.</dc:creator>
<dc:creator>Durmus, N. G.</dc:creator>
<dc:date>2022-11-04</dc:date>
<dc:identifier>doi:10.1101/2022.11.03.515127</dc:identifier>
<dc:title><![CDATA[Magnetic Levitation and Sorting of Neoplastic Circulating Cell Hybrids]]></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/10.1101/2022.11.08.515625v1?rss=1">
<title>
<![CDATA[
PathExpSurv: Pathway Expansion for Explainable Survival Analysis and Disease Gene Discovery 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.11.08.515625v1?rss=1</link>
<description><![CDATA[
MotivationIn the field of biology and medicine, the interpretability and accuracy are both important when designing predictive models. The interpretability of many machine learning models such as neural networks is still a challenge. Recently, many researchers utilized prior information such as biological pathways to develop bioinformatics methods based on neural networks, so that the prior information can provide some insights and interpretability for the models. However, the prior biological knowledge may be incomplete and there still exists some unknown information to be explored.

ResultsWe proposed a novel method, named PathExpSurv, to gain an insight into the black-box model of neural network for cancer survival analysis. We demonstrated that PathExpSurv could not only incorporate the known prior information into the model, but also explore the unknown possible expansion to the existing pathways. We performed downstream analyses based on the expanded pathways and successfully identified some key genes associated with the diseases and original pathways.

AvailabilityPython source code of PathExpSurv is freely available at https://github.com/Wu-Lab/PathExpSurv.

Contactlywu@amss.ac.cn

Supplementary informationSupplementary data are available at Bioinformatics online.
]]></description>
<dc:creator>Hou, Z.</dc:creator>
<dc:creator>Leng, J.</dc:creator>
<dc:creator>Yu, J.</dc:creator>
<dc:creator>Xia, Z.</dc:creator>
<dc:creator>Wu, L.-Y.</dc:creator>
<dc:date>2022-11-09</dc:date>
<dc:identifier>doi:10.1101/2022.11.08.515625</dc:identifier>
<dc:title><![CDATA[PathExpSurv: Pathway Expansion for Explainable Survival Analysis and Disease Gene Discovery]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-11-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.11.08.515705v1?rss=1">
<title>
<![CDATA[
Flexible, Scalable, High Channel Count Stereo-Electrode for Recording in the Human Brain 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.11.08.515705v1?rss=1</link>
<description><![CDATA[
Over the past decade, stereotactically placed electrodes have become the gold standard for deep brain recording and stimulation for a wide variety of neurological and psychiatric diseases. Current electrodes, however, are limited in their spatial resolution and ability to record from small populations of neurons, let alone individual neurons. Here, we report on a novel, reconfigurable, monolithically integrated human-grade flexible depth electrode capable of recording from up to 128 channels and able to record at a depth of 10 cm in brain tissue. This thin, stylet-guided depth electrode is capable of recording local field potentials and single unit neuronal activity (action potentials), validated across species. This device represents a major new advance in manufacturing and design approaches which extends the capabilities of a mainstay technology in clinical neurology.

One-Sentence SummaryA human-grade thin-film depth electrode offers new opportunities in spatial and temporal resolution for recording brain activity.
]]></description>
<dc:creator>Lee, K.</dc:creator>
<dc:creator>Paulk, A. C.</dc:creator>
<dc:creator>Ro, Y. G.</dc:creator>
<dc:creator>Cleary, D. R.</dc:creator>
<dc:creator>Tonsfeldt, K. J.</dc:creator>
<dc:creator>Kfir, Y.</dc:creator>
<dc:creator>Pezaris, J.</dc:creator>
<dc:creator>Tchoe, Y.</dc:creator>
<dc:creator>Lee, J.</dc:creator>
<dc:creator>Bourhis, A. M.</dc:creator>
<dc:creator>Vatsyayan, R.</dc:creator>
<dc:creator>Russman, S. M.</dc:creator>
<dc:creator>Yang, J. C.</dc:creator>
<dc:creator>Baohan, A.</dc:creator>
<dc:creator>Richardson, M.</dc:creator>
<dc:creator>Williams, Z. M.</dc:creator>
<dc:creator>Fried, S.</dc:creator>
<dc:creator>Raslan, A.</dc:creator>
<dc:creator>Halgren, E.</dc:creator>
<dc:creator>Cash, S. S.</dc:creator>
<dc:creator>Dayeh, S. A.</dc:creator>
<dc:date>2022-11-09</dc:date>
<dc:identifier>doi:10.1101/2022.11.08.515705</dc:identifier>
<dc:title><![CDATA[Flexible, Scalable, High Channel Count Stereo-Electrode for Recording in the Human Brain]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-11-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.11.14.516380v1?rss=1">
<title>
<![CDATA[
Septins and K63 chains form separate bacterial microdomains during autophagy of entrapped Shigella 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.11.14.516380v1?rss=1</link>
<description><![CDATA[
During host cell invasion, Shigella escapes to the cytosol and polymerizes actin for cell-to-cell spread. To restrict cell-to-cell spread, host cells employ cell-autonomous immune responses including antibacterial autophagy and septin cage entrapment. How septins interact with autophagy to target Shigella to destruction is poorly understood. Here, we employed a correlative light and cryo-soft X-ray tomography (cryo-SXT) pipeline to study Shigella septin cage entrapment in its near native state. Quantitative cryo-SXT showed that Shigella fragments mitochondria and enabled visualization of X-ray dense structures ([~]30 nm resolution) surrounding Shigella entrapped in septin cages. Using Airyscan confocal microscopy, we observed Lysine 63 (K63)-linked ubiquitin chains decorating septin caged entrapped Shigella. Remarkably, septins and K63 chains form separate bacterial microdomains, indicating they are recruited separately during antibacterial autophagy. Cryo-SXT and live cell imaging revealed an interaction between septins and LC3B-positive membranes during autophagy of Shigella. Together, these findings demonstrate how septin caged Shigella are targeted to autophagy and provide fundamental insights into autophagy-cytoskeleton interactions.
]]></description>
<dc:creator>Lobato-Marquez, D.</dc:creator>
<dc:creator>Conesa, J. J.</dc:creator>
<dc:creator>Lopez-Jimenez, A. T.</dc:creator>
<dc:creator>Divine, M. E.</dc:creator>
<dc:creator>Pruneda, J. N.</dc:creator>
<dc:creator>Mostowy, S.</dc:creator>
<dc:date>2022-11-15</dc:date>
<dc:identifier>doi:10.1101/2022.11.14.516380</dc:identifier>
<dc:title><![CDATA[Septins and K63 chains form separate bacterial microdomains during autophagy of entrapped Shigella]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-11-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.11.20.517258v1?rss=1">
<title>
<![CDATA[
Computational Pipeline to Identify Gene signatures that Define Cancer Subtypes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.11.20.517258v1?rss=1</link>
<description><![CDATA[
MotivationThe heterogeneous nature of cancers with multiple subtypes makes them challenging to treat. However, multi-omics data can be used to identify new therapeutic targets and we established a computational strategy to improve data mining.

ResultsUsing our approach we identified genes and pathways specific to cancer subtypes that can serve as biomarkers and therapeutic targets. Using a TCGA breast cancer dataset we applied the ExtraTreesClassifier dimensionality reduction along with logistic regression to select a subset of genes for model training. Applying hyperparameter tuning, increased the model accuracy up to 92%. Finally, we identified 20 significant genes using differential expression. These targetable genes are associated with various cellular processes that impact cancer progression. We then applied our approach to a glioma dataset and again identified subtype specific targetable genes.

ConclusionOur research indicates a broader applicability of our strategy to identify specific cancer subtypes and targetable pathways for various cancers.
]]></description>
<dc:creator>Mittal, E.</dc:creator>
<dc:creator>Parikh, V.</dc:creator>
<dc:creator>Kirchgaessner, R.</dc:creator>
<dc:date>2022-11-22</dc:date>
<dc:identifier>doi:10.1101/2022.11.20.517258</dc:identifier>
<dc:title><![CDATA[Computational Pipeline to Identify Gene signatures that Define Cancer Subtypes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-11-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.11.22.517284v1?rss=1">
<title>
<![CDATA[
Adaptive nanopore sequencing for single cell characterization of cancer mutations and gene rearrangements 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.11.22.517284v1?rss=1</link>
<description><![CDATA[
In this proof-of-concept study, we developed a single cell method that identifies somatic alterations found in coding regions of mRNAs and integrates these mutation genotypes with their matching cell transcriptomes. We used nanopore adaptive sampling on single cell cDNA libraries, generated long read sequences from target gene transcripts and identified coding variants among individual cells. Short-read single cell transcriptomes characterized the cell types with mutations. We delineated CRISPR edits from a cancer cell line. From primary cancer samples, we targeted hundreds of cancer genes, identified somatic coding mutations and a gene rearrangement among individual tumor cells.
]]></description>
<dc:creator>Grimes, S. M.</dc:creator>
<dc:creator>Kim, H. S.</dc:creator>
<dc:creator>Roy, S.</dc:creator>
<dc:creator>Sathe, A.</dc:creator>
<dc:creator>Ayala, C. I.</dc:creator>
<dc:creator>Almeida-Notestine, A. F.</dc:creator>
<dc:creator>Haebe, S.</dc:creator>
<dc:creator>Shree, T.</dc:creator>
<dc:creator>Levy, R.</dc:creator>
<dc:creator>Lau, B. T.</dc:creator>
<dc:creator>Ji, H. P.</dc:creator>
<dc:date>2022-11-24</dc:date>
<dc:identifier>doi:10.1101/2022.11.22.517284</dc:identifier>
<dc:title><![CDATA[Adaptive nanopore sequencing for single cell characterization of cancer mutations and gene rearrangements]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-11-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.11.23.517739v1?rss=1">
<title>
<![CDATA[
Consensus Label Propagation with Graph Convolutional Networks for Single-Cell RNA Sequencing Cell Type Annotation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.11.23.517739v1?rss=1</link>
<description><![CDATA[
MotivationSingle-cell RNA sequencing (scRNA-seq) data, annotated by cell type, is useful in a variety of downstream biological applications, such as profiling gene expression at the single-cell level. However, manually assigning these annotations with known marker genes is both time-consuming and subjective.

ResultsWe present a Graph Convolutional Network (GCN) based approach to automate the annotation process. Our process builds upon existing labeling approaches, using state-of-the-art tools to find cells with highly confident label assignments through consensus and spreading these confident labels with a semi-supervised GCN. Using simulated data and two scRNA-seq data sets from different tissues, we show that our method improves accuracy over a simple consensus algorithm and the average of the underlying tools. We also compare our method to a non-parametric neighbor majority approach, showing comparable results. We then demonstrate that our GCN method allows for feature interpretation, identifying important genes for cell type classification. We present our completed pipeline, written in PyTorch, as an end-to-end tool for automating and interpreting the classification of scRNA-seq data.

AvailabilityOur code for conducting the experiments in this paper and using our model is available at https://github.com/lewinsohndp/scSHARP

Contactd_lewinsohn@coloradocollege.edu

Supplementary informationSupplementary data are available at Journal Name online.
]]></description>
<dc:creator>Lewinsohn, D. P.</dc:creator>
<dc:creator>Vigh-Conrad, K. A.</dc:creator>
<dc:creator>Conrad, D.</dc:creator>
<dc:creator>Scott, C. B.</dc:creator>
<dc:date>2022-11-25</dc:date>
<dc:identifier>doi:10.1101/2022.11.23.517739</dc:identifier>
<dc:title><![CDATA[Consensus Label Propagation with Graph Convolutional Networks for Single-Cell RNA Sequencing Cell Type Annotation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-11-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.11.29.518385v1?rss=1">
<title>
<![CDATA[
SARS-CoV-2 Vaccine Booster Elicits Robust Prolonged Maternal Antibody Responses and Passive Transfer Via The Placenta And Breastmilk 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.11.29.518385v1?rss=1</link>
<description><![CDATA[
BackgroundInfection during pregnancy can result in adverse outcomes for both pregnant persons and offspring. Maternal vaccination is an effective mechanism to protect both mother and neonate into post-partum. However, our understanding of passive transfer of antibodies elicited by maternal SARS-CoV-2 mRNA vaccination during pregnancy remains incomplete.

ObjectiveWe aimed to evaluate the antibody responses engendered by maternal SARS-CoV-2 vaccination following initial and booster doses in maternal circulation and breastmilk to better understand passive immunization of the newborn.

Study DesignWe collected longitudinal blood samples from 121 pregnant women who received SARS-CoV-2 mRNA vaccines spanning from early gestation to delivery followed by collection of blood samples and breastmilk between delivery and 12 months post-partum. During the study, 70% of the participants also received a booster post-partum. Paired maternal plasma, breastmilk, umbilical cord plasma, and newborn plasma samples were tested via enzyme-linked immunosorbent assays (ELISA) to evaluate SARS-CoV-2 specific IgG antibody levels.

ResultsVaccine-elicited maternal antibodies were detected in both cord blood and newborn blood, albeit at lower levels than maternal circulation, demonstrating transplacental passive immunization. Booster vaccination significantly increased spike specific IgG antibody titers in maternal plasma and breastmilk. Finally, SARS-CoV-2 specific IgG antibodies in newborn blood correlated negatively with days post initial maternal vaccine dose.

ConclusionVaccine-induced maternal SARS-CoV-2 antibodies were passively transferred to the offspring in utero via the placenta and after birth via breastfeeding. Maternal booster vaccination, regardless of gestational age at maternal vaccination, significantly increased antibody levels in breastmilk and maternal plasma, indicating the importance of this additional dose to maximize passive protection against SARS-CoV-2 infection for neonates and infants until vaccination eligibility.
]]></description>
<dc:creator>Marshall, N. E.</dc:creator>
<dc:creator>Blanton, M. B.</dc:creator>
<dc:creator>Doratt, B. M.</dc:creator>
<dc:creator>Malherbe, D. C.</dc:creator>
<dc:creator>Rincon, M.</dc:creator>
<dc:creator>True, H.</dc:creator>
<dc:creator>McDonald, T.</dc:creator>
<dc:creator>Beauregard, C.</dc:creator>
<dc:creator>Adatorwovor, R.</dc:creator>
<dc:creator>Messaoudi, I.</dc:creator>
<dc:date>2022-11-29</dc:date>
<dc:identifier>doi:10.1101/2022.11.29.518385</dc:identifier>
<dc:title><![CDATA[SARS-CoV-2 Vaccine Booster Elicits Robust Prolonged Maternal Antibody Responses and Passive Transfer Via The Placenta And Breastmilk]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-11-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.11.29.518431v1?rss=1">
<title>
<![CDATA[
Intermittent treatment with elamipretide preserves exercise tolerance in aged female mice 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.11.29.518431v1?rss=1</link>
<description><![CDATA[
The pathology of aging impacts multiple organ systems including the kidney, skeletal, and cardiac muscle. Long-term treatment with the mitochondrial targeted peptide elamipretide has previously been shown to improve in vivo mitochondrial function in aged mice that is associated with increased fatigue resistance and treadmill performance, improved cardiovascular diastolic function, and glomerular architecture of the kidney. However, elamipretide is a short tetrameric peptide that is not orally bioavailable limiting its routes of administration. This study tested whether twice weekly intermittent injections of elamipretide could recapitulate the same functional improvements as continuous long-term infusion. We found that intermittent treatment with elamipretide for 8 months preserved endurance running in mice, skeletal muscle force production, and left ventricular mass but did not affect heart or kidney function as previously reported using continuous treatment.
]]></description>
<dc:creator>Campbell, M. D.</dc:creator>
<dc:creator>Samuelson, A. T.</dc:creator>
<dc:creator>Chiao, Y. A.</dc:creator>
<dc:creator>Sweetwyne, M. T.</dc:creator>
<dc:creator>Ladiges, W. C.</dc:creator>
<dc:creator>Rabinovitch, P. S.</dc:creator>
<dc:creator>Marcinek, D.</dc:creator>
<dc:date>2022-12-02</dc:date>
<dc:identifier>doi:10.1101/2022.11.29.518431</dc:identifier>
<dc:title><![CDATA[Intermittent treatment with elamipretide preserves exercise tolerance in aged female mice]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-12-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.11.29.518436v1?rss=1">
<title>
<![CDATA[
Data mining and experimental approaches to identify combination of natural herbs against bacterial infections 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.11.29.518436v1?rss=1</link>
<description><![CDATA[
Various studies have identified that natural herbs can be repurposed to treat infectious and bacterial diseases. The purpose of this study is first to test the medicinal value of five herbs including asafoetida, cumin, fenugreek, neem, and turmeric as single agent and in pairs using the bacterial zone of inhibition assay. Second, we used target and network analyses to predict the best combinations. We found that all the herbs as single agent were effective against bacterial infection in the following descending order of efficacy: cumin > turmeric > neem > fenugreek > asafoetida as compared to vehicle (ethanol) treated control. Among all the tested combinations the turmeric and fenugreek combination had the best efficacy in inhibiting the bacterial growth. Next to understand the mechanism of action and to predict the effective combinations among available herbs, we used a data mining and computational analysis approach. Using NPASS, BindingDB, and pathway analysis tools, we identified the bioactive compounds for each herb, then identified the targets for each bioactive compound, and then identified associated pathways for these targets. Then we measured the target/pathway overlap for each herb and identified that the most effective combinations were those which have non-overlapping targets/pathways. For example, we showed as a proof-of-concept that turmeric and fenugreek have the least overlapping targets/pathways and thus is most effective in inhibiting bacteria growth. Our approach is applicable to treat bacterial infections and other human diseases such as cancer. Overall, the computational prediction along with experimental validation can help identify novel combinations that have significant antibacterial activity and may help prevent drug-resistant microbial diseases in human and plants.
]]></description>
<dc:creator>Mittal, E.</dc:creator>
<dc:creator>Duncan, S.</dc:creator>
<dc:creator>Chamberlin, S.</dc:creator>
<dc:date>2022-11-29</dc:date>
<dc:identifier>doi:10.1101/2022.11.29.518436</dc:identifier>
<dc:title><![CDATA[Data mining and experimental approaches to identify combination of natural herbs against bacterial infections]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-11-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.12.04.518984v1?rss=1">
<title>
<![CDATA[
Discordant results among MHC binding affinity prediction tools 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.12.04.518984v1?rss=1</link>
<description><![CDATA[
A large number of machine learning-based Major Histocompatibility Complex (MHC) binding affinity (BA) prediction tools have been developed and are widely used for both investigational and therapeutic applications, so it is important to explore differences in tool outputs. We examined predictions of four popular tools (netMHCpan, HLAthena, MHCflurry, and MHCnuggets) across a range of possible peptide sources (human, viral, and randomly generated) and MHC class I alleles. We uncovered inconsistencies in predictions of BA, allele promiscuity and the relationship between physical properties of peptides by source and BA predictions, as well as quality of training data. Our work raises fundamental questions about the fidelity of peptide-MHC binding prediction tools and their real-world implications.
]]></description>
<dc:creator>Nguyen, A.</dc:creator>
<dc:creator>Nellore, A.</dc:creator>
<dc:creator>Thompson, R. F.</dc:creator>
<dc:date>2022-12-07</dc:date>
<dc:identifier>doi:10.1101/2022.12.04.518984</dc:identifier>
<dc:title><![CDATA[Discordant results among MHC binding affinity prediction tools]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-12-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.12.07.519382v1?rss=1">
<title>
<![CDATA[
DNA damage-induced lncRNA MEG9 impacts angiogenesis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.12.07.519382v1?rss=1</link>
<description><![CDATA[
Endothelial cells are highly responsive to environmental changes that allow them to adapt to intrinsic and extrinsic stimuli and switch their transcriptome accordingly to go back to vascular homeostasis. Our previous data demonstrated that small non-coding-RNAs respond quickly to genotoxic stressors and determined endothelial cell fate and DNA damage response. To further understand the contribution of non-coding-RNAs, we profiled differentially expressed long non-coding RNAs in response to genotoxic stress and compared them to pro-angiogenic growth factor signaling. We identified the Maternally expressed gene 9 (MEG9) as a cytoprotective lncRNA in the endothelium. Gain and Loss-of-function studies indicate that MEG9 prevents endothelial cells from cell death, suggesting that MEG9 responses to genotoxic stress can be an adaptive and protective mechanism. Consistent with this phenotype, the knockdown of MEG9 decreases growth factor-dependent angiogenesis in a 3D fibrin gel angiogenesis assay. Deletion of the MEG9 ortholog, Mirg, in mice results in increased vascular leak in Matrigel plugs and a sex and age-dependent decrease in platelets. Mechanistically, we observed that both MEG9 knockdown in vitro and Mirg-deleted mice in vivo activated common pathways, including apoptosis, clotting, and inflammation. Indeed, the proinflammatory adhesion molecule ICAM1 was significantly increased in human and mouse endothelial cells in a MEG9-dependent manner, supporting the increased vascular permeability observed on MEG9 deficient cells. Taken together, our findings illustrate how genotoxic stress responses through dynamic modulation of lncRNAs, such as MEG9, trigger adaptive mechanisms to maintain endothelial function, while loss of these molecules contributes to maladaptive responses and endothelial cell dysfunction.
]]></description>
<dc:creator>Fraile-Bethencourt, E.</dc:creator>
<dc:creator>Khou, S.</dc:creator>
<dc:creator>Wilson, R.</dc:creator>
<dc:creator>Baris, A.</dc:creator>
<dc:creator>Ruhl, R.</dc:creator>
<dc:creator>Espinosa-Diez, C.</dc:creator>
<dc:creator>Anand, S.</dc:creator>
<dc:date>2022-12-10</dc:date>
<dc:identifier>doi:10.1101/2022.12.07.519382</dc:identifier>
<dc:title><![CDATA[DNA damage-induced lncRNA MEG9 impacts angiogenesis]]></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/10.1101/2022.12.08.519209v1?rss=1">
<title>
<![CDATA[
SARM1 detection in oligodendrocytes but not Schwann cells though sarm1/Sarm1 deletion does not perturb CNS nor PNS myelination in zebrafish and mice. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.12.08.519209v1?rss=1</link>
<description><![CDATA[
SARM1 is a central regulator of programmed axon death and is required to initiate axon self-destruction after traumatic and toxic insults to the nervous system. Abnormal activation of this axon degeneration pathway is increasingly recognized as a contributor to human neurological disease and SARM1 knockdown or inhibition has become an attractive therapeutic strategy to preserve axon loss in a variety of disorders of the peripheral and central nervous system. Despite this, it remains unknown whether Sarm1/SARM1 is present in myelinating glia and whether it plays a role in myelination in the PNS or CNS. It is important to answer these questions to understand whether future therapies inhibiting SARM1 function may have unintended deleterious impacts on myelination. Here we show that Sarm1 mRNA is present in oligodendrocytes in zebrafish but only detectable at low levels in Schwann cells in both zebrafish and mice. We find SARM1 protein is readily detectable in murine oligodendrocytes in vitro and in vivo and activation of endogenous SARM1 in oligodendrocytes induces cell death. In contrast, SARM1 protein is not detectable in Schwann cells and satellite glia in the adult murine nervous system. Cultured Schwann cells contain negligible functional SARM1 and are insensitive to specific SARM1 activators. Using zebrafish and mouse Sarm1 mutants, we show that SARM1 is not required for initiation of myelination nor myelin sheath maintenance by oligodendrocytes and Schwann cells. Thus, strategies to inhibit SARM1 function in the nervous system to treat neurological disease are unlikely to perturb myelination in humans.

Main PointsO_LISARM1 protein is detectable in oligodendrocytes but not in Schwann cells
C_LIO_LIOligodendrocytes but not Schwann cells die in response to endogenous SARM1 activation
C_LIO_LICNS nor PNS myelination, in zebrafish and mice, is hindered by loss of sarm1/Sarm1
C_LI
]]></description>
<dc:creator>Fazal, S. V.</dc:creator>
<dc:creator>Mutschler, C.</dc:creator>
<dc:creator>Chen, C.</dc:creator>
<dc:creator>Turmaine, M.</dc:creator>
<dc:creator>Chen, C.-Y.</dc:creator>
<dc:creator>Hsueh, Y.-P.</dc:creator>
<dc:creator>Loreto, A.</dc:creator>
<dc:creator>Casillas-Bajo, A.</dc:creator>
<dc:creator>Cabedo, H.</dc:creator>
<dc:creator>Franklin, R. J.</dc:creator>
<dc:creator>Barker, R. A.</dc:creator>
<dc:creator>Monk, K.</dc:creator>
<dc:creator>Steventon, B.</dc:creator>
<dc:creator>Coleman, M.</dc:creator>
<dc:creator>Gomez-Sanchez, J. A.</dc:creator>
<dc:creator>Arthur-Farraj, P.</dc:creator>
<dc:date>2022-12-09</dc:date>
<dc:identifier>doi:10.1101/2022.12.08.519209</dc:identifier>
<dc:title><![CDATA[SARM1 detection in oligodendrocytes but not Schwann cells though sarm1/Sarm1 deletion does not perturb CNS nor PNS myelination in zebrafish and mice.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-12-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.12.09.519791v1?rss=1">
<title>
<![CDATA[
Frequent first-trimester pregnancy loss in rhesus macaques infected with African-lineage Zika virus 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.12.09.519791v1?rss=1</link>
<description><![CDATA[
In the 2016 Zika virus (ZIKV) pandemic, a previously unrecognized risk of birth defects surfaced in babies whose mothers were infected with Asian-lineage ZIKV during pregnancy. Less is known about the impacts of gestational African-lineage ZIKV infections. Given high human immunodeficiency virus (HIV) burdens in regions where African-lineage ZIKV circulates, we evaluated whether pregnant rhesus macaques infected with simian immunodeficiency virus (SIV) have a higher risk of African-lineage ZIKV-associated birth defects. Remarkably, in both SIV+ and SIV-animals, ZIKV infection early in the first trimester caused a high incidence (78%) of spontaneous pregnancy loss within 20 days. These findings suggest a significant risk for early pregnancy loss associated with African-lineage ZIKV infection and provide the first consistent ZIKV-associated phenotype in macaques for testing medical countermeasures.
]]></description>
<dc:creator>Rosinski, J. R.</dc:creator>
<dc:creator>Raasch, L. E.</dc:creator>
<dc:creator>Barros Tiburcio, P.</dc:creator>
<dc:creator>Breitbach, M. E.</dc:creator>
<dc:creator>Shepherd, P. M.</dc:creator>
<dc:creator>Yamamoto, K.</dc:creator>
<dc:creator>Razo, E.</dc:creator>
<dc:creator>Krabbe, N. P.</dc:creator>
<dc:creator>Bliss, M. I.</dc:creator>
<dc:creator>Richardson, A. D.</dc:creator>
<dc:creator>Einwalter, M. A.</dc:creator>
<dc:creator>Weiler, A. M.</dc:creator>
<dc:creator>Sneed, E. L.</dc:creator>
<dc:creator>Fuchs, K. B.</dc:creator>
<dc:creator>Zeng, X.</dc:creator>
<dc:creator>Noguchi, K. K.</dc:creator>
<dc:creator>Morgan, T. K.</dc:creator>
<dc:creator>Alberts, A. J.</dc:creator>
<dc:creator>Antony, K. M.</dc:creator>
<dc:creator>Spanton, R. V.</dc:creator>
<dc:creator>Kabakov, S.</dc:creator>
<dc:creator>Ausderau, K. K.</dc:creator>
<dc:creator>Bohm, E. K.</dc:creator>
<dc:creator>Pritchard, J. C.</dc:creator>
<dc:creator>Ver Hoeve, J.</dc:creator>
<dc:creator>Spanton, R. V.</dc:creator>
<dc:creator>Kim, C.</dc:creator>
<dc:creator>Nork, T. M.</dc:creator>
<dc:creator>Katz, A. W.</dc:creator>
<dc:creator>Rasmussen, C. A.</dc:creator>
<dc:creator>Hartman, A.</dc:creator>
<dc:creator>Mejia, A.</dc:creator>
<dc:creator>Basu, P.</dc:creator>
<dc:creator>Simmons, H. A.</dc:creator>
<dc:creator>Eickhoff, J. C.</dc:creator>
<dc:creator>Friedrich, T. C.</dc:creator>
<dc:creator>Aliota, M. T.</dc:creator>
<dc:creator>Mohr, E. L.</dc:creator>
<dc:creator>Dudley, D. M.</dc:creator>
<dc:creator>O'Connor, D. H.</dc:creator>
<dc:creator>N</dc:creator>
<dc:date>2022-12-09</dc:date>
<dc:identifier>doi:10.1101/2022.12.09.519791</dc:identifier>
<dc:title><![CDATA[Frequent first-trimester pregnancy loss in rhesus macaques infected with African-lineage Zika virus]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-12-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.12.09.519807v1?rss=1">
<title>
<![CDATA[
3D multiplexed tissue imaging reconstruction and optimized region-of-interest (ROI) selection through deep learning model of channels embedding 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.12.09.519807v1?rss=1</link>
<description><![CDATA[
Tissue-based sampling and diagnosis are defined as the extraction of information from certain limited spaces and its diagnostic significance of a certain object. Pathologists deal with issues related to tumor heterogeneity since analyzing a single sample does not necessarily capture a representative depiction of cancer, and a tissue biopsy usually only presents a small fraction of the tumor. Many multiplex tissue imaging platforms (MTIs) make the assumption that tissue microarrays (TMAs) containing small core samples of 2-dimensional (2D) tissue sections are a good approximation of bulk tumors although tumors are not 2D. However, emerging whole slide imaging (WSI) or 3D tumor atlases that employ MTIs like cyclic immunofluorescence (CyCIF) strongly challenge this assumption. In spite of the additional insight gathered by measuring the tumor microenvironment in WSI or 3D, it can be prohibitively expensive and time-consuming to process tens or hundreds of tissue sections with CyCIF. Even when resources are not limited, the criteria for region-of-interest (ROI) selection in tissues for downstream analysis remain largely qualitative and subjective as stratified sampling requires the knowledge of objects and evaluates their features. Despite the fact TMAs fail to adequately approximate whole tissue features, a theoretical subsampling of tissue exists that can best represent the tumor in the whole slide image. To address these challenges, we propose deep learning approaches to learn multi-modal image translation tasks from two aspects: 1) generative modeling approach to reconstruct 3D CyCIF representation and 2) co-embedding CyCIF image and Hematoxylin and Eosin (H&E) section to learn multi-modal mappings by a cross-domain translation for minimum representative ROI selection. We demonstrate that generative modeling enables a 3D virtual CyCIF reconstruction of a colorectal cancer specimen given a small subset of the imaging data at training time. By co-embedding histology and MTI features, we propose a simple convex optimization for objective ROI selection. We demonstrate the potential application of ROI selection and the efficiency of its performance with respect to cellular heterogeneity.
]]></description>
<dc:creator>Burlingame, E.</dc:creator>
<dc:creator>Ternes, L.</dc:creator>
<dc:creator>Lin, J.-R.</dc:creator>
<dc:creator>Chen, Y.-A.</dc:creator>
<dc:creator>Kim, E. N.</dc:creator>
<dc:creator>Gray, J. W.</dc:creator>
<dc:creator>Santagata, S.</dc:creator>
<dc:creator>Sorger, P. K.</dc:creator>
<dc:creator>Chang, Y. H.</dc:creator>
<dc:date>2022-12-12</dc:date>
<dc:identifier>doi:10.1101/2022.12.09.519807</dc:identifier>
<dc:title><![CDATA[3D multiplexed tissue imaging reconstruction and optimized region-of-interest (ROI) selection through deep learning model of channels embedding]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-12-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.12.14.520515v1?rss=1">
<title>
<![CDATA[
A set of orthogonal versatile interacting peptide tags for imaging cellular proteins 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.12.14.520515v1?rss=1</link>
<description><![CDATA[
Genetic tags are transformative tools for investigating the function, localization, and interactions of cellular proteins. Most studies today are reliant on selective labeling of more than one protein to obtain comprehensive information on a proteins behavior in situ. Some proteins can be analyzed by fusion to protein tag, such as green fluorescent protein, HaloTag, or SNAP-Tag. Other proteins benefit from labeling via small peptide tags, such as the recently reported versatile interacting peptide (VIP) tags. VIP tags enable observations of protein localization and trafficking with bright fluorophores or nanoparticles. Here we expand the VIP toolkit by presenting two new tags: TinyVIPER and PunyVIPER. These two tags were designed for use with MiniVIPER for labeling up to three distinct proteins at once in living cells. Labeling is mediated by the formation of a high affinity, biocompatible heterodimeric coiled coil. Each tag was validated by fluorescence microscopy, including observation of transferrin receptor 1 trafficking in live cells. We verified that labeling via each tag is highly specific, with no cross-reactivity between the three VIP tags under cellular conditions. Lastly, the self-sorting tags were used for simultaneous labeling of three protein targets (i.e., TOMM20, histone 2B, and actin), highlighting their utility for multicolor microscopy. MiniVIPER, TinyVIPER, and PunyVIPER are small and robust peptide tags for selective labeling of cellular proteins.
]]></description>
<dc:creator>Suyama, A.</dc:creator>
<dc:creator>Devlin, K. L.</dc:creator>
<dc:creator>Macias Contreras, M.</dc:creator>
<dc:creator>Doh, J. K.</dc:creator>
<dc:creator>Shinde, U.</dc:creator>
<dc:creator>Beatty, K. E.</dc:creator>
<dc:date>2022-12-15</dc:date>
<dc:identifier>doi:10.1101/2022.12.14.520515</dc:identifier>
<dc:title><![CDATA[A set of orthogonal versatile interacting peptide tags for imaging cellular proteins]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-12-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.12.16.520295v1?rss=1">
<title>
<![CDATA[
Predicting transcription factor activity using prior biological information 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.12.16.520295v1?rss=1</link>
<description><![CDATA[
Transcription factors are critical regulators of cellular gene expression programs. Disruption of normal transcription factor regulation is associated with a broad range of diseases. In order to understand the mechanisms that underly disease pathogenesis, it is critical to detect aberrant transcription factor activity. We have developed Priori, a computational method to predict transcription factor activity from RNA sequencing data. Priori has several key advantages over existing methods. Priori utilizes literature-supported regulatory relationship information to identify known transcription factor target genes. Using these transcriptional relationships, Priori uses linear models to determine the impact and direction of transcription factor regulation on the expression of its target genes. In our work, we evaluated the ability of Priori and 16 other methods to detect aberrant activity from 124 single-gene perturbation experiments. We show that Priori identifies perturbed transcription factors with greater sensitivity and specificity than other methods. Furthermore, our work demonstrates that Priori can be used to discover significant determinants of survival in breast cancer as well as identify mediators of drug response in leukemia from primary patient samples.
]]></description>
<dc:creator>Estabrook, J.</dc:creator>
<dc:creator>Yashar, W. M.</dc:creator>
<dc:creator>Holly, H. D.</dc:creator>
<dc:creator>Somers, J.</dc:creator>
<dc:creator>Nikolova, O.</dc:creator>
<dc:creator>Barbur, O.</dc:creator>
<dc:creator>Braun, T. P.</dc:creator>
<dc:creator>Demir, E.</dc:creator>
<dc:date>2022-12-19</dc:date>
<dc:identifier>doi:10.1101/2022.12.16.520295</dc:identifier>
<dc:title><![CDATA[Predicting transcription factor activity using prior biological information]]></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/10.1101/2022.12.16.520769v1?rss=1">
<title>
<![CDATA[
Identification of novel PGRP3 protein -protein interactions using yeast two hybrid system 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.12.16.520769v1?rss=1</link>
<description><![CDATA[
BackgroundPeptidoglycan recognition protein-3 (PGRP3) is a pattern recognition receptor that binds peptidoglycan to elicit an immune response. How PGRP3 mediates these immunomodulatory effects remains unknown. Identifying the proteins that interact with PGRP3 will help begin to address this important knowledge gap. Therefore, in this study our objective was to identify and validate novel PGRP3-protein interactions.

MethodsPGRP3-protein interactions were identified using a yeast two-hybrid system. PGRP3 cloned into a pGBKT7 DNA-BD vector and transformed into a Y2HGold yeast strain served as the bait. A normalized universal human cDNA library cloned in a pGADT7 AD vector and pre-transformed into an Y187 yeast strain served as the prey. Y2HGold and Y187 yeast strains were mated and diploids were plated on selective media. True positive clones were sent in for DNA sequencing and corresponding proteins were identified through NCBI Blast. Two proteins were selected for validation. His-tagged PGRP3 expressing 293T cells were then co-transfected with expression vectors containing the identified proteins tagged with c-Myc. Lysates from co-transfected cells were subjected to anti-His and anti-Myc co-immunoprecipitation and analyzed with western blots.

ResultsFour unique proteins-- MBNL3, RBP5, ATXN2, and GPATCH8--were identified from our yeast two-hybrid screen. Co-transfection and co-immunoprecipitation assays successfully confirmed MBNL3 and RBP5 as positive interactors with PGRP3.

ConclusionsBased on the known functions of MBNL3, RBP5, ATXN2, and GPATCH8, we hypothesize that PGRP3 may be involved in RNA processing, endocytic trafficking, and PPAR{gamma} pathways related to innate immunity.
]]></description>
<dc:creator>Lin, L.-j.</dc:creator>
<dc:creator>Lee, C.</dc:creator>
<dc:creator>Merritt, J.</dc:creator>
<dc:creator>Kreth, J.</dc:creator>
<dc:creator>Ranganathan, S.</dc:creator>
<dc:creator>Prakasam, S.</dc:creator>
<dc:date>2022-12-18</dc:date>
<dc:identifier>doi:10.1101/2022.12.16.520769</dc:identifier>
<dc:title><![CDATA[Identification of novel PGRP3 protein -protein interactions using yeast two hybrid system]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-12-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.12.16.520797v1?rss=1">
<title>
<![CDATA[
Motion Impact Score for Detecting Spurious Brain-Behavior Associations 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.12.16.520797v1?rss=1</link>
<description><![CDATA[
Between-participant differences in head motion introduce systematic bias to resting state fMRI brain-wide association studies (BWAS) that is not completely removed by denoising algorithms. Researchers who study traits, or phenotypes associated with in-scanner head motion (e.g. psychiatric disorders) need to know if trait-functional connectivity (FC) effects are biased by residual motion artifact in order to avoid reporting false positive results. We devised an adaptable method, Split Half Analysis of Motion Associated Networks (SHAMAN), to assign a motion impact score to specific trait-FC effects. The SHAMAN approach distinguishes between motion artifact causing overestimation or underestimation of trait-FC effects. SHAMAN was > 95% specific at sample sizes of n = 100 and above. SHAMAN was powered to detect motion overestimation scores 80% of the time at sample sizes of n = 5,000 but could detect motion underestimation scores only 50% of the time at n = 5000, making it most useful for researchers seeking to avoid overestimating trait-FC effects in large BWAS. We computed motion impact scores for trait-FC effect with 45 demographic, biophysical, cognitive, and personality traits from n = 7,270 participants in the Adolescent Brain Cognitive Development (ABCD) Study. After standard denoising with ABCD-BIDS and without motion censoring, 42% (19/45) of traits had significant (p < 0.05) motion overestimation scores and 38% (17/45) of traits had significant motion underestimation scores. Censoring at framewise displacement (FD) < 0.2 mm reduced the proportion of traits with significant motion overestimation scores from 42% to 2% (1/45) but did not decrease the number of traits with significant motion underestimation scores.
]]></description>
<dc:creator>Kay, B. P.</dc:creator>
<dc:creator>Montez, D. F.</dc:creator>
<dc:creator>Marek, S.</dc:creator>
<dc:creator>Tervo-Clemmens, B.</dc:creator>
<dc:creator>Siegel, J. S.</dc:creator>
<dc:creator>Adeyemo, B.</dc:creator>
<dc:creator>Laumann, T. O.</dc:creator>
<dc:creator>Metoki, A.</dc:creator>
<dc:creator>Chauvin, R. J.</dc:creator>
<dc:creator>Van, A. N.</dc:creator>
<dc:creator>Krimmel, S. R.</dc:creator>
<dc:creator>Miller, R.</dc:creator>
<dc:creator>Newbold, D. J.</dc:creator>
<dc:creator>Zheng, A.</dc:creator>
<dc:creator>Seider, N. A.</dc:creator>
<dc:creator>Scheidter, K. M.</dc:creator>
<dc:creator>Monk, J.</dc:creator>
<dc:creator>Feczko, E. J.</dc:creator>
<dc:creator>Randolph, A.</dc:creator>
<dc:creator>Miranda Dominguez, O.</dc:creator>
<dc:creator>Moore, L. A.</dc:creator>
<dc:creator>Perrone, A. J.</dc:creator>
<dc:creator>Conan, G. M.</dc:creator>
<dc:creator>Earl, E. A.</dc:creator>
<dc:creator>Malone, S. M.</dc:creator>
<dc:creator>Cordova, M.</dc:creator>
<dc:creator>Doyle, O.</dc:creator>
<dc:creator>Lynch, B. J.</dc:creator>
<dc:creator>Wilgenbusch, J. C.</dc:creator>
<dc:creator>Pengo, T.</dc:creator>
<dc:creator>Graham, A. M.</dc:creator>
<dc:creator>Roland, J. L.</dc:creator>
<dc:creator>Gordon, E. M.</dc:creator>
<dc:creator>Snyder, A. Z.</dc:creator>
<dc:creator>Barch, D.</dc:creator>
<dc:creator>Fair, D. A.</dc:creator>
<dc:creator>Dosenbach, N. U.</dc:creator>
<dc:date>2022-12-17</dc:date>
<dc:identifier>doi:10.1101/2022.12.16.520797</dc:identifier>
<dc:title><![CDATA[Motion Impact Score for Detecting Spurious Brain-Behavior Associations]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-12-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.12.17.520865v1?rss=1">
<title>
<![CDATA[
A versatile and interoperable computational framework for the analysis and modeling of COVID-19 disease mechanisms 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.12.17.520865v1?rss=1</link>
<description><![CDATA[
The COVID-19 Disease Map project is a large-scale community effort uniting 277 scientists from 130 Institutions around the globe. We use high-quality, mechanistic content describing SARS-CoV-2-host interactions and develop interoperable bioinformatic pipelines for novel target identification and drug repurposing. Community-driven and highly interdisciplinary, the project is collaborative and supports community standards, open access, and the FAIR data principles. The coordination of community work allowed for an impressive step forward in building interfaces between Systems Biology tools and platforms. Our framework links key molecules highlighted from broad omics data analysis and computational modeling to dysregulated pathways in a cell-, tissue- or patient-specific manner. We also employ text mining and AI-assisted analysis to identify potential drugs and drug targets and use topological analysis to reveal interesting structural features of the map. The proposed framework is versatile and expandable, offering a significant upgrade in the arsenal used to understand virus-host interactions and other complex pathologies.
]]></description>
<dc:creator>Niarakis, A.</dc:creator>
<dc:creator>Ostaszewski, M.</dc:creator>
<dc:creator>Mazein, A.</dc:creator>
<dc:creator>Kuperstein, I.</dc:creator>
<dc:creator>Gillespie, M. E.</dc:creator>
<dc:creator>Kutmon, M.</dc:creator>
<dc:creator>Funahashi, A.</dc:creator>
<dc:creator>Acencio, M. L.</dc:creator>
<dc:creator>Hemedan, A.</dc:creator>
<dc:creator>Aichem, M.</dc:creator>
<dc:creator>Klein, K.</dc:creator>
<dc:creator>Czauderna, T.</dc:creator>
<dc:creator>Burtscher, F.</dc:creator>
<dc:creator>Yamada, T. G.</dc:creator>
<dc:creator>Hiki, Y.</dc:creator>
<dc:creator>Hiroi, N. F.</dc:creator>
<dc:creator>Hu, F.</dc:creator>
<dc:creator>Pham, N.</dc:creator>
<dc:creator>Ehrhart, F.</dc:creator>
<dc:creator>Willighagen, E. L.</dc:creator>
<dc:creator>Valdeolivas, A.</dc:creator>
<dc:creator>Dugourd, A.</dc:creator>
<dc:creator>Messina, F.</dc:creator>
<dc:creator>Esteban-Medina, M.</dc:creator>
<dc:creator>Pena-Chilet, M.</dc:creator>
<dc:creator>Rian, K.</dc:creator>
<dc:creator>Soliman, S.</dc:creator>
<dc:creator>Aghamiri, S. S.</dc:creator>
<dc:creator>Puniya, B. L.</dc:creator>
<dc:creator>Naldi, A.</dc:creator>
<dc:creator>Helikar, T.</dc:creator>
<dc:creator>Singh, V.</dc:creator>
<dc:creator>Farinas Fernandez, M.</dc:creator>
<dc:creator>Bermudez, V.</dc:creator>
<dc:creator>Tsirvouli, E.</dc:creator>
<dc:creator>Montagud, A.</dc:creator>
<dc:creator>Noel, V.</dc:creator>
<dc:creator>Ponce de Leon, M.</dc:creator>
<dc:creator>Maier, D.</dc:creator>
<dc:creator>Bauch, A.</dc:creator>
<dc:creator>Gyori, B. M.</dc:creator>
<dc:creator>Bachman, J. A.</dc:creator>
<dc:creator>Luna, A</dc:creator>
<dc:date>2022-12-19</dc:date>
<dc:identifier>doi:10.1101/2022.12.17.520865</dc:identifier>
<dc:title><![CDATA[A versatile and interoperable computational framework for the analysis and modeling of COVID-19 disease mechanisms]]></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/10.1101/2022.12.18.520962v1?rss=1">
<title>
<![CDATA[
Control of stereocilia length during development of hair bundles 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.12.18.520962v1?rss=1</link>
<description><![CDATA[
Assembly of the hair bundle, the sensory organelle of the inner ear, depends on differential growth of actin-based stereocilia. Separate rows of stereocilia, labeled 1 through 3 from tallest to shortest, lengthen or shorten during discrete time intervals during development. We used lattice structured illumination microscopy and surface rendering of mouse apical inner hair cells to measure stereocilia dimensions during early postnatal development; these measurements revealed a sharp transition at postnatal day 8 between stage III (row 1 and 2 widening; row 2 shortening) and stage IV (final row 1 lengthening and widening). Tip proteins that determine row 1 lengthening did not accumulate simultaneously during stages III and IV; while the actin-bundling protein EPS8 peaked at the end of stage III, GNAI3 peaked several days later--in early stage IV--and GPSM2 peaked near the end of stage IV. To establish the contributions of key macromolecular assemblies to bundle structure, we examined mouse mutants that eliminated tip links (Cdh23v2J or Pcdh15av3J), transduction channels (TmieKO), or the row 1 tip complex (Myo15ash2). Cdh23v2J/v2J and Pcdh15av3J/av3J bundles had adjacent stereocilia in the same row that were not matched in length, revealing that a major role of these cadherins is to synchronize lengths of side-by-side stereocilia. Use of the tip-link mutants also allowed us to distinguish the role of transduction from effects of transduction proteins themselves. While levels of GNAI3 and GPSM2, which stimulate stereocilia elongation, were greatly attenuated at the tips of TmieKO/KO row 1 stereocilia, they accumulated normally in Cdh23v2J/v2J and Pcdh15av3J/av3J stereocilia. These results reinforced the suggestion that the transduction proteins themselves facilitate localization of proteins in the row 1 complex. By contrast, EPS8 concentrates at tips of all TmieKO/KO, Cdh23v2J/v2J and Pcdh15av3J/av3J stereocilia, correlating with the less polarized distribution of stereocilia lengths in these bundles. These latter results indicated that in wild-type hair cells, the transduction complex prevents accumulation of EPS8 at the tips of shorter stereocilia, causing them to shrink (row 2 and 3) or disappear (row 4 and microvilli). Reduced rhodamine-actin labeling at row 2 stereocilia tips of tip-link and transduction mutants suggests that transductions role is to destabilize actin filaments there. These results suggest that regulation of stereocilia length occurs through EPS8, and that CDH23 and PCDH15 regulate stereocilia lengthening beyond their role in gating mechanotransduction channels.
]]></description>
<dc:creator>Krey, J. F.</dc:creator>
<dc:creator>Chatterjee, P.</dc:creator>
<dc:creator>Halford, J.</dc:creator>
<dc:creator>Cunningham, C. L.</dc:creator>
<dc:creator>Perrin, B. J.</dc:creator>
<dc:creator>Barr-Gillespie, P. G.</dc:creator>
<dc:date>2022-12-19</dc:date>
<dc:identifier>doi:10.1101/2022.12.18.520962</dc:identifier>
<dc:title><![CDATA[Control of stereocilia length during development of hair bundles]]></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/10.1101/2022.12.19.521083v1?rss=1">
<title>
<![CDATA[
Silencing neuroinflammatory reactive astrocyte activating factors ameliorates disease outcomes in perinatal white matter injury 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.12.19.521083v1?rss=1</link>
<description><![CDATA[
The role of reactive astrocytes in perinatal white matter injury (WMI) is unclear. In a mouse model of WMI, we provide evidence that impairing the formation of a C3-expressing neuroinflammatory reactive astrocyte sub-state rescues myelination and behavioral deficits. We further demonstrate the presence of C3-expressing reactive astrocytes in human WMI. Our data point to these cells as putative drivers of myelination failure in WMI and a potentially promising therapeutic target.
]]></description>
<dc:creator>Renz, P. V.</dc:creator>
<dc:creator>Surbek, D.</dc:creator>
<dc:creator>Haesler, V.</dc:creator>
<dc:creator>Tscherrig, V.</dc:creator>
<dc:creator>Huang, E. J.</dc:creator>
<dc:creator>Chavali, M.</dc:creator>
<dc:creator>Liddelow, S. A.</dc:creator>
<dc:creator>Rowitch, D.</dc:creator>
<dc:creator>Schoeberlein, A.</dc:creator>
<dc:creator>Brosius Lutz, A. R.</dc:creator>
<dc:date>2022-12-20</dc:date>
<dc:identifier>doi:10.1101/2022.12.19.521083</dc:identifier>
<dc:title><![CDATA[Silencing neuroinflammatory reactive astrocyte activating factors ameliorates disease outcomes in perinatal white matter injury]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-12-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.12.21.520137v1?rss=1">
<title>
<![CDATA[
Multi-center integrated analysis of non-coding CRISPR screens 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.12.21.520137v1?rss=1</link>
<description><![CDATA[
The ENCODE Consortiums efforts to annotate non-coding, cis-regulatory elements (CREs) have advanced our understanding of gene regulatory landscapes which play a major role in health and disease. Pooled, non-coding CRISPR screens are a promising approach for systematically investigating gene regulatory mechanisms. Here, the ENCODE Functional Characterization Centers report 109 screens comprising 346,970 individual perturbations across 13.3Mb of the genome, using a variety of methods, readouts, and statistical analyses. Across 332 functionally confirmed CRE-gene links, we identify principles for screening endogenous, non-coding elements for causal regulatory mechanisms. Nearly all CREs show strong evidence of open chromatin, and targeting accessibility peak summits is a critical component of our proposed sgRNA design rules. We provide experimental guidelines to accurately detect CREs with variable, often low, transcriptional effects. We discover a previously undescribed DNA strand-bias for CRISPRi in transcribed regions with implications for screen design and analysis. Benchmarking five screen analysis tools, we find CASA produces the most conservative CRE calls and is robust to artifacts of low-specificity sgRNAs. Together, we provide an accessible data resource, predesigned sgRNAs targeting 3,275,697 ENCODE SCREEN candidate CREs, and screening guidelines to accelerate functional characterization of the non-coding genome.
]]></description>
<dc:creator>Yao, D.</dc:creator>
<dc:creator>Tycko, J.</dc:creator>
<dc:creator>Oh, W.</dc:creator>
<dc:creator>Bounds, L. R.</dc:creator>
<dc:creator>Gosai, S. J.</dc:creator>
<dc:creator>Lataniotis, L.</dc:creator>
<dc:creator>Mackay-Smith, A.</dc:creator>
<dc:creator>Doughty, B. R.</dc:creator>
<dc:creator>Gabdank, I.</dc:creator>
<dc:creator>Schmidt, H.</dc:creator>
<dc:creator>Youngworth, I.</dc:creator>
<dc:creator>Andreeva, K.</dc:creator>
<dc:creator>Ren, X.</dc:creator>
<dc:creator>Barrera, A.</dc:creator>
<dc:creator>Luo, Y.</dc:creator>
<dc:creator>Siklenka, K.</dc:creator>
<dc:creator>Yardimci, G. G.</dc:creator>
<dc:creator>The ENCODE4 Consortium,</dc:creator>
<dc:creator>Tewhey, R.</dc:creator>
<dc:creator>Kundaje, A.</dc:creator>
<dc:creator>Greenleaf, W. J.</dc:creator>
<dc:creator>Sabeti, P. C.</dc:creator>
<dc:creator>Leslie, C.</dc:creator>
<dc:creator>Pritykin, Y.</dc:creator>
<dc:creator>Moore, J. E.</dc:creator>
<dc:creator>Beer, M. A.</dc:creator>
<dc:creator>Gersbach, C.</dc:creator>
<dc:creator>Reddy, T. E.</dc:creator>
<dc:creator>Shen, Y.</dc:creator>
<dc:creator>Engreitz, J. M.</dc:creator>
<dc:creator>Bassik, M. C.</dc:creator>
<dc:creator>Reilly, S. K.</dc:creator>
<dc:date>2022-12-22</dc:date>
<dc:identifier>doi:10.1101/2022.12.21.520137</dc:identifier>
<dc:title><![CDATA[Multi-center integrated analysis of non-coding CRISPR screens]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-12-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.12.21.521495v1?rss=1">
<title>
<![CDATA[
Tumor cell intrinsic RIG-I activation is sufficient to drive immune mediated tumor rejection 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.12.21.521495v1?rss=1</link>
<description><![CDATA[
Targeting cytosolic nucleic acid sensors is a potent approach to drive type I interferon responses and anti-tumor immunity. Recent evidence suggests that activation of retinoic acid inducible gene-I (RIG-I) using synthetic hairpin RNA agonists decreases tumor progression in multiple preclinical models. However, the role of tumor cell intrinsic RIG-I in shaping tumor cell fates and the host immune microenvironment remains unclear. Here, we show that RIG-I expression is correlated with better overall survival and a distinct immune gene signature in specific human cancers including colorectal cancer. Activation of RIG-I in breast and colorectal cancer cells is sufficient to drive tumor cell death in vitro and significantly delay tumor growth in vivo in multiple preclinical models. Importantly, the efficacy of tumor cell RIG-I activation is lost in immune deficient mice suggesting the requirement of immune responses for this effect. We observe that tumor cell intrinsic RIG-I activation elicits a robust cellular and molecular immune response. We show that tumor cell RIG-I activation also leads to induction of specific immune checkpoints including PD-L1. Using a publicly available database, we found that RIG-I expression serves as an excellent prognostic marker for responders to checkpoint immunotherapy, particularly PD-L1/PD-1 across cancers. Finally, combination of tumor cell intrinsic RIG-I activation with anti-PD-L1 led to a synergistic decrease in tumor growth in a colorectal tumor model. Our findings suggest that tumor cell intrinsic RIG-I can be targeted to enhance anti-tumor immune responses and highlights a potential strategy for anti-cancer vaccines that can invigorate the immune system.
]]></description>
<dc:creator>Fraile-Bethencourt, E.</dc:creator>
<dc:creator>Khou, S.</dc:creator>
<dc:creator>Baris, A.</dc:creator>
<dc:creator>Ruhl, R.</dc:creator>
<dc:creator>Anand, S.</dc:creator>
<dc:date>2022-12-22</dc:date>
<dc:identifier>doi:10.1101/2022.12.21.521495</dc:identifier>
<dc:title><![CDATA[Tumor cell intrinsic RIG-I activation is sufficient to drive immune mediated tumor rejection]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-12-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.12.27.521482v1?rss=1">
<title>
<![CDATA[
Discovery of oncogenic ROS1 missense mutations with sensitivity to tyrosine kinase inhibitors 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.12.27.521482v1?rss=1</link>
<description><![CDATA[
Chromosomal rearrangements of ROS1 generate ROS1 tyrosine kinase fusion proteins that are established oncogenes predicting effectiveness of tyrosine kinase inhibitors (TKI) treatment. The cancer genome reveals nonsynonymous missense mutations in ROS1, however, their oncogenic potential remains unknown. We nominated thirty-four tumor-associated missense mutations in ROS1 kinase domain for functional interrogation. Immunoblotting revealed diverse impact of the mutations on the kinase, ranging from loss of function to significant increase in catalytic activity. Notably, Asn and Gly substitutions at the Asp-2113 position in ROS1 kinase domain were TKI- sensitive hyper-activating mutations, and transformative oncogenes in independent cell models. Molecular modeling revealed drastic alterations in the activation loop of ROS1D2113N compared to wildtype kinase. Proteomics studies showed that ROS1D2113N increases phosphorylation of known effectors akin to ROS1 fusions, and upregulates pathways not previously linked to ROS1, including mTORC2, JNK1/2, AP-1, TGFB1 and CCN1/2. In vivo, ROS1D2113N drove tumor formation that was sensitive to inhibition by crizotinib and lorlatinib. Taken together, these data show that select point mutations within ROS1 RTK are oncogenic, and maybe therapeutically targetable with FDA-approved TKI.
]]></description>
<dc:creator>Iyer, S. R.</dc:creator>
<dc:creator>Nusser, K.</dc:creator>
<dc:creator>Jones, K.</dc:creator>
<dc:creator>Shinde, P.</dc:creator>
<dc:creator>Beach, C. Z.</dc:creator>
<dc:creator>Keddy, C.</dc:creator>
<dc:creator>Aguero, E.</dc:creator>
<dc:creator>Force, J.</dc:creator>
<dc:creator>Shinde, U.</dc:creator>
<dc:creator>Davare, M. A.</dc:creator>
<dc:date>2022-12-27</dc:date>
<dc:identifier>doi:10.1101/2022.12.27.521482</dc:identifier>
<dc:title><![CDATA[Discovery of oncogenic ROS1 missense mutations with sensitivity to tyrosine kinase inhibitors]]></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/10.1101/2023.01.03.522674v1?rss=1">
<title>
<![CDATA[
Adaptation of prokaryotic toxins for negative selection and cloning-independent markerless mutagenesis (CIMM) in Streptococcus species 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.01.03.522674v1?rss=1</link>
<description><![CDATA[
The Streptococcus mutans genetic system offers a variety of strategies to rapidly engineer targeted chromosomal mutations. Previously, we reported the first S. mutans negative selection system that functions in a wild-type background. This system utilizes induced sensitivity to the toxic amino acid analog p-chlorophenylalanine (4-CP) as a negative selection mechanism, and was developed for counterselection-based cloning-independent markerless mutagenesis (CIMM). While we have employed this system extensively for our ongoing genetic studies, we have encountered a couple limitations with the system, mainly its narrow host range and the requirement for selection on a toxic substrate. Here, we report the development of a new negative selection system that addresses both limitations, while still retaining the utility of the previous 4-CP-based markerless mutagenesis system. We placed a variety of toxin-encoding genes under the control of the xylose-inducible Xyl-S expression cassette and found the Fst-sm and ParE toxins to be suitable candidates for inducible negative selection. We combined the inducible toxins with an antibiotic resistance gene to create several different counterselection cassettes. The most broadly useful of these contained a wild-type fst-sm open reading frame transcriptionally fused to a point mutant form of the Xyl-S expression system, which we subsequently named as IFDC4. IFDC4 was shown to exhibit exceptionally low background resistance, with 3 - 4 log reductions in cell number observed when plating on xylose-supplemented media. IFDC4 also functioned similarly in multiple strains of S. mutans as well as with S. gordonii and S. sanguinis. We performed CIMM with IFDC4 and successfully engineered a variety of different types of markerless mutations in all three species. The counterselection strategy described here provides a template approach that should be adaptable for the creation of similar counterselection systems in many other bacteria.
]]></description>
<dc:creator>Li, L.</dc:creator>
<dc:creator>Qin, H.</dc:creator>
<dc:creator>Zou, Z.</dc:creator>
<dc:creator>Kreth, J.</dc:creator>
<dc:creator>Merritt, J.</dc:creator>
<dc:date>2023-01-07</dc:date>
<dc:identifier>doi:10.1101/2023.01.03.522674</dc:identifier>
<dc:title><![CDATA[Adaptation of prokaryotic toxins for negative selection and cloning-independent markerless mutagenesis (CIMM) in Streptococcus species]]></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/10.1101/2023.01.09.523291v1?rss=1">
<title>
<![CDATA[
Acute psilocybin enhances cognitive flexibility in rats 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.01.09.523291v1?rss=1</link>
<description><![CDATA[
Psilocybin has been shown to improve symptoms of depression and anxiety when combined with psychotherapy or other clinician-guided interventions. To understand the neural basis for this pattern of clinical efficacy, experimental and conceptual approaches that are different than traditional laboratory models of anxiety and depression are needed. A potential novel mechanism is that acute psilocybin improves cognitive flexibility, which then enhances the impact of clinician-assisted interventions. Consistent with this idea, we find that acute psilocybin robustly improves cognitive flexibility in male and female rats using a task where animals switched between previously learned strategies in response to uncued changes in the environment. Psilocybin did not influence Pavlovian reversal learning, suggesting that its cognitive effects are selective to enhanced switching between previously learned behavioral strategies. The serotonin (5HT) 2A receptor antagonist ketanserin blocked psilocybins effect on set-shifting, while a 5HT2C-selective antagonist did not. Ketanserin alone also improved set-shifting performance, suggesting a complex relationship between psilocybins pharmacology and its impact on flexibility. Further, the psychedelic drug 2,5-Dimethoxy-4-iodoamphetamine (DOI) impaired cognitive flexibility in the same task, suggesting that this effect of psilocybin does not generalize to all other serotonergic psychedelics. We conclude that the acute impact of psilocybin on cognitive flexibility provides a useful behavioral model to investigate its neuronal effects relevant to its positive clinical outcome.
]]></description>
<dc:creator>Torrado Pacheco, A.</dc:creator>
<dc:creator>Olson, R. J.</dc:creator>
<dc:creator>Garza, G.</dc:creator>
<dc:creator>Moghaddam, B.</dc:creator>
<dc:date>2023-01-09</dc:date>
<dc:identifier>doi:10.1101/2023.01.09.523291</dc:identifier>
<dc:title><![CDATA[Acute psilocybin enhances cognitive flexibility in rats]]></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/10.1101/2023.01.11.523512v1?rss=1">
<title>
<![CDATA[
Neuronal activity-driven O-GlcNAcylation promotes mitochondrial plasticity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.01.11.523512v1?rss=1</link>
<description><![CDATA[
Neuronal activity is an energy-intensive process that is largely sustained by instantaneous fuel utilization and ATP synthesis. However, how neurons couple ATP synthesis rate to fuel availability is largely unknown. Here, we demonstrate that the metabolic sensor enzyme O-GlcNAc transferase regulates neuronal activity-driven mitochondrial bioenergetics. We show that neuronal activity upregulates O-GlcNAcylation mainly in mitochondria. Mitochondrial O-GlcNAcylation is promoted by activity-driven fuel consumption, which allows neurons to compensate for high energy expenditure based on fuel availability. To determine the proteins that are responsible for these adjustments, we mapped the mitochondrial O-GlcNAcome of neurons. Finally, we determine that neurons fail to meet activity-driven metabolic demand when O-GlcNAcylation dynamics are prevented. Our findings suggest that O-GlcNAcylation provides a fuel-dependent feedforward control mechanism in neurons to optimize mitochondrial performance based on neuronal activity. This mechanism thereby couples neuronal metabolism to mitochondrial bioenergetics and plays a key role in sustaining energy homeostasis.
]]></description>
<dc:creator>Yu, S. B.</dc:creator>
<dc:creator>Sanchez, R. G.</dc:creator>
<dc:creator>Papich, Z. D.</dc:creator>
<dc:creator>Whisenant, T. C.</dc:creator>
<dc:creator>Ghassemian, M.</dc:creator>
<dc:creator>Koberstein, J. N.</dc:creator>
<dc:creator>Stewart, M. L.</dc:creator>
<dc:creator>Pekkurnaz, G.</dc:creator>
<dc:date>2023-01-11</dc:date>
<dc:identifier>doi:10.1101/2023.01.11.523512</dc:identifier>
<dc:title><![CDATA[Neuronal activity-driven O-GlcNAcylation promotes mitochondrial plasticity]]></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/10.1101/2023.01.16.524157v1?rss=1">
<title>
<![CDATA[
The absence of a neurogenic response to a repeated concussive-like injury in mice 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.01.16.524157v1?rss=1</link>
<description><![CDATA[
In response to traumatic brain injury (TBI), the brain increases its generation of new neurons (neurogenesis) within the hippocampus, a brain region critical for learning and memory. Because neurogenesis plays important roles in learning and memory, post-traumatic neurogenesis may represent an adaptive response contributing to cognitive recovery. In contrast to increases in neurogenesis acutely after injury, levels of neurogenesis become impaired long after TBI. And although chronic deficits in neurogenesis after TBI have been reported by multiple groups, it is unknown whether the hippocampus remains capable of eliciting another neurogenic response to a repeated injury. To address this lack of knowledge, we used a closed head injury model that reflects a concussive-like injury or a mild TBI (mTBI) and assessed levels of neurogenesis in male and female adult mice. Mice received one or two mTBI or sham treatments 3 weeks apart. Compared to mice with a single mTBI, proliferation and neurogenesis were blunted in mice that received a second mTBI. This impaired response was unlikely due to a short recovery time between the two mTBIs as the proliferative response to a second mTBI was also impaired when two months were allowed between injuries. We further found that proliferation was impaired in the radial-glia like cells despite an intact pool. The mice that received two mTBIs also had a blunted intensity in their GFAP staining. In contrast to reports of aberrant post-TBI neurogenesis, we found that the neurons born after mTBI had normal dendritic branches. Lastly, we found that impairments in the inability to mount a neurogenic response after a second mTBI were associated with deficits in neurogenesisstrategy flexibility in the reversal water maze task. Our data suggests that a loss in the neurogenic response could in part contribute to worse cognitive recovery after a repeated concussion. These data may expose a novel target to help improve long-term cognitive outcome following repeated brain injury.
]]></description>
<dc:creator>Weingarten, A.</dc:creator>
<dc:creator>Bah, T. M.</dc:creator>
<dc:creator>Yeturu, S.</dc:creator>
<dc:creator>Samudrala, N.</dc:creator>
<dc:creator>Villasana, L. E.</dc:creator>
<dc:date>2023-01-19</dc:date>
<dc:identifier>doi:10.1101/2023.01.16.524157</dc:identifier>
<dc:title><![CDATA[The absence of a neurogenic response to a repeated concussive-like injury in mice]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-01-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.01.24.525282v1?rss=1">
<title>
<![CDATA[
UL135 and UL136 Epistasis Controls Reactivation of Human Cytomegalovirus 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.01.24.525282v1?rss=1</link>
<description><![CDATA[
Human cytomegalovirus (HCMV) is beta herpesvirus that persists indefinitely in the human host through a protracted, latent infection. The polycistronic UL133-UL138 gene locus of HCMV encodes genes regulating latency and reactivation. While UL138 is pro-latency, restricting virus replication in CD34+ hematopoietic progenitor cells (HPCs), UL135 overcomes this restriction for reactivation. By contrast, UL136 is expressed with later kinetics and encodes multiple protein isoforms with differential roles in latency and reactivation. Like UL135, the largest UL136 isoform, UL136p33, is required for reactivation from latency in hematopoietic cells. Furthermore, UL136p33 is unstable, and its instability is important for the establishment of latency and sufficient accumulation of UL136p33 is a checkpoint for reactivation. We hypothesized that stabilizing UL136p33 might overcome the requirement of UL135 for reactivation. To test this, we generated recombinant viruses lacking UL135 that expressed a stabilized variant of UL136p33. Stabilizing UL136p33 did not impact replication of the UL135-mutant virus in fibroblasts. However, in the context of infection in hematopoietic cells, stabilization of UL136p33 strikingly compensated for the loss of UL135, resulting in increased replication in CD34+ HPCs and in humanized NOD-scid IL2R{gamma}cnull (NSG) mice. This finding suggests that while UL135 is essential for reactivation, it functions at steps preceding the accumulation of UL136p33 and that stabilized expression of UL136p33 largely overcomes the requirement for UL135 in reactivation. Taken together, our genetic evidence indicates an epistatic relationship between UL136p33 and UL135 whereby UL135 may initiate events early in reactivation that will result in the accumulation of UL136p33 to a threshold required for productive reactivation.

SIGNIFICANCEHuman cytomegalovirus (HCMV) is one of nine human herpesviruses and a significant human pathogen. While HCMV establishes a life-long latent infection that is typically asymptomatic in healthy individuals, its reactivation from latency can have devastating consequences in the immune compromised. Defining virus-host and virus-virus interactions important for HCMV latency, reactivation and replication is critical to defining the molecular basis of latent and replicative states and in controlling infection and CMV disease. Here we define a genetic relationship between two viral genes in controlling virus reactivation from latency using primary human hematopoietic progenitor cell and humanized mouse models.
]]></description>
<dc:creator>Moy, M. A.</dc:creator>
<dc:creator>Collins-McMillen, D. K.</dc:creator>
<dc:creator>Crawford, L.</dc:creator>
<dc:creator>Parkins, C.</dc:creator>
<dc:creator>Zeltzer, S.</dc:creator>
<dc:creator>Caviness, K.</dc:creator>
<dc:creator>Caposio, P.</dc:creator>
<dc:creator>Goodrum, F.</dc:creator>
<dc:date>2023-01-24</dc:date>
<dc:identifier>doi:10.1101/2023.01.24.525282</dc:identifier>
<dc:title><![CDATA[UL135 and UL136 Epistasis Controls Reactivation of Human Cytomegalovirus]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-01-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.01.29.526113v1?rss=1">
<title>
<![CDATA[
In vivo Modulation of Intraocular and Intracranial Pressures Causes Nonlinear and Non-monotonic Deformations of the Lamina Cribrosa and Scleral Canal 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.01.29.526113v1?rss=1</link>
<description><![CDATA[
PurposeTo evaluate changes in monkey optic nerve head (ONH) morphology under acutely controlled intraocular pressure (IOP) and intracranial pressure (ICP).

MethodsSeven ONHs from six monkeys were imaged via optical coherence tomography while IOP and ICP were maintained at one of 16 conditions. These conditions were defined by 4 levels for each pressure: low, baseline, high and very high. Images were processed to determine scleral canal area, aspect ratio, and planarity and anterior lamina cribrosa (ALC) shape index and curvature. Linear mixed effect models were utilized to investigate the effects of IOP, ICP and their interactions on ONH morphological features. The IOP-ICP interaction model was compared with one based on translaminar pressure difference (TLPD).

ResultsWe observed complex, eye-specific, non-linear patterns of ONH morphological changes with changes in IOP and ICP. For all ONH morphological features, linear mixed effects models demonstrated significant interactions between IOP and ICP that were unaccounted for by TLPD. Interactions indicate that the effects of IOP and ICP depend on the other pressure. The IOP-ICP interaction model was a higher quality predictor of ONH features than a TLPD model.

ConclusionsIn vivo modulation of IOP and ICP causes nonlinear and non-monotonic changes in monkey ONH morphology that depend on both pressures and is not accounted for by a simplistic TLPD. These results support and extend prior findings.

Translational Relevance: A better understanding of ICPs influence on the effects of IOP can help inform the highly variable presentations of glaucoma and effective treatment strategies.
]]></description>
<dc:creator>Zhu, Z.</dc:creator>
<dc:creator>Waxman, S.</dc:creator>
<dc:creator>Wang, B.</dc:creator>
<dc:creator>Wallace, J.</dc:creator>
<dc:creator>Schmitt, S. E.</dc:creator>
<dc:creator>Tyler-Kabara, E.</dc:creator>
<dc:creator>Ishikawa, H.</dc:creator>
<dc:creator>Schuman, J. S.</dc:creator>
<dc:creator>Smith, M. A.</dc:creator>
<dc:creator>Wollstein, G.</dc:creator>
<dc:creator>Sigal, I. A.</dc:creator>
<dc:date>2023-01-31</dc:date>
<dc:identifier>doi:10.1101/2023.01.29.526113</dc:identifier>
<dc:title><![CDATA[In vivo Modulation of Intraocular and Intracranial Pressures Causes Nonlinear and Non-monotonic Deformations of the Lamina Cribrosa and Scleral Canal]]></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/10.1101/2023.01.30.526357v1?rss=1">
<title>
<![CDATA[
Rewiring of the host cell metabolome and lipidome during lytic gammaherpesvirus infection is essential for infectious virus production 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.01.30.526357v1?rss=1</link>
<description><![CDATA[
Oncogenic virus infections are estimated to cause [~]15% of all cancers. Two prevalent human oncogenic viruses are members of the gammaherpesvirus family: Epstein Barr Virus (EBV) and Kaposis Sarcoma Herpesvirus (KSHV). We use murine herpesvirus 68 (MHV-68), which shares significant homology with KSHV and EBV, as a model system to study gammaherpesvirus lytic replication. Viruses implement distinct metabolic programs to support their life cycle, such as increasing the supply of lipids, amino acids, and nucleotide materials necessary to replicate. Our data define the global changes in the host cell metabolome and lipidome during gammaherpesvirus lytic replication. Our metabolomics analysis found that MHV-68 lytic infection induces glycolysis, glutaminolysis, lipid metabolism, and nucleotide metabolism. We additionally observed an increase in glutamine consumption and glutamine dehydrogenase protein expression. While both glucose and glutamine starvation of host cells decreased viral titers, glutamine starvation led to a greater loss in virion production. Our lipidomics analysis revealed a peak in triacylglycerides early during infection and an increase in free fatty acids and diacylglyceride later in the viral life cycle. Furthermore, we observed an increase in the protein expression of multiple lipogenic enzymes during infection. Interestingly, pharmacological inhibitors of glycolysis or lipogenesis resulted in decreased infectious virus production. Taken together, these results illustrate the global alterations in host cell metabolism during lytic gammaherpesvirus infection, establish essential pathways for viral production, and recommend targeted mechanisms to block viral spread and treat viral induced tumors.

IMPORTANCEViruses are intracellular parasites which lack their own metabolism, so they must hijack host cell metabolic machinery in order to increase the production of energy, proteins, fats, and genetic material necessary to replicate. Using murine herpesvirus 68 (MHV-68) as a model system to understand how similar human gammaherpesviruses cause cancer, we profiled the metabolic changes that occur during lytic MHV-68 infection and replication. We found MHV-68 infection of host cells increases glucose, glutamine, lipid, and nucleotide metabolic pathways. We also showed inhibition or starvation of glucose, glutamine or lipid metabolic pathways results in an inhibition of virus production. Ultimately, targeting changes in host cell metabolism due to viral infection can be used to treat gammaherpesvirus induced cancers and infections in humans.
]]></description>
<dc:creator>Clark, S. A.</dc:creator>
<dc:creator>Vazquez, A.</dc:creator>
<dc:creator>Furiya, K.</dc:creator>
<dc:creator>Splattstoesser, M. K.</dc:creator>
<dc:creator>Bashmail, A. K.</dc:creator>
<dc:creator>Schwartz, H.</dc:creator>
<dc:creator>Russell, M.</dc:creator>
<dc:creator>Bhark, S.-J.</dc:creator>
<dc:creator>Moreno, O. K.</dc:creator>
<dc:creator>McGovern, M.</dc:creator>
<dc:creator>Owsley, E. R.</dc:creator>
<dc:creator>Nelson, T. A.</dc:creator>
<dc:creator>Sanchez, E.</dc:creator>
<dc:creator>Delgado, T.</dc:creator>
<dc:date>2023-01-31</dc:date>
<dc:identifier>doi:10.1101/2023.01.30.526357</dc:identifier>
<dc:title><![CDATA[Rewiring of the host cell metabolome and lipidome during lytic gammaherpesvirus infection is essential for infectious virus production]]></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/10.1101/2023.01.31.525753v1?rss=1">
<title>
<![CDATA[
Robust biomarker discovery through multiplatform multiplex image analysis of breast cancer clinical cohorts 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.01.31.525753v1?rss=1</link>
<description><![CDATA[
AO_SCPLOWBSTRACTC_SCPLOWSpatial profiling of tissues promises to elucidate tumor-microenvironment interactions and enable development of spatial biomarkers to predict patient response to immunotherapy and other therapeutics. However, spatial biomarker discovery is often carried out on a single patient cohort or imaging technology, limiting statistical power and increasing the likelihood of technical artifacts. In order to analyze multiple patient cohorts profiled on different platforms, we developed methods for comparative data analysis from three disparate multiplex imaging technologies: 1) cyclic immunofluorescence data we generated from 102 breast cancer patients with clinical follow-up, in addition to publicly available 2) imaging mass cytometry and 3) multiplex ion-beam imaging data. We demonstrate similar single-cell phenotyping results across breast cancer patient cohorts imaged with these three technologies and identify cellular abundance and proximity-based biomarkers with prognostic value across platforms. In multiple platforms, we identified lymphocyte infiltration as independently associated with longer survival in triple negative and high-proliferation breast tumors. Then, a comparison of nine spatial analysis methods revealed robust spatial biomarkers. In estrogen receptor-positive disease, quiescent stromal cells close to tumor were more abundant in good prognosis tumors while tumor neighborhoods of mixed fibroblast phenotypes were enriched in poor prognosis tumors. In triple-negative breast cancer (TNBC), macrophage proximity to tumor and B cell proximity to T cells were greater in good prognosis tumors, while tumor neighborhoods of vimentin-positive fibroblasts were enriched in poor prognosis tumors. We also tested previously published spatial biomarkers in our ensemble cohort, reproducing the positive prognostic value of isolated lymphocytes and lymphocyte occupancy and failing to reproduce the prognostic value of tumor-immune mixing score in TNBC. In conclusion, we demonstrate assembly of larger clinical cohorts from diverse platforms to aid in prognostic spatial biomarker identification and validation.

SO_SCPLOWTATEMENTC_SCPLOWO_SCPCAP C_SCPCAPO_SCPLOWOFC_SCPLOWO_SCPCAP C_SCPCAPO_SCPLOWSIGNIFICANCEC_SCPLOWOur single-cell spatial analysis of multiple clinical cohorts uncovered novel biomarkers of patient outcome in breast cancer. Additionally, our data, software, and methods will help advance spatial characterization of the tumor microenvironment.
]]></description>
<dc:creator>Eng, J. R.</dc:creator>
<dc:creator>Bucher, E.</dc:creator>
<dc:creator>Hu, Z.</dc:creator>
<dc:creator>Sanders, M.</dc:creator>
<dc:creator>Chakravarthy, B.</dc:creator>
<dc:creator>Pietenpol, J. A.</dc:creator>
<dc:creator>Gibbs, S. L.</dc:creator>
<dc:creator>Chin, K.</dc:creator>
<dc:date>2023-02-03</dc:date>
<dc:identifier>doi:10.1101/2023.01.31.525753</dc:identifier>
<dc:title><![CDATA[Robust biomarker discovery through multiplatform multiplex image analysis of breast cancer clinical cohorts]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-02-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.02.07.527452v1?rss=1">
<title>
<![CDATA[
Human Cytomegalovirus UL138 Interaction with USP1 Activates STAT1 in infection 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.02.07.527452v1?rss=1</link>
<description><![CDATA[
Innate immune responses are crucial for limiting virus infection. However, viruses often hijack our best defenses for viral objectives. Human Cytomegalovirus (HCMV) is a beta herpesvirus which establishes a life-long latent infection. Defining the virus-host interactions controlling latency and reactivation is vital to the control of viral disease risk posed by virus reactivation. We defined an interaction between UL138, a pro-latency HCMV gene, and the host deubiquintase complex, UAF1-USP1. UAF1 is a scaffold protein pivotal for the activity of ubiquitin specific peptidases (USP), including USP1. UAF1-USP1 sustains an innate immune response through the phosphorylation and activation of signal transducer and activator of transcription-1 (pSTAT1), as well as regulates the DNA damage response. After the onset of viral DNA synthesis, pSTAT1 levels are elevated and this depends upon UL138 and USP1. pSTAT1 localizes to viral centers of replication, binds to the viral genome, and influences UL138 expression. Inhibition of USP1 results in a failure to establish latency, marked by increased viral genome replication and production of viral progeny. Inhibition of Jak-STAT signaling also results in increased viral genome synthesis in hematopoietic cells, consistent with a role for USP1-mediated regulation of STAT1 signaling in the establishment of latency. These findings demonstrate the importance of the UL138-UAF1-USP1 virus-host interaction in regulating HCMV latency establishment through the control of innate immune signaling. It will be important going forward to distinguish roles of UAF1-USP1 in regulating pSTAT1 relative to its role in the DNA damage response in HCMV infection.

ImportanceHuman cytomegalovirus (HCMV) is one of nine herpesviruses that infect humans. Following a primary infection, HCMV establishes a life-long latent infection that is marked by sporadic, and likely frequent reactivation events. While these reactivation events are asymptomatic in the immune competent host, they pose important disease risks for the immune compromised, including solid organ or stem cell transplant recipients. Its complex interactions with host biology and deep coding capacity make it an excellent model for defining mechanisms important for viral latency and reactivation. Here we define an interaction with host proteins that commandeer typically antiviral innate immune signaling for the establishment of latency.
]]></description>
<dc:creator>Zarrella, K.</dc:creator>
<dc:creator>Longmire, P.</dc:creator>
<dc:creator>Zeltzer, S.</dc:creator>
<dc:creator>Collins-McMillen, D.</dc:creator>
<dc:creator>Hancock, M.</dc:creator>
<dc:creator>Buehler, J.</dc:creator>
<dc:creator>Reitsma, J. M.</dc:creator>
<dc:creator>Terhune, S. S.</dc:creator>
<dc:creator>Nelson, J. A.</dc:creator>
<dc:creator>Goodrum, F.</dc:creator>
<dc:date>2023-02-07</dc:date>
<dc:identifier>doi:10.1101/2023.02.07.527452</dc:identifier>
<dc:title><![CDATA[Human Cytomegalovirus UL138 Interaction with USP1 Activates STAT1 in infection]]></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/10.1101/2023.02.11.528140v1?rss=1">
<title>
<![CDATA[
Impact of aging on the immunological and microbial landscape of the lung during non-tuberculous mycobacterial infection 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.02.11.528140v1?rss=1</link>
<description><![CDATA[
Nontuberculous mycobacteria (NTM) are environmentally ubiquitous and predominately cause pulmonary disease (NTMPD). The incidence of NTMPD has steadily increased and is now more prevalent than that of Mycobacterium tuberculosis (M. tb) in the US. Moreover, the prevalence of NTMPD increases with age; therefore, it is likely that the burden of NTMPD will continue to increase in the coming decades as the number of those over the age of 65 increased in the U.S population. However, the mechanisms leading to higher susceptibility and severity of NTMPD with aging are poorly defined. Here, we used a rhesus macaque model of intrabronchial infection with M. avium complex in young and aged animals to address this knowledge gap. Unilateral infection resulted in a robust inflammatory response predominantly in the inoculated lung, however, immune cell infiltration and antigen-specific T cell responses were detected in both lungs. Nasal, oral, and fecal swabs, and BAL samples were profiled using 16S amplicon sequencing. These data suggested that decompartmentalization of the lower respiratory microbiome was occurring, evidenced by detection of bacterial DNA typically found in the gut and oral-pharyngeal cavity in bronchoalveolar samples following infection. Radiographic studies, gross pathology, and histopathology examination revealed increased disease severity in aged compared to young animals with pulmonary consolidation, edema, and lesions. Finally, single cell RNA sequencing indicated that aged animals generated a dysregulated macrophage and CD8 T cell response to MAC infection.
]]></description>
<dc:creator>Cinco, I. R.</dc:creator>
<dc:creator>Rhoades, N. S.</dc:creator>
<dc:creator>Napier, E. G.</dc:creator>
<dc:creator>Davies, M.</dc:creator>
<dc:creator>Allison, D. B.</dc:creator>
<dc:creator>Kohama, S. G.</dc:creator>
<dc:creator>Bermudez, L. E.</dc:creator>
<dc:creator>Winthrop, K.</dc:creator>
<dc:creator>Fuss, C.</dc:creator>
<dc:creator>Spindel, E. R.</dc:creator>
<dc:creator>Messaoudi, I.</dc:creator>
<dc:date>2023-02-13</dc:date>
<dc:identifier>doi:10.1101/2023.02.11.528140</dc:identifier>
<dc:title><![CDATA[Impact of aging on the immunological and microbial landscape of the lung during non-tuberculous mycobacterial infection]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-02-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.02.14.528504v1?rss=1">
<title>
<![CDATA[
Tertiary Lymphoid Structures Sustain Cutaneous B cell Activity in Hidradenitis Suppurativa 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.02.14.528504v1?rss=1</link>
<description><![CDATA[
Structured AbstractO_ST_ABSBackgroundC_ST_ABSHidradenitis suppurativa (HS) skin lesions are highly inflammatory and characterized by a large immune infiltrate. While B cells and plasma cells comprise a major component of this immune milieu the biology and contribution of these cells in HS pathogenesis is unclear.

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

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

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

ConclusionOur data definitively demonstrate the presence of TLS in lesional HS skin and point to ongoing cutaneous B cell maturation through class switch recombination and affinity maturation during disease progression in this inflamed non-lymphoid tissue.
]]></description>
<dc:creator>Lowe, M. M.</dc:creator>
<dc:creator>Cohen, J. N.</dc:creator>
<dc:creator>Moss, M. I.</dc:creator>
<dc:creator>Clancy, S.</dc:creator>
<dc:creator>Adler, J.</dc:creator>
<dc:creator>Yates, A.</dc:creator>
<dc:creator>Naik, H. B.</dc:creator>
<dc:creator>Pauli, M.</dc:creator>
<dc:creator>Taylor, I.</dc:creator>
<dc:creator>McKay, A.</dc:creator>
<dc:creator>Harris, H.</dc:creator>
<dc:creator>Kim, E.</dc:creator>
<dc:creator>Hansen, S. L.</dc:creator>
<dc:creator>Rosenblum, M. D.</dc:creator>
<dc:creator>Moreau, J. M.</dc:creator>
<dc:date>2023-02-15</dc:date>
<dc:identifier>doi:10.1101/2023.02.14.528504</dc:identifier>
<dc:title><![CDATA[Tertiary Lymphoid Structures Sustain Cutaneous B cell Activity in Hidradenitis Suppurativa]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-02-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.02.16.528867v1?rss=1">
<title>
<![CDATA[
Regulated assembly and neurosteroid modulation constrain GABAA receptor pharmacology in vivo 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.02.16.528867v1?rss=1</link>
<description><![CDATA[
Type A GABA receptors (GABAARs) are the principal inhibitory receptors in the brain and the target of a wide range of clinical agents, including anesthetics, sedatives, hypnotics, and antidepressants. However, our understanding of GABAAR pharmacology has been hindered by the vast number of pentameric assemblies that can be derived from a total 19 different subunits and the lack of structural knowledge of clinically relevant receptors. Here, we isolate native murine GABAAR assemblies containing the widely expressed 1 subunit, and elucidate their structures in complex with drugs used to treat insomnia (zolpidem and flurazepam) and postpartum depression (the neurosteroid allopregnanolone). Using cryo-EM analysis and single-molecule photobleaching experiments, we uncover only three structural populations in the brain: the canonical 1{beta}2{gamma}2 receptor containing two 1 subunits and two unanticipated assemblies containing one 1 and either an 2, 3 or 5 subunit. Both of the noncanonical assemblies feature a more compact arrangement between the transmembrane and extracellular domains. Interestingly, allopregnanolone is bound at the transmembrane /{beta} subunit interface, even when not added to the sample, revealing an important role for endogenous neurosteroids in modulating native GABAARs. Together with structurally engaged lipids, neurosteroids produce global conformational changes throughout the receptor that modify both the pore diameter and binding environments for GABA and insomnia medications. Together, our data reveal that GABAAR assembly is a strictly regulated process that yields a small number of structurally distinct complexes, defining a structural landscape from which subtype-specific drugs can be developed.
]]></description>
<dc:creator>Sun, C.</dc:creator>
<dc:creator>Zhu, H.</dc:creator>
<dc:creator>Clark, S. A.</dc:creator>
<dc:creator>Gouaux, E.</dc:creator>
<dc:date>2023-02-16</dc:date>
<dc:identifier>doi:10.1101/2023.02.16.528867</dc:identifier>
<dc:title><![CDATA[Regulated assembly and neurosteroid modulation constrain GABAA receptor pharmacology in vivo]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-02-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.02.23.529718v1?rss=1">
<title>
<![CDATA[
Dual-modality imaging of immunofluorescence and imaging mass cytometry for whole slide imaging with accurate single-cell segmentation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.02.23.529718v1?rss=1</link>
<description><![CDATA[
Imaging mass cytometry (IMC) is a powerful multiplexed tissue imaging technology that allows simultaneous detection of more than 30 makers on a single slide. It has been increasingly used for singlecell-based spatial phenotyping in a wide range of samples. However, it only acquires a small, rectangle field of view (FOV) with a low image resolution that hinders downstream analysis. Here, we reported a highly practical dual-modality imaging method that combines high-resolution immunofluorescence (IF) and high-dimensional IMC on the same tissue slide. Our computational pipeline uses the whole slide image (WSI) of IF as a spatial reference and integrates small FOVs IMC into a WSI of IMC. The high-resolution IF images enable accurate single-cell segmentation to extract robust high-dimensional IMC features for downstream analysis. We applied this method in esophageal adenocarcinoma of different stages, identified the single-cell pathology landscape via reconstruction of WSI IMC images, and demonstrated the advantage of the dual-modality imaging strategy.

MotivationHighly multiplexed tissue imaging allows visualization of the spatially resolved expression of multiple proteins at the single-cell level. Although imaging mass cytometry (IMC) using metal isotope-conjugated antibodies has a significant advantage of low background signal and absence of autofluorescence or batch effect, it has a low resolution that hampers accurate cell segmentation and results in inaccurate feature extraction. In addition, IMC only acquires mm2-sized rectangle regions, which limits its application and efficiency when studying larger clinical samples with non-rectangle shapes. To maximize the research output of IMC, we developed the dual-modality imaging method based on a highly practical and technical improvement requiring no extra specialized equipment or agents and proposed a comprehensive computational pipeline that combines IF and IMC. The proposed method greatly improves the accuracy of cell segmentation and downstream analysis and is able to obtain whole slide image IMC to capture the comprehensive cellular landscape of large tissue sections.
]]></description>
<dc:creator>Kim, E. N.</dc:creator>
<dc:creator>Chen, P. Z.</dc:creator>
<dc:creator>Bressan, D.</dc:creator>
<dc:creator>Tripathi, M.</dc:creator>
<dc:creator>Miremadi, A.</dc:creator>
<dc:creator>di Pietro, M.</dc:creator>
<dc:creator>Coussens, L. M.</dc:creator>
<dc:creator>Hannon, G. J.</dc:creator>
<dc:creator>Fitzgerald, R. C.</dc:creator>
<dc:creator>Zhuang, L.</dc:creator>
<dc:creator>Chang, Y. H.</dc:creator>
<dc:date>2023-02-23</dc:date>
<dc:identifier>doi:10.1101/2023.02.23.529718</dc:identifier>
<dc:title><![CDATA[Dual-modality imaging of immunofluorescence and imaging mass cytometry for whole slide imaging with accurate single-cell segmentation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-02-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.02.23.529809v1?rss=1">
<title>
<![CDATA[
SEG: Segmentation Evaluation in absence of Ground truth labels 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.02.23.529809v1?rss=1</link>
<description><![CDATA[
Identifying individual cells or nuclei is often the first step in the analysis of multiplex tissue imaging (MTI) data. Recent efforts to produce plug-and-play, end-to-end MTI analysis tools such as MCMICRO1- though groundbreaking in their usability and extensibility - are often unable to provide users guidance regarding the most appropriate models for their segmentation task among an endless proliferation of novel segmentation methods. Unfortunately, evaluating segmentation results on a users dataset without ground truth labels is either purely subjective or eventually amounts to the task of performing the original, time-intensive annotation. As a consequence, researchers rely on models pre-trained on other large datasets for their unique tasks. Here, we propose a methodological approach for evaluating MTI nuclei segmentation methods in absence of ground truth labels by scoring relatively to a larger ensemble of segmentations. To avoid potential sensitivity to collective bias from the ensemble approach, we refine the ensemble via weighted average across segmentation methods, which we derive from a systematic model ablation study. First, we demonstrate a proof-of-concept and the feasibility of the proposed approach to evaluate segmentation performance in a small dataset with ground truth annotation. To validate the ensemble and demonstrate the importance of our method-specific weighting, we compare the ensembles detection and pixel-level predictions - derived without supervision - with the datas ground truth labels. Second, we apply the methodology to an unlabeled larger tissue microarray (TMA) dataset, which includes a diverse set of breast cancer phenotypes, and provides decision guidelines for the general user to more easily choose the most suitable segmentation methods for their own dataset by systematically evaluating the performance of individual segmentation approaches in the entire dataset.
]]></description>
<dc:creator>Sims, Z.</dc:creator>
<dc:creator>Strgar, L.</dc:creator>
<dc:creator>Thirumalaisamy, D.</dc:creator>
<dc:creator>Heussner, R.</dc:creator>
<dc:creator>Thibault, G.</dc:creator>
<dc:creator>Chang, Y. H.</dc:creator>
<dc:date>2023-02-24</dc:date>
<dc:identifier>doi:10.1101/2023.02.23.529809</dc:identifier>
<dc:title><![CDATA[SEG: Segmentation Evaluation in absence of Ground truth labels]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-02-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.02.28.530337v1?rss=1">
<title>
<![CDATA[
The acetylase activity of Cdu1 protects Chlamydia effectors from degradation to regulate bacterial exit from infected cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.02.28.530337v1?rss=1</link>
<description><![CDATA[
Many cellular processes are regulated by ubiquitin-mediated proteasomal degradation. Pathogens can regulate eukaryotic proteolysis through the delivery of proteins with de-ubiquitinating (DUB) activities. The obligate intracellular pathogen Chlamydia trachomatis secretes Cdu1 (ChlaDUB1), a dual deubiquitinase and Lys-acetyltransferase, that promotes Golgi remodeling and survival of infected host cells presumably by regulating the ubiquitination of host and bacterial proteins. Here we determined that Cdu1s acetylase but not its DUB activity is important to protect Cdu1 from ubiquitin-mediated degradation. We further identified three C. trachomatis proteins on the pathogen-containing vacuole (InaC, IpaM, and CTL0480) that required Cdu1 s acetylase activity for protection from degradation and determined that Cdu1 and these Cdu1-protected proteins are required for optimal egress of Chlamydia from host cells. These findings highlight a non-canonical mechanism of pathogen-mediated protection of virulence factors from degradation after their delivery into host cells and the coordinated regulation of secreted effector proteins.
]]></description>
<dc:creator>Bastidas, R. J.</dc:creator>
<dc:creator>Kedzior, M.</dc:creator>
<dc:creator>Dolat, L.</dc:creator>
<dc:creator>Sixt, B. S.</dc:creator>
<dc:creator>Pruneda, J. N.</dc:creator>
<dc:creator>Valdivia, R. H.</dc:creator>
<dc:date>2023-02-28</dc:date>
<dc:identifier>doi:10.1101/2023.02.28.530337</dc:identifier>
<dc:title><![CDATA[The acetylase activity of Cdu1 protects Chlamydia effectors from degradation to regulate bacterial exit from infected cells]]></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/10.1101/2023.02.28.530460v1?rss=1">
<title>
<![CDATA[
MEMO1 is a Metal Containing Regulator of Iron Homeostasis in Cancer Cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.02.28.530460v1?rss=1</link>
<description><![CDATA[
Mediator of ERBB2-driven Cell Motility 1 (MEMO1) is an evolutionary conserved protein implicated in many biological processes; however, its primary molecular function remains unknown. Importantly, MEMO1 is overexpressed in many types of cancer and was shown to modulate breast cancer metastasis through altered cell motility.

To better understand the function of MEMO1 in cancer cells, we analyzed genetic interactions of MEMO1 using gene essentiality data from 1028 cancer cell lines and found multiple iron-related genes exhibiting genetic relationships with MEMO1. We experimentally confirmed several interactions between MEMO1 and iron-related proteins in living cells or in vitro, most notably, the iron transporters transferrin (TF), transferrin receptor 2 (TFR2), and mitoferrin-2 (SLC25A28), and the global iron response regulator IRP1 (ACO1). These interactions indicate that cells with high MEMO1 expression levels are hypersensitive to the disruptions in iron distribution. Our data also indicate that MEMO1 is involved in ferroptosis and is linked to iron supply to mitochondria.

We have found that purified MEMO1 binds iron with high affinity under redox conditions mimicking intracellular environment and solved MEMO1 structures in complex with iron and copper. Our work reveals that the iron coordination mode in MEMO1 is very similar to that of iron-containing extradiol dioxygenases, which also display a similar structural fold. We conclude that MEMO1 is an iron-binding protein that regulates iron homeostasis in cancer cells.
]]></description>
<dc:creator>Dolgova, N.</dc:creator>
<dc:creator>Uhlemann, E.-M. E.</dc:creator>
<dc:creator>Boniecki, M. T.</dc:creator>
<dc:creator>Vizeacoumar, F. S.</dc:creator>
<dc:creator>Ralle, M.</dc:creator>
<dc:creator>Tonelli, M.</dc:creator>
<dc:creator>Abbas, S. A.</dc:creator>
<dc:creator>Patry, J.</dc:creator>
<dc:creator>Elhasasna, H.</dc:creator>
<dc:creator>Freywald, A.</dc:creator>
<dc:creator>Vizeacoumar, F.</dc:creator>
<dc:creator>Dmitriev, O. Y.</dc:creator>
<dc:date>2023-02-28</dc:date>
<dc:identifier>doi:10.1101/2023.02.28.530460</dc:identifier>
<dc:title><![CDATA[MEMO1 is a Metal Containing Regulator of Iron Homeostasis in Cancer Cells]]></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/10.1101/2023.05.08.539829v1?rss=1">
<title>
<![CDATA[
Monomethylation of Lysine 27 at Histone 3 Confers Lifelong Susceptibility to Stress 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.05.08.539829v1?rss=1</link>
<description><![CDATA[
Histone post-translational modifications are critical for mediating persistent alterations in gene expression. By combining unbiased proteomics profiling, and genome-wide approaches, we uncovered a role for mono-methylation of lysine 27 at histone H3 (H3K27me1) in the enduring effects of stress. Specifically, mice exposed to early life stress (ELS) or to chronic social defeat stress (CSDS) in adulthood displayed increased enrichment of H3K27me1, and transient decreases in H3K27me2, in the nucleus accumbens (NAc), a key brain-reward region. Stress induction of H3K27me1 was mediated by the VEFS domain of SUZ12, a core subunit of the polycomb repressive complex-2, which is induced by chronic stress and controls H3K27 methylation patterns. Overexpression of the VEFS domain led to social, emotional, and cognitive abnormalities, and altered excitability of NAc D1 mediums spiny neurons. Together, we describe a novel function of H3K27me1 in brain and demonstrate its role as a "chromatin scar" that mediates lifelong stress susceptibility.
]]></description>
<dc:creator>Torres-Berrio, A.</dc:creator>
<dc:creator>Estill, M.</dc:creator>
<dc:creator>Ramakrishnan, A.</dc:creator>
<dc:creator>Kronman, H.</dc:creator>
<dc:creator>Patel, V.</dc:creator>
<dc:creator>Minier-Toribio, A.</dc:creator>
<dc:creator>Issler, O.</dc:creator>
<dc:creator>Browne, C. J.</dc:creator>
<dc:creator>Parise, E.</dc:creator>
<dc:creator>van der Zee, Y.</dc:creator>
<dc:creator>Walker, D.</dc:creator>
<dc:creator>Martinez-Rivera, F. J.</dc:creator>
<dc:creator>Lardner, C. K.</dc:creator>
<dc:creator>Durand, R.</dc:creator>
<dc:creator>Russo, S.</dc:creator>
<dc:creator>Shen, L.</dc:creator>
<dc:creator>Sidoli, S.</dc:creator>
<dc:creator>Nestler, E. J.</dc:creator>
<dc:date>2023-05-08</dc:date>
<dc:identifier>doi:10.1101/2023.05.08.539829</dc:identifier>
<dc:title><![CDATA[Monomethylation of Lysine 27 at Histone 3 Confers Lifelong Susceptibility to Stress]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-05-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/204172v1?rss=1">
<title>
<![CDATA[
TRPV5, TRPV6, TRPM6, and TRPM7 do not contribute to hair-cell mechanotransduction 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/204172v1?rss=1</link>
<description><![CDATA[
The hair-cell mechanotransduction channel remains unidentified. We tested whether four transient receptor channel (TRP) family members, TRPV5, TRPV6, TRPM6, and TRPM7, participated in transduction. Using cysteine-substitution mouse knock-ins and methanethiosulfonate reagents selective for those alleles, we found that inhibition of TRPV5 or TRPV6 had no effect on transduction in mouse cochlear hair cells. TRPM6 and TRPM7 each interacted with the tip-link component PCDH15 in cultured eukaryotic cells, which suggested they could participate in transduction. Cochlear hair cell transduction was insensitive to shRNA knockdown of Trpm6 or Trpm7, however, and was not affected by manipulations of Mg2+, which normally perturbs TRPM6 and TRPM7. To definitively examine the role of these two channels in transduction, we showed that deletion of either or both of their genes selectively in hair cells had no effect on auditory function. We suggest that TRPV5, TRPV6, TRPM6, and TRPM7 are unlikely to be the pore-forming subunit of the hair-cell transduction channel.
]]></description>
<dc:creator>Morgan, C. P.</dc:creator>
<dc:creator>Zhao, H.</dc:creator>
<dc:creator>LeMasurier, M.</dc:creator>
<dc:creator>Xiong, W.</dc:creator>
<dc:creator>Pan, B.</dc:creator>
<dc:creator>Avenarius, M. R.</dc:creator>
<dc:creator>Bateschell, M.</dc:creator>
<dc:creator>Larisch, R.</dc:creator>
<dc:creator>Ricci, A. J.</dc:creator>
<dc:creator>Mueller, U.</dc:creator>
<dc:creator>Barr-Gillespie, P. G.</dc:creator>
<dc:date>2017-10-16</dc:date>
<dc:identifier>doi:10.1101/204172</dc:identifier>
<dc:title><![CDATA[TRPV5, TRPV6, TRPM6, and TRPM7 do not contribute to hair-cell mechanotransduction]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-10-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/204370v1?rss=1">
<title>
<![CDATA[
Germline Contamination and Leakage in Whole Genome Somatic Single Nucleotide Variant Detection 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/204370v1?rss=1</link>
<description><![CDATA[
BackgroundThe clinical sequencing of cancer genomes to personalize therapy is becoming routine across the world. However, concerns over patient re-identification from these data lead to questions about how tightly access should be controlled. It is not thought to be possible to re-identify patients from somatic variant data. However, somatic variant detection pipelines can mistakenly identify germline variants as somatic ones, a process called "germline leakage". The rate of germline leakage across different somatic variant detection pipelines is not well-understood, and it is uncertain whether or not somatic variant calls should be considered re-identifiable. To fill this gap, we quantified germline leakage across 259 sets of whole-genome somatic single nucleotide variant (SNVs) predictions made by 21 teams as part of the ICGC-TCGA DREAM Somatic Mutation Calling Challenge.nnResultsThe median somatic SNV prediction set contained 4,325 somatic SNVs and leaked one germline polymorphism. The level of germline leakage was inversely correlated with somatic SNV prediction accuracy and positively correlated with the amount of infiltrating normal cells. The specific germline variants leaked differed by tumour and algorithm. To aid in quantitation and correction of leakage, we created a tool, called GermlineFilter, for use in public-facing somatic SNV databases.nnConclusionsThe potential for patient re-identification from leaked germline variants in somatic SNV predictions has led to divergent open data access policies, based on different assessments of the risks. Indeed, a single, well-publicized re-identification event could reshape public perceptions of the values of genomic data sharing. We find that modern somatic SNV prediction pipelines have low germline-leakage rates, which can be further reduced, especially for cloud-sharing, using pre-filtering software.
]]></description>
<dc:creator>Sendorek, D. H.</dc:creator>
<dc:creator>Caloian, C.</dc:creator>
<dc:creator>Ellrott, K.</dc:creator>
<dc:creator>Bare, J. C.</dc:creator>
<dc:creator>Yamaguchi, T. N.</dc:creator>
<dc:creator>Ewing, A. D.</dc:creator>
<dc:creator>Houlahan, K. E.</dc:creator>
<dc:creator>Norman, T. C.</dc:creator>
<dc:creator>Margolin, A. A.</dc:creator>
<dc:creator>Stuart, J. M.</dc:creator>
<dc:creator>Boutros, P. C.</dc:creator>
<dc:date>2017-10-17</dc:date>
<dc:identifier>doi:10.1101/204370</dc:identifier>
<dc:title><![CDATA[Germline Contamination and Leakage in Whole Genome Somatic Single Nucleotide Variant Detection]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-10-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/204859v1?rss=1">
<title>
<![CDATA[
Structural basis for recognition of diverse antidepressants by the human serotonin transporter 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/204859v1?rss=1</link>
<description><![CDATA[
Selective serotonin reuptake inhibitors are clinically prescribed antidepressants that act by increasing the local concentration of neurotransmitter at synapses and in extracellular spaces via blockade of the serotonin transporter. Here we report x-ray structures of engineered thermostable variants of the human serotonin transporter bound to the antidepressants sertraline, fluvoxamine, and paroxetine. The drugs prevent serotonin binding by occupying the central substrate binding site and stabilizing the transporter in an outward-open conformation. These structures explain how residues within the central site orchestrate binding of chemically diverse inhibitors and mediate transporter-drug selectivity.
]]></description>
<dc:creator>Coleman, J.</dc:creator>
<dc:creator>Gouaux, E.</dc:creator>
<dc:date>2017-10-17</dc:date>
<dc:identifier>doi:10.1101/204859</dc:identifier>
<dc:title><![CDATA[Structural basis for recognition of diverse antidepressants by the human serotonin transporter]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-10-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/206037v1?rss=1">
<title>
<![CDATA[
Genetic analysis of de novo variants reveals sex differences in complex and isolated congenital diaphragmatic hernia and indicates MYRF as a candidate gene 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/206037v1?rss=1</link>
<description><![CDATA[
Congenital diaphragmatic hernia (CDH) is one of the most common and lethal birth defects. Previous studies using exome sequencing support a significant contribution of coding de novo variants in complex CDH cases with additional anomalies and likely gene-disrupting (LGD) variants in isolated CDH cases. To further investigate the genetic architecture of CDH, we performed exome or genome sequencing in 283 proband-parent trios. Combined with data from previous studies, we analyzed a total of 357 trios, including 148 complex and 209 isolated cases. Complex and isolated cases both have a significant burden of deleterious de novo coding variants (1.7~fold, p= 1.2x10-5 for complex, 1.5~fold, p= 9.0x10-5 for isolated). Strikingly, in isolated CDH, almost all of the burden is carried by female cases (2.1~fold, p=0.004 for likely gene disrupting and 1.8~fold, p= 0.0008 for damaging missense variants); whereas in complex CDH, the burden is similar in females and males. Additionally, de novo LGD variants in complex cases are mostly enriched in genes highly expressed in developing diaphragm, but distributed in genes with a broad range of expression levels in isolated cases. Finally, we identified a new candidate risk gene MYRF (4 de novo variants, p-value=2x10-10), a transcription factor intolerant of mutations. Patients with MYRF mutations have additional anomalies including congenital heart disease and genitourinary defects, likely representing a novel syndrome.
]]></description>
<dc:creator>Qi, H.</dc:creator>
<dc:creator>Yu, L.</dc:creator>
<dc:creator>Zhou, X.</dc:creator>
<dc:creator>Kitaygorodsky, A.</dc:creator>
<dc:creator>Wynn, J.</dc:creator>
<dc:creator>Zhu, N.</dc:creator>
<dc:creator>Aspelund, G.</dc:creator>
<dc:creator>Lim, F. Y.</dc:creator>
<dc:creator>Crombleholme, T.</dc:creator>
<dc:creator>Cusick, R.</dc:creator>
<dc:creator>Azarow, K.</dc:creator>
<dc:creator>Danko, M. E.</dc:creator>
<dc:creator>Chung, D.</dc:creator>
<dc:creator>Warner, B. W.</dc:creator>
<dc:creator>Mychaliska, G. B.</dc:creator>
<dc:creator>Potoka, D.</dc:creator>
<dc:creator>Wagner, A. J.</dc:creator>
<dc:creator>ElFiky, M.</dc:creator>
<dc:creator>Wilson, J. M.</dc:creator>
<dc:creator>High, F. A.</dc:creator>
<dc:creator>Longoni, M.</dc:creator>
<dc:creator>Donahoe, P.</dc:creator>
<dc:creator>Chung, W. K.</dc:creator>
<dc:creator>Shen, Y.</dc:creator>
<dc:date>2017-10-19</dc:date>
<dc:identifier>doi:10.1101/206037</dc:identifier>
<dc:title><![CDATA[Genetic analysis of de novo variants reveals sex differences in complex and isolated congenital diaphragmatic hernia and indicates MYRF as a candidate gene]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-10-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/213553v1?rss=1">
<title>
<![CDATA[
A multi-center study on factors influencing the reproducibility of in vitro drug-response studies 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/213553v1?rss=1</link>
<description><![CDATA[
Evidence that some influential biomedical results cannot be repeated has increased interest in practices that generate data meeting findable, accessible, interoperable and reproducible (FAIR) standards. Multiple papers have identified examples of irreproducibility, but practical steps for increasing reproducibility have not been widely studied. Here, seven research centers in the NIH LINCS Program Consortium investigate the reproducibility of a prototypical perturbational assay: quantifying the responsiveness of cultured cells to anti-cancer drugs. Such assays are important for drug development, studying cell biology, and patient stratification. While many experimental and computational factors have an impact on intra- and inter-center reproducibility, the factors most difficult to identify and correct are those with a strong dependency on biological context. These factors often vary in magnitude with the drug being analyzed and with growth conditions. We provide ways of identifying such context-sensitive factors, thereby advancing the conceptual and practical basis for greater experimental reproducibility.
]]></description>
<dc:creator>Niepel, M.</dc:creator>
<dc:creator>Hafner, M.</dc:creator>
<dc:creator>Williams, E. H.</dc:creator>
<dc:creator>Chung, M.</dc:creator>
<dc:creator>Barrette, A. M.</dc:creator>
<dc:creator>Stern, A. D.</dc:creator>
<dc:creator>Hu, B.</dc:creator>
<dc:creator>LINCS Consortium,</dc:creator>
<dc:creator>Gray, J. W.</dc:creator>
<dc:creator>Birtwistle, M. R.</dc:creator>
<dc:creator>Heiser, L. M.</dc:creator>
<dc:creator>Sorger, P. K.</dc:creator>
<dc:date>2017-11-03</dc:date>
<dc:identifier>doi:10.1101/213553</dc:identifier>
<dc:title><![CDATA[A multi-center study on factors influencing the reproducibility of in vitro drug-response studies]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-11-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/213561v1?rss=1">
<title>
<![CDATA[
Metagenomics for chronic meningitis: clarifying interpretation and diagnosis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/213561v1?rss=1</link>
<description><![CDATA[
ImportanceIdentifying infectious causes of subacute and chronic meningitis can be challenging. Enhanced, unbiased diagnostic approaches are needed.nnObjectiveTo present a case series of patients with diagnostically challenging subacute and chronic meningitis in whom metagenomic next-generation sequencing (mNGS) of cerebrospinal fluid (CSF), supported by a statistical framework generated from mNGS sequencing of non-infectious patients and environmental controls, identified a pathogen.nnDesignCase series. Using mNGS data from the CSF of 94 non-infectious neuroinflammatory cases and 24 water and reagent controls, we developed and implemented a weighted scoring metric based on z-scores at the species and genus level for both nucleotide and protein databases to prioritize and rank mNGS results. We performed mNGS on total RNA extracted from CSF of patients with subacute or chronic meningitis and highlight seven cases representing a diverse array of pathogens.nnSettingA multi-center study of mNGS pathogen discovery in patients with suspected neuroinflammatory conditions.nnParticipantsPatients with diagnostically challenging subacute or chronic meningitis enrolled in a research study of mNGS performed on CSF.nnInterventionmNGS was performed on total RNA extracted from CSF (0.25-0.5 mL). A weighted z-score was used to filter out environmental contaminants and facilitate efficient data triage and analysis.nnMain Outcomes1) Pathogens identified by mNGS and 2) ability of a statistical model to prioritize, rank, and simplify mNGS results.nnResultsmNGS identified parasitic worms, fungi and viruses in seven subjects: Taenia solium (n=2), Cryptococcus neoformans, human immunodeficiency virus-1, Aspergillus oryzae, Histoplasma capsulatum, and Candida dubliniensis. Evaluating mNGS data with a weighted z-score based scoring algorithm effectively separated bona fide pathogen sequences from spurious environmental sequences.nnConclusions and RelevancemNGS of CSF identified a diversity of microbial pathogens in patients with diagnostically challenging subacute or chronic meningitis, including a case of subarachnoid neurocysticercosis that defied diagnosis for one year, the first case of CNS vasculitis caused by Aspergillus oryzae, and the fourth reported case of Candida dubliniensis meningitis. Filtering metagenomic data with a scoring algorithm greatly clarified data interpretation and highlights the difficulties attributing biological significance to organisms that may be present in control samples used for metagenomic sequencing studies.nnKey PointsQuestion: How can metagenomic next-generation sequencing of cerebrospinal fluid be leveraged to aid in the diagnosis of patients with subacute or chronic meningitis?nnFindings: Metagenomic next-generation sequencing identified parasitic worms, fungi and viruses in a case series of seven subjects. A database of water-only and healthy patient controls enabled application of a z-score based scoring algorithm to effectively separate bona fide pathogen sequences from spurious environmental sequences.nnMeaning: Our scoring algorithm greatly simplified data interpretation in a series of patients with a wide range of challenging infectious causes of subacute or chronic meningitis identified by metagenomic next-generation sequencing.
]]></description>
<dc:creator>Wilson, M. R.</dc:creator>
<dc:creator>O'Donovan, B. D.</dc:creator>
<dc:creator>Gelfand, J. M.</dc:creator>
<dc:creator>Sample, H. A.</dc:creator>
<dc:creator>Chow, F. C.</dc:creator>
<dc:creator>Betjemann, J. P.</dc:creator>
<dc:creator>Shah, M. P.</dc:creator>
<dc:creator>Richie, M. B.</dc:creator>
<dc:creator>Gorman, M. P.</dc:creator>
<dc:creator>Hajj-Ali, R. A.</dc:creator>
<dc:creator>Calabrese, L. H.</dc:creator>
<dc:creator>Zorn, K. C.</dc:creator>
<dc:creator>Greenlee, J. E.</dc:creator>
<dc:creator>Blum, J. H.</dc:creator>
<dc:creator>Green, G.</dc:creator>
<dc:creator>Khan, L. M.</dc:creator>
<dc:creator>Banerji, D.</dc:creator>
<dc:creator>Langelier, C.</dc:creator>
<dc:creator>Bryson-Cahn, C.</dc:creator>
<dc:creator>Harrington, W.</dc:creator>
<dc:creator>Lingappa, J. R.</dc:creator>
<dc:creator>Shanbhag, N. M.</dc:creator>
<dc:creator>Green, A. J.</dc:creator>
<dc:creator>Brew, B. J.</dc:creator>
<dc:creator>Soldatos, A.</dc:creator>
<dc:creator>Strnad, L.</dc:creator>
<dc:creator>Doernberg, S. B.</dc:creator>
<dc:creator>Jay, C. A.</dc:creator>
<dc:creator>Douglas, V.</dc:creator>
<dc:creator>Josephson, S. A.</dc:creator>
<dc:creator>DeRisi, J. L.</dc:creator>
<dc:date>2017-11-07</dc:date>
<dc:identifier>doi:10.1101/213561</dc:identifier>
<dc:title><![CDATA[Metagenomics for chronic meningitis: clarifying interpretation and diagnosis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-11-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/216499v1?rss=1">
<title>
<![CDATA[
Aquaporin-4 dependent glymphatic solute transport in rodent brain 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/216499v1?rss=1</link>
<description><![CDATA[
The glymphatic system is a brain-wide metabolite clearance pathway, impairment of which in post-traumatic and ischemic brain or healthy aging is proposed to contribute to intracerebral accumulation of amyloid-{beta} and tau proteins. Glymphatic perivascular influx of cerebrospinal fluid (CSF) depends upon the expression and perivascular localization of the astroglial water channel aquaporin-4 (AQP4). Prompted by a recent publication that failed to find an effect of Aqp4 knockout on perivascular CSF tracer influx and interstitial fluid (ISF) tracer dispersion, four independent research groups have herein re-examined the importance of Aqp4 in glymphatic fluid transport. We concur in finding that CSF tracer influx, as well as fluorescently-tagged amyloid-{beta} efflux, are significantly faster in wild-type mice than in three different transgenic lines featuring disruption of the Aqp4 gene and one line in which AQP4 expression lacks the critical perivascular localization (Snta1 knockout). These data validate the role of AQP4 in supporting fluid and solute transport and efflux in brain in accordance with the glymphatic system model.
]]></description>
<dc:creator>Mestre, H.</dc:creator>
<dc:creator>Kress, B. T.</dc:creator>
<dc:creator>Zou, W.</dc:creator>
<dc:creator>Pu, T.</dc:creator>
<dc:creator>Murlidharan, G.</dc:creator>
<dc:creator>Castellanos Rivera, R. M.</dc:creator>
<dc:creator>Simon, M. J.</dc:creator>
<dc:creator>Pike, M. M.</dc:creator>
<dc:creator>Plog, B. A.</dc:creator>
<dc:creator>Xavier, A. L. R.</dc:creator>
<dc:creator>Thrane, A. S.</dc:creator>
<dc:creator>Lundgaard, I.</dc:creator>
<dc:creator>Thomas, J. H.</dc:creator>
<dc:creator>Xiao, M.</dc:creator>
<dc:creator>Asokan, A.</dc:creator>
<dc:creator>Iliff, J. J.</dc:creator>
<dc:creator>Nedergaard, M.</dc:creator>
<dc:date>2017-11-09</dc:date>
<dc:identifier>doi:10.1101/216499</dc:identifier>
<dc:title><![CDATA[Aquaporin-4 dependent glymphatic solute transport in rodent brain]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-11-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/226084v1?rss=1">
<title>
<![CDATA[
An adaptable chromosome preparation methodology for use in invertebrate research organisms 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/226084v1?rss=1</link>
<description><![CDATA[
BackgroundThe ability to efficiently visualize and manipulate chromosomes is fundamental to understand the genetic architecture of organisms. This can be challenging for many invertebrates because conventional chromosome preparation protocols deal primarily hypotonic conditions tailored for use with vertebrates and rely extensively on cultured cells which are rarely available for invertebrates. Hence, a simple and inexpensive chromosome preparation protocol, adaptable to multiple invertebrate model species is needed.nnResultsWe optimized a chromosome preparation protocol and applied it to several planarian species (Phylum: Platyhelminthes), the freshwater apple snail Pomacea canaliculata (Phylum: Mollusca), and the starlet sea anemone Nematostella vectensis (Phylum: Cnidaria). We showed that both mitotically active adult tissues and embryos can be used as sources of metaphase chromosomes, expanding the potential use of this technique to invertebrates lacking cell lines and/or with limited access to the complete life cycle. Simple hypotonic treatment with DI water was sufficient for karyotyping. The karyotypes we obtained allowed the identification of differences in ploidy and chromosome architecture among otherwise morphologically indistinguishable organisms, as in the case of a mixed population of planarians collected in the wild. Furthermore, we showed that in all tested organisms representing three different phyla, this protocol can be effectively coupled with downstream applications, such as chromosome fluorescent in situ hybridization.nnConclusionThe simple and inexpensive chromosome preparation protocol reported here can be readily adapted to new invertebrate research organisms in order to expand and accelerate the discovery of new biology in understudied branches of the tree of life.
]]></description>
<dc:creator>Guo, L.</dc:creator>
<dc:creator>Accorsi, A.</dc:creator>
<dc:creator>He, S.</dc:creator>
<dc:creator>Guerrero-Hernandez, C.</dc:creator>
<dc:creator>Sivagnanam, S.</dc:creator>
<dc:creator>McKinney, S.</dc:creator>
<dc:creator>Gibson, M.</dc:creator>
<dc:creator>Sanchez Alvarado, A.</dc:creator>
<dc:date>2017-11-29</dc:date>
<dc:identifier>doi:10.1101/226084</dc:identifier>
<dc:title><![CDATA[An adaptable chromosome preparation methodology for use in invertebrate research organisms]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-11-29</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/10.1101/232280v1?rss=1">
<title>
<![CDATA[
TRIF is a key inflammatory mediator of acute sickness behavior and cancer cachexia 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/232280v1?rss=1</link>
<description><![CDATA[
Hypothalamic inflammation is a key component of acute sickness behavior and cachexia, yet mechanisms of inflammatory signaling in the central nervous system remain unclear. We assessed the role of TRIF signaling in acute inflammation (lipopolysaccharide (LPS) challenge) and in a chronic inflammatory state (cancer cachexia). TRIFKO mice resisted anorexia and weight loss after peripheral (intraperitoneal, IP) or central (intracerebroventricular, ICV) LPS challenge and in a model of pancreatic cancer cachexia. Compared to WT mice, TRIFKO mice showed attenuated upregulation of Il6, Ccl2, Ccl5, Cxcl1, Cxcl2, and Cxcl10 in the hypothalamus after IP LPS treatment, as well as attenuated microglial activation and neutrophil infiltration into the brain after ICV LPS treatment. Our results show that TRIF is an important inflammatory signaling mediator of sickness behavior and cachexia and presents a novel therapeutic target for these conditions.
]]></description>
<dc:creator>Burfeind, K. G.</dc:creator>
<dc:creator>Zhu, X.</dc:creator>
<dc:creator>Levasseur, P. R.</dc:creator>
<dc:creator>Michaelis, K. A.</dc:creator>
<dc:creator>Norgard, M. A.</dc:creator>
<dc:creator>Marks, D. L.</dc:creator>
<dc:date>2017-12-11</dc:date>
<dc:identifier>doi:10.1101/232280</dc:identifier>
<dc:title><![CDATA[TRIF is a key inflammatory mediator of acute sickness behavior and cancer cachexia]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-12-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/238360v1?rss=1">
<title>
<![CDATA[
The genomic false shuffle: epigenetic maintenance of topological domains in the rearranged gibbon genome 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/238360v1?rss=1</link>
<description><![CDATA[
The relationship between evolutionary genome remodeling and the three-dimensional structure of the genome remain largely unexplored. Here we use the heavily rearranged gibbon genome to examine how evolutionary chromosomal rearrangements impact genome-wide chromatin interactions, topologically associating domains (TADs), and their epigenetic landscape. We use high-resolution maps of gibbon-human breaks of synteny (BOS), apply Hi-C in gibbon, measure an array of epigenetic features, and perform cross-species comparisons. We find that gibbon rearrangements occur at TAD boundaries, independent of the parameters used to identify TADs. This overlap is supported by a remarkable genetic and epigenetic similarity between BOS and TAD boundaries, namely presence of CpG islands and SINE elements, and enrichment in CTCF and H3K4me3 binding. Cross-species comparisons reveal that regions orthologous to BOS also correspond with boundaries of large (400-600kb) TADs in human and other mammalian species. The co-localization of rearrangement breakpoints and TAD boundaries may be due to higher chromatin fragility at these locations and/or increased selective pressure against rearrangements that disrupt TAD integrity. We also examine the small portion of BOS that did not overlap with TAD boundaries and gave rise to novel TADs in the gibbon genome. We postulate that these new TADs generally lack deleterious consequences. Lastly, we show that limited epigenetic homogenization occurs across breakpoints, irrespective of their time of occurrence in the gibbon lineage. Overall, our findings demonstrate remarkable conservation of chromatin interactions and epigenetic landscape in gibbons, in spite of extensive genomic shuffling.
]]></description>
<dc:creator>Lazar, N. H.</dc:creator>
<dc:creator>Nevonen, K. A.</dc:creator>
<dc:creator>O'Connell, B.</dc:creator>
<dc:creator>Green, R. E.</dc:creator>
<dc:creator>Meyer, T. J.</dc:creator>
<dc:creator>Okhovat, M.</dc:creator>
<dc:creator>Carbone, L.</dc:creator>
<dc:date>2017-12-22</dc:date>
<dc:identifier>doi:10.1101/238360</dc:identifier>
<dc:title><![CDATA[The genomic false shuffle: epigenetic maintenance of topological domains in the rearranged gibbon genome]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-12-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/240556v1?rss=1">
<title>
<![CDATA[
Task Engagement Enhances Population Encoding of Stimulus Meaning in Primary Auditory Cortex 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/240556v1?rss=1</link>
<description><![CDATA[
The main functions of primary sensory cortical areas are classically considered to be the extraction and representation of stimulus features. In contrast, higher cortical sensory association areas are thought to be responsible for combining these sensory representations with internal motivations and learnt associations. These regions generate appropriate neural responses that are maintained until a motor command is executed. Within this framework, responses of the primary sensory areas during task performance are expected to carry less information about the behavioral meaning of the stimulus than higher sensory, association, motor and frontal cortices. Here we demonstrate instead that the neuronal population responses in the early primary auditory cortex (A1) display many aspects of responses generally associated with higher-level areas. A1 activity was recorded in awake ferrets while they were either passively listening or actively discriminating two periodic click trains of different rates in a Go/No-Go paradigm. By applying population-level dimensionality reduction techniques, we found that task-engagement induced a shift in the nature of the encoding from a sensory-driven representation of the two stimuli to a behaviorally relevant representation of the two categories that specifically enhances the target stimulus. We demonstrate that this shift in encoding relies partly on a novel mechanism of change in spontaneous activity patterns upon engagement in the task. We show that this population-level representation of stimuli in A1 population activity bears strong similarities to responses in the frontal cortex, but appears earlier following stimulus presentation. Analysis of neural activity recorded in various Go/No-Go tasks, with different sounds and reinforcement paradigms, reveals that this striking population-level enhancement of target representation is a general property of task engagement. These findings indicate that primary sensory cortices play a highly flexible role in the processing of incoming stimuli and implement a crucial change in the structure of population activity in order to extract task-relevant information during behavior.
]]></description>
<dc:creator>Bagur, S.</dc:creator>
<dc:creator>Averseng, M.</dc:creator>
<dc:creator>Elgueda, D.</dc:creator>
<dc:creator>David, S.</dc:creator>
<dc:creator>Fritz, J.</dc:creator>
<dc:creator>Yin, P.</dc:creator>
<dc:creator>Shamma, S.</dc:creator>
<dc:creator>Boubenec, Y.</dc:creator>
<dc:creator>Ostojic, S.</dc:creator>
<dc:date>2017-12-28</dc:date>
<dc:identifier>doi:10.1101/240556</dc:identifier>
<dc:title><![CDATA[Task Engagement Enhances Population Encoding of Stimulus Meaning in Primary Auditory Cortex]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-12-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/241851v1?rss=1">
<title>
<![CDATA[
Chromosome Removal Via Cellular Fragmentation and Aneuploid Blastomere Exclusion in Primate Embryos 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/241851v1?rss=1</link>
<description><![CDATA[
Aneuploidy that arises during meiosis and/or mitosis is a major contributor to early embryo loss. We previously demonstrated that human preimplantation embryos encapsulate mis-segregated chromosomes into micronuclei while undergoing cellular fragmentation and that fragments can contain chromosomal material, but the source of this DNA was unknown. Here, we leveraged the use of a non-human primate model and single-cell DNA-sequencing (scDNA-seq) to examine the chromosomal content of 471 individual samples comprising 254 blastomeres, 42 polar bodies, and 175 cellular fragments from a large number (N=50) of disassembled rhesus cleavage-stage embryos. Our analysis revealed that the frequency of aneuploidy and micronucleation is conserved between humans and macaques and that cellular fragments encapsulate whole and/or partial chromosomes lost from blastomeres. Single-cell/fragment genotyping demonstrated that these chromosome-containing cellular fragments (CCFs) can be either maternal or paternal in origin and display DNA damage via double-stranded breaks. Chromosome breakage and abnormal cytokinesis resulted in reciprocal losses/gains at the terminal ends of chromosome arms, uniparental genome segregation, and mixoploidy between blastomeres. Combining time-lapse imaging with scDNA-seq, we also determined that multipolar divisions at the zygote or 2-cell stage generated chaotic aneuploidy encompassing a complex mixture of maternal and paternal chromosomes. Despite frequent chromosomal mis-segregation at the cleavage-stage, we show that CCFs and non-dividing aneuploid blastomeres exhibiting extensive DNA damage are prevented from incorporation at the blastocyst stage. These findings suggest that embryos respond to chromosomal errors by encapsulation into micronuclei, elimination by cellular fragmentation, and selection against highly aneuploid blastomeres to overcome chromosome instability during preimplantation development.
]]></description>
<dc:creator>Daughtry, B. L.</dc:creator>
<dc:creator>Rosenkrantz, J. L.</dc:creator>
<dc:creator>Lazar, N. H.</dc:creator>
<dc:creator>Fei, S. S.</dc:creator>
<dc:creator>Redmayne, N.</dc:creator>
<dc:creator>Torkenczy, K. A.</dc:creator>
<dc:creator>Adey, A.</dc:creator>
<dc:creator>Gao, L.</dc:creator>
<dc:creator>Park, B.</dc:creator>
<dc:creator>Nevonen, K. A.</dc:creator>
<dc:creator>Carbone, L.</dc:creator>
<dc:creator>Chavez, S. L.</dc:creator>
<dc:date>2018-01-02</dc:date>
<dc:identifier>doi:10.1101/241851</dc:identifier>
<dc:title><![CDATA[Chromosome Removal Via Cellular Fragmentation and Aneuploid Blastomere Exclusion in Primate Embryos]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-01-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/242685v1?rss=1">
<title>
<![CDATA[
Acute Hyperalgesia and Delayed Dry Eye After Corneal Abrasion Injury 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/242685v1?rss=1</link>
<description><![CDATA[
Corneal nerves mediate pain from the ocular surface, lacrimation, and blinking, all of which protect corneal surface homeostasis and help preserve vision. Corneal nerve density correlates with neuropathic pain states and is used as an assessment of small fiber neuropathies. Because pain, lacrimation and blinking are rarely assessed at the same time, it is not known if their regulatory mechanisms have similar temporal dynamics after acute corneal injury. We examined changes in corneal nerve density, evoked and spontaneous pain, and ocular homeostasis in Sprague-Dawley male rats after a superficial epithelial injury with heptanol that acutely abolished nerve endings within the central cornea. Despite a profound loss of epithelial nerve endings, pain was transiently enhanced after abrasion injury, while basal tear production was normal. We found no relationship between epithelial nerve density and pain or homeostatic responses. Axotomy following corneal abrasion increased expression of both ATF3 (a nerve injury marker) and CGRP (a nociceptive peptide) in trigeminal ganglia 24 hours after injury. These molecular changes were absent on the contralateral side, despite reductions in corneal epithelial nerve density in the uninjured eye. ATF3 and CGRP levels in trigeminal ganglion were normal at one week post-injury when pain responses were normal. In contrast, CGRP was upregulated in peripheral corneal endings one week after injury, when dry eye symptoms emerged. Our results demonstrate dynamic trafficking of CGRP within trigeminal sensory nerves, with elevations in the ganglion correlated with pain behaviors and elevations in peripheral endings correlated with dry eye symptoms.
]]></description>
<dc:creator>Hegarty, D. M.</dc:creator>
<dc:creator>Hermes, S. M.</dc:creator>
<dc:creator>Morgan, M. M.</dc:creator>
<dc:creator>Aicher, S. A.</dc:creator>
<dc:date>2018-01-04</dc:date>
<dc:identifier>doi:10.1101/242685</dc:identifier>
<dc:title><![CDATA[Acute Hyperalgesia and Delayed Dry Eye After Corneal Abrasion Injury]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-01-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/242933v1?rss=1">
<title>
<![CDATA[
Differential regulation of microRNA-15a by radiation affects angiogenesis and tumor growth via modulation of acid sphingomyelinase 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/242933v1?rss=1</link>
<description><![CDATA[
Activation of acid sphingomyelinase (SMPD1) and the generation of ceramide is a critical regulator of apoptosis in response to cellular stress including radiation. Endothelial SMPD1 has been shown to regulate tumor responses to radiation therapy. We show here that the SMPD1 gene is regulated by a microRNA (miR), miR-15a, in endothelial cells (ECs). Standard low dose radiation (2 Gy) upregulates miR-15a and decreases SMPD1 levels. In contrast, high dose radiation (10 Gy and above) decreases miR-15a and increases SMPD1. Ectopic expression of miR-15a decreases both mRNA and protein levels of SMPD1. Mimicking the effects of high dose radiation with a miR-15a inhibitor decreases cell proliferation and increases active Caspase-3 & 7. Mechanistically, inhibition of miR-15a increases inflammatory cytokines, such as IP10, activates caspase-1 inflammasome and increases Gasdermin D, an effector of pyroptosis. Importantly, both systemic and vascular-targeted delivery of miR-15a inhibitor decreases angiogenesis and tumor growth in a CT26 murine colorectal carcinoma model. Taken together, our findings highlight a novel role for miR mediated regulation of SMPD1 during radiation responses and establish proof-of-concept that this pathway can be targeted with a miR inhibitor.
]]></description>
<dc:creator>Rana, S.</dc:creator>
<dc:creator>Espinosa-Diez, C.</dc:creator>
<dc:creator>Ruhl, R.</dc:creator>
<dc:creator>Thomas, C. R.</dc:creator>
<dc:creator>Anand, S.</dc:creator>
<dc:date>2018-01-04</dc:date>
<dc:identifier>doi:10.1101/242933</dc:identifier>
<dc:title><![CDATA[Differential regulation of microRNA-15a by radiation affects angiogenesis and tumor growth via modulation of acid sphingomyelinase]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-01-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/252387v1?rss=1">
<title>
<![CDATA[
Cellular tolerance at the μ-opioid receptor is phosphorylation dependent 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/252387v1?rss=1</link>
<description><![CDATA[
The role of phosphorylation of the -opioid receptor (MOR) in desensitization, internalization and long-term tolerance was examined in locus coeruleus (LC) neurons. Viral expression of wild type (exWT) and mutant MORs, where all phosphorylation sites on the C-terminus (Total Phosphorylation Deficient (TPD)) were mutated to alanine, were examined in a MOR knockout rat. Both expressed receptors acutely activate potassium conductance similar to endogenous receptors in wild type animals. The exWT receptors, like endogenous receptors, displayed signs of tolerance after chronic morphine treatment. There was however a loss of agonist-induced desensitization and internalization in experiments with the TPD receptors. In addition the development of tolerance was not observed in the TPD receptors following chronic morphine treatment. The results indicate a key role of C-terminal phosphorylation in the expression of acute desensitization, trafficking and long-term tolerance to morphine.
]]></description>
<dc:creator>Williams, J. T.</dc:creator>
<dc:creator>Arttamangkul, S.</dc:creator>
<dc:creator>Heinze, D. A.</dc:creator>
<dc:creator>Bunzow, J. R.</dc:creator>
<dc:creator>Song, X.</dc:creator>
<dc:date>2018-01-23</dc:date>
<dc:identifier>doi:10.1101/252387</dc:identifier>
<dc:title><![CDATA[Cellular tolerance at the μ-opioid receptor is phosphorylation dependent]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-01-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/255265v1?rss=1">
<title>
<![CDATA[
A saturation mutagenesis approach to understanding PTEN lipid phosphatase activity and genotype-phenotypes relationships 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/255265v1?rss=1</link>
<description><![CDATA[
Phosphatase and tensin homolog (PTEN) is a tumor suppressor frequently mutated in diverse cancers. Germline PTEN mutations are also associated with a range of clinical outcomes, including PTEN hamartoma tumor syndrome (PHTS) and autism spectrum disorder (ASD). To empower new insights into PTEN function and clinically relevant genotype-phenotype relationships, we systematically evaluated the effect of PTEN mutations on lipid phosphatase activity in vivo. Using a massively parallel approach that leverages an artificial humanized yeast model, we derived high-confidence estimates of functional impact for 7,244 single amino acid PTEN variants (86% of possible). These data uncovered novel insights into PTEN protein structure, biochemistry, and mutation tolerance. Variant functional scores can reliably discriminate likely pathogenic from benign alleles. Further, 32% of ClinVar unclassified missense variants are phosphatase deficient in our assay, supporting their reclassification. ASD associated mutations generally had less severe fitness scores relative to PHTS associated mutations (p = 7.16x10-5) and a higher fraction of hypomorphic mutations, arguing for continued genotype-phenotype studies in larger clinical datasets that can further leverage these rich functional data.
]]></description>
<dc:creator>Mighell, T. L.</dc:creator>
<dc:creator>Evans-Dutson, S.</dc:creator>
<dc:creator>O'Roak, B.</dc:creator>
<dc:date>2018-01-29</dc:date>
<dc:identifier>doi:10.1101/255265</dc:identifier>
<dc:title><![CDATA[A saturation mutagenesis approach to understanding PTEN lipid phosphatase activity and genotype-phenotypes relationships]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-01-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/256040v1?rss=1">
<title>
<![CDATA[
RNAi screening of the tyrosine kinome in primary patient samples of acute myeloid leukemia 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/256040v1?rss=1</link>
<description><![CDATA[
Targeted therapy has proven to be successful in improving outcomes across multiple cancer types. However, many challenges still remain for implementation of this strategy in most patient cohorts, especially with the challenges of identifying the specific mutations or abnormalities in a heterogeneous tumor that are functionally significant. Previously, we developed a functional screening assay, RNAi-assisted protein target identification (RAPID) technology, which evaluates the viability of tumor cells after exposure to siRNAs against members of the tyrosine kinome and NRAS/KRAS. Here, we publish the comprehensive results of this screen for 332 primary AML patient samples. Data from these screening efforts have already helped identify previously unknown therapeutic targets, and will continue to provide insights into better treatment strategies for these patients.
]]></description>
<dc:creator>Edwards, D.</dc:creator>
<dc:creator>Eryildiz, F.</dc:creator>
<dc:creator>Tyner, J. W.</dc:creator>
<dc:date>2018-01-30</dc:date>
<dc:identifier>doi:10.1101/256040</dc:identifier>
<dc:title><![CDATA[RNAi screening of the tyrosine kinome in primary patient samples of acute myeloid leukemia]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-01-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/258855v1?rss=1">
<title>
<![CDATA[
Causal interactions from proteomic profiles: molecular data meets pathway knowledge 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/258855v1?rss=1</link>
<description><![CDATA[
Measurement of changes in protein levels and in post-translational modifications, such as phosphorylation, can be highly informative about the phenotypic consequences of genetic differences or about the dynamics of cellular processes. Typically, such proteomic profiles are interpreted intuitively or by simple correlation analysis. Here, we present a computational method to generate causal explanations for proteomic profiles using prior mechanistic knowledge in the literature, as recorded in cellular pathway maps. To demonstrate its potential, we use this method to analyze the cascading events after EGF stimulation of a cell line, to discover new pathways in platelet activation, to identify influential regulators of oncoproteins in breast cancer, to describe signaling characteristics in predefined subtypes of ovarian and breast cancers, and to highlight which pathway relations are most frequently activated across 32 cancer types. Causal pathway analysis, that combines molecular profiles with prior biological knowledge captured in computational form, may become a powerful discovery tool as the amount and quality of cellular profiling rapidly expands. The method is freely available at http://causalpath.org.
]]></description>
<dc:creator>Babur, O.</dc:creator>
<dc:creator>Luna, A.</dc:creator>
<dc:creator>Korkut, A.</dc:creator>
<dc:creator>Durupinar, F.</dc:creator>
<dc:creator>Siper, M. C.</dc:creator>
<dc:creator>Dogrusoz, U.</dc:creator>
<dc:creator>Aslan, J. E.</dc:creator>
<dc:creator>Sander, C.</dc:creator>
<dc:creator>Demir, E.</dc:creator>
<dc:date>2018-02-02</dc:date>
<dc:identifier>doi:10.1101/258855</dc:identifier>
<dc:title><![CDATA[Causal interactions from proteomic profiles: molecular data meets pathway knowledge]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-02-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/274316v1?rss=1">
<title>
<![CDATA[
Cryo-EM visualization of an active high open probability CFTR ion channel 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/274316v1?rss=1</link>
<description><![CDATA[
The Cystic fibrosis transmembrane conductance regulator (CFTR) anion channel, crucial to epithelial salt and water homeostasis, and defective due to mutations in its gene in patients with cystic fibrosis is a unique member of the large family of ATP-binding cassette transport proteins. Regulation of CFTR channel activity is stringently controlled by phosphorylation and nucleotide binding. Structural changes that underlie transitions between active and inactive functional states are not yet fully understood. Indeed the first 3D structures of dephosphorylated, ATP-free and phosphorylated ATP-bound states were only recently reported. Here we have determined the structure of inactive and active states of a thermally stabilized CFTR with very high channel open probability, confirmed after reconstitution into proteoliposomes. The unique repositioning of the TMHs and R domain density that we observe provide insights into the structural transition between active and inactive functional states of CFTR.nnHighlightsO_LIStructures of thermostabilized avian CFTR in dephosphorylated or phosphorylated forms at 4.3 [A] and 6.6 [A] resolution, respectively.nC_LIO_LIConformational differences of transmembrane helices 7 & 8 compared to zebra fish and human CFTR structures reveal an extracellular vestibule that may provide anion access to the pore.nC_LIO_LIR-domain density appears to "plug" the intercellular vestibule in the dephosphorylated avian CFTR cryo-EM map.nC_LI
]]></description>
<dc:creator>Fay, J. F.</dc:creator>
<dc:creator>Aleksandrov, L. A.</dc:creator>
<dc:creator>Jensen, T. J.</dc:creator>
<dc:creator>Cui, L. L.</dc:creator>
<dc:creator>Kousouros, J. N.</dc:creator>
<dc:creator>He, L.</dc:creator>
<dc:creator>Aleksandrov, A. A.</dc:creator>
<dc:creator>Gingerich, D. S.</dc:creator>
<dc:creator>Riordan, J.</dc:creator>
<dc:creator>Chen, J. Z.</dc:creator>
<dc:date>2018-03-02</dc:date>
<dc:identifier>doi:10.1101/274316</dc:identifier>
<dc:title><![CDATA[Cryo-EM visualization of an active high open probability CFTR ion channel]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-03-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/282830v1?rss=1">
<title>
<![CDATA[
A Measure of Open Data: A Metric and Analysis of Reusable Data Practices in Biomedical Data Resources 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/282830v1?rss=1</link>
<description><![CDATA[
Data is the foundation of science, and there is an increasing focus on how data can be reused and enhanced to drive scientific discoveries. However, most seemingly "open data" do not provide legal permissions for reuse and redistribution. Not being able to integrate and redistribute our collective data resources blocks innovation, and stymies the creation of life-improving diagnostic and drug selection tools. To help the biomedical research and research support communities (e.g. libraries, funders, repositories, etc.) understand and navigate the data licensing landscape, the (Re)usable Data Project (RDP) (http://reusabledata.org) assesses the licensing characteristics of data resources and how licensing behaviors impact reuse. We have created a ruleset to determine the reusability of data resources and have applied it to 56 scientific data resources (i.e. databases) to date. The results show significant reuse and interoperability barriers. Inspired by game-changing projects like Creative Commons, the Wikipedia Foundation, and the Free Software movement, we hope to engage the scientific community in the discussion regarding the legal use and reuse of scientific data, including the balance of openness and how to create sustainable data resources in an increasingly competitive environment.
]]></description>
<dc:creator>Carbon, S.</dc:creator>
<dc:creator>Champieux, R.</dc:creator>
<dc:creator>McMurry, J.</dc:creator>
<dc:creator>Winfree, L.</dc:creator>
<dc:creator>Wyatt, L. R.</dc:creator>
<dc:creator>Haendel, M.</dc:creator>
<dc:date>2018-03-16</dc:date>
<dc:identifier>doi:10.1101/282830</dc:identifier>
<dc:title><![CDATA[A Measure of Open Data: A Metric and Analysis of Reusable Data Practices in Biomedical Data Resources]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-03-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/286005v1?rss=1">
<title>
<![CDATA[
Retinal Ganglion Cell Axon Sorting at the Optic Chiasm Requires Dystroglycan 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/286005v1?rss=1</link>
<description><![CDATA[
In the developing visual system, retinal ganglion cell (RGC) axons project from the retina to several distal retinorecipient regions in the brain. Several molecules have been implicated in guiding RGC axons in vivo, but the role of extracellular matrix molecules in this process remains poorly understood. Dystroglycan is a laminin-binding transmembrane protein important for formation and maintenance of the extracellular matrix and basement membranes and has previously been implicated in axon guidance in the developing spinal cord. Using two genetic models of functional dystroglycan loss, we show that dystroglycan is necessary for correct sorting of contralateral and ipsilateral RGC axons at the optic chiasm. Missorted axons still target retinorecipient brain regions and persist in adult mice, even after axon pruning is complete. Our results highlight the importance of the extracellular matrix for axon sorting at an intermediate choice point in the developing visual circuit.nnSummary StatementAbnormal retinal ganglion cell axon sorting in the optic chiasm in the absence of functional dystroglycan results in profound defects in retinorecipient innervation.
]]></description>
<dc:creator>Clements, R.</dc:creator>
<dc:creator>Wright, K. M.</dc:creator>
<dc:date>2018-03-21</dc:date>
<dc:identifier>doi:10.1101/286005</dc:identifier>
<dc:title><![CDATA[Retinal Ganglion Cell Axon Sorting at the Optic Chiasm Requires Dystroglycan]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-03-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/287763v1?rss=1">
<title>
<![CDATA[
Thrombospondins 1 and 2 affect lysyl oxidase protein and collagen matrix maturation in cortical bone of growing male and female mice via non-redundant pathways 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/287763v1?rss=1</link>
<description><![CDATA[
Thrombospondin-2-deficiency is associated with impaired matrix maturation in osteoblasts and cortical bone of growing mice. Here we addressed the possibility that lysyl oxidase (LOX) contributes to this phenotype. After overnight serum starvation, pro-LOX levels were elevated compared to wild-type in marrow-derived osteoblasts from male and female TSP2-/- mice. The liberated LOX pro-peptide (LOPP) was faintly visible in serum-starved cultures. When serum was maintained, pro-LOX content was not affected by TSP2 status, but relative LOPP levels were elevated in cultures from female TSP2-/- mice. Two isoforms of pro-LOX at 75 kDa and 50 kDa were detected in detergent soluble protein extracts of diaphyseal tissue from growing mice. In female mice, TSP2 status did not affect detergent soluble pro-LOX content or the relative contribution of each band to the total signal. Instead, levels of the 50 kDa band were reduced in female TSP1-/- samples. In male diaphyseal tissue, total pro-LOX content and the contribution each isoform made to the total signal was not affected by TSP1 or TSP2 status. We did not detect 32 kDa mature LOX in detergent soluble preparations of cells or whole bone tissue. Detergent insoluble hydroxyproline content was reduced in diaphyseal tissue obtained from female TSP1-/- and TSP2-/- mice. In male diaphyseal cortical samples, TSP2 but not TSP1 deficiency was associated with reduced insoluble hydroxyproline content. Our data suggest that the trimeric thrombospondins contribute to bone matrix quality via non-redundant mechanisms that are dependent on the unique tissue milieu of the male and female skeleton.
]]></description>
<dc:creator>Shearer, D.</dc:creator>
<dc:creator>Mervis, M. O.</dc:creator>
<dc:creator>Manley, E.</dc:creator>
<dc:creator>Reddy, A. B.</dc:creator>
<dc:creator>Alford, A. I.</dc:creator>
<dc:date>2018-03-23</dc:date>
<dc:identifier>doi:10.1101/287763</dc:identifier>
<dc:title><![CDATA[Thrombospondins 1 and 2 affect lysyl oxidase protein and collagen matrix maturation in cortical bone of growing male and female mice via non-redundant pathways]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-03-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/301416v1?rss=1">
<title>
<![CDATA[
Autoregulation of the S. mutans SloR metalloregulator is constitutive and driven by an independent promoter 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/301416v1?rss=1</link>
<description><![CDATA[
Streptococcus mutans, one of [~]600 bacterial species in the human oral cavity, is among the most acidogenic constituents of the plaque biofilm. Considered to be the primary causative agent of dental caries, S. mutans harbors a 25kDa SloR metalloregulatory protein which controls metal ion transport across the bacterial cell membrane to maintain essential metal ion homeostasis. The expression of SloR derives, in part, from transcriptional readthrough of the sloABC operon which encodes a Mn2+/Fe2+ ABC transport system. Herein, we describe the details of the sloABC promoter that drives this transcription, as well as a novel independent promoter in an intergenic region (IGR) that contributes to downstream sloR expression. RT-PCR studies support sloR transcription that is independent of sloABC expression, and the results of 5' RACE revealed a sloR transcription start site in the IGR from which the -10 and -35 promoter regions were predicted. The results of gel mobility shift assays support direct SloR binding to the IGR, albeit with lower affinity than SloR binding to the sloABCR promoter. Function of the sloR promoter was validated in qRT-PCR experiments. Interestingly, sloR expression was not significantly impacted when grown in the presence of high manganese, whereas expression of the sloABC operon was repressed under these conditions. The results of in vitro transcription studies support SloR-mediated transcriptional-activation of sloR and -repression of sloABC. Taken together, these findings implicate SloR as a bifunctional regulator that represses sloABC promoter activity and encourages sloR transcription from an independent promoter.nnImportanceTooth decay is a ubiquitous infectious disease that is especially pervasive in underserved communities worldwide. S. mutans-induced carious lesions cause functional, physical, and/or aesthetic impairment in the vast majority of adults, and in 60-90% of schoolchildren in industrialized countries. Billions of dollars are spent annually on caries treatment, and productivity losses due to absenteeism from the workplace are significant. Research aimed at alleviating S. mutans-induced tooth decay is important because it can address the socioeconomic disparity that is associated with dental cavities and improve overall general health which is inextricably linked to oral health. Research focused on the S. mutans SloR metalloregulatory protein can guide the development of novel therapeutics and so alleviate the burden of dental cavities.
]]></description>
<dc:creator>Monette, P.</dc:creator>
<dc:creator>Brach, R.</dc:creator>
<dc:creator>Cowan, A.</dc:creator>
<dc:creator>Winters, R.</dc:creator>
<dc:creator>Weisman, J.</dc:creator>
<dc:creator>Seybert, F.</dc:creator>
<dc:creator>Goguen, K.</dc:creator>
<dc:creator>Chen, J.</dc:creator>
<dc:creator>Glasfeld, A.</dc:creator>
<dc:creator>Spatafora, G. A.</dc:creator>
<dc:date>2018-04-14</dc:date>
<dc:identifier>doi:10.1101/301416</dc:identifier>
<dc:title><![CDATA[Autoregulation of the S. mutans SloR metalloregulator is constitutive and driven by an independent promoter]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-04-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/303479v1?rss=1">
<title>
<![CDATA[
Efficient sorting of single-unit activity from midbrain cells using KiloSort is as accurate as manual sorting. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/303479v1?rss=1</link>
<description><![CDATA[
Extracting single-unit activity from in vivo extracellular neural electrophysiology data requires sorting spikes from background noise and action potentials from multiple cells in order to identify the activity of individual neurons. Typically this has been achieved by algorithms that employ principal component analyses followed by manual allocation of spikes to individual clusters based on visual inspection of the waveform shape. This method of manual sorting can give varying results between human operators and is highly time-consuming, especially in recordings with higher levels of background noise. To address these problems, automatic sorting algorithms have begun to gain popularity as viable methods for sorting electrophysiological data, although little is known about the use of these algorithms with neural data from midbrain recordings. KiloSort is a relatively new algorithm that automatically clusters raw data which can then be manually curated. In this report, we compare results of manually-sorted and KiloSort-processed recordings from the ventral tegmental area (VTA) and substantia nigra pars compacta (SNc). Sorting with KiloSort required substantially less time to complete, while yielding comparable and consistent results. We conclude that the use of KiloSort to identify single units from multi-channel recording in the VTA and SNc is accurate and efficient.
]]></description>
<dc:creator>Allen, M.</dc:creator>
<dc:creator>Chowdhury, T.</dc:creator>
<dc:creator>Wegener, M.</dc:creator>
<dc:creator>Moghaddam, B.</dc:creator>
<dc:date>2018-04-18</dc:date>
<dc:identifier>doi:10.1101/303479</dc:identifier>
<dc:title><![CDATA[Efficient sorting of single-unit activity from midbrain cells using KiloSort is as accurate as manual sorting.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-04-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/310425v1?rss=1">
<title>
<![CDATA[
Creating Standards for Evaluating Tumour Subclonal Reconstruction 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/310425v1?rss=1</link>
<description><![CDATA[
Tumours evolve through time and space. Computational techniques have been developed to infer their evolutionary dynamics from DNA sequencing data. A growing number of studies have used these approaches to link molecular cancer evolution to clinical progression and response to therapy. There has not yet been a systematic evaluation of methods for reconstructing tumour subclonality, in part due to the underlying mathematical and biological complexity and to difficulties in creating gold-standards. To fill this gap, we systematically elucidated the key algorithmic problems in subclonal reconstruction and developed mathematically valid quantitative metrics for evaluating them. We then created approaches to simulate realistic tumour genomes, harbouring all known mutation types and processes both clonally and subclonally. We then simulated 580 tumour genomes for reconstruction, varying tumour read-depth and benchmarking somatic variant detection and subclonal reconstruction strategies. The inference of tumour phylogenies is rapidly becoming standard practice in cancer genome analysis; this study creates a baseline for its evaluation.
]]></description>
<dc:creator>Boutros, P. C.</dc:creator>
<dc:creator>Salcedo, A.</dc:creator>
<dc:creator>Tarabichi, M.</dc:creator>
<dc:creator>Espiritu, S. M. G.</dc:creator>
<dc:creator>Deshwar, A. G.</dc:creator>
<dc:creator>David, M.</dc:creator>
<dc:creator>Wilson, N. M.</dc:creator>
<dc:creator>Dentro, S.</dc:creator>
<dc:creator>Wintersinger, J. A.</dc:creator>
<dc:creator>Liu, L. Y.</dc:creator>
<dc:creator>Ko, M.</dc:creator>
<dc:creator>Sivanandan, S.</dc:creator>
<dc:creator>Zhang, H.</dc:creator>
<dc:creator>Zhu, K.</dc:creator>
<dc:creator>Yang, T.-H.</dc:creator>
<dc:creator>Chilton, J. M.</dc:creator>
<dc:creator>Buchanan, A.</dc:creator>
<dc:creator>Lalansingh, C. M.</dc:creator>
<dc:creator>P'ng, C.</dc:creator>
<dc:creator>Anghel, C. V.</dc:creator>
<dc:creator>Umar, I.</dc:creator>
<dc:creator>Lo, B.</dc:creator>
<dc:creator>Simpson, J. T.</dc:creator>
<dc:creator>Stuart, J. M.</dc:creator>
<dc:creator>Anastassiou, D.</dc:creator>
<dc:creator>Guan, Y.</dc:creator>
<dc:creator>Ewing, A.</dc:creator>
<dc:creator>Ellrott, K.</dc:creator>
<dc:creator>Wedge, D. C.</dc:creator>
<dc:creator>Morris, Q. D.</dc:creator>
<dc:creator>Van Loo, P.</dc:creator>
<dc:creator>DREAM SMC-Het Participants,</dc:creator>
<dc:date>2018-04-28</dc:date>
<dc:identifier>doi:10.1101/310425</dc:identifier>
<dc:title><![CDATA[Creating Standards for Evaluating Tumour Subclonal Reconstruction]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-04-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/310730v1?rss=1">
<title>
<![CDATA[
An RNAi screen in human cell lines reveals conserved DNA damage repair pathways that mitigate formaldehyde sensitivity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/310730v1?rss=1</link>
<description><![CDATA[
Formaldehyde is a ubiquitous DNA damaging agent, with human exposures occuring from both exogenous and endogenous sources. Formaldehyde can also form DNA-protein crosslinks and is representative of other such DNA damaging agents including ionizing radiation, metals, aldehydes, chemotherapeutics, and cigarette smoke. In order to identify genetic determinants of cell proliferation in response to continuous formaldehyde exposure, we quantified cell proliferation after siRNA-depletion of a comprehensive array of over 300 genes representing all of the major DNA damage response pathways. Three unrelated human cell lines (SW480, U-2 OS and GM00639) were used to identify common or cell line-specific mechanisms. Four cellular pathways were determined to mitigate formaldehyde toxicity in all three cell lines: homologous recombination, double-strand break repair, ionizing radiation response, and DNA replication. Differences between cell lines were further investigated by using exome sequencing and Cancer Cell Line Encyclopedia genomic data. Our results reveal major genetic determinants of formaldehyde toxicity in human cells and provide evidence for the conservation of these formaldehyde responses between human and budding yeast.
]]></description>
<dc:creator>Juarez, E.</dc:creator>
<dc:creator>Chambwe, N.</dc:creator>
<dc:creator>Tang, W.</dc:creator>
<dc:creator>Mitchell, A. D.</dc:creator>
<dc:creator>Owen, N.</dc:creator>
<dc:creator>Kumari, A.</dc:creator>
<dc:creator>Monnat, R. J.</dc:creator>
<dc:creator>McCullough, A.</dc:creator>
<dc:date>2018-04-30</dc:date>
<dc:identifier>doi:10.1101/310730</dc:identifier>
<dc:title><![CDATA[An RNAi screen in human cell lines reveals conserved DNA damage repair pathways that mitigate formaldehyde sensitivity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-04-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/311696v1?rss=1">
<title>
<![CDATA[
A crowdsourced analysis to identify ab initio molecular signatures predictive of susceptibility to viral infection 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/311696v1?rss=1</link>
<description><![CDATA[
Respiratory viruses are highly infectious; however, the variation of individuals physiologic responses to viral exposure is poorly understood. Most studies examining molecular predictors of response focus on late stage predictors, typically near the time of peak symptoms. To determine whether pre- or early post-exposure factors could predict response, we conducted a community-based analysis to identify predictors of resilience or susceptibility to several respiratory viruses (H1N1, H3N2, Rhinovirus, and RSV) using peripheral blood gene expression profiles collected from healthy subjects prior to viral exposure, as well as up to 24 hours following exposure. This analysis revealed that it is possible to construct models predictive of symptoms using profiles even prior to viral exposure. Analysis of predictive gene features revealed little overlap among models; however, in aggregate, these genes were enriched for common pathways. Heme Metabolism, the most significantly enriched pathway, was associated with higher risk of developing symptoms following viral exposure.
]]></description>
<dc:creator>Fourati, S.</dc:creator>
<dc:creator>Talla, A.</dc:creator>
<dc:creator>Mahmoudian, M.</dc:creator>
<dc:creator>Burkhart, J. G.</dc:creator>
<dc:creator>Klen, R.</dc:creator>
<dc:creator>Henao, R.</dc:creator>
<dc:creator>Aydin, Z.</dc:creator>
<dc:creator>Yeung, K. Y.</dc:creator>
<dc:creator>Ahsen, M. E.</dc:creator>
<dc:creator>Almugbel, R.</dc:creator>
<dc:creator>Jahandideh, S.</dc:creator>
<dc:creator>Liang, X.</dc:creator>
<dc:creator>Nordling, T. E. M.</dc:creator>
<dc:creator>Shiga, M.</dc:creator>
<dc:creator>Stanescu, A.</dc:creator>
<dc:creator>Vogel, R.</dc:creator>
<dc:creator>The Respiratory Viral DREAM Challenge Consortium,</dc:creator>
<dc:creator>Pandey, G.</dc:creator>
<dc:creator>Chiu, C.</dc:creator>
<dc:creator>McClain, M. T.</dc:creator>
<dc:creator>Woods, C. W.</dc:creator>
<dc:creator>Ginsburg, G. S.</dc:creator>
<dc:creator>Elo, L. L.</dc:creator>
<dc:creator>Tsalik, E. L.</dc:creator>
<dc:creator>Mangravite, L. M.</dc:creator>
<dc:creator>Sieberts, S. K.</dc:creator>
<dc:date>2018-04-30</dc:date>
<dc:identifier>doi:10.1101/311696</dc:identifier>
<dc:title><![CDATA[A crowdsourced analysis to identify ab initio molecular signatures predictive of susceptibility to viral infection]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-04-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/318832v1?rss=1">
<title>
<![CDATA[
Expression and purification of a functional heteromeric GABAA receptor for structural studies 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/318832v1?rss=1</link>
<description><![CDATA[
The multi-subunit GABA-gated chloride channels of the Cys-loop receptor family, known as GABAA receptors, function as the primary gatekeepers of fast inhibitory neurotransmission in the central nervous system. In addition to their role in controlling synaptic tone, these receptors are the targets of a vast array of therapeutic compounds that potentiate channel gating. Importantly, functional activity and pharmacological efficacy of GABAA receptors is coupled directly to the subunit composition. However, the absence of high resolution structural information precludes an explicit determination of the molecular mechanism of ligand binding to ion channel gating and modulation. Efforts to obtain this data are hindered largely by the lack of heterologous expression and purification protocols for high expressing receptor constructs. To address this issue, we describe a unique approach to identify bona fide functional GABAA receptor subunit combinations by using the Xenopus oocyte as an expression host in combination with fluorescence detection size exclusion chromatography. The results demonstrate that formation of a defined pentameric species is dependent on subunit composition. Furthermore, receptor subunits can tolerate large truncations in non-conserved M3/M4 cytoplasmic loop, although removal of N-linked glycosylation sites is negatively correlated with expression level. Additionally, we report methods to improve GABAA receptor expression in mammalian cell culture that employ recombinant baculovirus transduction. From these methods we have identified a well-behaving minimal functional construct for the 1/{beta}1 GABAA receptor subtype that can be purified in milligram quantities while retaining high affinity agonist binding activity.
]]></description>
<dc:creator>Claxton, D. P.</dc:creator>
<dc:creator>Gouaux, E.</dc:creator>
<dc:date>2018-05-09</dc:date>
<dc:identifier>doi:10.1101/318832</dc:identifier>
<dc:title><![CDATA[Expression and purification of a functional heteromeric GABAA receptor for structural studies]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-05-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/329854v1?rss=1">
<title>
<![CDATA[
Controlling for contaminants in low biomass 16S rRNA gene sequencing experiments 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/329854v1?rss=1</link>
<description><![CDATA[
BackgroundMicrobial communities are commonly studied using culture-independent methods such as 16S rRNA gene sequencing. However, one challenge in accurately characterizing microbial communities is exogenous bacterial DNA contamination. This is particularly problematic for sites of low microbial biomass such as the urinary tract, placenta, and lower airway. Computational approaches have been proposed as a post-processing step to identify and remove potential contaminants, but their performance has not been independently evaluated.nnTo identify the impact of decreasing microbial biomass on polymicrobial 16S rRNA gene sequencing experiments, we used a serial dilution of a mock microbial community. We evaluated two computational approaches to identify and remove contaminants: 1) identifying sequences that have an inverse correlation with DNA concentration implemented in Decontam and 2) predicting the proportion of experimental sample arising from defined contaminant sources implemented in SourceTracker.nnResultsAs expected, the proportion of contaminant bacterial DNA increased with decreasing starting microbial biomass, with 79.12% of the most dilute sample arising from contaminant sequences. Inclusion of contaminant sequences in analyses leads to overinflated diversity estimates (up to 12 times greater than the expected values) and distorts microbiome composition. SourceTracker successfully removed over 98% of contaminants when the experimental environments are well defined. However, SourceTracker performed poorly when the experimental environment is unknown, failing to remove the majority of contaminants. Decontam successfully removed 74-91% of contaminants regardless of prior knowledge of the experimental environment.nnConclusionsOur study indicates that computational methods can reduce the amount of contaminants in 16S rRNA gene sequencing experiments. The appropriate computational approach for removing contaminant sequences from an experiment depends on the prior knowledge about the microbial environment under investigation and can be evaluated with a dilution series of a mock microbial community.
]]></description>
<dc:creator>Karstens, L.</dc:creator>
<dc:creator>Asquith, M.</dc:creator>
<dc:creator>Davin, S.</dc:creator>
<dc:creator>Fair, D.</dc:creator>
<dc:creator>Gregory, W. T.</dc:creator>
<dc:creator>Wolfe, A. J.</dc:creator>
<dc:creator>Braun, J.</dc:creator>
<dc:creator>McWeeney, S.</dc:creator>
<dc:date>2018-05-25</dc:date>
<dc:identifier>doi:10.1101/329854</dc:identifier>
<dc:title><![CDATA[Controlling for contaminants in low biomass 16S rRNA gene sequencing experiments]]></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/10.1101/330647v1?rss=1">
<title>
<![CDATA[
Clarification and confocal imaging of the non-human primate placental micro-anatomy 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/330647v1?rss=1</link>
<description><![CDATA[
Placental function is essential for the development of the fetus, and is - in part - related to the 3D arrangement of the villous and vascular geometry. Recent advances in tissue clarification techniques allow for deep high-resolution imaging with confocal microscopy without altering the spatial characteristics of the tissue. These image stacks can be analyzed quantitatively to provide insights regarding the villous and vascular micro-anatomy as well as the interrelationships between the two. However, such analyses require optimization of the tissue preparation, immuno-labeling, and clarification protocol in order to provide reliable results suitable for the detection of subtle differences in pathologic pregnancies. Placental and fetal development are similar between human and non-human primate pregnancies, with the latter serving as a reliable, validated, highly-controlled, well-characterized translational model for the former.nnWe present a protocol for the preparation, immuno-labeling, and clarification of the non-human primate placenta optimized for confocal microscopy and subsequent quantification of the micro-anatomic structures.
]]></description>
<dc:creator>Sargent, J.</dc:creator>
<dc:creator>Roberts, V.</dc:creator>
<dc:creator>Gaffney, J.</dc:creator>
<dc:creator>Frias, A.</dc:creator>
<dc:date>2018-05-24</dc:date>
<dc:identifier>doi:10.1101/330647</dc:identifier>
<dc:title><![CDATA[Clarification and confocal imaging of the non-human primate placental micro-anatomy]]></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/10.1101/331991v1?rss=1">
<title>
<![CDATA[
Genetic Modifiers of Pathogenic LRRK2 G2019S Neurodegeneration in Drosophila 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/331991v1?rss=1</link>
<description><![CDATA[
Disease phenotypes can be highly variable among individuals with the same pathogenic mutation. There is increasing evidence that background genetic variation is a strong driver of disease variability in addition to the influence of environment. To understand the genotype-phenotype relationship that determines the expressivity of a pathogenic mutation, a large number of backgrounds must be studied. This can be efficiently achieved using model organism collections such as the Drosophila Genetic Reference Panel (DGRP). Here, we used the DGRP to assess the variability of locomotor dysfunction in a LRRK2 G2019S Drosophila melanogaster model of Parkinsons disease. We find substantial variability in the LRRK2 G2019S locomotor phenotype in different DGRP backgrounds. A genome-wide association study for candidate genetic modifiers reveals 177 genes that drive wide phenotypic variation, including 19 top association genes. Genes involved in the outgrowth and regulation of neuronal projections are enriched in these candidate modifiers. RNAi functional testing of the top association and neuronal projection-related genes reveals that pros, pbl, ct and CG33506 significantly modify age-related dopamine neuron loss and associated locomotor dysfunction in the Drosophila LRRK2 G2019S model. These results demonstrate how natural genetic variation can be used as a powerful tool to identify genes that modify disease-related phenotypes. We report novel candidate modifier genes for LRRK2 G2019S that may be used to interrogate the link between LRRK2, neurite regulation and neuronal degeneration in Parkinsons disease.
]]></description>
<dc:creator>Lavoy, S.</dc:creator>
<dc:creator>Chittoor-Vinod, V.</dc:creator>
<dc:creator>Chow, C.</dc:creator>
<dc:creator>Martin, I.</dc:creator>
<dc:date>2018-05-30</dc:date>
<dc:identifier>doi:10.1101/331991</dc:identifier>
<dc:title><![CDATA[Genetic Modifiers of Pathogenic LRRK2 G2019S Neurodegeneration in Drosophila]]></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/10.1101/335968v1?rss=1">
<title>
<![CDATA[
Thermostabilization and purification of the human dopamine transporter (hDAT) in an inhibitor and allosteric ligand bound conformation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/335968v1?rss=1</link>
<description><![CDATA[
The human dopamine transporter(hDAT) plays a major role in dopamine homeostasis and regulation of neurotransmission by clearing dopamine from the extracellular space using secondary active transport. Dopamine is an essential monoamine chemical messenger that regulates reward seeking behavior, motor control, hormonal release, and emotional response in humans. Psychostimulants such as cocaine primarily target the central binding site of hDAT and lock the transporter in an outward-facing conformation, thereby inhibiting dopamine reuptake. The inhibition of dopamine reuptake leads to accumulation of dopamine in the synapse causing heightened signaling. In addition, hDAT is implicated in various neurological disorders and disease-associated neurodegeneration. Despite its significance, the molecular architecture of hDAT and its various conformational states are poorly understood. Instability of hDAT in detergent micelles has been a limiting factor in its successful biochemical, biophysical, and structural characterization. To overcome this hurdle, first we identified ligands that stabilize hDAT in detergent micelles. Then, we screened [~]200 single residue mutants of hDAT using high-throughput scintillation proximity assay, and identified a thermostable variant(I248Y). Here we report a robust strategy to overexpress and successfully purify a thermostable variant of hDAT in an inhibitor and allosteric ligand bound conformation.
]]></description>
<dc:creator>Navratna, V.</dc:creator>
<dc:creator>Tosh, D. K.</dc:creator>
<dc:creator>Jacobson, K. A.</dc:creator>
<dc:creator>Gouaux, E.</dc:creator>
<dc:date>2018-05-31</dc:date>
<dc:identifier>doi:10.1101/335968</dc:identifier>
<dc:title><![CDATA[Thermostabilization and purification of the human dopamine transporter (hDAT) in an inhibitor and allosteric ligand bound conformation]]></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/10.1101/337360v1?rss=1">
<title>
<![CDATA[
Correction of respiratory artifacts in MRI head motion estimates 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/337360v1?rss=1</link>
<description><![CDATA[
Head motion represents one of the greatest technical obstacles for brain MRI. Accurate detection of artifacts induced by head motion requires precise estimation of movement. However, this estimation may be corrupted by factitious effects owing to main field fluctuations generated by body motion. In the current report, we examine head motion estimation in multiband resting state functional connectivity MRI (rs-fcMRI) data from the Adolescent Brain and Cognitive Development (ABCD) Study and a comparison  single-shot dataset from Oregon Health & Science University. We show unequivocally that respirations contaminate movement estimates in functional MRI and that respiration generates apparent head motion not associated with degraded quality of functional MRI. We have developed a novel approach using a band-stop filter that accurately removes these respiratory effects. Subsequently, we demonstrate that utilizing this filter improves post-processing data quality. Lastly, we demonstrate the real-time implementation of motion estimate filtering in our FIRMM (Framewise Integrated Real-Time MRI Monitoring) software package.
]]></description>
<dc:creator>Fair, D. A.</dc:creator>
<dc:creator>Miranda-Dominguez, O.</dc:creator>
<dc:creator>Snyder, A. Z.</dc:creator>
<dc:creator>Perrone, A. A.</dc:creator>
<dc:creator>Earl, E. A.</dc:creator>
<dc:creator>Van, A. N.</dc:creator>
<dc:creator>Koller, J. M.</dc:creator>
<dc:creator>Feczko, E.</dc:creator>
<dc:creator>Klein, R. L.</dc:creator>
<dc:creator>Mirro, A. E.</dc:creator>
<dc:creator>Hampton, J. M.</dc:creator>
<dc:creator>Adeyemo, B.</dc:creator>
<dc:creator>Laumann, T. O.</dc:creator>
<dc:creator>Gratton, C.</dc:creator>
<dc:creator>Greene, D. J.</dc:creator>
<dc:creator>Schlaggar, B.</dc:creator>
<dc:creator>Hagler, D.</dc:creator>
<dc:creator>Watts, R.</dc:creator>
<dc:creator>Garavan, H.</dc:creator>
<dc:creator>Barch, D. M.</dc:creator>
<dc:creator>Nigg, J. T.</dc:creator>
<dc:creator>Petersen, S. E.</dc:creator>
<dc:creator>Dale, A.</dc:creator>
<dc:creator>Feldstein-Ewing, S. W.</dc:creator>
<dc:creator>Nagel, B. J.</dc:creator>
<dc:creator>Dosenbach, N. U. F.</dc:creator>
<dc:date>2018-06-07</dc:date>
<dc:identifier>doi:10.1101/337360</dc:identifier>
<dc:title><![CDATA[Correction of respiratory artifacts in MRI head motion estimates]]></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/10.1101/337659v1?rss=1">
<title>
<![CDATA[
Cryo-EM structure of the benzodiazepine-sensitive α1β1γ2 heterotrimeric GABAA receptor in complex with GABA illuminates mechanism of receptor assembly and agonist binding 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/337659v1?rss=1</link>
<description><![CDATA[
Fast inhibitory neurotransmission in the mammalian nervous system is largely mediated by GABAA receptors, chloride-selective members of the superfamily of pentameric Cys-loop receptors. Native GABAA receptors are heteromeric assemblies sensitive to many important drugs, from sedatives to anesthetics and anticonvulsive agents, with mutant forms of GABAA receptors implicated in multiple neurological diseases, including epilepsy. Despite the profound importance of heteromeric GABAA receptors in neuroscience and medicine, they have proven recalcitrant to structure determination. Here we present the structure of the triheteromeric 1{beta}1{gamma}2EM GABAA receptor in complex with GABA, determined by single particle cryo-EM at 3.1-3.8 [A] resolution, elucidating the molecular principles of receptor assembly and agonist binding. Remarkable N-linked glycosylation on the 1 subunit occludes the extracellular vestibule of the ion channel and is poised to modulate receptor assembly and perhaps ion channel gating. Our work provides a pathway to structural studies of heteromeric GABAA receptors and a framework for the rational design of novel therapeutic agents.
]]></description>
<dc:creator>Phulera, S.</dc:creator>
<dc:creator>Zhu, H.</dc:creator>
<dc:creator>Yu, J.</dc:creator>
<dc:creator>Claxton, D.</dc:creator>
<dc:creator>Yoder, N.</dc:creator>
<dc:creator>Yoshioka, C.</dc:creator>
<dc:creator>Gouaux, E.</dc:creator>
<dc:date>2018-06-03</dc:date>
<dc:identifier>doi:10.1101/337659</dc:identifier>
<dc:title><![CDATA[Cryo-EM structure of the benzodiazepine-sensitive α1β1γ2 heterotrimeric GABAA receptor in complex with GABA illuminates mechanism of receptor assembly and agonist binding]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-06-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/338053v1?rss=1">
<title>
<![CDATA[
High-throughput mapping of meiotic crossover and chromosome mis-segregation events in interspecific hybrid mice 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/338053v1?rss=1</link>
<description><![CDATA[
We developed "sci-LIANTI", a high-throughput, high-coverage single-cell DNA sequencing method that combines single-cell combinatorial indexing ("sci") and linear amplification via transposon insertion ("LIANTI"). To characterize rare chromosome mis-segregation events in male meiosis and their relationship to the landscape of meiotic crossovers, we applied sci-LIANTI to profile the genomes of 6,928 sperm and sperm precursors from infertile, interspecific F1 male mice. From 1,663 haploid and 292 diploid cells, we mapped 24,672 crossover events and identified genomic and epigenomic contexts that influence crossover hotness. Surprisingly, we observed frequent mitotic chromosome segregation during meiosis. Moreover, segregation during meiosis in individual cells was highly biased towards either mitotic or meiotic events. We anticipate that sci-LIANTI can be applied to fully characterize various recombination landscapes, as well as to other fields requiring high-throughput, high-coverage single-cell genome sequencing.nnOne Sentence SummarySingle-cell genome sequencing maps crossover and non-meiotic chromosome segregation during spermatogenesis in interspecific hybrid mice.
]]></description>
<dc:creator>Yin, Y.</dc:creator>
<dc:creator>Jiang, Y.</dc:creator>
<dc:creator>Berletch, J. B.</dc:creator>
<dc:creator>Disteche, C. M.</dc:creator>
<dc:creator>Noble, W. S.</dc:creator>
<dc:creator>Steemers, F. J.</dc:creator>
<dc:creator>Adey, A. C.</dc:creator>
<dc:creator>Shendure, J. A.</dc:creator>
<dc:date>2018-06-04</dc:date>
<dc:identifier>doi:10.1101/338053</dc:identifier>
<dc:title><![CDATA[High-throughput mapping of meiotic crossover and chromosome mis-segregation events in interspecific hybrid mice]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-06-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/342931v1?rss=1">
<title>
<![CDATA[
Fbxw7 is a critical regulator of Schwann cell myelinating potential 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/342931v1?rss=1</link>
<description><![CDATA[
Myelin insulates and protects axons in vertebrate nervous systems. In the central nervous system (CNS), oligodendrocytes (OLs) make numerous myelin sheaths on multiple axons, whereas in the peripheral nervous system (PNS) myelinating Schwann cells (SCs) make just one myelin sheath on a single axon. Why the myelinating potentials of OLs and SCs are so fundamentally different is unclear. Here, we find that loss of Fbxw7, an E3 ubiquitin ligase component, enhances the myelinating potential of SCs. Fbxw7 mutant SCs are seen myelinating multiple axons in a fashion reminiscent of OLs as well as aberrantly myelinating large axons while simultaneously ensheathing small unmyelinated axons - typically distinct roles of myelinating SCs and non-myelinating Remak SCs, respectively. We found that several of the Fbxw7 mutant phenotypes, including the ability to generate thicker myelin sheaths, were due to dysregulation of mTOR. However, the remarkable ability of mutant SCs to either myelinate multiple axons or myelinate some axons while simultaneously encompassing other unmyelinated axons is independent of mTOR signaling. This indicates distinct roles for Fbxw7 in regulating multiple aspects of SC behavior and that novel Fbxw7-regulated mechanisms control modes of myelination previously thought to fundamentally distinguish myelinating SCs from non-myelinating SCs and OLs. Our data reveal unexpected plasticity in the myelinating potential of SCs, which may have important implications for our understanding of both PNS and CNS myelination and myelin repair.
]]></description>
<dc:creator>Harty, B. L.</dc:creator>
<dc:creator>Coelho, F.</dc:creator>
<dc:creator>Ackerman, S. D.</dc:creator>
<dc:creator>Herbert, A. L.</dc:creator>
<dc:creator>Lyons, D. A.</dc:creator>
<dc:creator>Monk, K. R.</dc:creator>
<dc:date>2018-06-08</dc:date>
<dc:identifier>doi:10.1101/342931</dc:identifier>
<dc:title><![CDATA[Fbxw7 is a critical regulator of Schwann cell myelinating potential]]></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/10.1101/363879v1?rss=1">
<title>
<![CDATA[
Evolution of Endothelin signaling and diversification of adult pigment pattern in Danio fishes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/363879v1?rss=1</link>
<description><![CDATA[
Fishes of the genus Danio exhibit diverse pigment patterns that serve as useful models for understanding the genes and cell behaviors underlying the evolution of adult form. Among these species, zebrafish D. rerio exhibit several dark stripes of melanophores with sparse iridophores that alternate with light interstripes of dense iridophores and xanthophores. By contrast, the closely related species D. nigrofasciatus has an attenuated pattern with fewer melanophores, stripes and interstripes. Here we demonstrate species differences in iridophore development that presage the fully formed patterns. Using genetic and transgenic approaches we identify the secreted peptide Endothelin-3 (Edn3)--a known melanogenic factor of tetrapods--as contributing to reduced iridophore proliferation and fewer stripes and interstripes in D. nigrofasciatus. We further show the locus encoding this factor is expressed at lower levels in D. nigrofasciatus owing to cis-regulatory differences between species. Finally, we show that functions of two paralogous loci encoding Edn3 have been partitioned between skin and non-skin iridophores. Our findings reveal genetic and cellular mechanisms contributing to pattern differences between these species and suggest a model for evolutionary changes in Edn3 requirements across vertebrates.nnAuthor SummaryNeural crest derived pigment cells generate the spectacular variation in skin pigment patterns among vertebrates. Mammals and birds have just a single skin pigment cell, the melanocyte, whereas ectothermic vertebrates have several pigment cells including melanophores, iridophores and xanthophores, that together organize into a diverse array of patterns. In the teleost zebrafish, Danio rerio, an adult pattern of stripes depends on interactions between pigment cell classes and between pigment cells and their tissue environment. The close relative, D. nigrofasciatus has fewer stripes and prior analyses suggested a difference between these species that lies extrinsic to the pigment cells themselves. A candidate for mediating this difference is Endothelin-3 (Edn3), essential for melanocyte development in warm-blooded animals, and required by all three classes of pigment cells in an amphibian. We show that Edn3 specifically promotes iridophore development in Danio, and that differences in Edn3 expression contribute to differences in iridophore complements, and striping, between D. rerio and D. nigrofasciatus. Our study reveals a novel function for Edn3 and provides new insights into how changes in gene expression yield morphogenetic outcomes to effect diversification of adult form.
]]></description>
<dc:creator>Spiewak, J.</dc:creator>
<dc:creator>Bain, E.</dc:creator>
<dc:creator>Liu, J.</dc:creator>
<dc:creator>Kou, K.</dc:creator>
<dc:creator>Sturiale, S.</dc:creator>
<dc:creator>Patterson, L.</dc:creator>
<dc:creator>Diba, P.</dc:creator>
<dc:creator>Eisen, J.</dc:creator>
<dc:creator>Braasch, I.</dc:creator>
<dc:creator>Ganz, J.</dc:creator>
<dc:creator>Parichy, D. M.</dc:creator>
<dc:date>2018-07-06</dc:date>
<dc:identifier>doi:10.1101/363879</dc:identifier>
<dc:title><![CDATA[Evolution of Endothelin signaling and diversification of adult pigment pattern in Danio fishes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-07-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/372615v1?rss=1">
<title>
<![CDATA[
Caspase-mediated cleavage of Murine Norovirus NS1/2 potentiates apoptosis and is required for persistent infection 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/372615v1?rss=1</link>
<description><![CDATA[
Human norovirus (HuNoV) is the leading cause of acute gastroenteritis and is spread by fecal shedding that can often persist for weeks to months after infection. Murine norovirus (MNV) is also shed persistently in the feces and provides a tractable model to study molecular mechanisms of enteric persistence. Previous studies have identified non-structural protein 1 (NS1) from the persistent MNV strain CR6 as critical for persistent infection in intestinal epithelial cells (IECs), but its mechanism of action remains unclear. We now find that the function of NS1 in promoting persistence is regulated by apoptosis. Following induction of apoptosis in infected cells, a minority of NS1 is cleaved from the precursor NS1/2 protein, and this cleavage is prevented by mutation of caspase target motifs. MNV strain CR6 with these mutations (CR6{triangleup}casp) is profoundly compromised in infection of IECs and persistence in the intestine. Conversely, replication in tissues outside of the intestine, or in a cultured macrophage cell line, is unchanged, indicating that the requirement of NS1/2 cleavage is intestine-specific. Intriguingly, we also find that cleavage of CR6 NS1/2 potentiates apoptosis, suggesting that regulation of cell death is a novel function of this viral protein. Together, these data indicate that the ability of NS1 to promote MNV persistence in IECs is regulated by host caspases, and suggest that potentiation of apoptosis plays a role in viral tropism in the intestine.nnAuthor SummaryHuman Norovirus infection is highly contagious and the most common cause of acute gastroenteritis. Norovirus can persist and be shed for months after infection, leading to continued outbreaks. There are many unanswered questions as to host and viral components of norovirus pathogenesis that can be addressed within the murine norovirus (MNV) model system. We previously identified a critical role for a viral protein, NS1, for intestinal persistence. Herein we describe how the regulation of NS1 is critical for persistent infection in intestinal epithelial cells, but is not required for acute infection of non-epithelial cells, or infection of tissues outside of the gut. Additionally, we demonstrate that NS1 is both regulated by the host cell death machinery, and also reciprocally regulates that machinery to promote cell death during MNV infection, and found that this is specific to persistent strain of MNV. Altogether these data identify a role for how NS1 in a new pathway involved in establishing a persistent norovirus infection in the intestine.
]]></description>
<dc:creator>Robinson, B. A.</dc:creator>
<dc:creator>McCune, B. T.</dc:creator>
<dc:creator>Peters, A. M.</dc:creator>
<dc:creator>Nice, T.</dc:creator>
<dc:date>2018-07-19</dc:date>
<dc:identifier>doi:10.1101/372615</dc:identifier>
<dc:title><![CDATA[Caspase-mediated cleavage of Murine Norovirus NS1/2 potentiates apoptosis and is required for persistent infection]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-07-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/373993v1?rss=1">
<title>
<![CDATA[
Accurate transmission of IGF-I into AKT signaling activity in individual cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/373993v1?rss=1</link>
<description><![CDATA[
Cells sense and respond to their environment by activating distinct intracellular signaling pathways, however an individual cells ability to faithfully transmit and discriminate environmental signals is thought to be limited. To assess the fidelity of signal transmission in the PI3K-AKT signaling pathway, we first developed an optimized genetically encoded sensor that had an increased dynamic range and reduced variation under basal conditions. We then used this reporter to track responses to varying doses of IGF-I in live cells and found that signaling responses from individual cells overlapped across a wide range of IGF-I doses, suggesting limited transmission accuracy. However, further analysis of individual cell traces revealed that responses were constant over time without stochastic fluctuations. We devised a new information theoretic approach to calculate the channel capacity using variance of the single cell time course data--rather than population-level variance as has been previously used--and predicted that cells were capable of discriminating multiple growth factor doses. We validated these predictions by tracking individual cell responses to multiple IGF-I doses and found that cells can accurately distinguish at least four different IGF-I concentrations, as demonstrated by their distinct responses. Furthermore, we found a similar discriminatory ability to pathway inhibition, as assessed by responses to the PI3K inhibitor alpelisib. Our studies indicate that cells can faithfully transmit an IGF-I input into a down-stream signaling response and that heterogeneous responses result from variation in the input-output relation across the population. These observations reveal the importance of viewing each cell as having its own communication channel and underscore the importance of understanding responses at the single cell level.
]]></description>
<dc:creator>Gross, S. M.</dc:creator>
<dc:creator>Dane, M. A.</dc:creator>
<dc:creator>Bucher, E.</dc:creator>
<dc:creator>Heiser, L. M.</dc:creator>
<dc:date>2018-07-21</dc:date>
<dc:identifier>doi:10.1101/373993</dc:identifier>
<dc:title><![CDATA[Accurate transmission of IGF-I into AKT signaling activity in individual cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-07-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/377994v1?rss=1">
<title>
<![CDATA[
Opposing effects of impulsivity and mindset on sources of science self-efficacy and STEM interest in adolescents 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/377994v1?rss=1</link>
<description><![CDATA[
Impulsivity has been linked to academic performance in the context of Attention Deficit Hyperactivity Disorder, though its influence on a wider spectrum of students remains largely unexplored, particularly in the context of STEM learning (i.e. science, technology, engineering, and math). STEM learning was hypothesized to be more challenging for impulsive students, since it requires the practice and repetition of tasks as well as concerted attention to task performance. Impulsivity was assessed in a cross-sectional sample of 2,476 students in grades 6-12. Results show impulsivity affects a larger population of students, not limited to students with learning disabilities. Impulsivity was associated with lower sources of science self-efficacy (SSSE) scores, interest in all STEM domains (particularly math), and self-reported STEM skills. The large negative effect observed for impulsivity was opposed by "growth" mindset, which describes a students belief in the importance of effort when learning is difficult. Mindset had a large positive effect, which was associated with greater SSSE, STEM interest, and STEM skills. When modeled together, results suggest that mindset interventions may benefit impulsive students who struggle with STEM. Together, these data suggest important interconnected roles for impulsivity and mindset that can influence secondary students STEM trajectories.
]]></description>
<dc:creator>Marriott, L. K.</dc:creator>
<dc:creator>Coppola, L. K.</dc:creator>
<dc:creator>Mitchell, S. H.</dc:creator>
<dc:creator>Bouwma-Gearhart, J.</dc:creator>
<dc:creator>Chen, Z.</dc:creator>
<dc:creator>Shifrer, D.</dc:creator>
<dc:creator>Shannon, J. K.</dc:creator>
<dc:date>2018-07-26</dc:date>
<dc:identifier>doi:10.1101/377994</dc:identifier>
<dc:title><![CDATA[Opposing effects of impulsivity and mindset on sources of science self-efficacy and STEM interest in adolescents]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-07-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/378422v1?rss=1">
<title>
<![CDATA[
Mechanisms for zinc and proton inhibition of the GluN1/GluN2A NMDA receptor 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/378422v1?rss=1</link>
<description><![CDATA[
N-Methyl-D-aspartate receptors (NMDARs) play essential roles in memory formation, neuronal plasticity and brain development with their dysfunction linked to a range of disorders from ischemia to schizophrenia. Zinc and pH are physiological allosteric modulators of NMDARs with GluN2A containing receptors inhibited by nanomolar concentrations of divalent zinc and by excursions to low pH. Despite the widespread importance of zinc and proton modulation of NMDARs, the molecular mechanism by which these ions modulate receptor activity has proven elusive. Here, we use cryo-electron microscopy to elucidate the structure of the GluN1/GluN2A NMDAR in a large ensemble of conformations under a range of physiologically relevant zinc and proton concentrations. We show how zinc binding to the amino terminal domain elicits structural changes that are transduced though the ligand-binding domain and result in constriction of the ion channel gate.
]]></description>
<dc:creator>Jalali-Yazdi, F.</dc:creator>
<dc:creator>Chowdhury, S.</dc:creator>
<dc:creator>Yoshioka, C.</dc:creator>
<dc:creator>Gouaux, E.</dc:creator>
<dc:date>2018-07-26</dc:date>
<dc:identifier>doi:10.1101/378422</dc:identifier>
<dc:title><![CDATA[Mechanisms for zinc and proton inhibition of the GluN1/GluN2A NMDA receptor]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-07-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/379685v1?rss=1">
<title>
<![CDATA[
Divalent cation and chloride ion sites of chicken acid sensing ion channel 1a elucidated by x-ray crystallography 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/379685v1?rss=1</link>
<description><![CDATA[
Acid sensing ion channels (ASICs) are proton-gated ion channels that are members of the degenerin/epithelial sodium channel superfamily and are expressed throughout central and peripheral nervous systems. ASICs have been implicated in multiple physiological processes and are subject to numerous forms of endogenous and exogenous regulation that include modulation by Ca2+ and Cl- ions. However, the mapping of ion binding sites as well as a structure-based understanding of the mechanisms underlying ionic modulation of ASICs have remained elusive. Here we present ion binding sites of chicken ASIC1a in resting and desensitized states at high and low pH, respectively, determined by anomalous diffraction x-ray crystallography. The acidic pocket serves as a nexus for divalent cation binding at both low and high pH, while we observe divalent cation binding within the central vestibule on the resting channel at high pH only. Moreover, neutralization of residues positioned to coordinate divalent cations via individual and combined Glu to Gln substitutions reduced, but did not extinguish, modulation of proton-dependent gating by Ca2+. Additionally, we demonstrate that anion binding at the canonical thumb domain site is state-dependent and present a previously undetected anion site at the mouth of the extracellular fenestrations on the resting channel. Our results map anion and cation sites on ASICs across multiple functional states, informing possible mechanisms of modulation and providing a blueprint for the design of therapeutics targeting ASICs.
]]></description>
<dc:creator>Yoder, N.</dc:creator>
<dc:creator>Gouaux, E.</dc:creator>
<dc:date>2018-07-30</dc:date>
<dc:identifier>doi:10.1101/379685</dc:identifier>
<dc:title><![CDATA[Divalent cation and chloride ion sites of chicken acid sensing ion channel 1a elucidated by x-ray crystallography]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-07-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/381087v1?rss=1">
<title>
<![CDATA[
Effects of cochlear synaptopathy on spontaneous and sound-evoked activity in the mouse inferior colliculus 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/381087v1?rss=1</link>
<description><![CDATA[
Tinnitus and hyperacusis are life-disrupting perceptual abnormalities that are often preceded by acoustic overexposure. Animal models of overexposure have suggested a link between these phenomena and neural hyperactivity, i.e. elevated spontaneous rates (SRs) and sound-evoked responses. Prior work has focused on changes in central auditory responses, with less attention paid to the exact nature of the associated peripheral damage. The demonstration that acoustic overexposure can cause cochlear nerve damage without permanent threshold elevation suggests this type of peripheral damage may be a key elicitor of tinnitus and hyperacusis in humans with normal audiograms. We addressed this idea by recording responses in the mouse inferior colliculus (IC) following a bilateral, neuropathic noise exposure. Two wks post-exposure, mean SRs were unchanged in mice recorded while awake, or under anesthesia. SRs were also unaffected by more intense, or unilateral exposures. These results suggest that neither neuropathy nor hair cell loss are sufficient to raise SRs in the IC, at least in mice. However, its not clear whether our mice had tinnitus. Tone-evoked rate-level functions at the CF were steeper following exposure, specifically in the region of maximal neuropathy. Furthermore, suppression driven by off-CF tones and by ipsilateral noise were also reduced. Both changes were especially pronounced in neurons of awake mice. These findings align with prior reports of elevated acoustic startle in neuropathic mice, and indicate that neuropathy may initiate a compensatory response in the central auditory system leading to the genesis of hyperacusis.
]]></description>
<dc:creator>Shaheen, L.</dc:creator>
<dc:creator>Liberman, M. C.</dc:creator>
<dc:date>2018-08-06</dc:date>
<dc:identifier>doi:10.1101/381087</dc:identifier>
<dc:title><![CDATA[Effects of cochlear synaptopathy on spontaneous and sound-evoked activity in the mouse inferior colliculus]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-08-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/385104v1?rss=1">
<title>
<![CDATA[
Isolation and Characterization of a Myoviridae phage that lyses two Escherichia coli pathotypes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/385104v1?rss=1</link>
<description><![CDATA[
PurposeTo identify therapeutic a bacteriophage that kills diarrheagenic enteroaggregative Escherichia coli (EAEC) while leaving the human microbiome intact.nnMethodologyPhages from wastewater in Portland, OR, were screened for bacteriolytic activity using an overlay assay, and isolated in a sequential procedure to enrich for the recognition of core bacterial antigens. Electron microscopy and genome sequencing were performed to classify the isolated phage, and the host range was determined by spot tests and plaque assays. One-step growth curves and time-kill assays were conducted to characterize the life cycle of the phage, and to interrogate the multiplicity of infection (MOI) necessary for killing. A mouse model of infection was used to determine whether the phage could be used therapeutically against EAEC in vivo. Anaerobic culture in the presence of human fecal bacteria determined whether the phage could kill EAEC in vitro, and to assess whether the microbiome had been altered.nnResultsThe isolated phage, termed Escherichia virus PDX, is a member of the strictly lytic Myoviridae family of viruses. Phage PDX killed EAEC isolate EN1E-0227, a case-associated isolate from a child in rural Tennessee, in a dose-dependent manner, and also formed plaques on case-associated clinical EAEC isolates from Columbian children suffering from diarrhea. A single dose of PDX, at a MOI of 100, one day post infection, reduced the population of recovered EAEC isolate EN1E-0227 bacteria in fecal pellets in a mouse model of colonization, over a five-day period. Phage PDX also killed EAEC EN1E-0227 when cultured anaerobically in vitro in the presence of human fecal bacteria. While the addition of EAEC EN1E-0227 reduced the -diversity of the human microbiota, that of the cultures with either feces alone, feces with EAEC and PDX, or with just the PDX phage were not different statistically, as measured by Chao1 and Shannon diversity indices. Additionally, {beta}-diversity and differential abundance analyses show that conditions with PDX added were not different from feces alone, but all groups were significantly different from feces + EAEC.nnConclusionsThe strictly bacteriolytic, Myoviridae Escherichia virus PDX killed EAEC isolate EN1E-0227 bacteria both in vivo and in vitro, while simultaneously not altering the diversity of normal human microbiota in anaerobic culture. Thus, the PDX phage could be part of an effective therapeutic intervention for children in developing countries who suffer from acute, or persistent EAEC-mediated diarrhea, and to help reduce the serious effects of environmental enteropathy. Because the emerging pathogen EAEC is now the second leading cause of travelers diarrhea, PDX could also provide therapeutic relief for these individuals, particularly in light of the growing crisis of antibiotic resistances.
]]></description>
<dc:creator>Cepko, L.</dc:creator>
<dc:creator>Garling, E.</dc:creator>
<dc:creator>Dinsdale, M.</dc:creator>
<dc:creator>Bandy, L.</dc:creator>
<dc:creator>Mellies, J.</dc:creator>
<dc:date>2018-08-04</dc:date>
<dc:identifier>doi:10.1101/385104</dc:identifier>
<dc:title><![CDATA[Isolation and Characterization of a Myoviridae phage that lyses two Escherichia coli pathotypes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-08-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/393330v1?rss=1">
<title>
<![CDATA[
Putative pore-forming subunits of the mechano-electrical transduction channel, Tmc1/2b, require Tmie to localize to the site of mechanotransduction in zebrafish sensory hair cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/393330v1?rss=1</link>
<description><![CDATA[
Mutations in transmembrane inner ear (TMIE) cause deafness in humans; previous studies suggest involvement in the mechano-electrical transduction (MET) complex in sensory hair cells, but TMIEs precise role is unclear. In tmie zebrafish mutants, we observed that GFP-tagged Tmc1 and Tmc2b, which are putative subunits of the MET channel, fail to target to the hair bundle. In contrast, overexpression of Tmie strongly enhances the targeting of Tmc2b-GFP to stereocilia. To identify the motifs of Tmie underlying the regulation of the Tmcs, we systematically deleted or replaced peptide segments. We then assessed localization and functional rescue of each mutated/chimeric form of Tmie in tmie mutants. We determined that the first putative helix was dispensable and identified a novel critical region of Tmie, the extracellular region and transmembrane domain, which mediates both mechanosensitivity and Tmc2b-GFP expression in bundles. Collectively, our results suggest that Tmies role in sensory hair cells is to target and stabilize Tmc subunits to the site of MET.nnAuthor summaryHair cells mediate hearing and balance through the activity of a pore-forming channel in the cell membrane. The transmembrane inner ear (TMIE) protein is an essential component of the protein complex that gates this so-called mechanotransduction channel. While it is known that loss of TMIE results in deafness, the function of TMIE within the complex is unclear. Using zebrafish as a deafness model, Pacentine and Nicolson demonstrate that Tmie is required for the localization of other essential complex members, the transmembrane channel-like (Tmc) proteins, Tmc1/2b. They then evaluate twelve unique versions of Tmie, each containing mutations to different domains of Tmie. This analysis reveals that some mutations in Tmie cause dysfunctional gating of the channel as demonstrated through reduced hair cell activity, and that these same dysfunctional versions also display reduced Tmc expression at the normal site of the channel. These findings link hair cell activity with the levels of Tmc in the bundle, reinforcing the currently-debated notion that the Tmcs are the pore-forming subunits of the mechanotransduction channel. The authors conclude that Tmie, through distinct regions, is involved in both trafficking and stabilizing the Tmcs at the site of mechanotransduction.
]]></description>
<dc:creator>Pacentine, I.</dc:creator>
<dc:creator>Nicolson, T.</dc:creator>
<dc:date>2018-08-16</dc:date>
<dc:identifier>doi:10.1101/393330</dc:identifier>
<dc:title><![CDATA[Putative pore-forming subunits of the mechano-electrical transduction channel, Tmc1/2b, require Tmie to localize to the site of mechanotransduction in zebrafish sensory hair cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-08-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/396358v1?rss=1">
<title>
<![CDATA[
Probabilistic modeling of personalized drug combinations from integrated chemical screen and molecular data in sarcoma 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/396358v1?rss=1</link>
<description><![CDATA[
Cancer patients with advanced disease exhaust available clinical regimens and lack actionable genomic medicine results, leaving a large patient population without effective treatments options when their disease inevitably progresses. To address the unmet clinical need for evidence-based therapy assignment when standard clinical approaches have failed, we have developed a probabilistic computational modeling approach which integrates sequencing data with functional assay data to develop patient-specific combination cancer treatments. This computational modeling approach addresses three major challenges in personalized cancer therapy, which we validate across multiple species via computationally-designed personalized synergistic drug combination predictions, identification of unifying therapeutic targets to overcome intra-tumor heterogeneity, and mitigation of cancer cell resistance and rewiring mechanisms. These proof-of-concept studies support the use of an integrative functional approach to personalized combination therapy prediction for the population of high-risk cancer patients lacking viable clinical options and without actionable DNA sequencing-based therapy.
]]></description>
<dc:creator>Berlow, N. E.</dc:creator>
<dc:creator>Rikhi, R.</dc:creator>
<dc:creator>Geltzeiler, M. N.</dc:creator>
<dc:creator>Abraham, J.</dc:creator>
<dc:creator>Svalina, M. N.</dc:creator>
<dc:creator>Davis, L. E.</dc:creator>
<dc:creator>Wise, E.</dc:creator>
<dc:creator>Mancini, M.</dc:creator>
<dc:creator>Noujaim, J.</dc:creator>
<dc:creator>Mansoor, A.</dc:creator>
<dc:creator>Quist, M. J.</dc:creator>
<dc:creator>Matlock, K. L.</dc:creator>
<dc:creator>Goros, M. W.</dc:creator>
<dc:creator>Hernandez, B. S.</dc:creator>
<dc:creator>Duong, Y. C.</dc:creator>
<dc:creator>Thway, K.</dc:creator>
<dc:creator>Tsukahara, T.</dc:creator>
<dc:creator>Nishio, J.</dc:creator>
<dc:creator>Huang, E. C.</dc:creator>
<dc:creator>Airhart, S.</dc:creator>
<dc:creator>Bult, C. J.</dc:creator>
<dc:creator>Gandour-Edwards, R.</dc:creator>
<dc:creator>Maki, R. G.</dc:creator>
<dc:creator>Jones, R. L.</dc:creator>
<dc:creator>Michalek, J. E.</dc:creator>
<dc:creator>Milovancev, M.</dc:creator>
<dc:creator>Ghosh, S.</dc:creator>
<dc:creator>Pal, R.</dc:creator>
<dc:creator>Keller, C.</dc:creator>
<dc:date>2018-08-21</dc:date>
<dc:identifier>doi:10.1101/396358</dc:identifier>
<dc:title><![CDATA[Probabilistic modeling of personalized drug combinations from integrated chemical screen and molecular data in sarcoma]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-08-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/399410v1?rss=1">
<title>
<![CDATA[
Epigenetic factors coordinate intestinal development 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/399410v1?rss=1</link>
<description><![CDATA[
Intestinal epithelium development depends on epigenetic modifications, but whether that is also the case for other intestinal tract cell types remains unclear. We found that functional loss of a DNA methylation machinery component, ubiquitin-like protein containing PHD and RING finger domains 1 (uhrf1), leads to reduced enteric neuron number, changes in neuronal morphology, and severe intestinal smooth muscle disruption. Genetic chimeras revealed that Uhrf1 functions both cell-autonomously in enteric neuron progenitors and cell-non-autonomously in surrounding intestinal cells. Uhrf1 recruits the DNA methyltransferase Dnmt1 to unmethylated DNA during replication. Dnmt1 is also expressed in enteric neuron and smooth muscle progenitors. dnmt1 mutants show a strong reduction in enteric neuron number and disrupted intestinal smooth muscle. Because dnmt1;uhrf1 double mutants have a similar phenotype to dnmt1 and uhrf1 single mutants, Dnmt1 and Uhrf1 must function together during enteric neuron and intestinal muscle development. This work shows that genes controlling epigenetic modifications are important in coordinating intestinal tract development, provides the first demonstration that these genes are important in ENS development, and advances uhrf1 and dnmt1 as potential new Hirschsprung disease candidates.nnSummaryThis work provides evidence that DNA methylation factors are important in all cell types that contribute to development of a functional intestine.
]]></description>
<dc:creator>Ganz, J.</dc:creator>
<dc:creator>Melancon, E.</dc:creator>
<dc:creator>Wilson, C.</dc:creator>
<dc:creator>Amores, A.</dc:creator>
<dc:creator>Batzel, P.</dc:creator>
<dc:creator>Strader, M.</dc:creator>
<dc:creator>Braasch, I.</dc:creator>
<dc:creator>Diba, P.</dc:creator>
<dc:creator>Kuhlman, J. A.</dc:creator>
<dc:creator>Postlethwait, J. H.</dc:creator>
<dc:creator>Eisen, J. S.</dc:creator>
<dc:date>2018-08-23</dc:date>
<dc:identifier>doi:10.1101/399410</dc:identifier>
<dc:title><![CDATA[Epigenetic factors coordinate intestinal development]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-08-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/405811v1?rss=1">
<title>
<![CDATA[
Genetic control of the HDL proteome 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/405811v1?rss=1</link>
<description><![CDATA[
High-density lipoproteins (HDL) are nanoparticles with >80 associated proteins, phospholipids, cholesterol and cholesteryl esters. A comprehensive genetic analysis of the regulation of proteome of HDL isolated from a panel of 100 diverse inbred strains of mice, Hybrid Mouse Diversity Panel (HMDP), revealed widely varied HDL protein levels across the strains. Some of this variation was explained by local, cis-acting regulation, termed cis-protein quantitative trait loci. Variations in apolipoprotein A-II and apolipoprotein C-3 affected the abundance of multiple HDL proteins indicating a coordinated regulation. We identified modules of co-varying proteins and define a protein-protein interaction network describing the protein composition of the naturally occurring subspecies of HDL in mice. Sterol efflux capacity varied up to 3-fold across the strains and HDL proteins displayed distinct correlation patterns with macrophage and ABCA1 specific cholesterol efflux capacity and cholesterol exchange, suggesting that subspecies of HDL participate in discrete functions. The baseline and stimulated sterol efflux capacity phenotypes associated with distinct QTLs with smaller effect size suggesting a multi genetic regulation. Our results highlight the complexity of HDL particles by revealing high degree of heterogeneity and intercorrelation, some of which is associated with functional variation, supporting the concept that HDL-cholesterol alone is not an accurate measure of HDLs properties such as protection against CAD.
]]></description>
<dc:creator>Pamir, N.</dc:creator>
<dc:creator>Pan, C.</dc:creator>
<dc:creator>Plubell, D. L.</dc:creator>
<dc:creator>Hutchins, P. M.</dc:creator>
<dc:creator>Tang, C.</dc:creator>
<dc:creator>Wimberger, J.</dc:creator>
<dc:creator>Irwin, A.</dc:creator>
<dc:creator>Vallim, T. Q. d. A.</dc:creator>
<dc:creator>Heinecke, J. W.</dc:creator>
<dc:creator>Lusis, A. J.</dc:creator>
<dc:date>2018-08-31</dc:date>
<dc:identifier>doi:10.1101/405811</dc:identifier>
<dc:title><![CDATA[Genetic control of the HDL proteome]]></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/10.1101/406108v1?rss=1">
<title>
<![CDATA[
Aperture Phase Modulation with Adaptive Optics: A Novel Approach for Speckle Reduction and Structure Extraction in Optical Coherence Tomography 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/406108v1?rss=1</link>
<description><![CDATA[
AbstractSpeckle is an inevitable consequence of the use of coherent light in optical coherence tomography (OCT), and often acts as noise that obscures micro-structures of biological tissue. We here present a novel method of suppressing speckle noise intrinsically compatible with adaptive optics (AO) in OCT system: by modulating the phase inside the imaging system pupil aperture with a segmented deformable mirror, thus producing minor perturbations in the point spread function (PSF) to create un-correlated speckle pattern between B-scans, and further averaging to wash out the speckle but maintain the structures. It is a well-controlled and universal method which can efficiently determine the optimal range of phase modulation that minimizing speckle noise while maximizing image resolution and signal strength for different systems and/or samples. As an active method, its effectiveness and efficiency were demonstrated by both ex-vivo non-biological and in-vivo biological applications.
]]></description>
<dc:creator>Zhang, P.</dc:creator>
<dc:creator>Manna, S. K.</dc:creator>
<dc:creator>Miller, E. B.</dc:creator>
<dc:creator>Jian, Y.</dc:creator>
<dc:creator>Meleppat, R. K.</dc:creator>
<dc:creator>Sarunic, M. V.</dc:creator>
<dc:creator>Pugh, E. N.</dc:creator>
<dc:creator>Zawadzki, R. J.</dc:creator>
<dc:date>2018-08-31</dc:date>
<dc:identifier>doi:10.1101/406108</dc:identifier>
<dc:title><![CDATA[Aperture Phase Modulation with Adaptive Optics: A Novel Approach for Speckle Reduction and Structure Extraction in Optical Coherence Tomography]]></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/10.1101/407668v1?rss=1">
<title>
<![CDATA[
The accessible chromatin landscape of the hippocampus at single-cell resolution 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/407668v1?rss=1</link>
<description><![CDATA[
Here we present a comprehensive map of the accessible chromatin landscape of the mouse hippocampus at single-cell resolution. Substantial advances of this work include the optimization of single-cell combinatorial indexing assay for transposase accessible chromatin (sci-ATAC-seq), a software suite, scitools, for the rapid processing and visualization of single-cell combinatorial indexing datasets, and a valuable resource of hippocampal regulatory networks at single-cell resolution. We utilized sci-ATAC-seq to produce 2,346 high-quality single-cell chromatin accessibility maps with a mean unique read count per cell of 29,201 from both fresh and frozen hippocampi, observing little difference in accessibility patterns between the preparations. Using this dataset, we identified eight distinct major clusters of cells representing both neuronal and non-neuronal cell types and characterized the driving regulatory factors and differentially accessible loci that define each cluster. We then applied a recently described co-accessibility framework, Cicero, which identified 146,818 links between promoters and putative distal regulatory DNA. Identified co-accessibility networks showed cell-type specificity, shedding light on key dynamic loci that reconfigure to specify hippocampal cell lineages. Lastly, we carried out an additional sci-ATAC-seq preparation from cultured hippocampal neurons (899 high-quality cells, 43,532 mean unique reads) that revealed substantial alterations in their epigenetic landscape compared to nuclei from hippocampal tissue. This dataset and accompanying analysis tools provide a new resource that can guide subsequent studies of the hippocampus.
]]></description>
<dc:creator>Sinnamon, J. R.</dc:creator>
<dc:creator>Torkenczy, K. A.</dc:creator>
<dc:creator>Linhoff, M. W.</dc:creator>
<dc:creator>Vitak, S. A.</dc:creator>
<dc:creator>Pliner, H. A.</dc:creator>
<dc:creator>Trapnell, C.</dc:creator>
<dc:creator>Steemers, F. J.</dc:creator>
<dc:creator>Mandel, G.</dc:creator>
<dc:creator>Adey, A. C.</dc:creator>
<dc:date>2018-09-04</dc:date>
<dc:identifier>doi:10.1101/407668</dc:identifier>
<dc:title><![CDATA[The accessible chromatin landscape of the hippocampus at single-cell resolution]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-09-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/409219v1?rss=1">
<title>
<![CDATA[
Specific detection of cell-free DNA derived from intestinal epithelial cells using methylation patterns 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/409219v1?rss=1</link>
<description><![CDATA[
Epithelial cells of the intestine undergo rapid turnover and are thought to be cleared via stool. Disruption of tissue architecture, as occurs in colorectal cancer (CRC), results in the release of material from dying intestinal epithelial cells to blood. This phenomenon could be utilized for diagnosis and monitoring of intestinal diseases, if circulating cell-free DNA (cfDNA) derived from intestinal cells could be specifically identified. Here we describe two genomic loci that are unmethylated specifically in intestinal epithelial cells, allowing for sensitive and specific detection of DNA derived from such cells. As expected, intestinal DNA is found in stool, but not in plasma, of healthy individuals. Patients with inflammatory bowel disease (IBD) have minimal amounts of intestinal cfDNA in the plasma, whereas patients with advanced CRC show a strong signal. The intestinal markers are not elevated in plasma samples from patients with pancreatic ductal adenocarcinoma (PDAC), and a combination of intestine- and pancreas-specific markers allowed for robust differentiation between plasma cfDNA derived from CRC and PDAC patients. Intestinal DNA markers provide a mutation-independent tool for monitoring intestinal dynamics in health and disease.
]]></description>
<dc:creator>Lehmann-Werman, R.</dc:creator>
<dc:creator>Zick, A.</dc:creator>
<dc:creator>Paweletz, C.</dc:creator>
<dc:creator>Welch, M.</dc:creator>
<dc:creator>Hubert, A.</dc:creator>
<dc:creator>Maoz, M.</dc:creator>
<dc:creator>Davidy, T.</dc:creator>
<dc:creator>Magenheim, J.</dc:creator>
<dc:creator>Piyanzin, S.</dc:creator>
<dc:creator>Neiman, D.</dc:creator>
<dc:creator>Moss, J.</dc:creator>
<dc:creator>Golan, H.</dc:creator>
<dc:creator>Israeli, E.</dc:creator>
<dc:creator>Fischer, M.</dc:creator>
<dc:creator>Segal, E.</dc:creator>
<dc:creator>Grompe, M.</dc:creator>
<dc:creator>Pikarsky, A.</dc:creator>
<dc:creator>Golan, T.</dc:creator>
<dc:creator>Glaser, B.</dc:creator>
<dc:creator>Shemer, R.</dc:creator>
<dc:creator>Wolpin, B.</dc:creator>
<dc:creator>Dor, Y.</dc:creator>
<dc:date>2018-09-05</dc:date>
<dc:identifier>doi:10.1101/409219</dc:identifier>
<dc:title><![CDATA[Specific detection of cell-free DNA derived from intestinal epithelial cells using methylation patterns]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-09-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/409854v1?rss=1">
<title>
<![CDATA[
Maternal Circulating MicroRNAs Control the Placental Response to Alcohol 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/409854v1?rss=1</link>
<description><![CDATA[
Prenatal Alcohol exposure (PAE), like other pregnancy complications, can result in placental insufficiency and fetal growth restriction, though the linking causal mechanisms are unclear. We previously identified 11 gestationally-elevated maternal circulating miRNAs that predicted infant growth deficits following PAE. Here, we investigated whether these HEamiRNAs contribute to the pathology of PAE, by inhibiting trophoblast epithelial-mesenchymal transition (EMT), a pathway critical for placental development. We now report for the first time, that PAE inhibits expression of placental pro-EMT pathway members in both rodents and primates, and that HEamiRNAs collectively, but not individually, mediate placental EMT inhibition. HEamiRNAs collectively, but not individually, also inhibited cell proliferation and the EMT pathway in cultured trophoblasts, while inducing cell stress, and following trophoblast syncytialization, aberrant endocrine maturation. Moreover, a single intra-vascular administration of the pooled murine-expressed HEamiRNAs, to pregnant mice, decreased placental and fetal growth and inhibited expression of pro-EMT transcripts in placenta. Our data suggests that HEamiRNAs collectively interfere with placental development, contributing to the pathology of PAE, and perhaps also, to other causes of fetal growth restriction.

SummaryMaternal gestational circulating microRNAs, predictive of adverse infant outcomes including growth deficits, following prenatal alcohol exposure, contribute to placental pathology by impairing the EMT pathway in trophoblasts.
]]></description>
<dc:creator>Tseng, A. M.</dc:creator>
<dc:creator>Mahnke, A. H.</dc:creator>
<dc:creator>Wells, A. B.</dc:creator>
<dc:creator>Salem, N. A.</dc:creator>
<dc:creator>Allan, A. M.</dc:creator>
<dc:creator>Roberts, V. H. J.</dc:creator>
<dc:creator>Newman, N.</dc:creator>
<dc:creator>Walter, N. A. R.</dc:creator>
<dc:creator>Kroenke, C. D.</dc:creator>
<dc:creator>Grant, K. A.</dc:creator>
<dc:creator>Akison, L. K.</dc:creator>
<dc:creator>Moritz, K. M.</dc:creator>
<dc:creator>Chambers, C. D.</dc:creator>
<dc:creator>Miranda, R. C.</dc:creator>
<dc:date>2018-09-06</dc:date>
<dc:identifier>doi:10.1101/409854</dc:identifier>
<dc:title><![CDATA[Maternal Circulating MicroRNAs Control the Placental Response to Alcohol]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-09-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/410944v1?rss=1">
<title>
<![CDATA[
Dystroglycan is a scaffold for extracellular axon guidance decisions 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/410944v1?rss=1</link>
<description><![CDATA[
Axon guidance requires interactions between extracellular signaling molecules and transmembrane receptors, but how appropriate context-dependent decisions are coordinated outside the cell remains unclear. Here we show that the transmembrane glycoprotein Dystroglycan interacts with a changing set of environmental cues that regulate the trajectories of extending axons throughout the brain and spinal cord. Dystroglycan operates primarily as an extracellular scaffold during axon guidance, as it functions non-cell autonomously and does not require signaling through its intracellular domain. We identify the transmembrane receptor Celsr3/Adgrc3 as a binding partner for Dystroglycan, and show that this interaction is critical for specific axon guidance events in vivo. These findings establish Dystroglycan as a multifunctional scaffold that coordinates extracellular matrix proteins, secreted cues, and transmembrane receptors to regulate axon guidance.
]]></description>
<dc:creator>Lindenmaier, L. B.</dc:creator>
<dc:creator>Parmentier, N.</dc:creator>
<dc:creator>Guo, C.</dc:creator>
<dc:creator>Tissir, F.</dc:creator>
<dc:creator>Wright, K.</dc:creator>
<dc:date>2018-09-06</dc:date>
<dc:identifier>doi:10.1101/410944</dc:identifier>
<dc:title><![CDATA[Dystroglycan is a scaffold for extracellular axon guidance decisions]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-09-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/418129v1?rss=1">
<title>
<![CDATA[
neoepiscope Improves Neoepitope Prediction with Multi-variant Phasing 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/418129v1?rss=1</link>
<description><![CDATA[
The vast majority of tools for neoepitope prediction from DNA sequencing of complementary tumor and normal patient samples do not consider germline context or the potential for co-occurrence of two or more somatic variants on the same mRNA transcript. Without consideration of these phenomena, existing approaches are likely to produce both false positive and false negative results, resulting in an inaccurate and incomplete picture of the cancer neoepitope landscape. We developed neoepiscope chiefly to address this issue for single nucleotide variants (SNVs) and insertions/deletions (indels), and herein illustrate how germline and somatic variant phasing affects neoepitope prediction across multiple datasets. We estimate that up to [~]5% of neoepitopes arising from SNVs and indels may require variant phasing for their accurate assessment. neoepiscope is performant, flexible, and supports several major histocompatibility complex binding affinity prediction tools. We have released neoepiscope as open-source software (MIT license, https://github.com/pdxgx/neoepiscope) for broad use.nnKEY POINTSO_LIGermline context and somatic variant phasing are important for neoepitope predictionnC_LIO_LIMany popular neoepitope prediction tools have issues of performance and reproducibilitynC_LIO_LIWe describe and provide performant software for accurate neoepitope prediction from DNA-seq datanC_LI
]]></description>
<dc:creator>Wood, M. A.</dc:creator>
<dc:creator>Nguyen, A.</dc:creator>
<dc:creator>Struck, A. J.</dc:creator>
<dc:creator>Ellrott, K.</dc:creator>
<dc:creator>Nellore, A.</dc:creator>
<dc:creator>Thompson, R. F.</dc:creator>
<dc:date>2018-09-14</dc:date>
<dc:identifier>doi:10.1101/418129</dc:identifier>
<dc:title><![CDATA[neoepiscope Improves Neoepitope Prediction with Multi-variant Phasing]]></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/10.1101/419606v1?rss=1">
<title>
<![CDATA[
Mutual transcriptional repression between Gli3 and Hox13 genes determines the anterior-posterior asymmetry of the autopod 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/419606v1?rss=1</link>
<description><![CDATA[
In the present study we have investigated the molecular causes of the absence of digit 1 in the Hoxa13 mutant and why the absence of Hoxa13 protein, whose expression spans the entire autopod, specifically impacts the anterior-most digit. We show that in the absence of Hoxa13, the expression of Hoxd13 does not extend into the anterior mesoderm consequently leaving the presumptive territory of digit1 devoid of distal Hox expression and providing an explanation for the agenesis of digit 1. We provide compelling evidence that the lack of Hoxd13 transcription in the anterior mesoderm is due to increased Gli3R activity, in turn resulting from the loss of transcriptional repression exerted by Hoxa13 on Gli3. Our results are compatible with a mutual transcriptional repression between Gli3 and Hox13 genes that determines the anterior-posterior asymmetry of the autopod.
]]></description>
<dc:creator>Bastida, M. F.</dc:creator>
<dc:creator>Perez-Gomez, R.</dc:creator>
<dc:creator>Trofka, A.</dc:creator>
<dc:creator>Sheth, R.</dc:creator>
<dc:creator>Stadler, H. S.</dc:creator>
<dc:creator>Mackem, S.</dc:creator>
<dc:creator>Ros, M. A.</dc:creator>
<dc:date>2018-09-17</dc:date>
<dc:identifier>doi:10.1101/419606</dc:identifier>
<dc:title><![CDATA[Mutual transcriptional repression between Gli3 and Hox13 genes determines the anterior-posterior asymmetry of the autopod]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-09-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/420406v1?rss=1">
<title>
<![CDATA[
Bidirectional Control of Coronary Vascular Resistance by Eicosanoids via a Novel GPCR 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/420406v1?rss=1</link>
<description><![CDATA[
Arachidonic acid metabolites epoxyeicosatrienoates (EETs) and hydroxyeicosatetraenoates (HETEs) are important regulators of myocardial blood flow and coronary vascular resistance (CVR), but their mechanisms of action are not fully understood. We identified G protein-coupled receptor 39 (GPR39) as a microvascular smooth muscle cell (mVSMC) receptor antagonistically regulated by two endogenous eicosanoids: 15-HETE, which stimulates GPR39 to increase mVSMC intracellular calcium and augment microvascular CVR, and 14,15-EET, which inhibits these actions. Furthermore, zinc ion acts as an allosteric modulator of GPR39 to potentiate the efficacy of the two ligands. Our findings will have a major impact on understanding the roles of eicosanoids in cardiovascular physiology and disease, and provide an opportunity for the development of novel GPR39-targeting therapies for cardiovascular disease.nnOne Sentence SummaryGPR39 is a microvascular smooth muscle cell receptor regulated by two vasoactive eicosanoids with opposing actions.
]]></description>
<dc:creator>Alkayed, N. J.</dc:creator>
<dc:creator>Cao, Z.</dc:creator>
<dc:creator>Qian, Z. Y.</dc:creator>
<dc:creator>Nagarajan, S.</dc:creator>
<dc:creator>Liu, X.</dc:creator>
<dc:creator>Nelson, J.</dc:creator>
<dc:creator>Xie, F.</dc:creator>
<dc:creator>Li, B.</dc:creator>
<dc:creator>Fan, W.</dc:creator>
<dc:creator>Liu, L.</dc:creator>
<dc:creator>Grafe, M. R.</dc:creator>
<dc:creator>Xiao, X.</dc:creator>
<dc:creator>Barnes, A. P.</dc:creator>
<dc:creator>Kaul, S.</dc:creator>
<dc:date>2018-09-19</dc:date>
<dc:identifier>doi:10.1101/420406</dc:identifier>
<dc:title><![CDATA[Bidirectional Control of Coronary Vascular Resistance by Eicosanoids via a Novel GPCR]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-09-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/424432v1?rss=1">
<title>
<![CDATA[
A specific Ret receptor isoform is required for pioneer axon outgrowth and growth cone dynamics 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/424432v1?rss=1</link>
<description><![CDATA[
In many cases, axon growth and guidance are driven by pioneer axons, the first axons to grow in a particular region. Despite their dynamic pathfinding capabilities and developmental importance, there are very few pioneer neuron specific markers and thus their in vivo identification and functional interrogation have been difficult. We found that a Ret receptor isoform, Ret51, is highly enriched in peripheral sensory pioneer neurons and is required for pioneer axon outgrowth. Ret null mutant pioneer neurons differentiate normally; however, they displayed defects in growth cone morphology and formation of filopodia before pioneer axon extension prematurely halts. We also demonstrate loss-of-function of a retrograde cargo adaptor, JNK-interacting protein 3 (Jip3), phenocopied many of these axonal defects. We further found that loss of Jip3 led to accumulation of activated Ret receptor in pioneer growth cones, indicating a failure in the clearance of activated Ret from growth cones. Using an axon sever approach as well as in vivo analysis of axonal transport, we showed Jip3 specifically mediates retrograde, but not anterograde, transport of activated Ret51. Finally, live imaging revealed that Jip3 and Ret51 were retrogradely co-transported in pioneer axons, suggesting Jip3 functions as an adapter for retrograde transport of Ret51. Taken together, these results identify Ret51 as a molecular marker of pioneer neurons and elucidate an important isoform-specific role for Ret51 in axon growth and growth cone dynamics during development.
]]></description>
<dc:creator>Tuttle, A. M.</dc:creator>
<dc:creator>Drerup, C. M.</dc:creator>
<dc:creator>Marra, M. H.</dc:creator>
<dc:creator>Nechiporuk, A. V.</dc:creator>
<dc:date>2018-09-21</dc:date>
<dc:identifier>doi:10.1101/424432</dc:identifier>
<dc:title><![CDATA[A specific Ret receptor isoform is required for pioneer axon outgrowth and growth cone dynamics]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-09-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/424952v1?rss=1">
<title>
<![CDATA[
Middle-way flexible docking: Pose prediction using mixed-resolution Monte Carlo in estrogen receptor α 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/424952v1?rss=1</link>
<description><![CDATA[
There is a vast gulf between the two primary strategies for simulating protein-ligand interactions. Docking methods significantly limit or eliminate protein flexibility to gain great speed at the price of uncontrolled inaccuracy, whereas fully flexible atomistic molecular dynamics simulations are expensive and often suffer from limited sampling. We have developed a flexible docking approach geared especially for highly flexible or poorly resolved targets based on mixed-resolution Monte Carlo (MRMC), which is intended to offer a balance among speed, protein flexibility, and sampling power. The binding region of the protein is treated with a standard atomistic force field, while the remainder of the protein is modeled at the residue level with a G[o] model that permits protein flexibility while saving computational cost. Implicit solvation is used. Here we assess three facets of the MRMC approach with implications for other docking studies: (i) the role of receptor flexibility in cross-docking pose prediction; (ii) the use of non-equilibrium candidate Monte Carlo (NCMC) and (iii) the use of pose-clustering in scoring. We examine 61 co-crystallized ligands of estrogen receptor , an important cancer target known for its flexibility. We also compare the performance of the MRMC approach with Autodock smina, a docking program. [1] Adding protein flexibility, not surprisingly, leads to significantly lower total energies and stronger interactions between protein and ligand, but notably we document the important role of backbone flexibility in the improvement. The improved backbone flexibility also leads to improved performance relative to smina. Somewhat unexpectedly, our implementation of NCMC leads to only modestly improved sampling of ligand poses. Overall, the addition of protein flexibility improves the performance of docking, as measured by energy-ranked poses, but we do not find significant improvements based on cluster information or the use of NCMC.
]]></description>
<dc:creator>Spiriti, J.</dc:creator>
<dc:creator>Subramanian, S. R.</dc:creator>
<dc:creator>Palli, R.</dc:creator>
<dc:creator>Wu, M.</dc:creator>
<dc:creator>Zuckerman, D. M.</dc:creator>
<dc:date>2018-09-24</dc:date>
<dc:identifier>doi:10.1101/424952</dc:identifier>
<dc:title><![CDATA[Middle-way flexible docking: Pose prediction using mixed-resolution Monte Carlo in estrogen receptor α]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-09-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/427393v1?rss=1">
<title>
<![CDATA[
Computational estimation of ms-sec atomistic folding times 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/427393v1?rss=1</link>
<description><![CDATA[
Despite the development of massively parallel computing hardware including inexpensive graphics processing units (GPUs), it has remained infeasible to simulate the folding of atomistic proteins at room temperature using conventional molecular dynamics (MD) beyond the s scale. Here we report the folding of atomistic, implicitly solvated protein systems with folding times {tau}f ranging from ~100 s to ~10s using the weighted ensemble (WE) strategy in combination with GPU computing. Starting from an initial structure or set of structures, WE organizes an ensemble of GPU-accelerated MD trajectory segments via intermittent pruning and replication events to generate statistically unbiased estimates of rate constants for rare events such as folding; no biasing forces are used. Although the variance among atomistic WE folding runs is significant, multiple independent runs are used to reduce and quantify statistical uncertainty. Three systems were examined: NTL9 at low solvent viscosity (yielding {tau}f ~ 5s), NTL9 at water-like viscosity ({tau}f ~ 40s), and Protein G at low viscosity ({tau}f ~ 10s). In all cases the folding time, uncertainty, and ensemble properties could be estimated from WE simulation; for protein G, this characterization required significantly less overall computing than would be required to observe a single folding event with conventional MD simulations. Our results suggest discrepancies with experimental folding times that should enable improvement of force fields and solvent models.
]]></description>
<dc:creator>Adhikari, U.</dc:creator>
<dc:creator>Mostofian, B.</dc:creator>
<dc:creator>Petersen, A.</dc:creator>
<dc:creator>Zuckerman, D. M.</dc:creator>
<dc:date>2018-09-29</dc:date>
<dc:identifier>doi:10.1101/427393</dc:identifier>
<dc:title><![CDATA[Computational estimation of ms-sec atomistic folding times]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-09-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/429373v1?rss=1">
<title>
<![CDATA[
Network stress test reveals novel drug potentiators in Mycobacterium tuberculosis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/429373v1?rss=1</link>
<description><![CDATA[
Deciphering molecular stress response is highly relevant to studies of microbes such as Mycobacterium tuberculosis (MTB), the causative pathogen of tuberculosis (TB) which sickens 10 million people and kills 1.8 million each year1. Prolonged therapy and unfavorable outcomes arise partially because MTB has evolved stress responses to achieve tolerance, wherein MTB persists in otherwise inhibitory drug concentrations by means independent of heritable resistance mutations2-4. Understanding these adaptations and how they are regulated can reveal new biology, including unexplored drug targets and treatment-enhancing strategies. Here, we present a novel network-based genetic screening approach: the Transcriptional Regulator Induced Phenotype (TRIP) screen, which we used to identify previously uncharacterized MTB network adaptations to the first-line drug isoniazid (INH). We found regulators that alter IN ...
]]></description>
<dc:creator>Ma, S.</dc:creator>
<dc:creator>Morrison, R.</dc:creator>
<dc:creator>Hobbs, S. J.</dc:creator>
<dc:creator>Farrow-Johnson, J.</dc:creator>
<dc:creator>Rustad, T. R.</dc:creator>
<dc:creator>Sherman, D. R.</dc:creator>
<dc:date>2018-09-27</dc:date>
<dc:identifier>doi:10.1101/429373</dc:identifier>
<dc:title><![CDATA[Network stress test reveals novel drug potentiators in Mycobacterium tuberculosis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-09-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/432252v1?rss=1">
<title>
<![CDATA[
Targeting mitochondria in cancer therapy could provide a basis for the selective anti-cancer activity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/432252v1?rss=1</link>
<description><![CDATA[
To determine the target of the recently identified lead compound NSC130362 that is responsible for its selective anti-cancer efficacy and safety in normal cells, structure-activity relationship (SAR) studies were conducted. First, NSC13062 was validated as a starting compound for the described SAR studies in a variety of cell-based viability assays. Then, a small library of 1,4-naphthoquinines (1,4-NQs) and quinoline-5,8-diones was tested in cell viability assays using pancreatic cancer MIA PaCa-2 cells and normal human hepatocytes. The obtained data allowed us to select a set of both non-toxic compounds that preferentially induced apoptosis in cancer cells and toxic compounds that induced apoptosis in both cancer and normal cells. Anti-cancer activity of the selected non-toxic compounds was confirmed in viability assays using breast cancer HCC1187 cells. Consequently, the two sets of compounds were tested in multiple cell-based and in vitro activity assays to identify key factors responsible for the observed activity. Inhibition of the mitochondrial electron transfer chain (ETC) is a key distinguishing activity between the non-toxic and toxic compounds. Finally, we developed a mathematical model that was able to distinguish these two sets of compounds. The development of this model supports our conclusion that appropriate quantitative SAR (QSAR) models have the potential to be employed to develop anticancer compounds with improved potency while maintaining non-toxicity to normal cells.
]]></description>
<dc:creator>Rozanov, D.</dc:creator>
<dc:creator>Cheltsov, A.</dc:creator>
<dc:creator>Nilsen, A.</dc:creator>
<dc:creator>Boniface, C.</dc:creator>
<dc:creator>Korkola, J.</dc:creator>
<dc:creator>Gray, J.</dc:creator>
<dc:creator>Tyner, J.</dc:creator>
<dc:creator>Tognon, C. E.</dc:creator>
<dc:creator>Mills, G. B.</dc:creator>
<dc:creator>Spellman, P.</dc:creator>
<dc:date>2018-10-01</dc:date>
<dc:identifier>doi:10.1101/432252</dc:identifier>
<dc:title><![CDATA[Targeting mitochondria in cancer therapy could provide a basis for the selective anti-cancer activity]]></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/10.1101/435784v1?rss=1">
<title>
<![CDATA[
TrkB activation during a critical period mimics the protective effects of early visual experience on the stability of receptive fields in adult superior colliculus 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/435784v1?rss=1</link>
<description><![CDATA[
During a critical period in postnatal development, spontaneous and evoked retinal activity shape nascent visual pathways in an adaptive fashion. Visual experience increases transcription of the neurotrophin BDNF, activating the BDNF receptor TrkB, which promotes maturation of parvalbumin (PV) positive inhibitory interneurons, a process thought to open a critical period for ocular dominance plasticity in visual cortex. Development of perineuronal nets around PV neurons limits plasticity, ending the critical period and restricting adult plasticity. Another form of critical period plasticity is receptive field (RF) refinement. Spontaneous activity alone is sufficient for spatial refinement of visual receptive fields in superior colliculus (SC) and visual cortex (V1), but visual experience during an early critical period is necessary to maintain inhibitory synapses and stabilize RFs in adulthood (Carrasco et al. 2005, 2011; Carrasco & Pallas 2006; Balmer & Pallas 2015a). We report here that deprivation-induced RF enlargement in adulthood has a behavioral consequence; it impairs fear responses to looming objects in mice and hamsters. The mechanism through which early experience protects RFs from deprivation-induced loss of inhibition in adulthood is unknown. Given that the loss of RF refinement in SC does not occur until adulthood, and that inhibitory PV neurons and perineuronal nets are rare in SC, we asked whether or not BDNF-TrkB signaling was involved. We find that early TrkB activation is necessary and sufficient to maintain visual RF refinement in adulthood, suggesting a common signaling pathway for maturation of inhibition across neuronal subtypes and locations within the visual pathway.nnSignificance StatementReceptive field refinement in superior colliculus (SC) differs from more commonly studied examples of critical period plasticity in visual pathways in that it does not require visual experience to occur; rather spontaneous activity is sufficient. Maintenance of refinement requires brief, early exposure to light to stabilize inhibition beyond puberty. This type of inhibitory plasticity must not depend on parvalbumin (PV)-containing GABAergic interneurons or on the formation of perineuronal nets, because these are very uncommon in SC. Nonetheless, we find that TrkB activation during a critical period can substitute for visual experience in maintaining receptive field refinement into adulthood, and that this maintenance is beneficial to visual survival behaviors. Thus, multiple types of plasticity converge on the same neurotrophin-dependent signaling cascade.
]]></description>
<dc:creator>Mudd, D. B.</dc:creator>
<dc:creator>Balmer, T. S.</dc:creator>
<dc:creator>Kim, S. Y.</dc:creator>
<dc:creator>Machhour, N.</dc:creator>
<dc:creator>Pallas, S. L.</dc:creator>
<dc:date>2018-10-04</dc:date>
<dc:identifier>doi:10.1101/435784</dc:identifier>
<dc:title><![CDATA[TrkB activation during a critical period mimics the protective effects of early visual experience on the stability of receptive fields in adult superior colliculus]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-10-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/442699v1?rss=1">
<title>
<![CDATA[
DNA damage dependent hypomethylation regulates the pro-angiogenic LncRNA MEG9 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/442699v1?rss=1</link>
<description><![CDATA[
Changes in gene expression are key for the cells to adapt and response to intrinsic and extrinsic stimulus. It has been shown that genotoxic stress induces global hypomethylation as a result of decreased expression of DNA methyl transferases (DNMT). We hypothesized that DNA damage suppresses long non-coding RNA expression in the vasculature via DNA methylation leading to more robust DNA repair/survival or cellular senescence/death cell fate decisions. We show here that ionizing radiation reduces the expression of DNMTs in the vascular endothelium and this leads to increased expression of the anti-apoptotic lncRNA MEG9. MEG9 is a lncRNA from the DLK1-DIO3 ncRNA cluster. Loss-of-function studies using RNA gapmers indicate that MEG9 protects endothelial cells from DNA damage induced cell death. Consistent with this phenotype, knockdown of MEG9 decreases growth factor dependent angiogenesis in a 3D fibrin gel angiogenesis assay. Mechanistically, we observed that MEG9 knockdown decreased the expression of cell survival genes including survivin and induced the expression of pro-apoptotic genes such as Bad/Bax. Taken together, our findings illustrate how DNA methylation at selective lncRNA loci can regulate their expression and drive endothelial cell fate decisions.
]]></description>
<dc:creator>Espinosa-Diez, C.</dc:creator>
<dc:creator>Wilson, R.</dc:creator>
<dc:creator>Mukherjee, R.</dc:creator>
<dc:creator>Feltham, M.</dc:creator>
<dc:creator>Hudson, C.</dc:creator>
<dc:creator>Ruhl, R.</dc:creator>
<dc:creator>Anand, S.</dc:creator>
<dc:date>2018-10-14</dc:date>
<dc:identifier>doi:10.1101/442699</dc:identifier>
<dc:title><![CDATA[DNA damage dependent hypomethylation regulates the pro-angiogenic LncRNA MEG9]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-10-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/452862v1?rss=1">
<title>
<![CDATA[
Dopamine modulation of prefrontal cortex activity is manifold and operates at multiple temporal and spatial scales 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/452862v1?rss=1</link>
<description><![CDATA[
While the function of dopamine in subcortical structures is largely limited to reward and movement, dopamine neurotransmission in the prefrontal cortex (PFC) is critical to a multitude of temporally and functionally diverse processes such as attention, working memory, behavioral flexibility, action selection, and stress adaptation. How does dopamine influence PFC computation of multiple temporally diverse functions? Here we find causation between sustained and burst patterns of phasic dopamine neuron activation and contemporaneous modulation of PFC neuronal activity at multiple spatio-temporal scales. These include a multidirectional and weak impact on individual PFC neuron rate activity and a robust influence on coordinated ensemble activity, gamma oscillations, and gamma-theta coupling that persisted for minutes. In addition, PFC network responses to burst pattern of dopamine firing were selectively strengthened in behaviorally active states. Thus, dopamine modulation of PFC is spatiotemporally diverse and is dictated by the pattern of dopamine neuron activation and behavioral state. These findings provide insight on the multiplex pattern of modulation by dopamine that may influence PFC computation of temporally diverse functions.
]]></description>
<dc:creator>Lohani, S.</dc:creator>
<dc:creator>Martig, A. K.</dc:creator>
<dc:creator>Deisseroth, K.</dc:creator>
<dc:creator>Witten, I.</dc:creator>
<dc:creator>Moghaddam, B.</dc:creator>
<dc:date>2018-10-25</dc:date>
<dc:identifier>doi:10.1101/452862</dc:identifier>
<dc:title><![CDATA[Dopamine modulation of prefrontal cortex activity is manifold and operates at multiple temporal and spatial scales]]></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/10.1101/453746v1?rss=1">
<title>
<![CDATA[
Cyclic, condition-independent activity in primary motor cortex predicts corrective movement behavior 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/453746v1?rss=1</link>
<description><![CDATA[
Reaching movements are known to have large condition-independent neural activity and cyclic neural dynamics. A new precision center-out task was performed by rhesus macaques to test the hypothesis that cyclic, condition-independent neural activity in the primary motor cortex (M1) occurs not only during initial reaching movements but also during subsequent corrective movements. Corrective movements were observed to be discrete with time courses and bell-shaped speed profiles similar to the initial movements. Condition-independent cyclic neural trajectories were similar and repeated for initial and each additional corrective submovement. The phase of the cyclic condition-independent neural activity predicted the time of peak movement speed more accurately than regression of instantaneous firing rate, even when the subject made multiple corrective movements. Rather than being controlled as continuations of the initial reach, a discrete cycle of motor cortex activity encodes each corrective submovement.

Significance StatementDuring a precision center-out task, initial and subsequent corrective movements occur as discrete submovements with bell-shaped speed profiles. A cycle of condition-independent activity in primary motor cortex neuron populations corresponds to each submovement, such that the phase of this cyclic activity predicts the time of peak speeds--both initial and corrective. These submovements accompanied by cyclic neural activity offer important clues into how we successfully execute precise, corrective reaching movements and may have implications for optimizing control of brain-computer interfaces.
]]></description>
<dc:creator>Rouse, A. G.</dc:creator>
<dc:date>2018-10-25</dc:date>
<dc:identifier>doi:10.1101/453746</dc:identifier>
<dc:title><![CDATA[Cyclic, condition-independent activity in primary motor cortex predicts corrective movement behavior]]></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/10.1101/457739v1?rss=1">
<title>
<![CDATA[
Image processing and analysis methods for the Adolescent Brain Cognitive Development Study 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/457739v1?rss=1</link>
<description><![CDATA[
The Adolescent Brain Cognitive Development (ABCD) Study is an ongoing, nationwide study of the effects of environmental influences on behavioral and brain development in adolescents. The ABCD Study is a collaborative effort, including a Coordinating Center, 21 data acquisition sites across the United States, and a Data Analysis and Informatics Center (DAIC). The main objective of the study is to recruit and assess over eleven thousand 9-10-year-olds and follow them over the course of 10 years to characterize normative brain and cognitive development, the many factors that influence brain development, and the effects of those factors on mental health and other outcomes. The study employs state-of-the-art multimodal brain imaging, cognitive and clinical assessments, bioassays, and careful assessment of substance use, environment, psychopathological symptoms, and social functioning. The data will provide a resource of unprecedented scale and depth for studying typical and atypical development. Here, we describe the baseline neuroimaging processing and subject-level analysis methods used by the ABCD DAIC in the centralized processing and extraction of neuroanatomical and functional imaging phenotypes. Neuroimaging processing and analyses include modality-specific corrections for distortions and motion, brain segmentation and cortical surface reconstruction derived from structural magnetic resonance imaging (sMRI), analysis of brain microstructure using diffusion MRI (dMRI), task-related analysis of functional MRI (fMRI), and functional connectivity analysis of resting-state fMRI.nnHighlightsO_LIAn overview of the MRI processing pipeline for the ABCD StudynC_LIO_LIA discussion on the challenges of large, multisite population studiesnC_LIO_LIA methodological reference for users of publicly shared data from the ABCD StudynC_LI
]]></description>
<dc:creator>Hagler, D. J.</dc:creator>
<dc:creator>Hatton, S. N.</dc:creator>
<dc:creator>Makowski, C.</dc:creator>
<dc:creator>Cornejo, M. D.</dc:creator>
<dc:creator>Fair, D. A.</dc:creator>
<dc:creator>Dick, A. S.</dc:creator>
<dc:creator>Sutherland, M. T.</dc:creator>
<dc:creator>Casey, B. J.</dc:creator>
<dc:creator>Barch, D. M.</dc:creator>
<dc:creator>Harms, M. P.</dc:creator>
<dc:creator>Watts, R.</dc:creator>
<dc:creator>Bjork, J. M.</dc:creator>
<dc:creator>Garavan, H. P.</dc:creator>
<dc:creator>Hilmer, L.</dc:creator>
<dc:creator>Pung, C. J.</dc:creator>
<dc:creator>Sicat, C. S.</dc:creator>
<dc:creator>Kuperman, J.</dc:creator>
<dc:creator>Bartsch, H.</dc:creator>
<dc:creator>Xue, F.</dc:creator>
<dc:creator>Heitzeg, M. M.</dc:creator>
<dc:creator>Laird, A. R.</dc:creator>
<dc:creator>Trinh, T. T.</dc:creator>
<dc:creator>Gonzalez, R.</dc:creator>
<dc:creator>Tapert, S. F.</dc:creator>
<dc:creator>Riedel, M. C.</dc:creator>
<dc:creator>Squeglia, L. M.</dc:creator>
<dc:creator>Hyde, L. W.</dc:creator>
<dc:creator>Rosenberg, M. D.</dc:creator>
<dc:creator>Earl, E. A.</dc:creator>
<dc:creator>Howlett, K. D.</dc:creator>
<dc:creator>Baker, F. C.</dc:creator>
<dc:creator>Soules, M.</dc:creator>
<dc:creator>Diaz, J.</dc:creator>
<dc:creator>Ruiz de Leon, O.</dc:creator>
<dc:creator>Thompson, W. K.</dc:creator>
<dc:creator>Neale, M. C.</dc:creator>
<dc:creator>Herting, M.</dc:creator>
<dc:creator>Sowell, E. R.</dc:creator>
<dc:creator>Alvarez, R. P.</dc:creator>
<dc:creator>Hawes, S. W.</dc:creator>
<dc:creator>Sanchez, M.</dc:creator>
<dc:creator>Bodurka, J.</dc:creator>
<dc:creator>Bre</dc:creator>
<dc:date>2018-11-04</dc:date>
<dc:identifier>doi:10.1101/457739</dc:identifier>
<dc:title><![CDATA[Image processing and analysis methods for the Adolescent Brain Cognitive Development Study]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-11-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/466300v1?rss=1">
<title>
<![CDATA[
Early detonation by sprouted mossy fibers enables aberrant dentate network activity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/466300v1?rss=1</link>
<description><![CDATA[
In temporal lobe epilepsy, sprouting of hippocampal mossy fiber axons onto dentate granule cell dendrites creates a recurrent excitatory network. However, unlike mossy fibers projecting to CA3, sprouted mossy fiber synapses depress upon repetitive activation. Thus, despite their proximal location, large presynaptic terminals, and ability to excite target neurons, the impact of sprouted mossy fiber synapses on hippocampal hyperexcitability is unclear. We find that despite their short-term depression, single episodes of sprouted mossy fiber activation in hippocampal slices initiated bursts of recurrent polysynaptic excitation. Consistent with a contribution to network hyperexcitability, optogenetic activation of sprouted mossy fibers reliably triggered action potential firing in postsynaptic dentate granule cells after single light pulses. This pattern resulted in a shift in network recruitment dynamics to an "early detonation" mode and an increased probability of release compared to mossy fiber synapses in CA3. A lack of tonic adenosine-mediated inhibition contributed to the higher probability of glutamate release thus facilitating reverberant circuit activity.nnSignificance StatementSprouted mossy fibers are one of the hallmark histopathological findings in temporal lobe epilepsy. These fibers form recurrent excitatory synapses onto other dentate granule cells that display profound short-term depression. Here, however, we show that although these sprouted mossy fibers weaken substantially during repetitive activation, their initial high probability of glutamate release can activate reverberant network activity. Furthermore, we find that a lack of tonic adenosine inhibition enables this high probability of release and, consequently, recurrent network activity.
]]></description>
<dc:creator>Hendricks, W. D.</dc:creator>
<dc:creator>Westbrook, G. L.</dc:creator>
<dc:creator>Schnell, E.</dc:creator>
<dc:date>2018-11-08</dc:date>
<dc:identifier>doi:10.1101/466300</dc:identifier>
<dc:title><![CDATA[Early detonation by sprouted mossy fibers enables aberrant dentate network activity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-11-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/467902v1?rss=1">
<title>
<![CDATA[
Anti-NMDA receptor encephalitis in mice induced by active immunization with conformationally-stabilized holoreceptors 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/467902v1?rss=1</link>
<description><![CDATA[
Autoimmunity to membrane proteins in the central nervous system has been increasingly recognized as a cause of neuropsychiatric disease. A key recent development was the discovery of antibodies to NMDA receptors in limbic encephalitis, characterized by cognitive changes, memory loss, seizures and sometimes long-term morbidity or mortality. Treatment approaches and experimental studies have largely focused on the pathogenic role of these autoantibodies. Passive antibody transfer to mice has provided useful insights, but does not produce the full spectrum of the human disease. Here we describe a de novo autoimmune mouse model of anti-NMDA receptor encephalitis. Active immunization of immune competent mice with conformationally-stabilized, native-like NMDA receptors induced a fulminant encephalitis that was strikingly similar to the behavioral and pathologic characteristics of human cases. Our results provide evidence of neuroinflammation and immune cell infiltration as early and robust features of the autoimmune response. Use of transgenic mice indicated that mature T cells as well as antibody-producing cells were required for disease induction. Our results provide new insights into disease pathogenesis as well as a platform for testing mechanisms of disease initiation and therapeutic approaches.nnOne Sentence SummaryWe report an active immunization model of anti-NMDA receptor encephalitis in mice that recapitulates the features of the clinical disease, provides new insights into the pathophysiology, and offers a platform for investigation of new therapeutic interventions.
]]></description>
<dc:creator>Jones, B. E.</dc:creator>
<dc:creator>Tovar, K. R.</dc:creator>
<dc:creator>Goehring, A.</dc:creator>
<dc:creator>Okada, N. J.</dc:creator>
<dc:creator>Gouaux, E.</dc:creator>
<dc:creator>Westbrook, G. L.</dc:creator>
<dc:date>2018-11-10</dc:date>
<dc:identifier>doi:10.1101/467902</dc:identifier>
<dc:title><![CDATA[Anti-NMDA receptor encephalitis in mice induced by active immunization with conformationally-stabilized holoreceptors]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-11-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/491068v1?rss=1">
<title>
<![CDATA[
Identification of A Disintegrin and Metalloproteinase 9 domain (ADAM9) required in the early stages of encephalomyocarditis virus infection 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/491068v1?rss=1</link>
<description><![CDATA[
Encephalomyocarditis virus (EMCV) is a picornavirus that produces lytic infections in murine and human cells. Employing a genome-wide CRISPR-Cas9 knockout screen to find host factors required for EMCV infection, we identified a role for ADAM9 in EMCV infection. CRISPR-mediated deletion of ADAM9 in multiple human cell lines rendered the cells highly resistant to EMCV infection and cell death. Primary fibroblasts from ADAM9 KO mice were also strongly resistant to EMCV infection and cell death. In contrast, ADAM9 KO and WT cells were equally susceptible to infection with other viruses, including the picornavirus Coxsackie virus B. ADAM9 KO cells failed to produce viral progeny when incubated with EMCV. However, bypassing EMCV entry into cells through delivery of viral RNA directly to the cytosol yielded infectious EMCV virions from ADAM9 KO cells, suggesting that ADAM9 is not required for EMCV replication post-entry. These findings establish that ADAM9 is required for the early stage of EMCV infection, likely for virus entry or viral genome delivery to the cytosol.nnImportanceViral myocarditis is a leading cause of death in the U.S., contributing to numerous unexplained deaths in people [&le;] 35 years old. Enteroviruses contribute to many cases of human myocarditis. Encephalomyocarditis virus (EMCV) infection causes viral myocarditis in rodent models but its receptor requirements have not been fully identified. CRISPR-Cas9 screens can identify host dependency factors essential for EMCV infection and enhance our understanding of key events that follow viral infection, potentially leading to new strategies for preventing viral myocarditis. Using a CRISPR-Cas9 screen, we identified A Disintegrin and Metalloproteinase 9 Domain (ADAM9) as a major factor required for the early stages of EMCV infection in both human and murine infection.
]]></description>
<dc:creator>Bazzone, L. E.</dc:creator>
<dc:creator>King, M.</dc:creator>
<dc:creator>MacKay, C. R.</dc:creator>
<dc:creator>Kyawe, P. P.</dc:creator>
<dc:creator>Meraner, P.</dc:creator>
<dc:creator>Lindstrom, D.</dc:creator>
<dc:creator>Rojas-Quintero, J.</dc:creator>
<dc:creator>Owen, C. A.</dc:creator>
<dc:creator>Wang, J. P.</dc:creator>
<dc:creator>Brass, A. L.</dc:creator>
<dc:creator>Kurt-Jones, E. A.</dc:creator>
<dc:creator>Finberg, R. W.</dc:creator>
<dc:date>2018-12-09</dc:date>
<dc:identifier>doi:10.1101/491068</dc:identifier>
<dc:title><![CDATA[Identification of A Disintegrin and Metalloproteinase 9 domain (ADAM9) required in the early stages of encephalomyocarditis virus infection]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-12-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/504290v1?rss=1">
<title>
<![CDATA[
Convergent Allostery in Ribonucleotide Reductase 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/504290v1?rss=1</link>
<description><![CDATA[
Ribonucleotide reductases (RNRs) use a conserved radical-based mechanism to catalyze the conversion of ribonucleotides to deoxyribonucleotides. Within the RNR family, class Ib RNRs are notable for being largely restricted to bacteria, including many pathogens, and for lacking an evolutionarily mobile ATP-cone domain that allosterically controls overall activity. In this study, we report the emergence of a new and unexpected mechanism of activity regulation in the sole RNR of the model organism Bacillus subtilis. Using a hypothesis-driven structural approach that combines the strengths of small-angle X-ray scattering (SAXS), crystallography, and cryo-electron microscopy (cryo-EM), we describe the reversible interconversion of six unique structures, including a flexible, active tetramer and two novel, inhibited filaments. These structures reveal the conformational gymnastics necessary for RNR activity and the molecular basis for its control via an evolutionarily convergent form of allostery.
]]></description>
<dc:creator>Thomas, W. C.</dc:creator>
<dc:creator>Brooks, F. P.</dc:creator>
<dc:creator>Burnim, A. A.</dc:creator>
<dc:creator>Bacik, J.-P.</dc:creator>
<dc:creator>Stubbe, J.</dc:creator>
<dc:creator>Kaelber, J. T.</dc:creator>
<dc:creator>Chen, J. Z.</dc:creator>
<dc:creator>Ando, N.</dc:creator>
<dc:date>2018-12-21</dc:date>
<dc:identifier>doi:10.1101/504290</dc:identifier>
<dc:title><![CDATA[Convergent Allostery in Ribonucleotide Reductase]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-12-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/514323v1?rss=1">
<title>
<![CDATA[
Prevention of heart failure in hypertension: disentangling the role of evolving left ventricular hypertrophy and blood pressure lowering: the ALLHAT study 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/514323v1?rss=1</link>
<description><![CDATA[
BackgroundHypertension (HTN) is a known risk factor for heart failure (HF), possibly via the mechanism of cardiac remodeling and left ventricular hypertrophy (LVH). We studied how much blood pressure (BP) change and evolving LVH contribute to the effect that lisinopril, doxazosin, amlodipine have on HF compared to chlorthalidone.nnMethodsWe conducted causal mediation analysis of Antihypertensive and Lipid-Lowering Treatment to Prevent Heart Attack Trial (ALLHAT) data. ALLHAT participants with available serial ECGs and BP measurements were included (n=29,892; mean age 67{+/-}4 y; 32% black; 56% men): 11,008 were randomized to chlorthalidone, 5,967 - to doxazosin, 6,593 - to amlodipine, and 6,324 - to lisinopril. Evolving ECG-LVH, and BP-lowering served as mediators. Incident symptomatic HF was the primary outcome. Linear regression (for mediator) and logistic regression (for outcome) models were adjusted for mediator-outcome confounders (demographic and clinical characteristics known to be associated both with both LVH/HTN and HF).nnResultsA large majority of participants (96%) had ECG-LVH status unchanged; 4% developed evolving ECG-LVH. On average, BP decreased by 11/7 mmHg. In adjusted Cox regression analyses, progressing ECG-LVH [HR 1.78(1.43-2.22)], resolving ECG-LVH [HR 1.33(1.03-1.70)], and baseline ECG-LVH [1.17(1.04-1.31)] carried risk of incident HF. After full adjustment, evolving ECG-LVH mediated 4% of the effect of doxazosin on HF. Systolic BP-lowering mediated 12% of the effect of doxazosin, and diastolic BP-lowering mediated 10% effect of doxazosin, 7% effect of amlodipine, and borderline 9% effect of lisinopril on HF.nnConclusionsEvolving ECG-LVH and BP change account for 4-13% of the mechanism by which antihypertensive medications prevent HF.
]]></description>
<dc:creator>Johnson, K.</dc:creator>
<dc:creator>Oparil, S.</dc:creator>
<dc:creator>Davis, B. R.</dc:creator>
<dc:creator>Tereshchenko, L. G.</dc:creator>
<dc:date>2019-01-12</dc:date>
<dc:identifier>doi:10.1101/514323</dc:identifier>
<dc:title><![CDATA[Prevention of heart failure in hypertension: disentangling the role of evolving left ventricular hypertrophy and blood pressure lowering: the ALLHAT study]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-01-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/514380v1?rss=1">
<title>
<![CDATA[
Dynamic Predictive Accuracy of Electrocardiographic Biomarkers of Sudden Cardiac Death within a Survival Framework: The Atherosclerosis Risk in Communities (ARIC) study 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/514380v1?rss=1</link>
<description><![CDATA[
BackgroundThe risk of sudden cardiac death (SCD) is known to be dynamic. However, an accuracy of a dynamic SCD prediction, and "expiration date" of ECG biomarkers is unknown. Our goal was to measure dynamic predictive accuracy of ECG biomarkers of SCD and competing outcomes.nnMethodsAtherosclerosis Risk In Community study participants with analyzable digital ECGs were included (n=15,768; 55% female, 73% white, age 54.2{+/-}5.8 y). ECGs of 5 follow-up visits were analyzed. Global electrical heterogeneity (GEH) and traditional ECG metrics were measured. Adjudicated SCD served as the primary outcome; non-sudden cardiac death served as competing outcome. Time-dependent area under the (receiver operating characteristic) curve (AUC) analysis was performed to assess prediction accuracy of a continuous biomarker in a period of 3,6,9 months, and 1,2,3,5,10, and 15 years, using survival analysis framework.nnResultsOver a median 24.4 y follow-up, there were 581 SCDs (incidence 1.77 (95%CI 1.63-1.92)/1,000 person-years), and 838 nonSCDs [2.55 (95%CI 2.39-2.73)]. Resting heart rate was the strongest (AUC 0.930) short-term (3-month) non-specific SCD predictor, whereas spatial peak QRS-T angle predicted specifically SCD 15 years after ECG recording (AUC 0.719). QRS duration (AUC 0.885) and QTc (AUC 0.711) short-term predicted advanced structural heart disease better than SCD. "Expiration date" for most ECG biomarkers was two years after ECG recording. GEH significantly improved reclassification of SCD risk beyond age, sex, race, diabetes, hypertension, coronary heart disease and stroke.nnConclusionShort-term predictors of SCD, nonSCD, and biomarkers of long-term SCD risk differed, reflecting differences in transient vs. persistent SCD substrates.
]]></description>
<dc:creator>Perez-Alday, E.</dc:creator>
<dc:creator>Bender, A.</dc:creator>
<dc:creator>Li-Pershing, Y.</dc:creator>
<dc:creator>German, D.</dc:creator>
<dc:creator>Mai, T.</dc:creator>
<dc:creator>Mukundan, S.</dc:creator>
<dc:creator>Thomas, J.</dc:creator>
<dc:creator>Hamilton, C.</dc:creator>
<dc:creator>Tereshchenko, L.</dc:creator>
<dc:date>2019-01-08</dc:date>
<dc:identifier>doi:10.1101/514380</dc:identifier>
<dc:title><![CDATA[Dynamic Predictive Accuracy of Electrocardiographic Biomarkers of Sudden Cardiac Death within a Survival Framework: The Atherosclerosis Risk in Communities (ARIC) study]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-01-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/523266v1?rss=1">
<title>
<![CDATA[
Selective targeting of unipolar brush cell subtypes by cerebellar mossy fibers 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/523266v1?rss=1</link>
<description><![CDATA[
In vestibular cerebellum, primary afferents carry signals from single vestibular end organs, whereas secondary afferents from vestibular nucleus carry integrated signals. Selective targeting of distinct mossy fibers to postsynaptic cells determines how the cerebellum processes vestibular signals. We focused on vestibular projections to ON and OFF classes of unipolar brush cells (UBCs), which transform single mossy fiber signals into long-lasting excitation or inhibition respectively, and impact the activity of ensembles of granule cells. To determine whether these contacts are indeed selective, connectivity was traced back from UBC to specific ganglion cell, hair cell and vestibular organ subtypes. We show that a specialized subset of primary afferents contacts ON UBCs, but not OFF UBCs, while secondary afferents contact both subtypes. Striking anatomical differences were observed between primary and secondary afferents, their synapses, and the UBCs they contact. Thus, each class of UBC functions to transform specific signals through distinct anatomical pathways.
]]></description>
<dc:creator>Balmer, T. S.</dc:creator>
<dc:creator>Trussell, L.</dc:creator>
<dc:date>2019-01-17</dc:date>
<dc:identifier>doi:10.1101/523266</dc:identifier>
<dc:title><![CDATA[Selective targeting of unipolar brush cell subtypes by cerebellar mossy fibers]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-01-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/525782v1?rss=1">
<title>
<![CDATA[
Synapse-specific Opioid Modulation of Thalamo-cortico-striatal Circuits 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/525782v1?rss=1</link>
<description><![CDATA[
The medial thalamus (MThal), anterior cingulate cortex (ACC) and striatum play important roles in affective-motivational pain processing and reward learning. Opioids affect both pain and reward through uncharacterized modulation of this circuitry. This study examined opioid actions on glutamate transmission between these brain regions in mouse. Mu-opioid receptor (MOR) agonists potently inhibited MThal inputs without affecting ACC inputs to individual striatal medium spiny neurons (MSNs). MOR activation also inhibited MThal inputs to the pyramidal neurons in the ACC. In contrast, delta-opioid receptor (DOR) agonists disinhibited ACC pyramidal neuron responses to MThal inputs by suppressing local feed-forward GABA signaling from parvalbumin-positive interneurons. As a result, DOR activation in the ACC facilitated poly-synaptic (thalamo-cortico-striatal) excitation of MSNs by MThal inputs. These results suggest that opioid effects on pain and reward may be shaped by the relative selectivity of opioid drugs to the specific circuit components.
]]></description>
<dc:creator>Birdsong, W. T.</dc:creator>
<dc:creator>Jongbloets, B. C.</dc:creator>
<dc:creator>Engeln, K. A.</dc:creator>
<dc:creator>Wang, D.</dc:creator>
<dc:creator>Scherrer, G.</dc:creator>
<dc:creator>Mao, T.</dc:creator>
<dc:date>2019-01-21</dc:date>
<dc:identifier>doi:10.1101/525782</dc:identifier>
<dc:title><![CDATA[Synapse-specific Opioid Modulation of Thalamo-cortico-striatal Circuits]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-01-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/531293v1?rss=1">
<title>
<![CDATA[
Inter-individual Variability of Functional Connectivity in Awake and Anesthetized Rhesus Monkeys 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/531293v1?rss=1</link>
<description><![CDATA[
BackgroundNonhuman primate models (NHP) are commonly used to advance our understanding of brain function and organization. However, to date, they have offered few insights into individual differences among NHPs. In large part, this is due to the logistical challenges of NHP research, which limit most studies to five subjects or fewer.

MethodsWe leveraged the availability of a large-scale open NHP imaging resource to provide an initial examination of individual differences in the functional organization of the nonhuman primate brain. Specifically, we selected one awake fMRI dataset (Newcastle: n = 10) and two anesthetized fMRI data sets (Oxford: n = 19; UC-Davis: n = 19) to examine individual differences in functional connectivity characteristics across the cortex, as well as potential state dependencies.

ResultsWe noted significant individual variations of functional connectivity across the macaque cortex. Similar to the findings in human, during the awake state, the primary sensory and motor cortices showed lower variability than the high-order association regions. This variability pattern was significantly correlated with T1w/T2w map, the degree of long-distance connectivity, but not short-distance connectivity. However, the inter-individual variability under anesthesia exhibited a very distinct pattern, with lower variability in medial frontal cortex, precuneus and somatomotor regions and higher variability in the lateral ventral frontal and insular cortices.

ConclusionsThis work has implications for our understanding of the evolutionary origins of individual variation in the human brain, as well as methodological implications that must be considered in any pursuit to study individual variation in NHP models.
]]></description>
<dc:creator>Xu, T.</dc:creator>
<dc:creator>Sturgeon, D.</dc:creator>
<dc:creator>Ramirez, J. S. B.</dc:creator>
<dc:creator>Froudist-Walsh, S.</dc:creator>
<dc:creator>Margulies, D. S.</dc:creator>
<dc:creator>Schroeder, C. E.</dc:creator>
<dc:creator>Fair, D. A.</dc:creator>
<dc:creator>Milham, M.</dc:creator>
<dc:date>2019-01-26</dc:date>
<dc:identifier>doi:10.1101/531293</dc:identifier>
<dc:title><![CDATA[Inter-individual Variability of Functional Connectivity in Awake and Anesthetized Rhesus Monkeys]]></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/10.1101/534537v1?rss=1">
<title>
<![CDATA[
Spectral tuning of adaptation supports coding of sensory context in auditory cortex 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/534537v1?rss=1</link>
<description><![CDATA[
Perception of vocalizations and other behaviorally relevant sounds requires integrating acoustic information over hundreds of milliseconds, but the latency of sound-evoked activity in auditory cortex typically has much shorter latency. It has been observed that the acoustic context, i.e., sound history, can modulate sound evoked activity. Contextual effects are attributed to modulatory phenomena, such as stimulus-specific adaption and contrast gain control. However, an encoding model that links context to natural sound processing has yet to be established. We tested whether a model in which spectrally tuned inputs undergo adaptation mimicking short-term synaptic plasticity can account for contextual effects during natural sound processing. Single-unit activity was recorded from primary auditory cortex of awake ferrets during presentation of noise with natural temporal dynamics and fully natural sounds. Encoding properties were characterized by a standard linear-nonlinear spectro-temporal receptive field model (LN STRF) and STRF variants that incorporated STP-like adaptation. In two models, STP was applied either globally across all spectral channels or locally to subsets of channels. For most neurons, STRFs incorporating locally tuned STP predicted neural activity as well or better than the LN and global STP STRF. The strength of nonlinear adaptation varied across neurons. Within neurons, adaptation was generally stronger for activation with excitatory than inhibitory gain. Neurons showing improved STP model performance also tended to undergo stimulus-specific adaptation, suggesting a common mechanism for these phenomena. When STP STRFs were compared between passive and active behavior conditions, response gain often changed, but average STP parameters were stable. Thus, spectrally and temporally heterogeneous adaptation, subserved by a mechanism with STP-like dynamics, may support representation of the diverse spectro-temporal patterns that comprise natural sounds.

Author summarySuccessfully discriminating between behaviorally relevant sounds such as vocalizations and environmental noise requires processing how acoustic information changes over many tens to hundreds of milliseconds. The sound-evoked activity measured for most auditory cortical neurons is relatively short (< 50 ms), so it is not clear how the auditory cortex encodes sound information over longer periods. In this study, we propose that nonlinear adaptation, mimicking the effects of short-term synaptic plasticity (STP), enables auditory neurons to encode longer and more complex spectro-temporal patterns. A model in which sound history is stored in the latent state of plastic synapses is able to describe responses of single cortical neurons to natural sounds better than a standard encoding model that does not include nonlinear adaptation. Moreover, STP-like adaptation can account for contextual effects on sound evoked activity that cannot be accounted for by standard encoding models.
]]></description>
<dc:creator>Lopez Espejo, M.</dc:creator>
<dc:creator>Schwartz, Z. P.</dc:creator>
<dc:creator>David, S. V.</dc:creator>
<dc:date>2019-01-29</dc:date>
<dc:identifier>doi:10.1101/534537</dc:identifier>
<dc:title><![CDATA[Spectral tuning of adaptation supports coding of sensory context in auditory cortex]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-01-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/545582v1?rss=1">
<title>
<![CDATA[
Exercise-induced enhancement of synaptic functiontriggered by the inverse BAR protein, Mtss1L. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/545582v1?rss=1</link>
<description><![CDATA[
Exercise is a potent enhancer of learning and memory, yet we know little of the underlying mechanisms that likely include alterations in synaptic efficacy in the hippocampus. To address this issue, we exposed mice to a single episode of voluntary exercise, and permanently marked mature hippocampal dentate granule cells that were specifically activated during exercise using conditional Fos-TRAP mice. Only a few dentate granule cells were active at baseline, but two hours of voluntary exercise markedly increased the number of activated neurons. Activated neurons (Fos-TRAPed) showed an input-selective increase in dendritic spines and excitatory postsynaptic currents at 3 days post-exercise, indicative of exercise-induced structural plasticity. Laser-capture microdissection and RNASeq of activated neurons revealed that the most highly induced transcript was Mtss1L, a little-studied gene in the adult brain. Overexpression of Mtss1L in neurons increased spine density, leading us to hypothesize that its I-BAR domain initiated membrane curvature and dendritic spine formation. shRNA-mediated Mtss1L knockdown in vivo prevented the exercise-induced increases in spines and excitatory postsynaptic currents. Our results link short-term effects of exercise to activity-dependent expression of Mtss1L, which we propose as a novel effector of activity-dependent rearrangement of synapses.

One Sentence SummarySingle episodes of voluntary exercise induced a functional increase in hippocampal synapses mediated by activity-dependent expression of the BAR protein Mtss1L, acting as a novel early effector of synapse formation.
]]></description>
<dc:creator>Chatzi, C.</dc:creator>
<dc:creator>Zhang, Y.</dc:creator>
<dc:creator>Hendricks, W. D.</dc:creator>
<dc:creator>Chen, Y.</dc:creator>
<dc:creator>Schnell, E.</dc:creator>
<dc:creator>Goodman, R. H.</dc:creator>
<dc:creator>Westbrook, G. L.</dc:creator>
<dc:date>2019-02-09</dc:date>
<dc:identifier>doi:10.1101/545582</dc:identifier>
<dc:title><![CDATA[Exercise-induced enhancement of synaptic functiontriggered by the inverse BAR protein, Mtss1L.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-02-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/549196v1?rss=1">
<title>
<![CDATA[
Variable Retention of Differentiation-specific DNA Replication Timing in Human Pediatric Leukemia 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/549196v1?rss=1</link>
<description><![CDATA[
Human B-lineage precursor acute lymphoid leukemias (BCP-ALLs) comprise a group of genetically and clinically distinct disease entities with features of differentiation arrest at known stages of normal B-lineage differentiation. We previously showed BCP-ALL cells display unique and clonally heritable DNA-replication timing (RT) programs; i.e., programs describing the variable order of replication of megabase-scale chromosomal units of DNA in different cell types. To determine the extent to which BCP-ALL RT programs mirror or deviate from specific stages of normal human B-cell differentiation, we transplanted immunodeficient mice with quiescent normal human CD34+ cord blood cells and obtained RT signatures of the regenerating B-lineage populations. We then compared these with RT signatures for leukemic cells from a large cohort of BCP-ALL patients. The results identify BCP-ALL subtype-specific features that resemble specific stages of B-cell differentiation and features that appear associated with relapse. These results suggest the genesis of BCP-ALL involves alterations in RT that reflect clinically relevant leukemia-specific genetic and/or epigenetic changes.

SUMMARYGenome-wide DNA replication timing profiles of >100 pediatric leukemic samples and normally differentiating human B-lineage cells isolated from xenografted immunodeficient mice were generated. Comparison of these identified potentially clinically relevant features that both match and deviate from the normal profiles.
]]></description>
<dc:creator>Rivera-Mulia, J. C.</dc:creator>
<dc:creator>Sasaki, T.</dc:creator>
<dc:creator>Trevilla-Garcia, C.</dc:creator>
<dc:creator>Nakamichi, N.</dc:creator>
<dc:creator>Knapp, D. J. H. F.</dc:creator>
<dc:creator>Hammond, C.</dc:creator>
<dc:creator>Chang, B.</dc:creator>
<dc:creator>Tyner, J. W.</dc:creator>
<dc:creator>Devidas, M.</dc:creator>
<dc:creator>Zimmerman, J.</dc:creator>
<dc:creator>Klein, K.</dc:creator>
<dc:creator>Somasundaram, V.</dc:creator>
<dc:creator>Druker, B.</dc:creator>
<dc:creator>Gruber, T.</dc:creator>
<dc:creator>Koren, A.</dc:creator>
<dc:creator>Eaves, C. J.</dc:creator>
<dc:creator>Gilbert, D. M.</dc:creator>
<dc:date>2019-02-13</dc:date>
<dc:identifier>doi:10.1101/549196</dc:identifier>
<dc:title><![CDATA[Variable Retention of Differentiation-specific DNA Replication Timing in Human Pediatric Leukemia]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-02-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/552075v1?rss=1">
<title>
<![CDATA[
High-throughput single-particle tracking reveals nested membrane nanodomain organization that dictates Ras diffusion and trafficking 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/552075v1?rss=1</link>
<description><![CDATA[
Membrane nanodomains have been implicated in Ras signaling, but what these domains are and how they interact with Ras remain obscure. Using high throughput single particle tracking with photoactivated localization microscopy and detailed trajectory analysis, here we show that distinct membrane domains dictate KRas diffusion and trafficking in U2OS cells. KRas exhibits an immobile state in domains [~]70 nm in size, each embedded in a larger domain ([~]200 nm) that confers intermediate mobility, while the rest of the membrane supports fast diffusion. Moreover, KRas is continuously removed from the membrane via the immobile state and replenished to the fast state, likely coupled to internalization and recycling. Importantly, both the diffusion and trafficking properties of KRas remain invariant over a broad range of protein expression levels. Our results reveal how membrane organization dictates KRas diffusion and trafficking and offer insight into how Ras signaling may be regulated through spatial mechanisms.
]]></description>
<dc:creator>Lee, Y.</dc:creator>
<dc:creator>Phelps, C.</dc:creator>
<dc:creator>Huang, T.</dc:creator>
<dc:creator>Mostofian, B.</dc:creator>
<dc:creator>Wu, L.</dc:creator>
<dc:creator>Zhang, Y.</dc:creator>
<dc:creator>Chang, Y. H.</dc:creator>
<dc:creator>Stork, P. J. S.</dc:creator>
<dc:creator>Gray, J. W.</dc:creator>
<dc:creator>Zuckerman, D. M.</dc:creator>
<dc:creator>Nan, X.</dc:creator>
<dc:date>2019-02-15</dc:date>
<dc:identifier>doi:10.1101/552075</dc:identifier>
<dc:title><![CDATA[High-throughput single-particle tracking reveals nested membrane nanodomain organization that dictates Ras diffusion and trafficking]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-02-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/565226v1?rss=1">
<title>
<![CDATA[
Sphingomyelin is critical in organizing phosphoinositide dynamics during phagocytic uptake of Mycobacterium tuberculosis. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/565226v1?rss=1</link>
<description><![CDATA[
Tuberculosis, the pathological result of infection by Mycobacterium tuberculosis (M.tb), represents a massive and persistent global health burden with vast overrepresentation in regions with poor access to healthcare. This pathogen survives intracellularly within lung macrophages, and has been shown to extensively reprogram the host lipid compartment to the effect of enhanced survival and immune evasion. Sphingolipids are a class of lipid with extensive functions in a wide variety of cellular contexts, including apoptosis, autophagy, stress response, and inflammation. There remains a significant knowledge gap surrounding the lipid-mediated interactions between M.tb and its host. Our investigation aims to explore the mechanisms through which M.tb harnesses host-derived sphingolipids to gain entry to the host and to survive intracellularly. To this end, we use chemical and genetic perturbations of the sphingolipid de novo synthesis pathway and fluorescence microscopy to show that sphingolipids are required for efficient phagocytosis of M.tb. We furthermore show that the dynamics of phosphoinositides at the phagosomal synapse are perturbed upon blockade of sphingolipid synthesis. Finally, we demonstrate that perturbing the homeostasis of a specific sphingolipid (sphingomyelin) through both enrichment and depletion at the cell surface reduces M.tb uptake. This work represents a first step in building comprehensive knowledge of the mechanisms of host-pathogen interactions in M.tb, and demonstrates a cohort of methodologies that may be applied to a variety of pathogenic contexts.
]]></description>
<dc:creator>Niekamp, P.</dc:creator>
<dc:creator>Guzman, G.</dc:creator>
<dc:creator>Leier, H.</dc:creator>
<dc:creator>Rashidfarrokhi, A.</dc:creator>
<dc:creator>Richina, V.</dc:creator>
<dc:creator>Holthuis, J.</dc:creator>
<dc:creator>Tafesse, F. G.</dc:creator>
<dc:date>2019-03-01</dc:date>
<dc:identifier>doi:10.1101/565226</dc:identifier>
<dc:title><![CDATA[Sphingomyelin is critical in organizing phosphoinositide dynamics during phagocytic uptake of Mycobacterium tuberculosis.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-03-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/572297v1?rss=1">
<title>
<![CDATA[
Mechanism of pharmacochaperoning in KATP channels revealed by cryo-EM 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/572297v1?rss=1</link>
<description><![CDATA[
ATP-sensitive potassium (KATP) channels composed of a pore-forming Kir6.2 potassium channel and a regulatory ABC transporter sulfonylurea receptor 1 (SUR1) regulate insulin secretion in pancreatic {beta}-cells to maintain glucose homeostasis. Mutations that impair channel folding or assembly prevent cell surface expression and cause congenital hyperinsulinism. Structurally diverse KATP inhibitors have been shown to act as pharmacochaperones to correct mutant channel expression, but the mechanism is unknown. Here, we compare cryoEM structures of KATP channels bound to pharmacochaperones glibenclamide, repaglinide, and carbamazepine. We found all three drugs bind within a common pocket in SUR1. Further, we found the N-terminus of Kir6.2 inserted within the central cavity of the SUR1 ABC core, adjacent the drug binding pocket. The findings reveal a common mechanism by which diverse compounds stabilize the Kir6.2 N-terminus within the SUR1 ABC core, allowing it to act as a firm "handle" for the assembly of metastable mutant SUR1-Kir6.2 complexes.
]]></description>
<dc:creator>Martin, G. M.</dc:creator>
<dc:creator>Sung, M. W.</dc:creator>
<dc:creator>Yang, Z.</dc:creator>
<dc:creator>Innes, L. M.</dc:creator>
<dc:creator>Kandasamy, B.</dc:creator>
<dc:creator>David, L.</dc:creator>
<dc:creator>Yoshioka, C.</dc:creator>
<dc:creator>Shyng, S.-L.</dc:creator>
<dc:date>2019-03-08</dc:date>
<dc:identifier>doi:10.1101/572297</dc:identifier>
<dc:title><![CDATA[Mechanism of pharmacochaperoning in KATP channels revealed by cryo-EM]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-03-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/581546v1?rss=1">
<title>
<![CDATA[
Sex differences in reward- and punishment-guided actions 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/581546v1?rss=1</link>
<description><![CDATA[
Differences in the prevalence and presentation of psychiatric illnesses in men and women suggest that neurobiological sex differences confer vulnerability or resilience in these disorders. Rodent behavioral models are critical for understanding the mechanisms of these differences. Reward processing and punishment avoidance are fundamental dimensions of the symptoms of psychiatric disorders. Here we explored sex differences along these dimensions using multiple and distinct behavioral paradigms. We found no sex difference in reward-guided associative learning but a faster punishment-avoidance learning in females. After learning, females were more sensitive than males to probabilistic punishment but less sensitive when punishment could be avoided with certainty. No sex differences were found in reward-guided cognitive flexibility. Thus, sex differences in goal-directed behaviors emerged selectively when there was an aversive context. These differences were critically sensitive to whether the punishment was certain or unpredictable. Our findings with these new paradigms provide conceptual and practical tools for investigating brain mechanisms that account for sex differences in susceptibility to anxiety and impulsivity. They may also provide insight for understanding the evolution of sex-specific optimal behavioral strategies in dynamic environments.
]]></description>
<dc:creator>Tara, C. G.</dc:creator>
<dc:creator>Wallin-Miller, K. G.</dc:creator>
<dc:creator>Rear, A. A.</dc:creator>
<dc:creator>Park, J.</dc:creator>
<dc:creator>Diaz, V.</dc:creator>
<dc:creator>Simon, N. W.</dc:creator>
<dc:creator>Moghaddam, B.</dc:creator>
<dc:date>2019-03-18</dc:date>
<dc:identifier>doi:10.1101/581546</dc:identifier>
<dc:title><![CDATA[Sex differences in reward- and punishment-guided actions]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-03-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/589184v1?rss=1">
<title>
<![CDATA[
Identification of PKC -dependent phosphoproteins in mouse retina 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/589184v1?rss=1</link>
<description><![CDATA[
Adjusting to a wide range of light intensities is an essential feature of retinal rod bipolar cell (RBC) function. While persuasive evidence suggests this modulation involves phosphorylation by protein kinase C-alpha (PKC), the targets of PKC phosphorylation in the retina have not been identified. PKC activity and phosphorylation in RBCs was examined by immunofluorescence confocal microscopy using a conformation-specific PKC antibody and antibodies to phosphorylated PKC motifs. PKC activity was dependent on light and expression of TRPM1, and RBC dendrites were the primary sites of light-dependent phosphorylation. PKC-dependent retinal phosphoproteins were identified using a phosphoproteomics approach to compare total protein and phosphopeptide abundance between phorbol ester-treated wild type and PKC knockout (PKC-KO) mouse retinas. Phosphopeptide mass spectrometry identified over 1100 phosphopeptides in mouse retina, with 12 displaying significantly greater phosphorylation in WT compared to PKC-KO samples. The differentially phosphorylated proteins fall into the following functional groups: cytoskeleton/trafficking (4 proteins), ECM/adhesion (2 proteins), signaling (2 proteins), transcriptional regulation (3 proteins), and homeostasis/metabolism (1 protein). Two strongly differentially expressed phosphoproteins, BORG4 and TPBG, were localized to the synaptic layers of the retina, and may play a role in PKC-dependent modulation of RBC physiology. Data are available via ProteomeXchange with identifier PXD012906.nnSignificanceRetinal rod bipolar cells (RBCs), the second-order neurons of the mammalian rod visual pathway, are able to modulate their sensitivity to remain functional across a wide range of light intensities, from starlight to daylight. Evidence suggests that this modulation requires the serine/threonine kinase, PKC, though the specific mechanism by which PKC modulates RBC physiology is unknown. This study examined PKC phosophorylation patterns in mouse rod bipolar cells and then used a phosphoproteomics approach to identify PKC-dependent phosphoproteins in the mouse retina. A small number of retinal proteins showed significant PKC-dependent phosphorylation, including BORG4 and TPBG, suggesting a potential contribution to PKC-dependent modulation of RBC physiology.nnHighlightsO_LIPKC is a major source of phosphorylation in retinal RBC dendrites and its activity in RBCs is light dependent.nC_LIO_LIProteins showing differential phosphorylation between phorbol ester-treated wild type and PKC-KO retinas belong to the following major functional groups: cytoskeleton/trafficking (4 proteins), ECM/adhesion (2 proteins), signaling (2 proteins), transcriptional regulation (3 proteins), and homeostasis/metabolism (1 protein).nC_LIO_LIThe PKC-dependent phosphoproteins, BORG4 and TPBG, are present in the synaptic layers of the retina and may be involved in PKC-dependent modulation of RBC physiology.nC_LI
]]></description>
<dc:creator>Wakeham, C. M.</dc:creator>
<dc:creator>Wilmarth, P. A.</dc:creator>
<dc:creator>Cunliffe, J. M.</dc:creator>
<dc:creator>Klimek, J. E.</dc:creator>
<dc:creator>Ren, G.</dc:creator>
<dc:creator>David, L. L.</dc:creator>
<dc:creator>Morgans, C. W.</dc:creator>
<dc:date>2019-03-27</dc:date>
<dc:identifier>doi:10.1101/589184</dc:identifier>
<dc:title><![CDATA[Identification of PKC -dependent phosphoproteins in mouse retina]]></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/10.1101/590216v1?rss=1">
<title>
<![CDATA[
Expression and distribution of Trophoblast Glycoprotein in the mouse retina 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/590216v1?rss=1</link>
<description><![CDATA[
We recently identified the leucine-rich repeat adhesion protein, trophoblast glycoprotein (TPBG), as a novel PKC-dependent phosphoprotein in retinal rod bipolar cells (RBCs). Since TPBG has not been thoroughly examined in the retina, this study characterizes the localization and expression patterns of TPBG in the developing and adult mouse retina using two antibodies, one against the N-terminal, leucine-rich domain and the other against the C-terminal PDZ-interacting motif. Both antibodies labeled dendrites and synaptic terminals of RBCs, as well as the cell bodies and dendrites of an uncharacterized class of amacrine cell. In transfected HEK293 cells, TPBG was localized to the plasma membrane and intracellular membranes and was associated with the tips of thin filopodia-like membrane projections. TPBG immunofluorescence in RBCs detected with the C-terminal antibody was strongly dependent on the activity state of the adult retina, with less labeling in dark-adapted compared to light-adapted retina, and less labeling in light-adapted PKC knockout and TRPM1 knockout retinas compared to wild type, despite no change in total TPBG detected by immunoblotting. These results suggest that the C-terminal epitope is blocked in the dark-adapted and knockout retinas compared to light-adapted wild type retinas, possibly through interaction with a PDZ domain protein. During development, TPBG expression increases dramatically just prior to eye opening with a time course closely correlated with that of TRPM1 expression. In the retina, leucine-rich repeat proteins like TPBG have been implicated in the development and maintenance of functional bipolar cell synapses, and TPBG may play a similar role in RBCs.
]]></description>
<dc:creator>Wakeham, C. M.</dc:creator>
<dc:creator>Ren, G.</dc:creator>
<dc:creator>Morgans, C. W.</dc:creator>
<dc:date>2019-03-27</dc:date>
<dc:identifier>doi:10.1101/590216</dc:identifier>
<dc:title><![CDATA[Expression and distribution of Trophoblast Glycoprotein in the mouse retina]]></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/10.1101/601542v1?rss=1">
<title>
<![CDATA[
The Dialysis Procedure Triggers Autonomic Imbalance and Cardiac Arrhythmias: Insights from Continuous 14-day ECG Monitoring 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/601542v1?rss=1</link>
<description><![CDATA[
BackgroundIn end-stage kidney disease the dialytic cycle relates to the rate of sudden cardiac death. We hypothesized that circadian, dialytic cycles, paroxysmal arrhythmias, and cardiovascular risk factors are associated with periodic changes in heart rate and heart rate variability (HRV) in incident dialysis patients.nnMethodsWe conducted a prospective ancillary study of the Predictors of Arrhythmic and Cardiovascular Risk in End Stage Renal Disease cohort (n=28; age 54{+/-}13 y; 57% men; 96% black; 33% with a history of structural heart disease; left ventricular ejection fraction 70{+/-}9%). Continuous ECG monitoring was performed using an ECG patch (Zio Patch, iRhythm) and short-term HRV was measured for three minutes every hour. HRV was measured by root mean square of the successive normal-to-normal intervals (rMSSD), high and low frequency power, Poincare plot, and sample and Renyi entropy.nnResultsArrhythmias were detected in 46% (n=13). Non-sustained ventricular tachycardia (VT) was more frequent during dialysis or within 6 hours post-dialysis, as compared to pre-or between-dialysis (63% vs. 37%, P=0.015), whereas supraventricular tachycardia was more frequent pre-/ between-dialysis, as compared to during-/ post-dialysis (84% vs. 16%, P=0.015). In adjusted for cardiovascular disease and its risk factors autoregressive conditional heteroscedasticity panel (ARCH) model, VT events were associated with increased heart rate by 11.2 (95%CI 10.1-12.3) bpm (P<0.0001). During regular dialytic cycle, rMSSD demonstrated significant circadian pattern (Mesor 10.6(0.9-11.2) ms; Amplitude 1.5(1.0-3.1) ms; Peak at 02:01(20:22-03:16) am; P<0.0001), which was abolished on a second day interdialytic extension (adjusted ARCH trend for rMSSD -1.41(-1.67 to -1.15) ms per 24h; P<0.0001).nnConclusionCardiac arrhythmias associate with dialytic phase. Regular dialytic schedule preserves physiological circadian rhythm, but the second day without dialysis is characterized by parasympathetic withdrawal and a steady increase in sympathetic predominance.nnSubject TermsArrhythmias, Autonomic Nervous System, Electrocardiology (ECG), Treatment.
]]></description>
<dc:creator>Rogovoy, N. M.</dc:creator>
<dc:creator>Howell, S. J.</dc:creator>
<dc:creator>Lee, T. L.</dc:creator>
<dc:creator>Hamilton, C.</dc:creator>
<dc:creator>Perez-Alday, E. A.</dc:creator>
<dc:creator>Kabir, M. M.</dc:creator>
<dc:creator>Li-Pershing, Y.</dc:creator>
<dc:creator>Zhang, Y.</dc:creator>
<dc:creator>Kim, E. D.</dc:creator>
<dc:creator>Fitzpatrick, J.</dc:creator>
<dc:creator>Monroy-Trujillo, J. M.</dc:creator>
<dc:creator>Estrella, M. M.</dc:creator>
<dc:creator>Sozio, S. M.</dc:creator>
<dc:creator>Jaar, B. G.</dc:creator>
<dc:creator>Parekh, R. S.</dc:creator>
<dc:creator>Tereshchenko, L. G.</dc:creator>
<dc:date>2019-04-12</dc:date>
<dc:identifier>doi:10.1101/601542</dc:identifier>
<dc:title><![CDATA[The Dialysis Procedure Triggers Autonomic Imbalance and Cardiac Arrhythmias: Insights from Continuous 14-day ECG Monitoring]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-04-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/604975v1?rss=1">
<title>
<![CDATA[
Post-ictal neuronal network remodeling and Wnt pathway dysregulation in the intra-hippocampal kainate mouse model of temporal lobe epilepsy 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/604975v1?rss=1</link>
<description><![CDATA[
Mouse models of mesial temporal lobe epilepsy recapitulate aspects of human epilepsy, which is characterized by neuronal network remodeling in the hippocampal dentate gyrus. Observational studies suggest that this remodeling is associated with altered Wnt pathway signaling, although this has not been experimentally examined. We used the well-characterized mouse intrahippocampal kainate model of temporal lobe epilepsy to examine associations between post-seizure hippocampal neurogenesis and altered Wnt signaling. Tissue was analyzed using immunohistochemistry and confocal microscopy, and transcriptome analysis was performed on RNA extracted from anatomically micro-dissected dentate gyri. Seizures increased neurogenesis and dendritic arborization of newborn hippocampal dentate granule cells in peri-ictal regions, and decreased neurogenesis in the ictal zone, 2-weeks after kainate injection. Interestingly, administration of the novel canonical Wnt pathway inhibitor XAV939 daily for 2-weeks after kainate injection further increased dendritic arborization in peri-ictal regions after seizure, without an effect on baseline neurogenesis in control animals. Transcriptome analysis of dentate gyri demonstrated significant canonical Wnt gene dysregulation in kainate-injected mice across all regions for Wnt3, 5a and 9a. Intriguingly, certain Wnt genes demonstrated differential patterns of dysregulation between the ictal and peri-ictal zones, most notably Wnt5B, 7B and DKK-1. Together, these results demonstrate regional variation in Wnt pathway dysregulation in the early post-ictal period, and surprisingly, suggest that some Wnt-mediated effects might actually temper aberrant neurogenesis after seizures. The Wnt pathway may therefore provide suitable targets for novel therapies that prevent network remodeling and the development of epileptic foci in high-risk patients.
]]></description>
<dc:creator>Gupta, K.</dc:creator>
<dc:creator>Schnell, E.</dc:creator>
<dc:date>2019-04-10</dc:date>
<dc:identifier>doi:10.1101/604975</dc:identifier>
<dc:title><![CDATA[Post-ictal neuronal network remodeling and Wnt pathway dysregulation in the intra-hippocampal kainate mouse model of temporal lobe epilepsy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-04-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/619619v1?rss=1">
<title>
<![CDATA[
Pupil-associated states modulate excitability but not stimulus selectivity in primary auditory cortex 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/619619v1?rss=1</link>
<description><![CDATA[
Recent research in mice indicates that luminance-independent fluctuations in pupil size predict variability in spontaneous and evoked activity of single neurons in auditory and visual cortex. These findings suggest that pupil is an indicator of large-scale changes in arousal state that affect sensory processing. However, it is not known whether pupil-related state also influences the selectivity of auditory neurons. We recorded pupil size and single-unit spiking activity in the primary auditory cortex (A1) of non-anesthetized male and female ferrets during presentation of natural vocalizations and tone stimuli that allow measurement of frequency and level tuning. Neurons showed a systematic increase in both spontaneous and sound-evoked activity when pupil was large, as well as desynchronization and a decrease in trial-to-trial variability. Relationships between pupil size and firing rate were non-monotonic in some cells. In most neurons, several measurements of tuning, including acoustic threshold, spectral bandwidth, and best frequency, remained stable across large changes in pupil size. Across the population, however, there was a small but significant decrease in acoustic threshold when pupil was dilated. In some recordings, we observed rapid, saccade-like eye movements during sustained pupil constriction, which may indicate sleep. Including the presence of this state as a separate variable in a regression model of neural variability accounted for some, but not all, of the variability and non-monotonicity associated with changes in pupil size.nnNew & NoteworthyCortical neurons vary in their response to repeated stimuli, and some portion of the variability is due to fluctuations in network state. By simultaneously recording pupil and single-neuron activity in auditory cortex of ferrets, we provide new evidence that network state affects the excitability of auditory neurons, but not sensory selectivity. In addition, we report the occurrence of possible sleep states, adding to evidence that pupil provides an index of both sleep and physiological arousal.
]]></description>
<dc:creator>Schwartz, Z. P.</dc:creator>
<dc:creator>Buran, B. N.</dc:creator>
<dc:creator>David, S. V.</dc:creator>
<dc:date>2019-04-26</dc:date>
<dc:identifier>doi:10.1101/619619</dc:identifier>
<dc:title><![CDATA[Pupil-associated states modulate excitability but not stimulus selectivity in primary auditory cortex]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-04-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/627802v1?rss=1">
<title>
<![CDATA[
Transformation and Integration of Microenvironment Microarray Data Improves Discovery of Latent Effects 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/627802v1?rss=1</link>
<description><![CDATA[
The immediate physical and bio-chemical surroundings of a cell, the cellular microenvironment, is an important component of many fundamental cell and tissue level processes and is implicated in many diseases and dysfunctions. Thus understanding the interaction of cells with their microenvironment can further both basic research and aid the discovery of therapeutic agents. To study perturbations of cellular microenvironments a novel image-based cell-profiling technology called the microenvironment microarray (MEMA) has been recently employed. In this paper we explore the effect of preprocessing transformations for MEMA data on the discovery of biological and technical latent effects. We find that Gaussianizing the data and carefully removing outliers can enhance discovery of important biological effects. In particular, these transformations help reveal a relationship between cell morphological features and the extra-cellular-matrix protein THBS1 in MCF10A breast tissue. More broadly, MEMAs are part of a recent and wide-spread adoption of image-based cell-profiling technologies in the quantification of phenotypic differences among cell populations (Caicedo et al., 2017). Thus we anticipate that the advantages of the proposed preprocessing transformations will likely also be realized in the analysis of data from other highly-multiplexed technologies like Cyclic Immunofluorescence. All code and supplementary analysis for this paper is available at gjhunt.github.io/rr.
]]></description>
<dc:creator>Hunt, G. J.</dc:creator>
<dc:creator>Dane, M. A.</dc:creator>
<dc:creator>Korkola, J. E.</dc:creator>
<dc:creator>Heiser, L. M.</dc:creator>
<dc:creator>Gagnon-Bartsch, J. A.</dc:creator>
<dc:date>2019-05-05</dc:date>
<dc:identifier>doi:10.1101/627802</dc:identifier>
<dc:title><![CDATA[Transformation and Integration of Microenvironment Microarray Data Improves Discovery of Latent Effects]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-05-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/637256v1?rss=1">
<title>
<![CDATA[
Improved single-cell ATAC-seq reveals chromatin dynamics of in vitro corticogenesis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/637256v1?rss=1</link>
<description><![CDATA[
Development is a complex process that requires the precise modulation of regulatory gene networks controlled through dynamic changes in the epigenome. Single-cell-omic technologies provide an avenue for understanding the mechanisms of these processes by capturing the progression of epigenetic cell states during the course of cellular differentiation using in vitro or in vivo models1. However, current single-cell epigenomic methods are limited in the information garnered per individual cell, which in turn limits their ability to measure chromatin dynamics and state shifts. Single-cell combinatorial indexing (sci-) has been applied as a strategy for identifying single-cell-omic originating libraries and removes the necessity of single-cell, single-compartment chemistry2. Here, we report an improved sci-assay for transposase accessible chromatin by sequencing (ATAC-seq), which utilizes the small molecule inhibitor Pitstop 2 (scip-ATAC-seq)3. We demonstrate that these improvements, which theoretically could be applied to any in situ transposition method for single-cell library preparation, significantly increase the ability of transposase to enter the nucleus and generate highly complex single-cell libraries, without altering biological signal. We applied sci-ATAC-seq and scip-ATAC-seq to characterize the chromatin dynamics of developing forebrain-like organoids, an in vitro model of human corticogenesis4. Using these data, we characterized novel putative regulatory elements, compared the epigenome of the organoid model to human cortex data, generated a high-resolution pseudotemporal map of chromatin accessibility through differentiation, and measured epigenomic changes coinciding with a neurogenic fate decision point. Finally, we combined transcription factor motif accessibility with gene activity (GA) scores to directly observe the dynamics of complex regulatory programs that regulate neurogenesis through developmental pseudotime. Overall, scip-ATAC-seq increases information content per cell and bolsters the potential for future single-cell studies into complex developmental processes.
]]></description>
<dc:creator>Mulqueen, R. M.</dc:creator>
<dc:creator>DeRosa, B. A.</dc:creator>
<dc:creator>Thornton, C. A.</dc:creator>
<dc:creator>Sayar, Z.</dc:creator>
<dc:creator>Torkenczy, K. A.</dc:creator>
<dc:creator>Fields, A. J.</dc:creator>
<dc:creator>Wright, K. M.</dc:creator>
<dc:creator>Nan, X.</dc:creator>
<dc:creator>Ramji, R.</dc:creator>
<dc:creator>Steemers, F. J.</dc:creator>
<dc:creator>O'Roak, B. J.</dc:creator>
<dc:creator>Adey, A. C.</dc:creator>
<dc:date>2019-05-15</dc:date>
<dc:identifier>doi:10.1101/637256</dc:identifier>
<dc:title><![CDATA[Improved single-cell ATAC-seq reveals chromatin dynamics of in vitro corticogenesis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-05-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/639617v1?rss=1">
<title>
<![CDATA[
Myeloid Lineage Enhancers Drive Oncogene Synergy in CEBPA/CSF3R Mutant Acute Myeloid Leukemia 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/639617v1?rss=1</link>
<description><![CDATA[
Acute Myeloid Leukemia (AML) develops due to the acquisition of mutations from multiple functional classes. Here, we demonstrate that activating mutations in the granulocyte colony stimulating factor receptor (CSF3R), cooperate with loss of function mutations in the transcription factor CEBPA to promote acute leukemia development. This finding of mutation-synergy is broadly applicable other mutations that activate the JAK/STAT pathway or disrupt CEBPA function (i.e. activating mutations in JAK3 and Core Binding Factor translocations). The interaction between these distinct classes of mutations occurs at the level of myeloid lineage enhancers where mutant CEBPA prevents activation of subset of differentiation associated enhancers. To confirm this enhancer-dependent mechanism, we demonstrate that CEBPA mutations must occur as the initial event in AML initiation, confirming predictions from clinical sequencing data. This improved mechanistic understanding will facilitate therapeutic development targeting the intersection of oncogene cooperativity.
]]></description>
<dc:creator>Braun, T. P.</dc:creator>
<dc:creator>Okhovat, M.</dc:creator>
<dc:creator>Coblentz, C.</dc:creator>
<dc:creator>Carratt, S. A.</dc:creator>
<dc:creator>Foley, A.</dc:creator>
<dc:creator>Schonrock, Z.</dc:creator>
<dc:creator>Nevonen, K.</dc:creator>
<dc:creator>Davis, B.</dc:creator>
<dc:creator>Garcia, B.</dc:creator>
<dc:creator>LaTocha, D.</dc:creator>
<dc:creator>Weeder, B. R.</dc:creator>
<dc:creator>Grzadkowski, M. R.</dc:creator>
<dc:creator>Estabrook, J. C.</dc:creator>
<dc:creator>Manning, H. G.</dc:creator>
<dc:creator>Watanabe-Smith, K.</dc:creator>
<dc:creator>Smith, J. L.</dc:creator>
<dc:creator>Leonti, A. R.</dc:creator>
<dc:creator>Ries, R. E.</dc:creator>
<dc:creator>Jeng, S.</dc:creator>
<dc:creator>McWeeney, S.</dc:creator>
<dc:creator>Di Genua, C.</dc:creator>
<dc:creator>Drissen, R.</dc:creator>
<dc:creator>Nerlov, C.</dc:creator>
<dc:creator>Meshinchi, S.</dc:creator>
<dc:creator>Carbone, L.</dc:creator>
<dc:creator>Druker, B. J.</dc:creator>
<dc:creator>Maxson, J. E.</dc:creator>
<dc:date>2019-05-15</dc:date>
<dc:identifier>doi:10.1101/639617</dc:identifier>
<dc:title><![CDATA[Myeloid Lineage Enhancers Drive Oncogene Synergy in CEBPA/CSF3R Mutant Acute Myeloid Leukemia]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-05-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/645374v1?rss=1">
<title>
<![CDATA[
HCMV miR-US22 down-regulation of EGR-1 regulates CD34+ hematopoietic progenitor cell proliferation and viral reactivation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/645374v1?rss=1</link>
<description><![CDATA[
Reactivation of latent Human Cytomegalovirus (HCMV) in CD34+ hematopoietic progenitor cells (HPCs) is closely linked to hematopoiesis. Viral latency requires maintenance of the progenitor cell quiescence, while reactivation initiates following mobilization of HPCs to the periphery and differentiation into CD14+ macrophages. Early growth response gene 1 (EGR-1) is a transcription factor activated by Epidermal growth factor receptor (EGFR) signaling that is essential for the maintenance of CD34+ HPC self-renewal in the bone marrow niche. Down-regulation of EGR-1 results in mobilization and differentiation of CD34+ HPC from the bone marrow to the periphery. In the current study we demonstrate that the transcription factor EGR-1 is directly targeted for down-regulation by HCMV miR-US22 that results in decreased proliferation of CD34+ HPCs and a decrease in total hematopoietic colony formation. We also show that an HCMV miR-US22 mutant fails to reactivate in CD34+ HPCs, indicating that expression of EGR-1 inhibits viral reactivation during latency. Since EGR-1 promotes CD34+ HPC self-renewal in the bone marrow niche, HCMV miR-US22 down-regulation of EGR-1 is a necessary step to block HPC self-renewal and proliferation to induce a cellular differentiation pathway necessary to promote reactivation of virus.nnAuthor summaryHuman cytomegalovirus (HCMV) is a widespread herpesvirus that persists in the host and remains a significant cause of morbidity and mortality in solid organ and stem cell transplant patients. HCMV latency is complex, and the molecular mechanisms for establishment, maintenance, and reactivation from latency are poorly understood.nnQuiescent stem cells in the bone marrow represent a critical reservoir of latent HCMV, and the mobilization and differentiation of these cells is closely linked to viral reactivation from latency. HCMV encodes small regulatory RNAs, called miRNAs that play important roles in the regulation of viral and cellular gene expression. In this study, we show that HCMV miR-US22 targets Early growth response gene 1 (EGR-1) a host transcription factor that is necessary for stem cell quiescence and self-renewal in the bone marrow. Expression of this miR-US22 down-regulates expression of EGR-1 that reduces CD34+ HPCs proliferation and total hematopoietic colony formation. An HCMV miR-US22 mutant is unable to reactivate from latency suggesting that the ability of the miRNA to disrupt CD34+ HPC renewal in the bone marrow niche to initiate a differentiation pathway is critical for viral reactivation.
]]></description>
<dc:creator>Mikell, I. A.</dc:creator>
<dc:creator>Crawford, L. B.</dc:creator>
<dc:creator>Hancock, M. H.</dc:creator>
<dc:creator>Mitchell, J.</dc:creator>
<dc:creator>Buehler, J.</dc:creator>
<dc:creator>Goodrum, F.</dc:creator>
<dc:creator>Nelson, J. A.</dc:creator>
<dc:date>2019-05-21</dc:date>
<dc:identifier>doi:10.1101/645374</dc:identifier>
<dc:title><![CDATA[HCMV miR-US22 down-regulation of EGR-1 regulates CD34+ hematopoietic progenitor cell proliferation and viral reactivation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-05-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/646083v1?rss=1">
<title>
<![CDATA[
Cataract-associated deamidations on the surface of γS-crystallin increase protein unfolding and flexibility at distant regions 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/646083v1?rss=1</link>
<description><![CDATA[
Deamidation is a major age-related modification in the human lens that is highly prevalent in crystallins isolated from cataractous lenses. However, the mechanism by which deamidation causes proteins to become insoluble is not known, because of only subtle structural changes observed in vitro. We have identified Asn14 and Asn76 of {gamma}S-crystallin as highly deamidated in insoluble proteins. These sites are on the surface of the N-terminal domain and were mimicked by replacing the Asn with Asp residues. We used heteronuclear NMR spectroscopy to measure their amide hydrogen exchange and 15N relaxation dynamics to identify regions with significantly increased dynamics compared to wildtype-{gamma}S. Changes in dynamics were localized to the C-terminal domain, particularly to helix and surface loops distant from the mutation sites. Thus, a potential mechanism for {gamma}S deamidation-induced insolubilization in cataractous lenses is altered dynamics due to local regions of unfolding and increased flexibility.
]]></description>
<dc:creator>Forsythe, H. M.</dc:creator>
<dc:creator>Vetter, C. J.</dc:creator>
<dc:creator>Jara, K. J.</dc:creator>
<dc:creator>Reardon, P. N.</dc:creator>
<dc:creator>David, L. L.</dc:creator>
<dc:creator>Barbar, E. J.</dc:creator>
<dc:creator>Lampi, K.</dc:creator>
<dc:date>2019-05-22</dc:date>
<dc:identifier>doi:10.1101/646083</dc:identifier>
<dc:title><![CDATA[Cataract-associated deamidations on the surface of γS-crystallin increase protein unfolding and flexibility at distant regions]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-05-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/648543v1?rss=1">
<title>
<![CDATA[
EGR1 transcriptional control of human cytomegalovirus latency 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/648543v1?rss=1</link>
<description><![CDATA[
Sustained phosphotinositide3-kinase (PI3K) signaling is critical to the maintenance of herpesvirus latency. We have previously shown that the beta-herpesvirus, human cytomegalovirus (CMV), regulates epidermal growth factor receptor (EGFR), upstream of PI3K, to control states of latency and reactivation. Inhibition of EGFR signaling enhances CMV reactivation from latency and increases viral replication, but the mechanisms by which EGFR impacts replication and latency is not known. We demonstrate that HCMV downregulates MEK/ERK and AKT phosphorylation, but not STAT3 or PLC{gamma} for productive replication. Similarly, inhibition of either MEK/ERK or PI3K/AKT, but not STAT or PLC{gamma}, pathways increases viral reactivation from latently infected CD34+ hematopoietic progenitor cells (HPCs), defining a role for these pathways in latency. We hypothesized that CMV modulation of EGFR signaling might impact viral transcription. Indeed, EGF-stimulation increased expression of the UL138 latency gene, but not immediate early or early viral genes, suggesting that EGFR signaling promotes latent gene expression. The early growth response-1 (EGR1) transcription factor is induced downstream of EGFR signaling through both PI3K/AKT and MEK/ERK pathways. EGR1 expression is important for the maintenance of HPC stemness and its downregulation drives HPC differentiation and mobilization. We demonstrate that EGR1 binds upstream of UL138 and is sufficient to promote UL138 expression. Further, disruption of EGR1 binding upstream of UL138 prevented CMV from establishing a latent infection in CD34+ HPCs. Our results indicate a model whereby UL138 modulation of EGFR signaling feeds back to promote UL138 expression and suppression of replication to establish or maintain viral quiescence.nnAUTHOR SUMMARYCMV regulates EGFR signaling to balance states of viral latency and replication. CMV blocks downstream PI3K/AKT and MEK/ERK signaling pathways through down-regulation of EGFR at the plasma membrane. PI3K/AKT and MEK/ERK signaling increases expression of the EGR1 transcription factor that is necessary for the maintenance of stem cell stemness. A decrease in EGR1 expression promotes HPC mobilization to the periphery and differentiation, a known stimulus for CMV reactivation. We identified functional EGR1 binding sites upstream of the UL138 gene and EGR-1 binding stimulates UL138 expression. Additionally, down-regulation of EGR1 by CMV miR-US22 decreases UL138 expression emphasizing the importance of this transcription factor in expression of this latency gene. Lastly, we demonstrate that a CMV mutant virus lacking an upstream EGR1 binding site is unable to establish latency in CD34+ HPCs. This study defines one mechanism by which EGFR signaling impacts viral gene expression to promote CMV latency.
]]></description>
<dc:creator>Buehler, J. C.</dc:creator>
<dc:creator>Carpenter, E.</dc:creator>
<dc:creator>Zeltzer, S.</dc:creator>
<dc:creator>Igarashi, S.</dc:creator>
<dc:creator>Rak, M.</dc:creator>
<dc:creator>Mikell, I.</dc:creator>
<dc:creator>Nelson, J. A.</dc:creator>
<dc:creator>Goodrum, F.</dc:creator>
<dc:date>2019-05-24</dc:date>
<dc:identifier>doi:10.1101/648543</dc:identifier>
<dc:title><![CDATA[EGR1 transcriptional control of human cytomegalovirus latency]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-05-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/653527v1?rss=1">
<title>
<![CDATA[
Mutually exclusive autism mutations point to the circadian clock and PI3K signaling pathways 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/653527v1?rss=1</link>
<description><![CDATA[
Mutual exclusivity analysis of genomic mutations has proven useful for detecting driver alterations in cancer patient cohorts. Here we demonstrate, for the first time, that this pattern is also present among de novo mutations in autism spectrum disorder. We analyzed three large whole genome sequencing studies and identified mutual exclusivity patterns within the most confident set of autism-related genes, as well as in the circadian clock and PI3K/AKT signaling pathways.
]]></description>
<dc:creator>Manning, H.</dc:creator>
<dc:creator>O'Roak, B. J.</dc:creator>
<dc:creator>Babur, O.</dc:creator>
<dc:date>2019-05-30</dc:date>
<dc:identifier>doi:10.1101/653527</dc:identifier>
<dc:title><![CDATA[Mutually exclusive autism mutations point to the circadian clock and PI3K signaling pathways]]></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/10.1101/654988v1?rss=1">
<title>
<![CDATA[
HER2 Cancer Protrusion Growth Signaling Regulated by Unhindered, Localized Filopodial Dynamics 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/654988v1?rss=1</link>
<description><![CDATA[
Protrusions are plasma membrane extensions that are found in almost every cell in the human body. Cancer cell filopodial and lamellipodial protrusions play key roles in the integral processes of cell motility and signaling underlying tumor invasion and metastasis. HER2 (ErbB-2) is overexpressed in diverse types of tumors and regulates PI3K-pathway-mediated protrusion growth. It is known that HER2 resides at breast cancer cell protrusions, but how protrusion-based HER2 spatiotemporal dynamics shape cancer signaling is unclear. Here, we study how HER2 location and motion regulate protrusion signaling and growth using quantitative spatio-temporal molecular imaging approaches. Our data highlight morphologically-segregated features of filopodial and lamellipodial protrusions, in in vitro 2D breast cancer cells and in vivo intact breast tumor. Functional-segregation parallels morphological-segregation, as HER2 and its activated downstream pAKT-PI3K signaling remain spatially-localized at protrusions, provoking new protrusion growth proximal to sites of HER2 activation. HER2 in SKBR3 breast cancer cell filopodia exhibits fast, linearly-directed motion that is distinct from lamellipodia and non-protrusion subcellular regions ([~]3-4 times greater diffusion constant, rapid speeds of 2-3 um2/s). Surprisingly, filopodial HER2 motion is passive, requiring no active energy sources. Moreover, while HER2 motion in lamellipodia and non-protrusion regions show hindered diffusion typical of membrane proteins, HER2 diffuses freely within filopodia. We conclude that HER2 activation, propagation, and functional protrusion growth is a local process in which filopodia have evolved to exploit Brownian thermal fluctuations within a barrier-free nanostructure to transduce rapid signaling. These results support the importance of developing filopodia and other protrusion-targeted strategies for cancer.
]]></description>
<dc:creator>Lam, W. Y.</dc:creator>
<dc:creator>Wang, Y.</dc:creator>
<dc:creator>Mostofian, B.</dc:creator>
<dc:creator>Jorgens, D.</dc:creator>
<dc:creator>Kwon, S.</dc:creator>
<dc:creator>Chin, K.</dc:creator>
<dc:creator>Carpenter, M.</dc:creator>
<dc:creator>Jacob, T.</dc:creator>
<dc:creator>Heiser, K.</dc:creator>
<dc:creator>Agrawal, A.</dc:creator>
<dc:creator>Wang, J.</dc:creator>
<dc:creator>Nan, X.</dc:creator>
<dc:creator>Chang, Y. H.</dc:creator>
<dc:creator>Zuckerman, D. M.</dc:creator>
<dc:creator>Gray, J.</dc:creator>
<dc:creator>Bruchez, M.</dc:creator>
<dc:creator>Lidke, K.</dc:creator>
<dc:creator>Vu, T. Q.</dc:creator>
<dc:date>2019-06-02</dc:date>
<dc:identifier>doi:10.1101/654988</dc:identifier>
<dc:title><![CDATA[HER2 Cancer Protrusion Growth Signaling Regulated by Unhindered, Localized Filopodial Dynamics]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-06-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/657833v1?rss=1">
<title>
<![CDATA[
Affective Pavlovian motivation is enhanced in obesity susceptible populations; implications for incentive motivation in obesity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/657833v1?rss=1</link>
<description><![CDATA[
Global obesity rates continue to rise, presenting a major challenge to human health. Efforts to uncover the drivers of this epidemic have highlighted the contribution of Pavlovian motivational processes to overeating. In humans, brain and behavioral reactivity to food related stimuli positively correlates with subsequent weight gain. In concordance with this, selectively bred obesity-prone rats exhibit stronger cue-triggered food-seeking via single outcome Pavlovian-to-instrumental transfer (SO PIT) than obesity-resistant rats. These data show that Pavlovian motivation is stronger in selectively bred obesity-prone groups. However, whether obesity susceptibility in outbred populations is associated with enhanced PIT is unknown. Moreover, PIT can arise via two neurobehaviorally dissociable processes, a sensory specific versus a general affective process that cannot be distinguished by SO PIT. Thus, it is unclear which PIT process is enhanced in obesity-prone groups. Therefore, we determined whether obesity susceptibility in outbred populations is associated with enhanced Sensory Specific (SS) PIT or General PIT and whether expression of these forms of PIT differs between selectively bred obesity-prone versus obesity-resistant rats. We find that in outbred rats, the magnitude of General PIT is positively correlated with subsequently determined obesity susceptibility. In selectively bred rats, the magnitude of General PIT was stronger in obesity-prone versus obesity-resistant groups. Jointly, these data show that enhanced affective Pavlovian motivation is tightly linked to obesity vulnerability, supporting a role for phenotypic differences in incentive motivation for the development of obesity. This has important implications for obesity prevention and for understanding the neurocircuitry mediating enhanced food-seeking in vulnerable individuals.
]]></description>
<dc:creator>Derman, R. C.</dc:creator>
<dc:creator>Ferrario, C.</dc:creator>
<dc:date>2019-06-02</dc:date>
<dc:identifier>doi:10.1101/657833</dc:identifier>
<dc:title><![CDATA[Affective Pavlovian motivation is enhanced in obesity susceptible populations; implications for incentive motivation in obesity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-06-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/659060v1?rss=1">
<title>
<![CDATA[
A distinct neutrophil population invades the central nervous system during pancreatic cancer 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/659060v1?rss=1</link>
<description><![CDATA[
Weight loss, fatigue, and cognitive dysfunction are common symptoms in cancer patients that occur prior to initiation of cancer therapy. Inflammation in the brain is a driver of these symptoms, yet cellular sources of neuroinflammation during malignancy are unknown. In a mouse model of pancreatic ductal adenocarcinoma (PDAC), we observed early and robust myeloid cell infiltration into the brain. Infiltrating immune cells were predominately neutrophils, which accumulated at a unique central nervous system entry portal called the velum interpositum, where they expressed CCR2. CCR2 knockout mice had significantly decreased brain-infiltrating neutrophils as well as attenuated anorexia and muscle catabolism during PDAC, without any changes in neutrophils in other organs. Lastly, intracerebroventricular blockade of the purinergic receptor P2RX7 during PDAC abolished neutrophil recruitment to the brain and attenuated anorexia. Our data demonstrate a novel function for the CCR2/CCL2 axis in recruiting neutrophils to the brain, which drives anorexia and muscle catabolism.
]]></description>
<dc:creator>Burfeind, K. G.</dc:creator>
<dc:creator>Zhu, X.</dc:creator>
<dc:creator>Norgard, M. A.</dc:creator>
<dc:creator>Levasseur, P. R.</dc:creator>
<dc:creator>Olson, B.</dc:creator>
<dc:creator>Michaelis, K. A.</dc:creator>
<dc:creator>Torres, E. R. S.</dc:creator>
<dc:creator>Jeng, S.</dc:creator>
<dc:creator>McWeeney, S.</dc:creator>
<dc:creator>Raber, J.</dc:creator>
<dc:creator>Marks, D. L.</dc:creator>
<dc:date>2019-06-03</dc:date>
<dc:identifier>doi:10.1101/659060</dc:identifier>
<dc:title><![CDATA[A distinct neutrophil population invades the central nervous system during pancreatic cancer]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-06-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/659409v1?rss=1">
<title>
<![CDATA[
Behavioral and neural signatures of working memory in childhood 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/659409v1?rss=1</link>
<description><![CDATA[
Working memory function changes across development and varies across individuals. The patterns of behavior and brain function that track individual differences in working memory during development, however, are not well understood. Here we establish associations between working memory, cognitive abilities, and functional MRI activation in data from over 4,000 9-10-year-olds enrolled in the Adolescent Brain Cognitive Development study, an ongoing longitudinal study in the United States. Behavioral analyses reveal robust relationships between working memory, short-term memory, language skills, and fluid intelligence. Analyses relating out-of-scanner working memory performance to memory-related fMRI activation in an emotional n-back task demonstrate that frontoparietal activity in response to an explicit memory challenge indexes working memory ability. Furthermore, this relationship is domain-specific, such that fMRI activation related to emotion processing during the emotional n-back task, inhibitory control during a stop-signal task, and reward processing during a monetary incentive delay task does not track memory abilities. Together these results inform our understanding of the emergence of individual differences in working memory and lay the groundwork for characterizing the ways in which they change across adolescence.
]]></description>
<dc:creator>Rosenberg, M. D.</dc:creator>
<dc:creator>Martinez, S. A.</dc:creator>
<dc:creator>Rapuano, K. M.</dc:creator>
<dc:creator>Conley, M. I.</dc:creator>
<dc:creator>Cohen, A. O.</dc:creator>
<dc:creator>Cornejo, M. D.</dc:creator>
<dc:creator>Hagler, D. J.</dc:creator>
<dc:creator>Anderson, K. M.</dc:creator>
<dc:creator>Wager, T. D.</dc:creator>
<dc:creator>Feczko, E.</dc:creator>
<dc:creator>Earl, E.</dc:creator>
<dc:creator>Fair, D. A.</dc:creator>
<dc:creator>Barch, D. M.</dc:creator>
<dc:creator>Watts, R.</dc:creator>
<dc:creator>Casey, B.</dc:creator>
<dc:date>2019-06-06</dc:date>
<dc:identifier>doi:10.1101/659409</dc:identifier>
<dc:title><![CDATA[Behavioral and neural signatures of working memory in childhood]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-06-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/665026v1?rss=1">
<title>
<![CDATA[
Burden of tumor mutations, neoepitopes, and other variants are cautionary predictors of cancer immunotherapy response and overall survival 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/665026v1?rss=1</link>
<description><![CDATA[
BackgroundTumor mutational burden (TMB, the quantity of aberrant nucleotide sequences a given tumor may harbor) has been associated with response to immune checkpoint inhibitor therapy and is gaining broad acceptance as a result. However, TMB harbors intrinsic variability across cancer types, and its assessment and interpretation are poorly standardized.

MethodsUsing a standardized approach, we quantify the robustness of TMB as a metric and its potential as a predictor of immunotherapy response and survival among a diverse cohort of cancer patients. We also explore the additive predictive potential of RNA-derived variants and neoepitope burden, incorporating several novel metrics of immunogenic potential.

ResultsWe find that TMB is a partial predictor of immunotherapy response in melanoma and non-small cell lung cancer, but not renal cell carcinoma. We find that TMB is predictive of overall survival in melanoma patients receiving immunotherapy, but not in an immunotherapy-naive population. We also find that it is an unstable metric with potentially problematic repercussions for clinical cohort classification. We finally note minimal additional predictive benefit to assessing neoepitope burden or its bulk derivatives, including RNA-derived sources of neoepitopes.

ConclusionsWe find sufficient cause to suggest that the predictive clinical value of TMB should not be overstated or oversimplified. While it is readily quantified, TMB is at best a limited surrogate biomarker of immunotherapy response. The data do not support isolated use of TMB in renal cell carcinoma.
]]></description>
<dc:creator>Wood, M. A.</dc:creator>
<dc:creator>Weeder, B. R.</dc:creator>
<dc:creator>David, J. K.</dc:creator>
<dc:creator>Nellore, A.</dc:creator>
<dc:creator>Thompson, R. F.</dc:creator>
<dc:date>2019-06-08</dc:date>
<dc:identifier>doi:10.1101/665026</dc:identifier>
<dc:title><![CDATA[Burden of tumor mutations, neoepitopes, and other variants are cautionary predictors of cancer immunotherapy response and overall survival]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-06-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/668277v1?rss=1">
<title>
<![CDATA[
The Eya1 phosphatase mediates Shh-driven symmetric cell division of cerebellar granule cell precursors 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/668277v1?rss=1</link>
<description><![CDATA[
During neural development, stem and precursor cells can divide either symmetrically or asymmetrically. The transition between symmetric and asymmetric cell divisions is a major determinant of precursor cell expansion and neural differentiation, but the underlying mechanisms that regulate this transition are not well understood. Here, we identify the Sonic hedgehog (Shh) pathway as a critical determinant regulating the mode of division of cerebellar granule cell precursors (GCPs). Using partial gain and loss of function mutations within the Shh pathway, we show that pathway activation determines spindle orientation of GCPs, and that mitotic spindle orientation directly correlates with the mode of division. Mechanistically, we show that the phosphatase Eya1 is essential for implementing Shh-dependent GCP spindle orientation. We identify atypical protein kinase C (aPKC) as a direct target of Eya1 activity and show that Eya1 dephosphorylates Threonine (T410) in the activation loop of this polarity complex component. Thus, Eya1 inactivates the cell polarity complex, resulting in reduced phosphorylation of Numb and other components that regulate the mode of division. This Eya1-dependent cascade is critical in linking spindle orientation, cell cycle exit and terminal differentiation. Together these findings demonstrate that a Shh-Eya1 regulatory axis selectively promotes symmetric cell divisions during cerebellar development by coordinating spindle orientation and cell fate determinants.nnSummary statementBiological responses to Shh signaling are specified by the magnitude of pathway activation and the cellular context. This study shows that potent Shh signaling regulates mitotic orientation and symmetric division of cerebellar granule cell precursors in a process that requires the phosphatase Eya1 and unequal distribution of cell fate determinants to daughter cells.
]]></description>
<dc:creator>Merk, D. J.</dc:creator>
<dc:creator>Zhou, P.</dc:creator>
<dc:creator>Cohen, S. M.</dc:creator>
<dc:creator>Pazyra-Murphy, M. F.</dc:creator>
<dc:creator>Hwang, G. H.</dc:creator>
<dc:creator>Rehm, K. J.</dc:creator>
<dc:creator>Alfaro, J.</dc:creator>
<dc:creator>Zhao, X.</dc:creator>
<dc:creator>Park, E.</dc:creator>
<dc:creator>Xu, P.-X.</dc:creator>
<dc:creator>Chan, J. A.</dc:creator>
<dc:creator>Eck, M. J.</dc:creator>
<dc:creator>Nazemi, K. J.</dc:creator>
<dc:creator>Segal, R. A.</dc:creator>
<dc:date>2019-06-11</dc:date>
<dc:identifier>doi:10.1101/668277</dc:identifier>
<dc:title><![CDATA[The Eya1 phosphatase mediates Shh-driven symmetric cell division of cerebellar granule cell precursors]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-06-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/671172v1?rss=1">
<title>
<![CDATA[
Extensive Evaluation of Weighted Ensemble Strategies for Calculating Rate Constants and Binding Affinities of Molecular Association/Dissociation Processes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/671172v1?rss=1</link>
<description><![CDATA[
The weighted ensemble (WE) path sampling strategy is highly efficient in generating pathways and rate constants for rare events using atomistic molecular dynamics simulations. Here we extensively evaluated the impact of several advances to the WE strategy on the efficiency of computing association and dissociation rate constants (kon, koff) as well as binding affinities (KD) for a set of benchmark systems, listed in order of increasing timescales of molecular association/dissociation processes: methane/methane, Na+/Cl-, and K+/18-crown-6 ether. In particular, we assessed the advantages of carrying out (i) a large set of "light-weight" WE simulations that each consist of a small number of trajectories vs. a single "heavy-weight" WE simulation that consists of a relatively large number of trajectories, (ii) equilibrium vs. steady-state WE simulations, (iii) history augmented Markov State Model (haMSM) post-simulation analysis of equilibrium sets of trajectories, and (iv) tracking of trajectory history (the state last visited) during the dynamics propagation of equilibrium WE simulations. Provided that state definitions are known in advance, our results reveal that heavy-weight, steady-state WE simulations are the most efficient protocol for calculating kon, koff, and KD values. If states are not strictly defined in advance, heavy-weight, equilibrium WE simulations are the most efficient protocol. This efficiency can be further improved with the inclusion of trajectory history during dynamics propagation. In addition, applying the haMSM post-simulation analysis enhances the efficiency of both steady-state and equilibrium WE simulations. Recommendations of appropriate WE protocols are made according to the goals of the simulations (e.g. to efficiently calculate rate constants and/or generate a diverse set of pathways).
]]></description>
<dc:creator>Pratt, A.</dc:creator>
<dc:creator>Suarez, E.</dc:creator>
<dc:creator>Zuckerman, D.</dc:creator>
<dc:creator>Chong, L.</dc:creator>
<dc:date>2019-06-14</dc:date>
<dc:identifier>doi:10.1101/671172</dc:identifier>
<dc:title><![CDATA[Extensive Evaluation of Weighted Ensemble Strategies for Calculating Rate Constants and Binding Affinities of Molecular Association/Dissociation Processes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-06-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/673186v1?rss=1">
<title>
<![CDATA[
How to build a fast and highly sensitive sound detector that remains robust to temperature shifts 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/673186v1?rss=1</link>
<description><![CDATA[
Frogs must have sharp hearing abilities during the warm summer months to successfully find mating partners. This study aims to understand how frog hair cell ribbon-type synapses preserve both sensitivity and temporal precision during temperature changes. We performed in vitro patch-clamp recordings of hair cells and their afferent fibers in bullfrog amphibian papillae under room (23-25{degrees}C) and high (30-33{degrees}C) temperature. Afferent fibers exhibited a wide heterogeneity in membrane input resistance (Rin) from 100 M{Omega} to 1000 M{Omega}, which may contribute to variations in spike threshold and firing frequency. At higher temperatures, most fibers increased their frequency of action potential firing due to an increase in spontaneous EPSC frequencies. Hair cell resting membrane potential (Vrest) remained surprisingly stable during temperature increases, although both inward Ca2+ current and outward K+ current increased in amplitude. This increase in Ca2+ current may explain the higher spontaneous EPSC frequencies. The larger "leak currents" at Vrest lowered Rin and produced higher electrical resonant frequencies. However, lower Rin should decrease sensitivity to sound detection via smaller receptor potentials. Using membrane capacitance measurements, we suggest that hair cells can partially compensate for this reduced sensitivity by increasing exocytosis efficiency and the size of the readily releasable pool of synaptic vesicles. Furthermore, paired recordings of hair cells and their afferent fibers showed that synaptic delays become shorter and multivesicular release becomes more synchronous at higher temperatures, which should improve temporal precision. Altogether, our results explain many previous in vivo observations on the temperature dependence of spikes in auditory nerves.nnSignificance StatementThe vertebrate inner ear detects and transmits auditory information over a broad dynamic range of sound frequency and intensity. It achieves remarkable sensitivity to soft sounds and precise frequency selectivity. How does the ear of cold-blooded vertebrates maintain its performance level as temperature changes? More specifically, how does the hair cell to afferent fiber synapse in bullfrog amphibian papilla adjust to a wide range of physiological temperatures without losing its sensitivity and temporal fidelity to sound signals? This study uses in vitro experiments to reveal the biophysical mechanisms that explain many observations made from in vivo auditory nerve fiber recordings. We find that higher temperature facilitates vesicle exocytosis and electrical tuning to higher sound frequencies, which benefits sensitivity and selectivity.
]]></description>
<dc:creator>Chen, M.</dc:creator>
<dc:creator>von Gersdorff, H.</dc:creator>
<dc:date>2019-06-16</dc:date>
<dc:identifier>doi:10.1101/673186</dc:identifier>
<dc:title><![CDATA[How to build a fast and highly sensitive sound detector that remains robust to temperature shifts]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-06-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/674689v1?rss=1">
<title>
<![CDATA[
Does Sex Modify an Association of Electrophysiological Substrate with Sudden Cardiac Death? The Atherosclerosis Risk in Communities (ARIC) Study 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/674689v1?rss=1</link>
<description><![CDATA[
BackgroundSex is a well-recognized risk factor for sudden cardiac death (SCD). Sex differences in electrophysiological (EP) substrate of SCD are known. However, it remains unknown whether sex can modify an association of EP substrate with SCD.nnMethodsParticipants from the Atherosclerosis Risk in Communities study with analyzable ECGs (n=14,725; age, 54.2{+/-}5.8 yrs; 55% female, 74% white) were included. EP substrate was characterized by traditional 12-lead ECG (heart rate, QRS, QTc, Cornell voltage), spatial ventricular gradient (SVG) and sum absolute QRST integral (SAI QRST) metrics. Two competing outcomes were adjudicated SCD and nonSCD. Interaction of ECG metrics with sex was studied in Cox proportional hazards and Fine-Gray competing risk models. Relative hazard ratio (RHR) and relative sub-hazard ratio (RSHR) with a 95% confidence interval for SCD and nonSCD risk for women relative to men were calculated. Model 1 was adjusted for prevalent cardiovascular disease (CVD) and risk factors. Time-updated model 2 was additionally adjusted for incident non-fatal CVD.nnResultsOver a median follow-up of 24.4 years, there were 530 SCDs (incidence 1.72 (1.58-1.88)/1000 person-years) and 2,178 nonSCDs (incidence 7.09; (6.80-7.39)/ 1000 person-years). Women experienced a greater than men risk of SCD associated with Cornell voltage (RHR 1.18(1.06-1.32); P=0.003), SAI QRST (RHR 1.16(1.04-1.30); P=0.007), area SVG magnitude (RHR 1.24(1.05-1.45); P=0.009), and peak SVG magnitude (RHR 1.22(1.04-1.44); P=0.018), independently from incident CVD. Greater risk of SCD for women than men associated with QRS duration (RHR 1.24(1.07-1.44); P=0.004) and QTc (RSHR 1.15(1.02-1.30); P=0.025) was explained by incident CVD. Furthermore, women had greater odds of SCD associated with heart rate (RSHR 1.19(1.01-1.40); P=0.036), independently of incident CVD.nnConclusionsSex modifies an association of EP substrate with SCD. In women, global EP substrate is associated with up to 27% greater risk of SCD than in men. Development of sex-specific risk scores of SCD is necessary. Further studies of mechanisms behind sex differences in EP substrate of SCD are warranted.
]]></description>
<dc:creator>Howell, S.</dc:creator>
<dc:creator>German, D.</dc:creator>
<dc:creator>Bender, A.</dc:creator>
<dc:creator>Phan, F.</dc:creator>
<dc:creator>Mukundan, S.</dc:creator>
<dc:creator>Perez-Alday, E.</dc:creator>
<dc:creator>Rogovoy, N.</dc:creator>
<dc:creator>Haq, K.</dc:creator>
<dc:creator>Yang, K.</dc:creator>
<dc:creator>Wirth, A.</dc:creator>
<dc:creator>Jensen, K.</dc:creator>
<dc:creator>Tereshchenko, L. G.</dc:creator>
<dc:date>2019-06-21</dc:date>
<dc:identifier>doi:10.1101/674689</dc:identifier>
<dc:title><![CDATA[Does Sex Modify an Association of Electrophysiological Substrate with Sudden Cardiac Death? The Atherosclerosis Risk in Communities (ARIC) Study]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-06-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/675371v1?rss=1">
<title>
<![CDATA[
A workflow for visualizing human cancer biopsies using large-format electron microscopy 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/675371v1?rss=1</link>
<description><![CDATA[
Recent developments in large format electron microscopy have enabled generation of images that provide detailed ultrastructural information on normal and diseased cells and tissues. Analyses of these images increase our understanding of cellular organization and interactions and disease-related changes therein. In this manuscript, we describe a workflow for two-dimensional (2D) and three-dimensional (3D) imaging, including both optical and scanning electron microscopy (SEM) methods, that allow pathologists and cancer biology researchers to identify areas of interest from human cancer biopsies. The protocols and mounting strategies described in this workflow are compatible with 2D large format EM mapping, 3D focused ion beam-SEM and serial block face-SEM. The flexibility to use diverse imaging technologies available at most academic institutions makes this workflow useful and applicable for most life science samples. Volumetric analysis of the biopsies studied here revealed morphological, organizational and ultrastructural aspects of the tumor cells and surrounding environment that cannot be revealed by conventional 2D EM imaging. Our results indicate that although 2D EM is still an important tool in many areas of diagnostic pathology, 3D images of ultrastructural relationships between both normal and cancerous cells, in combination with their extracellular matrix, enables cancer researchers and pathologists to better understand the progression of the disease and identify potential therapeutic targets.
]]></description>
<dc:creator>Riesterer, J. L.</dc:creator>
<dc:creator>Lopez, C. S.</dc:creator>
<dc:creator>Stempinski, E. S.</dc:creator>
<dc:creator>Williams, M.</dc:creator>
<dc:creator>Loftis, K.</dc:creator>
<dc:creator>Stoltz, K.</dc:creator>
<dc:creator>Thibault, G.</dc:creator>
<dc:creator>Lanicault, C.</dc:creator>
<dc:creator>Williams, T.</dc:creator>
<dc:creator>Gray, J. W.</dc:creator>
<dc:date>2019-06-19</dc:date>
<dc:identifier>doi:10.1101/675371</dc:identifier>
<dc:title><![CDATA[A workflow for visualizing human cancer biopsies using large-format electron microscopy]]></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/10.1101/675538v1?rss=1">
<title>
<![CDATA[
Host mucin is subverted by Pseudomonas aeruginosa during infection to provide free glycans required for successful colonization 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/675538v1?rss=1</link>
<description><![CDATA[
The mucosal barrier, found lining epithelial cells, serves multiple functions in a range of animals. The major structural components of mucus are mucins, which are heavily glycosylated proteins that are either membrane bound or secreted by the epithelial cells. Mucins are key components of the innate immune system, as they are involved in the clearance of pathogens from the airways and intestines, and their expression is typically upregulated upon epithelial cell exposure to a variety of pathogens. In this study, we identified the mucin MUL-1 as an innate immune factor that appears to be utilized by P. aeruginosa to colonize hosts. We found that while the expression of several mucins, including MUL-1, increased upon P. aeruginosa infection of the nematode Caenorhabditis elegans, silencing of or deletion of mul-1 resulted in enhanced survival and reduced bacterial accumulation. P. aeruginosa required host sialidase CTSA-1.1 to use mucin-derived glycans to colonize the host, while sialidase-encoding bacteria required host MUL-1 but not CTSA-1.1 to cause a lethal infection. This role of mucins and free glycans in host-pathogen interaction appears to be conserved from C. elegans to humans, as P. aeruginosa binding to human lung epithelial cells was also enhanced in the presence of free glycans, and free glycans reversed the binding defect of P. aeruginosa to human lung cells lacking the mucin MUC1.nnAuthor SummaryThe gastrointestinal, respiratory, reproductive, and urinary tracts, are large surfaces exposed to the exterior environment and thus, these mucosal epithelial tissues serve as primary routes of infection. One of the first lines of defense present at these barriers is mucus, which is a highly viscous material formed by mucin glycoproteins. Mucins serve various functions, but importantly they aid in the clearance of pathogens and debris from epithelial barriers and serve as innate immune effectors. In this study, we describe the ability of Pseudomonas aeruginosa to utilize mucin-derived glycans to colonize the intestine and ultimately cause death in Caenorhabditis elegans. We also show conserved mechanisms of P. aeruginosa virulence traits, by demonstrating that free glycans alter the ability of the bacteria to bind to human lung alveolar epithelial cells. Over the course of host-pathogen evolution, pathogens seem to have evolved to use mucins for their own advantage, and thus one of the biggest questions is which party benefits from pathogen-mucin binding. By gaining a better understanding of pathogen-mucin interactions, we can better protect against pathogen infection.
]]></description>
<dc:creator>Hoffman, C. L.</dc:creator>
<dc:creator>Aballay, A.</dc:creator>
<dc:date>2019-06-20</dc:date>
<dc:identifier>doi:10.1101/675538</dc:identifier>
<dc:title><![CDATA[Host mucin is subverted by Pseudomonas aeruginosa during infection to provide free glycans required for successful colonization]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-06-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/683672v1?rss=1">
<title>
<![CDATA[
Using a bar-coded AAV capsid library to select for novel clinically relevant gene therapy vectors 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/683672v1?rss=1</link>
<description><![CDATA[
ABSTRACTWhile gene transfer using recombinant adeno-associated viral (rAAV) vectors have shown success in some clinical trials, there remain many tissues that are not well transduced. Because of the recent success in reprogramming islet derived cells into functional {beta}-cells in animal models, we constructed two highly complex barcoded replication competent capsid shuffled libraries and selected for high transducing variants on primary human islets. We describe a chimeric capsid (AAV-KP1) that penetrated and transduced primary human islet cells and human embryonic stem cell derived {beta}-cells with up to 10-fold higher efficiency compared to previously studied best in class AAV vectors. Remarkably, this chimeric capsid was also able to transduce both mouse and human hepatocytes at very high levels in a humanized-chimeric mouse model, thus providing a versatile vector which has the potential to be used in both preclinical testing and human clinical trials for both liver-based diseases and diabetes.
]]></description>
<dc:creator>Pekrun, K.</dc:creator>
<dc:creator>De Alencastro, G.</dc:creator>
<dc:creator>Luo, Q.-J.</dc:creator>
<dc:creator>Kim, Y.</dc:creator>
<dc:creator>Nygaard, S.</dc:creator>
<dc:creator>Galivo, F.</dc:creator>
<dc:creator>Zhang, F.</dc:creator>
<dc:creator>Song, R.</dc:creator>
<dc:creator>Tiffany, M.</dc:creator>
<dc:creator>Xu, J.</dc:creator>
<dc:creator>Hebron, M.</dc:creator>
<dc:creator>Grompe, M.</dc:creator>
<dc:creator>Kay, M. A.</dc:creator>
<dc:date>2019-06-27</dc:date>
<dc:identifier>doi:10.1101/683672</dc:identifier>
<dc:title><![CDATA[Using a bar-coded AAV capsid library to select for novel clinically relevant gene therapy vectors]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-06-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/684563v1?rss=1">
<title>
<![CDATA[
Should virus capsids assemble perfectly? A simple equilibrium model for defects 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/684563v1?rss=1</link>
<description><![CDATA[
Although published structural models of viral capsids generally exhibit a high degree of regularity or symmetry, structural defects might be expected because of the fluctuating environment in which capsids assemble and the requirement of some capsids for disassembly prior to genome delivery. Defective structures are observed in computer simulations, and are evident in single-particle cryoEM studies. Here, we quantify the conditions under which defects might be expected, using a statistical mechanics model allowing for ideal, defective, and vacant sites. The model displays a threshold in affinity parameters below which there is an appreciable population of defective capsids. Even when defective sites are not allowed, there is generally some population of vacancies. Analysis of single particles in cryoEM micrographs yields a confirmatory {gtrsim}15% of defective particles. Our findings suggest structural heterogeneity in virus capsids may be under-appreciated, and also points to a non-traditional strategy for assembly inhibition.
]]></description>
<dc:creator>Spiriti, J.</dc:creator>
<dc:creator>Conway, J.</dc:creator>
<dc:creator>Zuckerman, D. M.</dc:creator>
<dc:date>2019-06-27</dc:date>
<dc:identifier>doi:10.1101/684563</dc:identifier>
<dc:title><![CDATA[Should virus capsids assemble perfectly? A simple equilibrium model for defects]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-06-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/685818v1?rss=1">
<title>
<![CDATA[
α2δ-2 protein controls structure and function at the cerebellar climbing fiber synapse 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/685818v1?rss=1</link>
<description><![CDATA[
2{delta} proteins (Cacna2d1-4) are auxiliary subunits of voltage-dependent calcium channels that also drive synapse formation and maturation. Because cerebellar Purkinje cells (PCs) only express one isoform of this family, 2{delta}-2 (Cacna2d2), we used PCs as a model system to examine roles of 2{delta} in excitatory synaptic function in a Cacna2d2 knockout mouse. Whole-cell recordings of PCs from acute cerebellar slices revealed altered climbing fiber (CF)-evoked complex spike generation, as well as increased amplitude and faster decay of CF-evoked excitatory postsynaptic currents (EPSCs). CF terminals in the KO were localized more proximally on PC dendrites, as indicated by VGLUT2+ immunoreactive puncta, and computational modeling demonstrated that the increased EPSC amplitude can be partly attributed to the more proximal location of CF terminals. In addition, CFs in KO mice exhibited increased multivesicular transmission, corresponding to greater sustained responses during repetitive stimulation, despite a reduction in the measured probability of release. Electron microscopy demonstrated that mutant CF terminals had twice as many vesicle release sites, providing a morphologic explanation for the enhanced glutamate release. Though KO CFs evoked larger amplitude EPSCs, the charge transfer was the same as wildtype as a result of increased glutamate re-uptake, producing faster decay kinetics. Together, the larger, faster EPSCs in the KO explain the altered complex spike responses, which degrade information transfer from PCs and likely contribute to ataxia in Cacna2d2 KO mice. Our results also illustrate the multidimensional synaptic roles of 2{delta} proteins.

Significance Statement2{delta} proteins (Cacna2d1-4) regulate synaptic transmission and synaptogenesis, but co-expression of multiple 2{delta} isoforms has obscured a clear understanding of how various 2{delta} proteins control synaptic function. We focused on roles of the 2{delta}-2 protein (Cacna2d2), whose deletion causes cerebellar ataxia and epilepsy in mice and humans. Because cerebellar Purkinje cells only expresses this single isoform, we studied excitatory climbing fiber synaptic function onto Purkinje cells in Cacna2d2 knockout mice. Using optical and electrophysiological analysis, we provide a detailed description of the changes in Purkinje cells lacking 2{delta}-2, and provide a comprehensive mechanistic explanation for how functional synaptic phenotypes contribute to the altered cerebellar output.
]]></description>
<dc:creator>Beeson, K. A.</dc:creator>
<dc:creator>Beeson, R.</dc:creator>
<dc:creator>Westbrook, G. L.</dc:creator>
<dc:creator>Schnell, E.</dc:creator>
<dc:date>2019-06-28</dc:date>
<dc:identifier>doi:10.1101/685818</dc:identifier>
<dc:title><![CDATA[α2δ-2 protein controls structure and function at the cerebellar climbing fiber synapse]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-06-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/685933v1?rss=1">
<title>
<![CDATA[
What is that molecular machine really doing? Automated exploration of alternative transporter mechanisms 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/685933v1?rss=1</link>
<description><![CDATA[
Motivated by growing evidence for pathway heterogeneity and alternative functions of molecular machines, we demonstrate a computational approach for investigating two questions: (1) Are there multiple mechanisms (state-space pathways) by which a machine can perform a given function, such as cotransport across a membrane? (2) How can additional functionality, such as proofreading/error-correction, be built into machine function using standard biochemical processes? Answers to these questions will aid both the understanding of molecular-scale cell biology and the design of synthetic machines. Focusing on transport in this initial study, we sample a variety of mechanisms by employing Metropolis Markov chain Monte Carlo. Trial moves adjust transition rates among an automatically generated set of conformational and binding states while maintaining fidelity to thermodynamic principles and a user-supplied fitness/functionality goal. Each accepted move generates a new model. The simulations yield both single and mixed reaction pathways for cotransport in a simple environment with a single substrate along with a driving ion. In a "competitive" environment including an additional decoy substrate, several qualitatively distinct reaction pathways are found which are capable of extremely high discrimination coupled to a leak of the driving ion, akin to proofreading. The array of functional models would be difficult to find by intuition alone in the complex state-spaces of interest.

Author summaryMolecular machines, which operate on the nanoscale, are proteins/complexes that perform remarkable tasks such as the selective absorption of nutrients into the cell by transporters. These complex machines are often described using a fairly simple set of states and transitions that may not account for the stochasticity and heterogeneity generally expected at the nanoscale at body temperature. New tools are needed to study the full array of possibilities. This study presents a novel in silico method to systematically generate testable molecular-machine kinetic models and explore alternative mechanisms, applied first to membrane transport proteins. Our initial results suggest these transport machines may contain mechanisms which  detoxify the cell of an unwanted toxin, as well as significantly discriminate against the import of the toxin. This novel approach should aid the experimental study of key physiological processes such as renal glucose re-absorption, rational drug design, and potentially the development of synthetic machines.
]]></description>
<dc:creator>George, A.</dc:creator>
<dc:creator>Bisignano, P.</dc:creator>
<dc:creator>Rosenberg, J.</dc:creator>
<dc:creator>Grabe, M.</dc:creator>
<dc:creator>Zuckerman, D. M.</dc:creator>
<dc:date>2019-06-28</dc:date>
<dc:identifier>doi:10.1101/685933</dc:identifier>
<dc:title><![CDATA[What is that molecular machine really doing? Automated exploration of alternative transporter mechanisms]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-06-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/687756v1?rss=1">
<title>
<![CDATA[
ELQ-331 as a prototype for extremely durable chemoprotection against malaria 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/687756v1?rss=1</link>
<description><![CDATA[
BackgroundThe potential benefits of long-acting injectable chemoprotection (LAI-C) against malaria have been recently recognized, prompting a call for suitable candidate drugs to help meet this need. On the basis of its known pharmacodynamic and pharmacokinetic profiles after oral dosing, ELQ-331, a prodrug of the parasite mitochondrial electron transport inhibitor ELQ-300, was selected for study of pharmacokinetics and efficacy as LAI-C in mice.nnMethodsFour trials were conducted in which mice were injected with a single intramuscular dose of ELQ-331 or other ELQ-300 prodrugs in sesame oil with 1.2% benzyl alcohol; the ELQ-300 content of the doses ranged from 2.5 to 30 mg/kg. Initial blood stage challenges with Plasmodium yoelii were used to establish the model, but the definitive study measure of efficacy was outcome after sporozoite challenge with a luciferase-expressing P.yoelii, assessed by whole-body live animal imaging. Snapshot determinations of plasma ELQ-300 concentration ([ELQ-300]) were made after all prodrug injections; after the highest dose of ELQ-331 (equivalent to 30 mg/kg ELQ-300), both [ELQ-331] and [ELQ-300] were measured at a series of timepoints from 6 hours to 5 [1/2] months after injection.nnResultsA single intramuscular injection of ELQ-331 outperformed four other ELQ-300 prodrugs and, at a dose equivalent to 30 mg/kg ELQ-300, protected mice against challenge with P. yoelii sporozoites for at least 4 [1/2] months. Pharmacokinetic evaluation revealed rapid and essentially complete conversion of ELQ-331 to ELQ-300, a rapidly achieved (< 6 hours) and sustained (4-5 months) effective plasma ELQ-300 concentration, maximum ELQ-300 concentrations far below the estimated threshold for toxicity, and a distinctive ELQ-300 concentration vs. time profile. Pharmacokinetic modeling indicates a high-capacity, slow-exchange tissue compartment which serves to accumulate and then slowly redistribute ELQ-300 into blood, and this property facilitates an extremely long period during which ELQ-300 concentration is sustained above a minimum fully-protective threshold (60-80 nM).nnConclusionsExtrapolation of these results to humans clearly predicts that ELQ-331 should be capable of meeting and far-exceeding currently published duration-of-effect goals for antimalarial LAI-C. Allometric scaling from mice to humans would predict a several-fold enhancement in the relationship between duration-of-effect and dose, and available drug engineering and formulation technologies would be expected to offer significant improvement over the simple powder in sesame oil used here. Furthermore, the distinctive pharmacokinetic profile of ELQ-300 after treatment with ELQ-331 may facilitate durable protection using a variety of delivery and formulation options, and may enable protection for far longer than 3 months. Particularly in light of the favorable pharmacodynamic profile of ELQ-300, ELQ-331 warrants consideration as a leading prototype for LAI-C.
]]></description>
<dc:creator>Smilkstein, M. J.</dc:creator>
<dc:creator>Pou, S.</dc:creator>
<dc:creator>Krollenbrock, A.</dc:creator>
<dc:creator>Bleyle, L. A.</dc:creator>
<dc:creator>Dodean, R. A.</dc:creator>
<dc:creator>Frueh, L.</dc:creator>
<dc:creator>Hinrichs, D. J.</dc:creator>
<dc:creator>Li, Y.</dc:creator>
<dc:creator>Martinson, T.</dc:creator>
<dc:creator>Munar, M. Y.</dc:creator>
<dc:creator>Winter, R. W.</dc:creator>
<dc:creator>Bruzual, I.</dc:creator>
<dc:creator>Whiteside, S.</dc:creator>
<dc:creator>Nilsen, A.</dc:creator>
<dc:creator>Koop, D. R.</dc:creator>
<dc:creator>Kelly, J. X.</dc:creator>
<dc:creator>Kappe, S. H. I.</dc:creator>
<dc:creator>Wilder, B. K.</dc:creator>
<dc:creator>Riscoe, M. K.</dc:creator>
<dc:date>2019-07-02</dc:date>
<dc:identifier>doi:10.1101/687756</dc:identifier>
<dc:title><![CDATA[ELQ-331 as a prototype for extremely durable chemoprotection against malaria]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-07-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/688309v1?rss=1">
<title>
<![CDATA[
Dendritic speeding of synaptic potentials in an auditory brainstem principal neuron 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/688309v1?rss=1</link>
<description><![CDATA[
Principal cells of the medial nucleus of the trapezoid body (MNTB) in the mammalian auditory brainstem receive most of their strong synaptic inputs directly on the cell soma. However, these neurons also grow extensive dendrites during the first four postnatal weeks. What are the functional roles of these dendrites? We studied the morphology and growth of the dendrites in the mouse MNTB using both electron microscopy and confocal fluorescence imaging from postnatal day 9 (P9; pre-hearing) to P30. The soma of principal cells sprouted 1 to 3 thin dendrites (diameter ~ 1.5 microns) by P21 to P30. Each dendrite bifurcated into 2-3 branches and spanned an overall distance of about 80 to 200 microns. By contrast, at P9-11 the soma had 1 to 2 dendrites that extended for only 25 microns on average. Patch clamp experiments revealed that the growth of dendrites during development correlates with a progressive decrease in the input resistance, whereas acute removal of dendrites during brain slicing leads to higher input resistances. Accordingly, recordings of excitatory postsynaptic potentials (EPSPs) evoked by afferent fiber stimulation show that EPSP decay is faster in P21-24 neurons with intact dendrites than in neurons without dendrites. This dendritic speeding of the EPSP reduces the decay time constant 5-fold, which will impact significantly synaptic current summation and the ability to fire high-frequency spike trains. These data suggest a novel role for dendrites in auditory brainstem neurons: the speeding of EPSPs for faster and more precise output signal transfer.nnSignificance StatementAuditory circuits that compute sound localization express different types of specialized synapses. Some are capable of fast, precise and sustained synaptic transmission. As the paradigm example, principal cells of the MNTB receive a single calyx-type nerve terminal on their soma and this large excitatory synapse produces fast and brief supra-threshold EPSPs that can trigger trains of high frequency spikes. However, these neurons also extend thin and long dendrites with unknown function. We examined the relationship between dendritic morphology, passive electrical properties and EPSP waveform. We found that more mature neurons with intact dendrites have lower input resistances and short EPSP waveforms, ideally suited for conveying precise timing information, whereas immature neurons with shorter dendrites and higher input resistance have longer lasting EPSPs.
]]></description>
<dc:creator>Srinivasan, G.</dc:creator>
<dc:creator>Dagostin, A.</dc:creator>
<dc:creator>Leao, R. N.</dc:creator>
<dc:creator>Balakrishnan, V.</dc:creator>
<dc:creator>Holcomb, P.</dc:creator>
<dc:creator>Jackson, D.</dc:creator>
<dc:creator>Spirou, G.</dc:creator>
<dc:creator>von Gersdorff, H.</dc:creator>
<dc:date>2019-07-02</dc:date>
<dc:identifier>doi:10.1101/688309</dc:identifier>
<dc:title><![CDATA[Dendritic speeding of synaptic potentials in an auditory brainstem principal neuron]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-07-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/689828v1?rss=1">
<title>
<![CDATA[
Predicting Primary Site of Secondary Liver Cancer with a Neural Estimator of Metastatic Origin (NEMO) 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/689828v1?rss=1</link>
<description><![CDATA[
Pathologists rely on clinical information, tissue morphology, and sophisticated molecular diagnostics to accurately infer the metastatic origin of secondary liver cancer. In this paper, we introduce a deep learning approach to identify spatially localized regions of cancerous tumor within hematoxylin and eosin stained tissue sections of liver cancer and to generate predictions of the cancers metastatic origin. Our approach achieves an accuracy of 90.2% when classifying metastatic origin of whole slide images into three distinct classes, which compares favorably to an established clinical benchmark by three board-certified pathologists whose accuracies ranged from 90.2% to 94.1% on the same prediction task. This approach illustrates the potential impact of deep learning systems to leverage morphological and structural features of H&E stained tissue sections to guide pathological and clinical determination of the metastatic origin of secondary liver cancers.
]]></description>
<dc:creator>Schau, G. F.</dc:creator>
<dc:creator>Burlingame, E. A.</dc:creator>
<dc:creator>Thibault, G.</dc:creator>
<dc:creator>Anekpuritanang, T.</dc:creator>
<dc:creator>Wang, Y.</dc:creator>
<dc:creator>Gray, J. W.</dc:creator>
<dc:creator>Corless, C.</dc:creator>
<dc:creator>Chang, Y. H.</dc:creator>
<dc:date>2019-07-02</dc:date>
<dc:identifier>doi:10.1101/689828</dc:identifier>
<dc:title><![CDATA[Predicting Primary Site of Secondary Liver Cancer with a Neural Estimator of Metastatic Origin (NEMO)]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-07-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/690875v1?rss=1">
<title>
<![CDATA[
Restructuring of amygdala subregion apportion across adolescence 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/690875v1?rss=1</link>
<description><![CDATA[
Total amygdala volumes develop in association with sex and puberty, and postmortem studies find neuronal numbers increase in a nuclei specific fashion across development. Thus, amygdala subregions and composition may evolve with age. Our goal was to examine if amygdala subregion absolute volumes and/or relative proportion varies as a function of age, sex, or puberty in a large sample of typically developing adolescents (N=408, 43% female, 10-17 years). Utilizing the in vivo CIT168 atlas, we quantified 9 subregions and implemented Generalized Additive Mixed Models to capture potential non-linear associations with age and pubertal status between sexes. Only males showed significant age associations with the basolateral ventral and paralaminar subdivision (BLVPL), central nucleus (CEN), and amygdala transition area (ATA). Again, only males showed relative differences in the proportion of the BLVPL, CEN, ATA, along with lateral (LA) and amygdalostriatal transition area (ASTA), with age. Using a best-fit modeling approach, age, and not puberty, was found to drive these associations. The results suggest that amygdala subregions show unique developmental patterns with age in males across adolescence. Future research is warranted to determine if our findings may contribute to sex differences in mental health that emerge across adolescence.
]]></description>
<dc:creator>Campbell, C. E.</dc:creator>
<dc:creator>Mezher, A. F.</dc:creator>
<dc:creator>Eckel, S. P.</dc:creator>
<dc:creator>Tyszka, J. M.</dc:creator>
<dc:creator>Pauli, W. M.</dc:creator>
<dc:creator>Nagel, B. J.</dc:creator>
<dc:creator>Herting, M. M.</dc:creator>
<dc:date>2019-07-09</dc:date>
<dc:identifier>doi:10.1101/690875</dc:identifier>
<dc:title><![CDATA[Restructuring of amygdala subregion apportion across adolescence]]></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/10.1101/694489v1?rss=1">
<title>
<![CDATA[
Individual Variation in Control Network Topography Supports Executive Function in Youth 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/694489v1?rss=1</link>
<description><![CDATA[
The spatial distribution of large-scale functional networks on the anatomic cortex differs between individuals, and is particularly variable in networks responsible for executive function. However, it remains unknown how this functional topography evolves in development and supports cognition. Capitalizing upon advances in machine learning and a large sample of youth (n=693, ages 8-23y) imaged with 27 minutes of high-quality fMRI data, we delineate how functional topography evolves during youth. We found that the functional topography of association networks is refined with age, allowing accurate prediction of an unseen individuals brain maturity. Furthermore, the cortical representation of executive networks predicts individual differences in executive function. Finally, variability of functional topography is associated with fundamental properties of brain organization including evolutionary expansion, cortical myelination, and cerebral blood flow. Our results emphasize the importance of considering both the plasticity and diversity of functional neuroanatomy during development, and suggest advances in personalized therapeutics.
]]></description>
<dc:creator>Cui, Z.</dc:creator>
<dc:creator>Li, H.</dc:creator>
<dc:creator>Xia, C. H.</dc:creator>
<dc:creator>Larsen, B.</dc:creator>
<dc:creator>Adebimpe, A.</dc:creator>
<dc:creator>Baum, G. L.</dc:creator>
<dc:creator>Cieslak, M.</dc:creator>
<dc:creator>Gur, R. E.</dc:creator>
<dc:creator>Gur, R. C.</dc:creator>
<dc:creator>Moore, T. M.</dc:creator>
<dc:creator>Oathes, D. J.</dc:creator>
<dc:creator>Alexander-Bloch, A. F.</dc:creator>
<dc:creator>Raznahan, A.</dc:creator>
<dc:creator>Roalf, D. R.</dc:creator>
<dc:creator>Shinohara, R. T.</dc:creator>
<dc:creator>Wolf, D. H.</dc:creator>
<dc:creator>Davatzikos, C.</dc:creator>
<dc:creator>Bassett, D. S.</dc:creator>
<dc:creator>Fair, D. A.</dc:creator>
<dc:creator>Fan, Y.</dc:creator>
<dc:creator>Satterthwaite, T. D.</dc:creator>
<dc:date>2019-07-08</dc:date>
<dc:identifier>doi:10.1101/694489</dc:identifier>
<dc:title><![CDATA[Individual Variation in Control Network Topography Supports Executive Function in Youth]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-07-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/695296v1?rss=1">
<title>
<![CDATA[
Alternative Splicing of MR1 regulates antigen presentation to MAIT cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/695296v1?rss=1</link>
<description><![CDATA[
Mucosal Associated Invariant T (MAIT) cells can sense intracellular infection by a broad array of pathogens. These cells are activated upon encountering microbial antigen(s) displayed by MR1 on the surface of an infected cell. Human MR1 undergoes alternative splicing. The full length isoform, MR1A, can activate MAIT cells, while the function of the isoforms, MR1B and MR1C, are not well characterized.nnIn this report, we sought to characterize these splice variants. Using a transcriptomic analysis in conjunction with qPCR, we find that that MR1A and MR1B transcripts are widely expressed. Despite the widespread expression of MR1A and MR1B, only MR1A can present mycobacterial antigen to MAIT cells. Coexpression of MR1B with MR1A serves to decrease MAIT cell activation following bacterial infection. However, expression of MR1B prior to MR1A lowers total MR1A abundance, suggesting competition between MR1A and MR1B for either ligands or chaperones required for folding and/or trafficking. Finally, we evaluated CD4/CD8 double positive thymocytes expressing surface MR1. Relative MR1A/MR1B expression in MR1-expressing thymocytes is associated with their prevalence.nnOur results suggest alternative splicing of MR1 represents a means of regulating MAIT activation in response to microbial ligand.nnFundingThis work was supported by NIH T32HL083808 (EK, GAN, EM), VA Merit Award I01CX001562 (MJH), NIH R01AI29976 (MJH), NIH R01AI048090 (DML), NIH R21AI124225-01A1 (FT) and VA Merit Award I01BX000533 (DML). The contents do not represent the views of the U.S. Department of Veterans Affairs or the United States Government.
]]></description>
<dc:creator>Narayanan, G. A.</dc:creator>
<dc:creator>Nellore, A.</dc:creator>
<dc:creator>Tran, J.</dc:creator>
<dc:creator>Worley, A. H.</dc:creator>
<dc:creator>Meermeier, E. W.</dc:creator>
<dc:creator>Karamooz, E.</dc:creator>
<dc:creator>Huber, M.</dc:creator>
<dc:creator>Kurapova, R.</dc:creator>
<dc:creator>Tafesse, F.</dc:creator>
<dc:creator>Harriff, M. J.</dc:creator>
<dc:creator>Lewinsohn, D. M.</dc:creator>
<dc:date>2019-07-11</dc:date>
<dc:identifier>doi:10.1101/695296</dc:identifier>
<dc:title><![CDATA[Alternative Splicing of MR1 regulates antigen presentation to MAIT cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-07-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/700120v1?rss=1">
<title>
<![CDATA[
Effects of hM4Di activation in CamKII basolateral amygdala neurons and CNO treatment on Sensory-Specific vs. General-PIT; refining PIT circuits and considerations for using CNO. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/700120v1?rss=1</link>
<description><![CDATA[
Pavlovian stimuli can influence instrumental behaviors, a phenomenon known as Pavlovian-to-instrumental transfer (PIT). PIT arises via psychologically and neurobiologically independent processes as Sensory-Specific-PIT (SS-PIT) and General-PIT. SS-, but not General-PIT, relies on the basolateral amygdala (BLA), however the specific BLA neuronal populations involved are unknown. Therefore, here we determined the contribution of glutamatergic BLA neurons to SS-PIT. The BLA was transduced with virus containing either GFP or hM4Di, driven by the CamKII promoter. Rats were then tested for SS- and General-PIT following Vehicle or Clozapine-n-oxide (CNO, the hM4Di-activating ligand) injection. CNO had no effect on SS-PIT in the GFP control group, but selectively blocked its expression in the hM4Di-expressing group. Furthermore, CNO did not alter the expression of Pavlovian outcome devaluation effects in GFP or hM4Di expressing groups, indicating that the hM4Di-mediated loss of SS-PIT did not result from an inability to recall the sensory-specific details of the Pavlovian stimulus-outcome associations. Unexpectedly, CNO disrupted General-PIT in both GFP and hM4Di expressing groups, indicating that CNO alone is sufficient to disrupt affective, but not sensory-specific processes. Disruption of General-PIT by CNO was not due to generalized motor effects, but instead may be related to shifts in internal state produced by CNO. Together these data identify BLA CamKII neurons as critical for the expression of SS-PIT, and reveal important considerations for using CNO to study general affective motivation.
]]></description>
<dc:creator>Derman, R. C.</dc:creator>
<dc:creator>Bass, C. E.</dc:creator>
<dc:creator>Ferrario, C.</dc:creator>
<dc:date>2019-07-14</dc:date>
<dc:identifier>doi:10.1101/700120</dc:identifier>
<dc:title><![CDATA[Effects of hM4Di activation in CamKII basolateral amygdala neurons and CNO treatment on Sensory-Specific vs. General-PIT; refining PIT circuits and considerations for using CNO.]]></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/10.1101/703256v1?rss=1">
<title>
<![CDATA[
Trace amine-associated receptor gene polymorphism increases drug craving 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/703256v1?rss=1</link>
<description><![CDATA[
BACKGROUNDMethamphetamine (MA) is a potent agonist at the trace amine-associated receptor 1 (TAAR1). This study evaluated a common variant (CV) in the human TAAR1 gene, synonymous single nucleotide polymorphism (SNP) V288V, to determine the involvement of TAAR1 in MA dependence.nnMETHODSParticipants (n = 106) with active MA dependence (MA-ACT), in remission from MA dependence (MA-REM), with active polysubstance dependence, in remission from polysubstance dependence, and with no history of substance dependence completed neuropsychiatric symptom questionnaires and provided blood samples. In vitro expression and function of CV and wild type TAAR1 receptors were also measured.nnRESULTSThe V288V polymorphism demonstrated a 40% increase in TAAR1 protein expression in cell culture, but message sequence and protein function were unchanged, suggesting an increase in translation efficiency. Principal components analysis resolved neuropsychiatric symptoms into four components, PC1 (depression, anxiety, memory, and fatigue), PC2 (pain), PC3 (drug and alcohol craving), and PC4 (sleep disturbances). Analyses of study group and TAAR1 genotype revealed a significant interaction for PC3 (craving response) (p = 0.003). The control group showed no difference in PC3 associated with TAAR1, while adjusted mean craving for the MA-ACT and MA-REM groups, among those with at least one copy of V288V, was estimated to be, respectively, 1.55 (p = 0.036) and 1.77 (p = 0.071) times the adjusted mean craving for those without the TAAR1 SNP.nnCONCLUSIONSNeuroadaptation to chronic MA use may be altered by TAAR1 genotype and result in increased dopamine signaling and craving in individuals with the V288V genotype.
]]></description>
<dc:creator>Loftis, J. M.</dc:creator>
<dc:creator>Lasarev, M.</dc:creator>
<dc:creator>Shi, X.</dc:creator>
<dc:creator>Lapidus, J.</dc:creator>
<dc:creator>Janowsky, A.</dc:creator>
<dc:creator>Hoffman, W.</dc:creator>
<dc:creator>Huckans, M.</dc:creator>
<dc:date>2019-07-15</dc:date>
<dc:identifier>doi:10.1101/703256</dc:identifier>
<dc:title><![CDATA[Trace amine-associated receptor gene polymorphism increases drug craving]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-07-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/709261v1?rss=1">
<title>
<![CDATA[
Acidic fibroblast growth factor underlies microenvironmental regulation of MYC in pancreatic cancer 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/709261v1?rss=1</link>
<description><![CDATA[
Despite a critical role for MYC as an effector of oncogenic RAS, strategies to target MYC activity in RAS-driven cancers are lacking. Oncogenic KRAS is insufficient to drive tumorigenesis, while addition of modest overexpression of MYC drives robust tumor formation, suggesting that mechanisms beyond the RAS pathway play key roles in MYC regulation and RAS-driven tumorigenesis. Here we show that acidic fibroblast growth factor (FGF1) derived from cancer-associated fibroblasts (CAFs) cooperates with cancer cell-autonomous signals to increase MYC level, promoter occupancy, and activity. FGF1 is necessary and sufficient for paracrine regulation of MYC protein stability, signaling through AKT and GSK-3{beta}. These signals cooperate with, but are distinct from, cell-autonomous signals from oncogenic KRAS which stabilize MYC. Human pancreatic cancer specimens reveal a strong correlation between stromal CAF content and MYC protein level in the neoplastic compartment, and identify CAFs as the specific source of FGF1 in the tumor microenvironment. Together, our findings demonstrate that MYC is coordinately regulated by cell-autonomous and microenvironmental signals, and establish CAF-derived FGF1 as a novel paracrine regulator of oncogenic transcription.nnStatement of significanceOur work highlights an unanticipated role for the tumor microenvironment in the regulation of MYC protein stability in pancreatic cancer cells and identifies CAF-derived FGF1 as a novel, specific paracrine regulator of MYC.
]]></description>
<dc:creator>Bhattacharyya, S.</dc:creator>
<dc:creator>Oon, C.</dc:creator>
<dc:creator>Kothari, A.</dc:creator>
<dc:creator>Horton, W.</dc:creator>
<dc:creator>Link, J.</dc:creator>
<dc:creator>Sears, R. C.</dc:creator>
<dc:creator>Sherman, M. H.</dc:creator>
<dc:date>2019-07-22</dc:date>
<dc:identifier>doi:10.1101/709261</dc:identifier>
<dc:title><![CDATA[Acidic fibroblast growth factor underlies microenvironmental regulation of MYC in pancreatic cancer]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-07-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/711895v1?rss=1">
<title>
<![CDATA[
BET inhibition induces an anti-apoptotic adaptive response and therapeutic vulnerability to MCL1 inhibitors in breast cancer 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/711895v1?rss=1</link>
<description><![CDATA[
The development of effective targeted therapies for the treatment of basal-like breast cancers remains challenging. Here, we demonstrate that BET inhibition induces a multi-faceted adaptive response program leading to MCL1 protein-driven evasion of apoptosis in breast cancers. Consequently, co-targeting MCL1 and BET is highly synergistic in in vitro and in vivo breast cancer models. Drug response and genomics analyses revealed that MCL1 copy number alterations, including low-level gains, are selectively enriched in basal-like breast cancers and associated with effective BET and MCL1 co-targeting. The mechanism of adaptive response to BET inhibition involves upregulation of critical lipid metabolism enzymes including the rate-limiting enzyme stearoyl-CoA desaturase (SCD). Changes in the lipid metabolism are associated with increases in cell motility and membrane fluidity as well as transitions in cell morphology and adhesion. The structural changes in the cell membrane leads to re-localization and activation of HER2/EGFR which can be interdicted by inhibiting SCD activity. Active HER2/EGFR, in turn, induces accumulation of MCL1 protein and therapeutic vulnerability to MCL1 inhibitors. The BET protein, lipid metabolism and receptor tyrosine kinase activation cascade is observed in patient cohorts of basal-like and HER2-amplified breast cancers. The high frequency of MCL1 chromosomal amplifications (>30%) and gains (>50%) in basal-like breast cancers suggests that BET and MCL1 co-inhibition may have therapeutic utility in this aggressive subtype.
]]></description>
<dc:creator>Yan, G.</dc:creator>
<dc:creator>Wang, H.</dc:creator>
<dc:creator>Luna, A.</dc:creator>
<dc:creator>Bozorgui, B.</dc:creator>
<dc:creator>Li, X.</dc:creator>
<dc:creator>Dereli, Z.</dc:creator>
<dc:creator>Chen, X.</dc:creator>
<dc:creator>Lu, Y.</dc:creator>
<dc:creator>Babur, O.</dc:creator>
<dc:creator>Sander, C.</dc:creator>
<dc:creator>Mills, G.</dc:creator>
<dc:creator>Korkut, A.</dc:creator>
<dc:date>2019-07-23</dc:date>
<dc:identifier>doi:10.1101/711895</dc:identifier>
<dc:title><![CDATA[BET inhibition induces an anti-apoptotic adaptive response and therapeutic vulnerability to MCL1 inhibitors in breast cancer]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-07-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/726760v1?rss=1">
<title>
<![CDATA[
Myelin and nodal plasticity modulate action potential conduction in the adult mouse brain 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/726760v1?rss=1</link>
<description><![CDATA[
Central nervous system myelination increases action potential conduction velocity, however, it is unclear how myelination is coordinated to ensure the temporally precise arrival of action potentials, and facilitate information processing within cortical and associative circuits. Here, we show that mature myelin remains plastic in the adult mouse brain and can undergo subtle structural modifications to influence action potential arrival times. Repetitive transcranial magnetic stimulation and spatial learning, two stimuli that modify neuronal activity, alter the length of the nodes of Ranvier and the size of the periaxonal space within active brain regions. This change in the axon-glial configuration is independent of oligodendrogenesis and tunes conduction velocity to increase the synchronicity of action potential transit.
]]></description>
<dc:creator>Cullen, C. L.</dc:creator>
<dc:creator>Pepper, R. E.</dc:creator>
<dc:creator>Clutterbuck, M. T.</dc:creator>
<dc:creator>Pitman, K. A.</dc:creator>
<dc:creator>Oorschot, V.</dc:creator>
<dc:creator>Auderset, L.</dc:creator>
<dc:creator>Tang, A. D.</dc:creator>
<dc:creator>Ramm, G.</dc:creator>
<dc:creator>Emery, B.</dc:creator>
<dc:creator>Rodger, J.</dc:creator>
<dc:creator>Jolivet, R. B.</dc:creator>
<dc:creator>Young, K. M.</dc:creator>
<dc:date>2019-08-06</dc:date>
<dc:identifier>doi:10.1101/726760</dc:identifier>
<dc:title><![CDATA[Myelin and nodal plasticity modulate action potential conduction in the adult mouse brain]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-08-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/727412v1?rss=1">
<title>
<![CDATA[
Single-cell proteomics reveals downregulation of TMSB4X to drive actin release for stereocilia assembly 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/727412v1?rss=1</link>
<description><![CDATA[
Hearing and balance rely on small sensory hair cells that reside in the inner ear. To explore dynamic changes in the abundant proteins present in differentiating hair cells, we used nanoliter-scale shotgun mass spectrometry of single cells, each [~]1 picoliter, from utricles of embryonic day 15 chickens. We identified unique constellations of proteins or protein groups from presumptive hair cells and from progenitor cells. The single-cell proteomes enabled the de novo reconstruction of a developmental trajectory. Inference of protein expression dynamics revealed that the actin monomer binding protein thymosin {beta}4 (TMSB4X) was present in progenitors but dropped precipitously during hair-cell differentiation. Complementary single-cell transcriptome profiling showed downregulation of TMSB4X mRNA during maturation of hair cells. We propose that most actin is sequestered by TMSB4X in progenitor cells, but upon differentiation to hair cells, actin is released to build the sensory hair bundle.
]]></description>
<dc:creator>Zhu, Y.</dc:creator>
<dc:creator>Scheibinger, M.</dc:creator>
<dc:creator>Ellwanger, D. C.</dc:creator>
<dc:creator>Krey, J. F.</dc:creator>
<dc:creator>Choi, D.</dc:creator>
<dc:creator>Kelly, R. T.</dc:creator>
<dc:creator>Heller, S.</dc:creator>
<dc:creator>Barr-Gillespie, P. G.</dc:creator>
<dc:date>2019-08-06</dc:date>
<dc:identifier>doi:10.1101/727412</dc:identifier>
<dc:title><![CDATA[Single-cell proteomics reveals downregulation of TMSB4X to drive actin release for stereocilia assembly]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-08-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/729475v1?rss=1">
<title>
<![CDATA[
Structured Reviews for Data and Knowledge Driven Research 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/729475v1?rss=1</link>
<description><![CDATA[
MotivationHypothesis generation is a critical step in research and a cornerstone in the rare disease field. Research is most efficient when those hypotheses are based on the entirety of knowledge known to date. Systematic review articles are commonly used in biomedicine to summarize existing knowledge and contextualize experimental data. But the information contained within review articles is typically only expressed as free-text, which is difficult to use computationally. Researchers struggle to navigate, collect and remix prior knowledge as it is scattered in several silos without seamless integration and access. This lack of a structured information framework hinders research by both experimental and computational scientists.nnResultsTo better organize knowledge and data, we built a structured review article that is specifically focused on NGLY1 Deficiency, an ultra-rare genetic disease first reported in 2012. We represented this structured review as a knowledge graph, and then stored this knowledge graph in a Neo4j database to simplify dissemination, querying, and visualization of the network. Relative to free-text, this structured review better promotes the principles of findability, accessibility, interoperability, and reusability (FAIR). In collaboration with domain experts in NGLY1 Deficiency, we demonstrate how this resource can improve the efficiency and comprehensiveness of hypothesis generation. We also developed a read-write interface that allows domain experts to contribute FAIR structured knowledge to this community resource. In contrast to traditional free-text review articles, this structured review exists as a living knowledge graph that is curated by humans and accessible to computational analyses. Finally, we have generalized this workflow into modular and repurposable components that can be applied to other domain areas. This NGLY1 Deficiency-focused network is publicly available at http://ngly1graph.org/.nnAvailability and implementationSource code and network data files are at: https://github.com/SuLab/ngly1-graph and https://github.com/SuLab/bioknowledge-reviewer.nnContactasu@scripps.edu
]]></description>
<dc:creator>Queralt-Rosinach, N.</dc:creator>
<dc:creator>Stupp, G. S.</dc:creator>
<dc:creator>Li, T. S.</dc:creator>
<dc:creator>Mayers, M.</dc:creator>
<dc:creator>Hoatlin, M. E.</dc:creator>
<dc:creator>Might, M.</dc:creator>
<dc:creator>Good, B. M.</dc:creator>
<dc:creator>Su, A. I.</dc:creator>
<dc:date>2019-08-12</dc:date>
<dc:identifier>doi:10.1101/729475</dc:identifier>
<dc:title><![CDATA[Structured Reviews for Data and Knowledge Driven Research]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-08-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/730309v1?rss=1">
<title>
<![CDATA[
SHIFT: speedy histological-to-immunofluorescent translation of whole slide images enabled by deep learning 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/730309v1?rss=1</link>
<description><![CDATA[
Spatially-resolved molecular profiling by immunostaining tissue sections is a key feature in cancer diagnosis, subtyping, and treatment, where it complements routine histopathological evaluation by clarifying tumor phenotypes. In this work, we present a deep learning-based method called speedy histological-to-immunofluorescent translation (SHIFT) which takes histologic images of hematoxylin and eosin-stained tissue as input, then in near-real time returns inferred virtual immunofluorescence (IF) images that accurately depict the underlying distribution of phenotypes without requiring immunostaining of the tissue being tested. We show that deep learning-extracted feature representations of histological images can guide representative sample selection, which improves SHIFT generalizability. SHIFT could serve as an efficient preliminary, auxiliary, or substitute for IF by delivering multiplexed virtual IF images for a fraction of the cost and in a fraction of the time required by nascent multiplexed imaging technologies.nnKEY POINTSO_LISpatially-resolved molecular profiling is an essential complement to histopathological evaluation of cancer tissues.nC_LIO_LIInformation obtained by immunofluorescence imaging is encoded by features in histological images.nC_LIO_LISHIFT leverages previously unappreciated features in histological images to facilitate virtual immunofluorescence staining.nC_LIO_LIFeature representations of images guide sample selection, improving model generalizability.nC_LI
]]></description>
<dc:creator>Burlingame, E. A.</dc:creator>
<dc:creator>McDonnell, M.</dc:creator>
<dc:creator>Schau, G. F.</dc:creator>
<dc:creator>Thibault, G.</dc:creator>
<dc:creator>Lanciault, C.</dc:creator>
<dc:creator>Morgan, T.</dc:creator>
<dc:creator>Johnson, B. E.</dc:creator>
<dc:creator>Corless, C.</dc:creator>
<dc:creator>Gray, J. W.</dc:creator>
<dc:creator>Chang, Y. H.</dc:creator>
<dc:date>2019-08-15</dc:date>
<dc:identifier>doi:10.1101/730309</dc:identifier>
<dc:title><![CDATA[SHIFT: speedy histological-to-immunofluorescent translation of whole slide images enabled by deep learning]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-08-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/731455v1?rss=1">
<title>
<![CDATA[
Mechanotransduction-dependent control of stereocilia dimensions and row identity in inner hair cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/731455v1?rss=1</link>
<description><![CDATA[
Actin-rich structures like stereocilia and microvilli are assembled with precise control of length, diameter, and relative spacing. We found that developmental widening of the second-tallest stereocilia rank (row 2) of mouse inner hair cells correlated with the appearance of mechanotransduction. Correspondingly, Tmc1KO/KO;Tmc2KO/KO or TmieKO/KO hair cells, which lack transduction, have significantly altered stereocilia lengths and diameters. EPS8 and the short splice isoform of MYO15A, identity markers for row 1 (tallest), lost their row exclusivity in transduction mutants, a result that was mimicked by block of transduction channels. Likewise, the heterodimeric capping protein subunit CAPZB and its partner TWF2 lost their row 2 tip localization in mutants, and GNAI3 failed to accumulate at row 1 tips. Redistribution of marker proteins was accompanied by increased variability in stereocilia height. Transduction channels thus specify and maintain row identity and control addition of new actin filaments to increase stereocilia diameter.
]]></description>
<dc:creator>Krey, J. F.</dc:creator>
<dc:creator>Chatterjee, P.</dc:creator>
<dc:creator>Dumont, R. A.</dc:creator>
<dc:creator>Choi, D.</dc:creator>
<dc:creator>Bird, J. E.</dc:creator>
<dc:creator>Barr-Gillespie, P. G.</dc:creator>
<dc:date>2019-08-13</dc:date>
<dc:identifier>doi:10.1101/731455</dc:identifier>
<dc:title><![CDATA[Mechanotransduction-dependent control of stereocilia dimensions and row identity in inner hair cells]]></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/10.1101/732784v1?rss=1">
<title>
<![CDATA[
Reciprocal monoallelic expression of ASAR lncRNA genes controls replication timing of human chromosome 6. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/732784v1?rss=1</link>
<description><![CDATA[
AbstractDNA replication occurs on mammalian chromosomes in a cell-type distinctive temporal order known as the replication timing program. We previously found that disruption of the noncanonical lncRNA genes ASAR6 and ASAR15 results in delayed replication timing and delayed mitotic chromosome condensation of human chromosome 6 and 15, respectively. ASAR6 and ASAR15 display random monoallelic expression, and display asynchronous replication between alleles that is coordinated with other random monoallelic genes on their respective chromosomes. Disruption of the expressed allele, but not the silent allele, of ASAR6 leads to delayed replication, activation of the previously silent alleles of linked monoallelic genes, and structural instability of human chromosome 6. In this report, we describe a second lncRNA gene (ASAR6-141) on human chromosome 6 that when disrupted results in delayed replication timing in cis. ASAR6-141 is subject to random monoallelic expression and asynchronous replication, and is expressed from the opposite chromosome 6 homolog as ASAR6. ASAR6-141 RNA, like ASAR6 and ASAR15 RNAs, contains a high L1 content and remains associated with the chromosome territory where it is transcribed. Three classes of cis-acting elements control proper chromosome function in mammals: origins of replication, centromeres; and telomeres, which are responsible for replication, segregation and stability of all chromosomes. Our work supports a fourth type of essential chromosomal element, "Inactivation/Stability Centers", which express ASAR lncRNAs responsible for proper replication timing, monoallelic expression, and structural stability of each chromosome.nnAuthor summaryMammalian cells replicate their chromosomes during a highly ordered and cell type-specific program. Genetic studies have identified two long non-coding RNA genes, ASAR6 and ASAR15, as critical regulators of the replication timing program of human chromosomes 6 and 15, respectively. There are several unusual characteristics of the ASAR6 and ASAR15 RNAs that distinguish them from other long non-coding RNAs, including: being very long (>200 kb), lacking splicing of the transcripts, lacking polyadenylation, and being retained in the nucleus on the chromosomes where they are made. ASAR6 and ASAR15 also have the unusual property of being expressed from only one copy of the two genes located on homologous chromosome pairs. Using these unusual characteristics shared between ASAR6 and ASAR15, we have identified a second ASAR lncRNA gene located on human chromosome 6, which we have named ASAR6-141. ASAR6-141 is expressed from the opposite chromosome 6 homolog as ASAR6, and disruption of the expressed allele results in delayed replication of chromosome 6. ASAR6-141 RNA had previously been annotated as vlinc273. The very long intergenic non-coding (vlinc)RNAs represent a recently annotated class of RNAs that are long (>50 kb), non-spliced, and non-polyadenlyated nuclear RNAs. There are currently >2,700 vlincRNAs expressed from every chromosome, are encoded by >15% of the human genome, and with a few exceptions have no known function. Our results suggest the intriguing possibility that the vlinc class of RNAs may be functioning to control the replication timing program of all human chromosomes.
]]></description>
<dc:creator>Heskett, M. B.</dc:creator>
<dc:creator>Smith, L.</dc:creator>
<dc:creator>Spellman, P.</dc:creator>
<dc:creator>Thayer, M. J.</dc:creator>
<dc:date>2019-08-12</dc:date>
<dc:identifier>doi:10.1101/732784</dc:identifier>
<dc:title><![CDATA[Reciprocal monoallelic expression of ASAR lncRNA genes controls replication timing of human chromosome 6.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-08-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/738583v1?rss=1">
<title>
<![CDATA[
Streaming of repeated noise in primary and secondary fields of auditory cortex 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/738583v1?rss=1</link>
<description><![CDATA[
Statistical regularities in natural sounds facilitate the perceptual segregation of auditory sources, or streams. Repetition is one cue that drives stream segregation in humans, but the neural basis of this perceptual phenomenon remains unknown. We demonstrated a similar perceptual ability in animals by training ferrets to detect a stream of repeating noise samples (foreground) embedded in a stream of random samples (background). During passive listening, we recorded neural activity in primary (A1) and secondary (PEG) fields of auditory cortex. We used two context-dependent encoding models to test for evidence of streaming of the repeating stimulus. The first was based on average evoked activity per noise sample and the second on the spectro-temporal receptive field (STRF). Both approaches tested whether changes in the neural response to repeating versus random stimuli were better modeled by scaling the response to both streams equally (global gain) or by separately scaling the response to the foreground versus background stream (stream-specific gain). Consistent with previous observations of adaptation, we found an overall reduction in global gain when the stimulus began to repeat. However, when we measured stream-specific changes in gain, responses to the foreground were enhanced relative to the background. This enhancement was stronger in PEG than A1. In A1, enhancement was strongest in units with low sparseness (i.e., broad sensory tuning) and with tuning selective for the repeated sample. Enhancement of responses to the foreground relative to the background provides evidence for stream segregation that emerges in A1 and is refined in PEG.
]]></description>
<dc:creator>Saderi, D.</dc:creator>
<dc:creator>Buran, B. N.</dc:creator>
<dc:creator>David, S. V.</dc:creator>
<dc:date>2019-08-19</dc:date>
<dc:identifier>doi:10.1101/738583</dc:identifier>
<dc:title><![CDATA[Streaming of repeated noise in primary and secondary fields of auditory cortex]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-08-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/748053v1?rss=1">
<title>
<![CDATA[
The tooth on-a-chip: a microphysiologic model system mimicking the pulp-dentin interface and its interaction with biomaterials 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/748053v1?rss=1</link>
<description><![CDATA[
The tooth has a unique configuration with respect to biomaterials that are used for its treatment. Cells inside of the dental pulp interface indirectly with biomaterials via a calcified permeable membrane, formed by a dentin barrier which is composed of several thousands of dentinal tubules (~2 {micro}m in diameter) connecting the dental pulp tissue to the outer surface of the tooth. Although the cytotoxic response of the dental pulp to biomaterials has been extensively studied, there is a shortage of in vitro model systems that mimic the dentin-pulp interface, enabling an improved understanding of the morphologic, metabolic and functional influence of biomaterials on live dental pulp cells. To address this shortage, here we developed an organ-on-a-chip model system which integrates cells cultured directly on a dentin wall within a microdevice which replicates some of the architecture and dynamics of the dentin-pulp interface. The tooth-on-a-chip is made out of molded polydimethylsiloxane (PDMS) with a design consisting of two chambers separated by a dentin fragment. To characterize pulp cell responses to dental materials on-chip, stem cell-derived odontoblasts were seeded onto the dentin surface, and observed using live-cell microscopy. Standard dental materials used clinically (2-hydroxyethylmethacrylate - HEMA, Phosphoric Acid - PA, and Adper-Scotchbond - SB) were tested for cytotoxicity, cell morphology and metabolic activity on-chip, and compared against standardized off-chip controls. All dental materials had cytotoxic effects in both on-chip and off-chip systems in the following order: HEMA>SB>PA (p<0.05), and cells presented consistently higher metabolic activity on-chip than off-chip (p<0.05). Furthermore, the tooth-on-a-chip enabled real-time tracking of odontoblast monolayer formation, remodeling, and death in response to biomaterial treatments, and gelatinolytic activity in a model hybrid layer (HL) formed in the microdevice. In conclusion, the tooth-on-a-chip is a novel platform that replicates near-physiologic conditions of the pulp-dentin interface, and enables live-cell imaging to study dental pulp cell response to biomaterials.
]]></description>
<dc:creator>Miranda Franca, C.</dc:creator>
<dc:creator>Tahayeri, A.</dc:creator>
<dc:creator>Sousa Rodrigues, N.</dc:creator>
<dc:creator>Fedorsian, S.</dc:creator>
<dc:creator>Puppin-Rontani, R.</dc:creator>
<dc:creator>Ferracane, J.</dc:creator>
<dc:creator>Bertassoni, L. E.</dc:creator>
<dc:date>2019-08-28</dc:date>
<dc:identifier>doi:10.1101/748053</dc:identifier>
<dc:title><![CDATA[The tooth on-a-chip: a microphysiologic model system mimicking the pulp-dentin interface and its interaction with biomaterials]]></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/10.1101/749390v1?rss=1">
<title>
<![CDATA[
Defective cell death of distinct microglial subsets contributes to ADHD-like behavior in mice 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/749390v1?rss=1</link>
<description><![CDATA[
Microglia are resident immune cells in the central nervous system that play essential roles to maintain homeostasis and neuronal function. Microglia are heterogeneous cells but the mechanisms by which they contribute to normal brain development remain unclear. Here,we show that microglia in the developing striatum and thalamus undergo pyroptosis,a type of lytic cell death that occurs as a result of Caspase-1 (CASP1) activation downstream of inflammasomes. We observe that pyroptosis occurs in a spatiotemporally regulated and Casp1-dependent manner during fetal brain development. Mice lacking Casp1 or the inflammasome regulating molecules, NLRP3, IL-1R, and Gasdermin D exhibit behavior changes characterized by hyperactivity, inattention, and impulsivity that are similar to attention-deficit/hyperactivity disorder (ADHD). Furthermore, re-expression of Casp1 in Cx3cr1+ cells including microglia restores normal behavior and cell death. We demonstrate that injection of an NLRP3 inhibitor into pregnant wild-type mice is sufficient to induce ADHD-like behaviors in offspring. These data suggest that microglial inflammasome activation and pyroptosis are essential for normal brain development and that genetic and pharmacological disruptions in this pathway may represent new ADHD risk factors.
]]></description>
<dc:creator>Chuang, H.-C.</dc:creator>
<dc:creator>Nichols, E. K.</dc:creator>
<dc:creator>Rauch, I.</dc:creator>
<dc:creator>Chang, W.-C.</dc:creator>
<dc:creator>Misra, R.</dc:creator>
<dc:creator>Lin, P. M.</dc:creator>
<dc:creator>Kitaoka, M.</dc:creator>
<dc:creator>Vance, R. E.</dc:creator>
<dc:creator>Saijo, K.</dc:creator>
<dc:date>2019-08-30</dc:date>
<dc:identifier>doi:10.1101/749390</dc:identifier>
<dc:title><![CDATA[Defective cell death of distinct microglial subsets contributes to ADHD-like behavior in mice]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-08-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/754044v1?rss=1">
<title>
<![CDATA[
Putatively cancer-specific alternative splicing is shared across patients and present in developmental and other non-cancer cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/754044v1?rss=1</link>
<description><![CDATA[
We compared cancer and non-cancer RNA sequencing (RNA-seq) data from The Cancer Genome Atlas (TCGA), the Genotype-Tissue Expression (GTEx) Project, and the Sequence Read Archive (SRA). We found that: 1) averaging across cancer types, 80.6% of exon-exon junctions thought to be cancer-specific based on comparison with tissue-matched samples are in fact present in other adult non-cancer tissues throughout the body; 2) 30.8% of junctions not present in any GTEx or TCGA normal tissues are shared by multiple samples within at least one cancer type cohort, and 87.4% of these distinguish between different cancer types; and 3) many of these junctions not found in GTEx or TCGA normal tissues (15.4% on average) are also found in embryological and other developmentally associated cells. This study probes the distribution of putatively cancer-specific junctions across a broad set of publicly available non-cancer human RNA-seq datasets. Overall, we identify a subset of shared cancer-specific junctions that could represent novel sources of cancer neoantigens. We further describe a framework for characterizing possible origins of these junctions, including potential developmental and embryological sources, as well as cell type-specific markers particularly related to cell types of cancer origin. These findings refine the meaning of RNA splicing event novelty, particularly with respect to the human neoepitope repertoire. Ultimately, cancer-specific exon-exon junctions may affect the anti-cancer immune response and may have a substantial causal relationship with the biology of disease.
]]></description>
<dc:creator>David, J. K.</dc:creator>
<dc:creator>Maden, S. K.</dc:creator>
<dc:creator>Weeder, B. R.</dc:creator>
<dc:creator>Thompson, R. F.</dc:creator>
<dc:creator>Nellore, A.</dc:creator>
<dc:date>2019-09-05</dc:date>
<dc:identifier>doi:10.1101/754044</dc:identifier>
<dc:title><![CDATA[Putatively cancer-specific alternative splicing is shared across patients and present in developmental and other non-cancer cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-09-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/765230v1?rss=1">
<title>
<![CDATA[
Co-option of the gibbon-specific LAVA retrotransposon in DNA repair pathways 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/765230v1?rss=1</link>
<description><![CDATA[
Co-option of transposable elements (TEs) to become part of existing or new enhancers is an important mechanism for evolution of gene regulation. However, contributions of lineage-specific TE insertions to recent regulatory adaptations remain poorly understood. Gibbons present a suitable model to study these contributions as they have evolved a lineage-specific TE called LAVA, which is still active in the gibbon genome. The LAVA retrotransposon is thought to have played a role in the emergence of the unusually rearranged structure of the gibbon genome by disrupting transcription of cell cycle genes. In this study, we investigated whether LAVA may have also contributed to the evolution of gene regulation by adopting enhancer function. We characterized fixed and polymorphic LAVA insertions across multiple gibbons and found 96 LAVA elements overlapping enhancer chromatin states. Moreover, LAVA was enriched in multiple transcription factor binding motifs, was bound by an important transcription factor (PU.1), and was associated with higher levels of gene expression in cis. We found gibbon-specific signatures of purifying/positive selection at 27 LAVA insertions. Two of these insertions were fixed in the gibbon lineage and overlapped with enhancer chromatin states, representing putative co-opted LAVA enhancers. These putative enhancers were located within genes encoding SETD2 and RAD9A, two proteins that facilitate accurate repair of DNA double-strand breaks and prevent chromosomal rearrangement mutations. Thus, LAVAs co-option in these genes may have influenced regulation of processes that preserve genome integrity. Our findings highlight the importance of considering lineage-specific TEs in studying evolution of novel gene regulatory elements.
]]></description>
<dc:creator>Okhovat, M.</dc:creator>
<dc:creator>Nevonen, K. A.</dc:creator>
<dc:creator>Davis, B.</dc:creator>
<dc:creator>Michener, P.</dc:creator>
<dc:creator>Ward, S.</dc:creator>
<dc:creator>Milhaven, M.</dc:creator>
<dc:creator>Harshman, L.</dc:creator>
<dc:creator>Sohota, A.</dc:creator>
<dc:creator>O'Neill, R. J.</dc:creator>
<dc:creator>Ahituv, N.</dc:creator>
<dc:creator>Veeramah, K. R.</dc:creator>
<dc:creator>Carbone, L.</dc:creator>
<dc:date>2019-09-11</dc:date>
<dc:identifier>doi:10.1101/765230</dc:identifier>
<dc:title><![CDATA[Co-option of the gibbon-specific LAVA retrotransposon in DNA repair pathways]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-09-11</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/10.1101/768192v1?rss=1">
<title>
<![CDATA[
Estimating The Lateral Margin Of Stability During Walking And Turning Using Inertial Sensors 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/768192v1?rss=1</link>
<description><![CDATA[
There is growing interest in using inertial sensors to continuously monitor gait during free-living mobility. Inertial sensors can provide many gait measures, but they struggle to capture the spatial stability of the center-of-mass due to limitations estimating sensor-to-sensor distance. While the margin of stability (MoS) is an established outcome describing the instantaneous mechanical stability of gait relating to fall-risk, methods to estimate the MoS from inertial sensors have been lacking. Here, we developed and tested a framework, based on centripetal acceleration, to determine a correlate for the lateral MoS using inertial sensors during walking with or without turning. Using three synchronized sensors located bilaterally on the feet and lumbar spine, the average centripetal acceleration over the subsequent step can be used as a correlate for lateral MoS. Relying only on a single sensor on the lumbar spine yielded similar results if the stance foot can be determined from other means. Additionally, the centripetal acceleration correlate of lateral MoS demonstrates clear differences between walking and turning, inside and outside turning limbs, and speed. While limitations and assumptions need to be considered when implemented in practice, this method presents a novel correlate for the lateral MoS during walking and turning using inertial sensors, although further validation is required for other activities and populations.
]]></description>
<dc:creator>Fino, P. C.</dc:creator>
<dc:creator>Horak, F. B.</dc:creator>
<dc:creator>Curtze, C.</dc:creator>
<dc:date>2019-09-23</dc:date>
<dc:identifier>doi:10.1101/768192</dc:identifier>
<dc:title><![CDATA[Estimating The Lateral Margin Of Stability During Walking And Turning Using Inertial Sensors]]></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/10.1101/773028v1?rss=1">
<title>
<![CDATA[
Mutation of CFAP57 causes primary ciliary dyskinesia by disrupting theasymmetric targeting of a subset of ciliary inner dynein arms 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/773028v1?rss=1</link>
<description><![CDATA[
Primary ciliary dyskinesia (PCD) is characterized by chronic airway disease, male infertility, and randomization of the left/right body axis, and is caused by defects of motile cilia and sperm flagella. We screened a cohort of affected individuals that lack an obvious TEM structural phenotype for pathogenic variants using whole exome capture and next generation sequencing. The population sampling probability (PSAP) algorithm identified one subject with a homozygous nonsense variant [(c.1762C>T) p.(Arg588*) exon 11] in the uncharacterized CFAP57 gene. In normal human nasal epithelial cells, CFAP57 localizes throughout the ciliary axoneme. Analysis of cells from the PCD patient shows a loss of CFAP57, reduced beat frequency, and an alteration in the ciliary waveform. Knockdown of CFAP57 in human tracheobronchial epithelial cells (hTECs) recapitulates these findings. Phylogenetic analysis showed that CFAP57 is conserved in organisms that assemble motile cilia, and CFAP57 is allelic with the BOP2 gene identified previously in Chlamydomonas. Two independent, insertional fap57 Chlamydomonas mutant strains show reduced swimming velocity and altered waveforms. Tandem mass spectroscopy showed that CFAP57 is missing, and the "g" inner dyneins (DHC7 and DHC3) and the "d" inner dynein (DHC2) are reduced. Our data demonstrate that the FAP57 protein is required for the asymmetric assembly of inner dyneins on only a subset of the microtubule doublets, and this asymmetry is essential for the generation of an effective axonemal waveform. Together, our data identifies mutations in CFAP57 as a cause of PCD with a specific defect in the inner dynein arm assembly process.nnSignificanceMotile cilia are found throughout eukaryotic organisms and performs essential functions. Primary ciliary dyskinesia (PCD) is a rare disease that affects the function of motile cilia. By applying a novel population sampling probability algorithm (PSAP) that uses large population sequencing databases and pathogenicity prediction algorithms, we identified a variant in an uncharacterized gene, CFAP57. This is the first reported example of PCD caused by a mutation that affects only a subset of the inner dynein arms, which are needed to generate the waveform. CFAP57 identifies an address for specific dynein arms. These findings demonstrate the effectiveness of the PSAP algorithm, expand our understanding of the positioning of dynein arms, and identify mutations in CFAP57 as a cause of PCD.
]]></description>
<dc:creator>Bustamante Marin, X.</dc:creator>
<dc:creator>Horani, A.</dc:creator>
<dc:creator>Stoyanova, M.</dc:creator>
<dc:creator>Charng, W.-L.</dc:creator>
<dc:creator>Bottier, M.</dc:creator>
<dc:creator>Sears, P.</dc:creator>
<dc:creator>Daniels, L. A.</dc:creator>
<dc:creator>Bowen, H.</dc:creator>
<dc:creator>Conrad, D.</dc:creator>
<dc:creator>Knowles, M.</dc:creator>
<dc:creator>Ostrowski, L. E.</dc:creator>
<dc:creator>Zariwala, M. A.</dc:creator>
<dc:creator>Dutcher, S. K.</dc:creator>
<dc:date>2019-09-24</dc:date>
<dc:identifier>doi:10.1101/773028</dc:identifier>
<dc:title><![CDATA[Mutation of CFAP57 causes primary ciliary dyskinesia by disrupting theasymmetric targeting of a subset of ciliary inner dynein arms]]></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/10.1101/773911v1?rss=1">
<title>
<![CDATA[
Exploring Integrative Analysis using the BioMedical Evidence Graph 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/773911v1?rss=1</link>
<description><![CDATA[
The analysis of cancer biology data involves extremely heterogeneous datasets including information from RNA sequencing, genome-wide copy number, DNA methylation data reporting on epigenomic regulation, somatic mutations from whole-exome or whole-genome analyses, pathology estimates from imaging sections or subtyping, drug response or other treatment outcomes, and various other clinical and phenotypic measurements. Bringing these different resources into a common framework, with a data model that allows for complex relationships as well as dense vectors of features, will unlock integrative analysis. We introduce a graph database and query engine for discovery and analysis of cancer biology, called the BioMedical Evidence Graph (BMEG). The BMEG is unique from other biological data graphs in that sample level molecular information is connected to reference knowledge bases. It combines gene expression and mutation data, with drug response experiments, pathway information databases and literature derived associations. The construction of the BMEG has resulted in a graph containing over 36M vertices and 29M edges. The BMEG system provides a graph query based API to enable analysis, with client code available for Python, Javascript and R, and a server online at bmeg.io. Using this system we have developed several forms of integrated analysis to demonstrate the utility of the system. The BMEG is an evolving resource dedicated to enabling integrative analysis. We have demonstrated queries on the system that illustrate mutation significance analysis, drug response machine learning, patient level knowledge base queries and pathway level analysis. We have compared the resulting graph to other available integrated graph systems, and demonstrated that it is unique in the scale of the graph and the type of data it makes available.nnHighlightsO_LIData resource connected extremely diverse set of cancer data setsnC_LIO_LIGraph query engine that can be easily deployed and used on new datasetsnC_LIO_LIEasily installed python clientnC_LIO_LIServer online at bmeg.ionC_LInnSummaryThe analysis of cancer biology data involves extremely heterogeneous datasets including information. Bringing these different resources into a common framework, with a data model that allows for complex relationships as well as dense vectors of features, will unlock integrative analysis. We introduce a graph database and query engine for discovery and analysis of cancer biology, called the BioMedical Evidence Graph (BMEG). The construction of the BMEG has resulted in a graph containing over 36M vertices and 29M edges. The BMEG system provides a graph query based API to enable analysis, with client code available for Python, Javascript and R, and a server online at bmeg.io. Using this system we have developed several forms of integrated analysis to demonstrate the utility of the system.
]]></description>
<dc:creator>Struck, A.</dc:creator>
<dc:creator>Walsh, B.</dc:creator>
<dc:creator>Buchanan, A.</dc:creator>
<dc:creator>Lee, J. A.</dc:creator>
<dc:creator>Spangler, R.</dc:creator>
<dc:creator>Stuart, J.</dc:creator>
<dc:creator>Ellrott, K.</dc:creator>
<dc:date>2019-09-25</dc:date>
<dc:identifier>doi:10.1101/773911</dc:identifier>
<dc:title><![CDATA[Exploring Integrative Analysis using the BioMedical Evidence Graph]]></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/10.1101/781658v1?rss=1">
<title>
<![CDATA[
G-OnRamp: Generating genome browsers to facilitate undergraduate-driven collaborative genome annotation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/781658v1?rss=1</link>
<description><![CDATA[
Scientists are sequencing new genomes at an increasing rate with the goal of associating genome contents with phenotypic traits. After a new genome is sequenced and assembled, structural gene annotation is often the first step in analysis. Despite advances in computational gene prediction algorithms, most eukaryotic genomes still benefit from manual gene annotation. Undergraduates can become skilled annotators, and in the process learn both about genes/genomes and about how to utilize large datasets. Data visualizations provided by a genome browser are essential for manual gene annotation, enabling annotators to quickly evaluate multiple lines of evidence (e.g., sequence similarity, RNA-Seq, gene predictions, repeats). However, creating genome browsers requires extensive computational skills; lack of the expertise required remains a major barrier for many biomedical researchers and educators.nnTo address these challenges, the Genomics Education Partnership (GEP; https://gep.wustl.edu/) has partnered with the Galaxy Project (https://galaxyproject.org) to develop G-OnRamp (http://g-onramp.org), a web-based platform for creating UCSC Assembly Hubs and JBrowse genome browsers. G-OnRamp can also convert a JBrowse instance into an Apollo instance for collaborative genome annotations in research and educational settings. G-OnRamp enables researchers to easily visualize their experimental results, educators to create Course-based Undergraduate Research Experiences (CUREs) centered on genome annotation, and students to participate in genomics research.nnDevelopment of G-OnRamp was guided by extensive user feedback from in-person workshops. Sixty-five researchers and educators from over 40 institutions participated in these workshops, which produced over 20 genome browsers now available for research and education. For example, genome browsers for four parasitoid wasp species were used in a CURE engaging 142 students taught by 13 faculty members -- producing a total of 192 gene models. G-OnRamp can be deployed on a personal computer or on cloud computing platforms, and the genome browsers produced can be transferred to the CyVerse Data Store for long-term access.
]]></description>
<dc:creator>Sargent, L.</dc:creator>
<dc:creator>Liu, Y.</dc:creator>
<dc:creator>Leung, W.</dc:creator>
<dc:creator>Mortimer, N. T.</dc:creator>
<dc:creator>Lopatto, D.</dc:creator>
<dc:creator>Goecks, J.</dc:creator>
<dc:creator>Elgin, S. C. R.</dc:creator>
<dc:date>2019-10-02</dc:date>
<dc:identifier>doi:10.1101/781658</dc:identifier>
<dc:title><![CDATA[G-OnRamp: Generating genome browsers to facilitate undergraduate-driven collaborative genome annotation]]></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/10.1101/781831v1?rss=1">
<title>
<![CDATA[
Optimizing auditory brainstem response acquisition using interleaved frequencies 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/781831v1?rss=1</link>
<description><![CDATA[
Auditory brainstem responses (ABRs) require averaging responses to hundreds or thousands of repetitions of a stimulus (e.g., tone pip) to obtain a measurable evoked response at the scalp. Fast repetition rates lead to changes in ABR amplitude and latency due to adaptation. To minimize the effect of adaptation, stimulus rates are sometimes as low as 10 to 13.3 stimuli per second, requiring long acquisition times. The trade-off between reducing acquisition time and minimizing the effect of adaptation on ABR responses is an especially important consideration for studies of cochlear synaptopathy, which use the amplitude of short latency responses (wave 1) to assess auditory nerve survival. It has been proposed that adaptation during ABR acquisition can be reduced by interleaving tones at different frequencies, rather than testing each frequency serially. With careful ordering of frequencies and levels in the stimulus train, adaptation in the auditory nerve can be minimized, thereby permitting an increase in the rate at which tone bursts are presented. However, widespread adoption of this stimulus design has been hindered by lack of available software. Here, we develop and validate an interleaved stimulus design to optimize the rate of ABR measurement while minimizing adaptation. We implement this method in an open-source data acquisition software tool that permits either serial or interleaved ABR measurements. The open-source software library, psiexperiment, is compatible with widely-used ABR hardware. Consistent with previous studies, careful design of an interleaved stimulus train can reduce ABR acquisition time by more than half, with minimal effect on ABR thresholds and wave 1 latency, while improving measures of wave 1 amplitude.
]]></description>
<dc:creator>Buran, B. N.</dc:creator>
<dc:creator>Elkins, S.</dc:creator>
<dc:creator>Kempton, J. B.</dc:creator>
<dc:creator>Porsov, E. V.</dc:creator>
<dc:creator>Brigande, J. V.</dc:creator>
<dc:creator>David, S. V.</dc:creator>
<dc:date>2019-09-25</dc:date>
<dc:identifier>doi:10.1101/781831</dc:identifier>
<dc:title><![CDATA[Optimizing auditory brainstem response acquisition using interleaved frequencies]]></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/10.1101/782938v1?rss=1">
<title>
<![CDATA[
Systematic profiling of full-length immunoglobulin and T-cell receptor repertoire diversity in rhesus macaque through long read transcriptome sequencing 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/782938v1?rss=1</link>
<description><![CDATA[
The diversity of immunoglobulin (Ig) and T-cell receptor (TCR) repertoires is a focal point of immunological studies. Rhesus macaques are key for modeling human immune responses, placing critical importance on the accurate annotation and quantification of their Ig and TCR repertoires. However, due to incomplete reference resources, the coverage and accuracy of the traditional targeted amplification strategies for profiling rhesus Ig and TCR repertoires are largely unknown. Here, using long read sequencing, we sequenced four Indian-origin rhesus macaque tissues and obtained high quality, full-length sequences for over 6,000 unique Ig and TCR transcripts, without the need for sequence assembly. We constructed the first complete reference set for the constant regions of all known isotypes and chain types of rhesus Ig and TCR repertoires. We show that sequence diversity exists across the entire variable regions of rhesus Ig and TCR transcripts. Consequently, existing strategies using targeted amplification of rearranged variable regions comprised of V(D)J gene segments miss a significant fraction (27% to 53% and 42% to 49%) of rhesus Ig/TCR diversity. To overcome these limitations, we designed new rhesus-specific assays that remove the need for primers conventionally targeting variable regions and allow single cell-level Ig and TCR repertoire analysis. Our improved approach will enable future studies to fully capture rhesus Ig and TCR repertoire diversity and is applicable for improving annotations in any model organism.
]]></description>
<dc:creator>Brochu, H.</dc:creator>
<dc:creator>Tseng, E.</dc:creator>
<dc:creator>Smith, E.</dc:creator>
<dc:creator>Thomas, M. J.</dc:creator>
<dc:creator>Jones, A.</dc:creator>
<dc:creator>Diveley, K.</dc:creator>
<dc:creator>Law, L.</dc:creator>
<dc:creator>Hansen, S. G.</dc:creator>
<dc:creator>Picker, L.</dc:creator>
<dc:creator>Gale, M.</dc:creator>
<dc:creator>Peng, X.</dc:creator>
<dc:date>2019-09-26</dc:date>
<dc:identifier>doi:10.1101/782938</dc:identifier>
<dc:title><![CDATA[Systematic profiling of full-length immunoglobulin and T-cell receptor repertoire diversity in rhesus macaque through long read transcriptome sequencing]]></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/10.1101/786632v1?rss=1">
<title>
<![CDATA[
Mechanism of gating and partial agonist action in the glycine receptor 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/786632v1?rss=1</link>
<description><![CDATA[
The glycine receptor is a pentameric, neurotransmitter-activated ion channel that transitions between closed/resting, open and desensitized states. Glycine, a full agonist, produces an open channel probability (Po) of [~]1.0 while partial agonists, such as taurine and {gamma}-amino butyric acid (GABA) yield submaximal Po values. Despite extensive studies of pentameric Cys-loop receptors, there is little knowledge of the molecular mechanisms underpinning partial agonist action and how the receptor transitions from the closed to open and to desensitized conformations. Here we use electrophysiology and molecular dynamics (MD) simulations, together with a large ensemble of single particle cryo-EM reconstructions, to show how agonists populate agonist-bound yet closed channel states, thus explaining their lesser efficacy, yet also populate agonist-bound open and desensitized states. Measurements within the neurotransmitter binding pocket, as a function of bound agonist, provide a metric to correlate the extent of agonist-induced conformational changes to open channel probability across the Cys-loop receptor family.
]]></description>
<dc:creator>Gouaux, E.</dc:creator>
<dc:creator>Yu, J.</dc:creator>
<dc:creator>Zhu, H.</dc:creator>
<dc:creator>Du, J.</dc:creator>
<dc:creator>Lu, W.</dc:creator>
<dc:creator>Tajkhorshid, E.</dc:creator>
<dc:creator>Shahoei, R.</dc:creator>
<dc:creator>Wang, Y.</dc:creator>
<dc:creator>Sivilotti, L.</dc:creator>
<dc:creator>Lape, R.</dc:creator>
<dc:creator>Greiner, T.</dc:creator>
<dc:date>2019-09-30</dc:date>
<dc:identifier>doi:10.1101/786632</dc:identifier>
<dc:title><![CDATA[Mechanism of gating and partial agonist action in the glycine receptor]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-09-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/788521v1?rss=1">
<title>
<![CDATA[
Pathway Commons: 2019 Update 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/788521v1?rss=1</link>
<description><![CDATA[
Pathway Commons (https://www.pathwaycommons.org) is an integrated resource of publicly available information about biological pathways including biochemical reactions, assembly of biomolecular complexes, transport and catalysis events and physical interactions involving proteins, DNA, RNA, and small molecules (e.g., metabolites and drug compounds). Data is collected from multiple providers in standard formats, including the Biological Pathway Exchange (BioPAX) language and the Proteomics Standards Initiative Molecular Interactions format, and then integrated. Pathway Commons provides biologists with (1) tools to search this comprehensive resource, (2) a download site offering integrated bulk sets of pathway data (e.g., tables of interactions and gene sets), (3) reusable software libraries for working with pathway information in several programming languages (Java, R, Python, and Javascript), and (4) a web service for programmatically querying the entire dataset. Visualization of pathways is supported using the Systems Biological Graphical Notation (SBGN). Pathway Commons currently contains data from 22 databases with 4,794 detailed human biochemical processes (i.e., pathways) and [~]2.3 million interactions. To enhance the usability of this large resource for end-users, we develop and maintain interactive web applications and training materials that enable pathway exploration and advanced analysis.
]]></description>
<dc:creator>Rodchenkov, I.</dc:creator>
<dc:creator>Babur, O.</dc:creator>
<dc:creator>Luna, A.</dc:creator>
<dc:creator>Aksoy, B. A.</dc:creator>
<dc:creator>Wong, J. V.</dc:creator>
<dc:creator>Fong, D.</dc:creator>
<dc:creator>Franz, M.</dc:creator>
<dc:creator>Siper, M. C.</dc:creator>
<dc:creator>Cheung, M.</dc:creator>
<dc:creator>Wrana, M.</dc:creator>
<dc:creator>Mistry, H.</dc:creator>
<dc:creator>Mosier, L.</dc:creator>
<dc:creator>Dlin, J.</dc:creator>
<dc:creator>Wen, Q.</dc:creator>
<dc:creator>O'Callaghan, C.</dc:creator>
<dc:creator>Li, W.</dc:creator>
<dc:creator>Elder, G.</dc:creator>
<dc:creator>Smith, P. T.</dc:creator>
<dc:creator>Dallago, C.</dc:creator>
<dc:creator>Cerami, E.</dc:creator>
<dc:creator>Gross, B. E.</dc:creator>
<dc:creator>Dogrusoz, U.</dc:creator>
<dc:creator>Demir, E.</dc:creator>
<dc:creator>Bader, G.</dc:creator>
<dc:creator>Sander, C.</dc:creator>
<dc:date>2019-10-01</dc:date>
<dc:identifier>doi:10.1101/788521</dc:identifier>
<dc:title><![CDATA[Pathway Commons: 2019 Update]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-10-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/788794v1?rss=1">
<title>
<![CDATA[
Long-term alterations in brain and behavior after postnatal Zika virus infections in infant macaques 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/788794v1?rss=1</link>
<description><![CDATA[
Considering the impact that Zika virus (ZIKV) infection has on the fetal nervous system and given that the postnatal period is also a time of rapid brain growth, it is important to understand the potential neurobehavioral consequences of ZIKV infection during infancy. Postnatal ZIKV infection in a rhesus macaque (RM) model resulted in long-term behavioral, motor, and cognitive changes, including increased emotional reactivity, decreased social contact, loss of balance, and deficits in visual recognition memory at one year of age. Structural and functional MRI showed that ZIKV-infected infant RMs had persistent enlargement of lateral ventricles, smaller volumes and altered functional connectivity between brain areas important for socioemotional behavior, cognitive, and motor function (e.g. amygdala, hippocampus, cerebellum). Neuropathological changes corresponded with neuroimaging results and were consistent with the behavioral and memory deficits. Overall, this study demonstrates that postnatal ZIKV infection of infants in this model has long lasting neurodevelopmental consequences.
]]></description>
<dc:creator>Raper, J.</dc:creator>
<dc:creator>Kovacs-Balint, Z.</dc:creator>
<dc:creator>Mavinger, M.</dc:creator>
<dc:creator>Gumber, S.</dc:creator>
<dc:creator>Burke, M. W.</dc:creator>
<dc:creator>Habib, J.</dc:creator>
<dc:creator>Mattingly, C.</dc:creator>
<dc:creator>Fair, D.</dc:creator>
<dc:creator>Earl, E.</dc:creator>
<dc:creator>Feczko, E.</dc:creator>
<dc:creator>Styner, M.</dc:creator>
<dc:creator>Jean, S.</dc:creator>
<dc:creator>Cohen, J.</dc:creator>
<dc:creator>Suthar, M.</dc:creator>
<dc:creator>Sanchez, M. M.</dc:creator>
<dc:creator>Alvarado, M. C.</dc:creator>
<dc:creator>Chahroudi, A.</dc:creator>
<dc:date>2019-10-01</dc:date>
<dc:identifier>doi:10.1101/788794</dc:identifier>
<dc:title><![CDATA[Long-term alterations in brain and behavior after postnatal Zika virus infections in infant macaques]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-10-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/792770v1?rss=1">
<title>
<![CDATA[
RESTORE: Robust intEnSiTy nORmalization mEthod for Multiplexed Imaging 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/792770v1?rss=1</link>
<description><![CDATA[
Recent advances in multiplexed imaging technologies promise to improve the understanding of the functional states of individual cells and the interactions between the cells in tissues. This often requires compilation of results from multiple samples. However, quantitative integration of information between samples is complicated by variations in staining intensity and background fluorescence that obscure biological variations. Failure to remove these unwanted artefacts will complicate downstream analysis and diminish the value of multiplexed imaging for clinical applications. Here, to compensate for unwanted variations, we automatically identify negative control cells for each marker within the same tissue and use their expression levels to infer background signal level. The intensity profile is normalized by the inferred level of the negative control cells to remove between-sample variation. Using a tissue microarray data and a pair of longitudinal biopsies sample, we demonstrated that the proposed approach can remove unwanted variations effectively and shows robust performance.
]]></description>
<dc:creator>Chang, Y. H.</dc:creator>
<dc:creator>Chin, K.</dc:creator>
<dc:creator>Thibault, G.</dc:creator>
<dc:creator>Eng, J.</dc:creator>
<dc:creator>Grace, L.</dc:creator>
<dc:creator>Gray, J. W.</dc:creator>
<dc:date>2019-10-03</dc:date>
<dc:identifier>doi:10.1101/792770</dc:identifier>
<dc:title><![CDATA[RESTORE: Robust intEnSiTy nORmalization mEthod for Multiplexed Imaging]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-10-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/793042v1?rss=1">
<title>
<![CDATA[
The lhfpl5 ohnologs lhfpl5a and lhfpl5b are required for mechanotransduction in distinct populations of sensory hair cells in zebrafish. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/793042v1?rss=1</link>
<description><![CDATA[
1Hair cells sense and transmit auditory, vestibular, and hydrodynamic information by converting mechanical stimuli into electrical signals. This process of mechano-electrical transduction (MET) requires a mechanically-gated channel localized in the apical stereocilia of hair cells. In mice, lipoma HMGIC fusion partner-like 5 (LHFPL5) acts as an auxiliary subunit of the MET channel whose primary role is to correctly localize PCDH15 and TMC1 to the mechanotransduction complex. Zebrafish have two lhfpl5 genes (lhfpl5a and lhfpl5b), but their individual contributions to MET channel assembly and function have not been analyzed.

Here we show that the zebrafish lhfpl5 genes are expressed in discrete populations of hair cells: lhfpl5a expression is restricted to auditory and vestibular hair cells in the inner ear, while lhfpl5b expression is specific to hair cells of the lateral line organ. Consequently, lhfpl5a mutants exhibit defects in auditory and vestibular function, while disruption of lhfpl5b affects hair cells only in the lateral line neuromasts. In contrast to previous reports in mice, localization of Tmc1 does not depend upon Lhfpl5 function in either the inner ear or lateral line organ. In both lhfpl5a and lhfpl5b mutants, GFP-tagged Tmc1 and Tmc2b proteins still localize to the stereocilia of hair cells. Using a stably integrated GFP-Lhfpl5a transgene, we show that the tip link cadherins Pcdh15a and Cdh23, along with the Myo7aa motor protein, are required for correct Lhfpl5a localization at the tips of stereocilia. Our work corroborates the evolutionarily conserved co-dependence between Lhfpl5 and Pcdh15, but also reveals novel requirements for Cdh23 and Myo7aa to correctly localize Lhfpl5a. In addition, our data suggest that targeting of Tmc1 and Tmc2b proteins to stereocilia in zebrafish hair cells occurs independently of Lhfpl5 proteins.
]]></description>
<dc:creator>Erickson, T.</dc:creator>
<dc:creator>Pacentine, I. V.</dc:creator>
<dc:creator>Venuto, A.</dc:creator>
<dc:creator>Clemens, R.</dc:creator>
<dc:creator>Nicolson, T.</dc:creator>
<dc:date>2019-10-07</dc:date>
<dc:identifier>doi:10.1101/793042</dc:identifier>
<dc:title><![CDATA[The lhfpl5 ohnologs lhfpl5a and lhfpl5b are required for mechanotransduction in distinct populations of sensory hair cells in zebrafish.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-10-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/796235v1?rss=1">
<title>
<![CDATA[
Herpes Simplex Virus type 1 Inflammasome Activation in Human Macrophages is Dependent on NLRP3, ASC, and Caspase-1 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/796235v1?rss=1</link>
<description><![CDATA[
The pro-inflammatory cytokines interleukin (IL)-1{beta} and IL-18 are products of activation of the inflammasome, an innate sensing system, and important in the pathogenesis of herpes simplex type 1 (HSV-1). The release of IL-18 and IL-1{beta} from monocytes/macrophages is critical for protection from HSV-1 based on animal models of encephalitis and genital infection, yet if and how HSV-1 activates inflammasomes in human macrophages is unknown. To investigate this, we utilized both primary human monocyte derived macrophages and human monocytic cell lines (THP-1 cells) with various inflammasome components knocked-out. We found that HSV-1 activates inflammasome signaling in pro-inflammatory primary human macrophages. Additionally, HSV-1 inflammasome activation is dependent on nucleotide-binding domain and leucine-rich repeat-containing receptor 3 (NLRP3), apoptosis-associated speck-like molecule containing a caspase recruitment domain (ASC), and caspase-1, but not on absent in melanoma 2 (AIM2), or gamma interferon-inducible protein 16 (IFI16). Ultraviolet irradiation of HSV-1 enhanced inflammasome activation, demonstrating that viral replication suppresses inflammasome activation. These results confirm that HSV-1 is capable of activating the inflammasome in human macrophages through an NLRP3 dependent process and that the virus has an NLRP3 specific mechanism to inhibit inflammasome activation in monocytes and macrophages.nnAuthor SummaryThe inflammasome is a multi-protein complex that forms in response to pathogens and cellular damage. Active inflammasomes recruit pro-caspase-1 via ASC and cleave the cytokine precursors pro-IL-1{beta} and pro-IL-18 into mature IL-1{beta} and IL-18. These cytokines serve to activate other immune cells that either repair the damage or attempt to clear the invading pathogen. Upon activation, the inflammasome also promotes an inflammatory form of cell death called pyroptosis. Herpes simplex virus type 1 (HSV-1) is a common human pathogen that can cause cold sores, genital ulcers, encephalitis, and blindness. HSV-1 infection leads to induction of IL-1{beta} and IL-18, but whether it is capable of activating inflammasomes in macrophages, which play a role in severe forms of HSV-1 infection, was unclear. Here, we infected both primary human macrophages and a macrophage/monocytic cell line, THP-1 cells, with HSV-1. We found that HSV-1 does activate inflammasome signaling in macrophages in a process dependent on NLRP3, ASC, and caspase-1. This is important because it illustrates the mechanism by which HSV-1 infection leads to inflammasome activation in macrophages, known to be crucial for protection from severe disease in mouse models.
]]></description>
<dc:creator>Karaba, A. H.</dc:creator>
<dc:creator>Figueroa, A.</dc:creator>
<dc:creator>Massaccesi, G.</dc:creator>
<dc:creator>Botto, S.</dc:creator>
<dc:creator>DeFilippis, V. R.</dc:creator>
<dc:creator>Cox, A.</dc:creator>
<dc:date>2019-10-07</dc:date>
<dc:identifier>doi:10.1101/796235</dc:identifier>
<dc:title><![CDATA[Herpes Simplex Virus type 1 Inflammasome Activation in Human Macrophages is Dependent on NLRP3, ASC, and Caspase-1]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-10-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/803346v1?rss=1">
<title>
<![CDATA[
Biallelic mutations in M1AP are a frequent cause of meiotic arrest leading to male infertility 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/803346v1?rss=1</link>
<description><![CDATA[
Male infertility affects [~]7% of men in Western societies, but its causes remain poorly understood. The most clinically severe form of male infertility is non-obstructive azoospermia (NOA), which is, in part, caused by an arrest at meiosis, but so far only few genes have been reported to cause germ cell arrest in males. To address this gap, whole exome sequencing was performed in 60 German men with complete meiotic arrest, and we identified in three unrelated men the same homozygous frameshift variant c.676dup (p.Trp226LeufsTer4) in M1AP, encoding meiosis 1 arresting protein. Then, with collaborators from the International Male Infertility Genomics Consortium (IMIGC), we screened a Dutch cohort comprising 99 infertile men and detected the same homozygous variant c.676dup in a man with hypospermatogenesis predominantly displaying meiotic arrest. We also identified two Portuguese men with NOA carrying likely biallelic loss-of-function (LoF) and missense variants in M1AP among men screened by the Genetics of Male Infertility Initiative (GEMINI). Moreover, we discovered a homozygous missense variant p.(Pro389Leu) in M1AP in a consanguineous Turkish family comprising five infertile men. M1AP is predominantly expressed in human and mouse spermatogonia up to secondary spermatocytes and previous studies have shown that knockout male mice are infertile due to meiotic arrest. Collectively, these findings demonstrate that both LoF and missense M1AP variants that impair its protein cause autosomal-recessive meiotic arrest, non-obstructive azoospermia and male infertility. In view of the evidence from several independent groups and populations, M1AP should be included in the growing list of validated NOA genes.
]]></description>
<dc:creator>Wyrwoll, M. J.</dc:creator>
<dc:creator>Temel, S. G.</dc:creator>
<dc:creator>Nagirnaja, L.</dc:creator>
<dc:creator>Oud, M. S.</dc:creator>
<dc:creator>Lopes, A. M.</dc:creator>
<dc:creator>van der Heijden, G. W.</dc:creator>
<dc:creator>Rotte, N.</dc:creator>
<dc:creator>Wistuba, J.</dc:creator>
<dc:creator>Wöste, M.</dc:creator>
<dc:creator>Ledig, S.</dc:creator>
<dc:creator>Krenz, H.</dc:creator>
<dc:creator>Smits, R. M.</dc:creator>
<dc:creator>Carvalho, F.</dc:creator>
<dc:creator>Goncalves, J.</dc:creator>
<dc:creator>Fietz, D.</dc:creator>
<dc:creator>Türkgenc, B.</dc:creator>
<dc:creator>Ergören, M. C.</dc:creator>
<dc:creator>Cetinkaya, M.</dc:creator>
<dc:creator>Basar, M.</dc:creator>
<dc:creator>Kahraman, S.</dc:creator>
<dc:creator>Pilatz, A.</dc:creator>
<dc:creator>Röpke, A.</dc:creator>
<dc:creator>Dugas, M.</dc:creator>
<dc:creator>Kliesch, S.</dc:creator>
<dc:creator>Neuhaus, N.</dc:creator>
<dc:creator>Aston, K. I.</dc:creator>
<dc:creator>Conrad, D. F.</dc:creator>
<dc:creator>Veltman, J. A.</dc:creator>
<dc:creator>Friedrich, C.</dc:creator>
<dc:creator>Tüttelmann, F.</dc:creator>
<dc:date>2019-10-15</dc:date>
<dc:identifier>doi:10.1101/803346</dc:identifier>
<dc:title><![CDATA[Biallelic mutations in M1AP are a frequent cause of meiotic arrest leading to male infertility]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-10-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/805309v1?rss=1">
<title>
<![CDATA[
Broad binaural fusion impairs segregation of speech based on voice pitch differences in a cocktail party environment 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/805309v1?rss=1</link>
<description><![CDATA[
In the normal auditory system, central auditory neurons are sharply tuned to the same frequency ranges for each ear. This precise tuning is mirrored behaviorally as the binaural fusion of tones evoking similar pitches across ears. In contrast, hearing-impaired listeners exhibit abnormally broad tuning of binaural pitch fusion, fusing sounds with pitches differing by up to 3-4 octaves across ears into a single object. Here we present evidence that such broad fusion may similarly impair the segregation and recognition of speech based on voice pitch differences in a  cocktail party environment. Speech recognition performance in a multi-talker environment was measured in four groups of adult subjects: normal-hearing (NH) listeners and hearing-impaired listeners with bilateral hearing aids (HAs), bimodal cochlear implant (CI) worn with a contralateral HA, or bilateral CIs. Performance was measured as the threshold target-to-masker ratio needed to understand a target talker in the presence of masker talkers either co-located or symmetrically spatially separated from the target. Binaural pitch fusion was also measured. Voice pitch differences between target and masker talkers improved speech recognition performance for the NH, bilateral HA, and bimodal CI groups, but not the bilateral CI group. Spatial separation only improved performance for the NH group, indicating an inability of the hearing-impaired groups to benefit from spatial release from masking. A moderate to strong negative correlation was observed between the benefit from voice pitch differences and the breadth of binaural pitch fusion in all groups except the bilateral CI group in the co-located spatial condition. Hence, tuning of binaural pitch fusion predicts the ability to segregate voices based on pitch when acoustic cues are available. The findings suggest that obligatory binaural fusion, with a concomitant loss of information from individual streams, may occur at a level of processing before auditory object formation and segregation.
]]></description>
<dc:creator>Oh, Y.</dc:creator>
<dc:creator>Hartling, C. L.</dc:creator>
<dc:creator>Srinivasan, N. K.</dc:creator>
<dc:creator>Eddolls, M.</dc:creator>
<dc:creator>Diedesch, A. C.</dc:creator>
<dc:creator>Gallun, F. J.</dc:creator>
<dc:creator>Reiss, L. A. J.</dc:creator>
<dc:date>2019-10-16</dc:date>
<dc:identifier>doi:10.1101/805309</dc:identifier>
<dc:title><![CDATA[Broad binaural fusion impairs segregation of speech based on voice pitch differences in a cocktail party environment]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-10-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/805515v1?rss=1">
<title>
<![CDATA[
Structural basis for adhesion G protein-coupled receptor Gpr126 function 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/805515v1?rss=1</link>
<description><![CDATA[
Many drugs target the extracellular regions (ECRs) of cell-surface receptors. The large and alternatively-spliced ECRs of adhesion G protein-coupled receptors (aGPCRs) have key functions in diverse biological processes including neurodevelopment, embryogenesis, and tumorigenesis. However, their structures and mechanisms of action remain unclear, hampering drug development. The aGPCR Gpr126/Adgrg6 regulates Schwann cell myelination, ear canal formation, and heart development; and GPR126 mutations cause myelination defects in human. Here, we determine the structure of the complete zebrafish Gpr126 ECR and reveal five domains including a previously-unknown domain. Strikingly, the Gpr126 ECR adopts a closed conformation that is stabilized by an alternatively spliced linker and a conserved calcium-binding site. Alternative splicing regulates ECR conformation and receptor signaling, while mutagenesis of the newly-characterized calcium-binding site abolishes Gpr126 function in vivo. These results demonstrate that Gpr126 ECR utilizes a multi-faceted dynamic approach to regulate receptor function and provide novel insights into ECR-targeted drug design.
]]></description>
<dc:creator>Leon, K.</dc:creator>
<dc:creator>Cunningham, R.</dc:creator>
<dc:creator>Riback, J.</dc:creator>
<dc:creator>Feldman, E.</dc:creator>
<dc:creator>Li, J.</dc:creator>
<dc:creator>Sosnick, T.</dc:creator>
<dc:creator>Zhao, M.</dc:creator>
<dc:creator>Monk, K.</dc:creator>
<dc:creator>Arac, D.</dc:creator>
<dc:date>2019-10-17</dc:date>
<dc:identifier>doi:10.1101/805515</dc:identifier>
<dc:title><![CDATA[Structural basis for adhesion G protein-coupled receptor Gpr126 function]]></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/10.1101/815720v1?rss=1">
<title>
<![CDATA[
Spatially-mapped single-cell chromatin accessibility 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/815720v1?rss=1</link>
<description><![CDATA[
High-throughput single-cell epigenomic assays can resolve the heterogeneity of cell types and states in complex tissues, however, spatial orientation within the network of interconnected cells is lost. Here, we present a novel method for highly scalable, spatially resolved, single-cell profiling of chromatin states. We use high-density multiregional sampling to perform single-cell combinatorial indexing on Microbiopsies Assigned to Positions for the Assay for Transposase Accessible Chromatin (sciMAP-ATAC) to produce single-cell data of an equivalent quality to non-spatially resolved single-cell ATAC-seq, where each cell is localized to a three-dimensional position within the tissue. A typical experiment comprises between 96 and 384 spatially mapped tissue positions, each producing 10s to over 100 individual single-cell ATAC-seq profiles, and a typical resolution of 214 cubic microns; with the ability to tune the resolution and cell throughput to suit each target application. We apply sciMAP-ATAC to the adult mouse primary somatosensory cortex, where we profile cortical lamination and demonstrate the ability to analyze data from a single tissue position or compare a single cell type in adjacent positions. We also profile the human primary visual cortex, where we produce spatial trajectories through the cortex. Finally, we characterize the spatially progressive nature of cerebral ischemic infarct in the mouse brain using a model of transient middle cerebral artery occlusion. We leverage the spatial information to identify novel and known transcription factor activities that vary by proximity to the ischemic infarction core with cell type specificity.
]]></description>
<dc:creator>Thornton, C. A.</dc:creator>
<dc:creator>Mulqueen, R. M.</dc:creator>
<dc:creator>Torkenczy, K. A.</dc:creator>
<dc:creator>Lowenstein, E. G.</dc:creator>
<dc:creator>Fields, A. J.</dc:creator>
<dc:creator>Steemers, F. J.</dc:creator>
<dc:creator>Wright, K. M.</dc:creator>
<dc:creator>Adey, A. C.</dc:creator>
<dc:date>2019-10-22</dc:date>
<dc:identifier>doi:10.1101/815720</dc:identifier>
<dc:title><![CDATA[Spatially-mapped single-cell chromatin accessibility]]></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/10.1101/818534v1?rss=1">
<title>
<![CDATA[
Multi-region exome sequencing of ovarian teratomas reveals 2N near-diploid genomes, paucity of somatic mutations, and extensive allelic imbalances shared across mature, immature, and disseminated components 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/818534v1?rss=1</link>
<description><![CDATA[
Immature teratoma is a subtype of malignant germ cell tumor of the ovary that occurs most commonly in the first three decades of life, frequently with bilateral ovarian disease. Despite being the second most common malignant germ cell tumor of the ovary, little is known about its genetic underpinnings. Here we performed multi-region whole exome sequencing to interrogate the genetic zygosity, clonal relationship, DNA copy number, and mutational status of 52 pathologically distinct tumor components from 10 females with ovarian immature teratomas, with bilateral tumors present in 5 cases and peritoneal dissemination in 7 cases. We found that ovarian immature teratomas are genetically characterized by 2N near-diploid genomes with extensive loss of heterozygosity and an absence of genes harboring recurrent somatic mutations or known oncogenic variants. All components within a single ovarian tumor (immature teratoma, mature teratoma with different histologic patterns of differentiation, and yolk sac tumor) were found to harbor an identical pattern of loss of heterozygosity across the genome, indicating a shared clonal origin. In contrast, the 4 analyzed bilateral teratomas showed distinct patterns of zygosity changes in the right versus left sided tumors, indicating independent clonal origins. All disseminated teratoma components within the peritoneum (including gliomatosis peritonei) shared a clonal pattern of loss of heterozygosity with either the right or left primary ovarian tumor. The observed genomic loss of heterozygosity patterns indicate that diverse meiotic errors contribute to the formation of ovarian immature teratomas, with 11 out of the 15 genetically distinct clones determined to result from the failure of meiosis I or II. Overall, these findings suggest that copy-neutral loss of heterozygosity resulting from meiotic abnormalities may be sufficient to generate ovarian immature teratomas from germ cells.
]]></description>
<dc:creator>Heskett, M. B.</dc:creator>
<dc:creator>Sanborn, J. Z.</dc:creator>
<dc:creator>Boniface, C.</dc:creator>
<dc:creator>Goode, B.</dc:creator>
<dc:creator>Chapman, J.</dc:creator>
<dc:creator>Garg, K.</dc:creator>
<dc:creator>Rabban, J. T.</dc:creator>
<dc:creator>Zaloudek, C.</dc:creator>
<dc:creator>Benz, S. C.</dc:creator>
<dc:creator>Spellman, P. T.</dc:creator>
<dc:creator>Solomon, D. A.</dc:creator>
<dc:creator>Cho, R. J.</dc:creator>
<dc:date>2019-10-28</dc:date>
<dc:identifier>doi:10.1101/818534</dc:identifier>
<dc:title><![CDATA[Multi-region exome sequencing of ovarian teratomas reveals 2N near-diploid genomes, paucity of somatic mutations, and extensive allelic imbalances shared across mature, immature, and disseminated components]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-10-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/823021v1?rss=1">
<title>
<![CDATA[
TGF-β signaling is critical for maintenance of the tendon cell fate 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/823021v1?rss=1</link>
<description><![CDATA[
Studies of cell fate focus on specification, but little is known about maintenance of the differentiated state. We find that TGF{beta} signaling plays an essential role in maintenance of the tendon cell fate. To examine the role TGF{beta} signaling in tenocytes TGF{beta} type II receptor was targeted in the Scleraxis cell lineage. Tendon development was not disrupted in mutant embryos, but shortly after birth tenocytes lost differentiation markers and reverted to a more stem/progenitor state. Targeting of Tgfbr2 using other Cre drivers did not cause tenocyte dedifferentiation suggesting a critical significance for the spatio-temporal activity of ScxCre. Viral reintroduction of Tgfbr2 to mutants was sufficient to prevent and even rescue mutant tenocytes suggesting a continuous and cell-autonomous role for TGF{beta} signaling in cell fate maintenance. These results uncover the critical importance of molecular pathways that maintain the differentiated cell fate and a key role for TGF{beta} signaling in these processes.
]]></description>
<dc:creator>Tan, G.-K.</dc:creator>
<dc:creator>Pryce, B. A.</dc:creator>
<dc:creator>Stabio, A.</dc:creator>
<dc:creator>Brigande, J. V.</dc:creator>
<dc:creator>Wang, C.</dc:creator>
<dc:creator>Xia, Z.</dc:creator>
<dc:creator>Tufa, S. F.</dc:creator>
<dc:creator>Keene, D. R.</dc:creator>
<dc:creator>Schweitzer, R.</dc:creator>
<dc:date>2019-10-29</dc:date>
<dc:identifier>doi:10.1101/823021</dc:identifier>
<dc:title><![CDATA[TGF-β signaling is critical for maintenance of the tendon cell fate]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-10-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/825737v1?rss=1">
<title>
<![CDATA[
A cryo-tomography-based volumetric model of the actin core of mouse vestibular hair cell stereocilia lacking plastin 1 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/825737v1?rss=1</link>
<description><![CDATA[
Electron cryo-tomography allows for high-resolution imaging of stereocilia in their native state. Because their actin filaments have a higher degree of order, we imaged stereocilia from mice lacking the actin crosslinker plastin 1 (PLS1). We found that while stereocilia actin filaments run 13 nm apart in parallel for long distances, there were gaps of significant size that were stochastically distributed throughout the actin core. Actin crosslinkers were distributed through the stereocilium, but did not occupy all possible binding sites. At stereocilia tips, protein density extended beyond actin filaments, especially on the side of the tip where a tip link should anchor. Along the shaft, repeating density was observed that corresponds to actin-to-membrane connectors. In the taper region, most actin filaments terminated near the plasma membrane. The remaining filaments twisted together to make a tighter bundle than was present in the shaft region; the spacing between them decreased from 13 nm to 9 nm. Our models illustrate detailed features of distinct structural domains that are present within the stereocilium.
]]></description>
<dc:creator>Song, J.</dc:creator>
<dc:creator>Patterson, R.</dc:creator>
<dc:creator>Krey, J. F.</dc:creator>
<dc:creator>Hao, S.</dc:creator>
<dc:creator>Wang, L.</dc:creator>
<dc:creator>Ng, B.</dc:creator>
<dc:creator>Sazzed, S.</dc:creator>
<dc:creator>Kovacs, J.</dc:creator>
<dc:creator>Wriggers, W.</dc:creator>
<dc:creator>He, J.</dc:creator>
<dc:creator>Barr-Gillespie, P. G.</dc:creator>
<dc:creator>Auer, M. G.</dc:creator>
<dc:date>2019-10-31</dc:date>
<dc:identifier>doi:10.1101/825737</dc:identifier>
<dc:title><![CDATA[A cryo-tomography-based volumetric model of the actin core of mouse vestibular hair cell stereocilia lacking plastin 1]]></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/10.1101/837161v1?rss=1">
<title>
<![CDATA[
Removal of high frequency contamination from motion estimates in single-band fMRI saves data without biasing functional connectivity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/837161v1?rss=1</link>
<description><![CDATA[
Denoising fMRI data requires assessment of frame-to-frame head motion and removal of the biases motion introduces. This is usually done through analysis of the parameters calculated during retrospective head motion correction (i.e.,  motion parameters). However, it is increasingly recognized that respiration introduces factitious head motion via perturbations of the main (B0) field. This effect appears as higher-frequency fluctuations in the motion parameters (> 0.1 Hz, here referred to as  HF-motion), primarily in the phase-encoding direction. This periodicity can sometimes be obscured in standard single-band fMRI (TR 2.0 - 2.5 s.) due to aliasing. Here we examined (1) how prevalent HF-motion effects are in seven single-band datasets with TR from 2.0 - 2.5 s and (2) how HF-motion affects functional connectivity. We demonstrate that HF-motion is relatively trait-like and more common in older adults, those with higher body mass index, and those with lower cardiorespiratory fitness. We propose a low-pass filtering approach to remove the contamination of high frequency effects from motion summary measures, such as framewise displacement (FD). We demonstrate that in most datasets this filtering approach saves a substantial amount of data from FD-based frame censoring, while at the same time reducing motion biases in functional connectivity measures. These findings suggest that filtering motion parameters is an effective way to improve the fidelity of head motion estimates, even in single band datasets. Particularly large data savings may accrue in datasets acquired in older and less fit participants.

Highlights- Single-band fMRI motion traces show factitious high-frequency content (HF-motion)
- The magnitude of HF-motion relates to age and other demographic factors
- HF-motion elevates framewise displacement (FD) and causes data loss
- Substantial fMRI data can be recovered from censoring by filtering motion traces
- Filtering motion traces reduces motion artifacts in functional connectivity
]]></description>
<dc:creator>Gratton, C.</dc:creator>
<dc:creator>Coalson, R. S.</dc:creator>
<dc:creator>Dworetsky, A.</dc:creator>
<dc:creator>Adeyemo, B.</dc:creator>
<dc:creator>Laumann, T. O.</dc:creator>
<dc:creator>Wig, G. S.</dc:creator>
<dc:creator>Kong, T. S.</dc:creator>
<dc:creator>Gratton, G.</dc:creator>
<dc:creator>Fabiani, M.</dc:creator>
<dc:creator>Barch, D. M.</dc:creator>
<dc:creator>Tranel, D.</dc:creator>
<dc:creator>Miranda-Dominguez, O.</dc:creator>
<dc:creator>Fair, D. A.</dc:creator>
<dc:creator>Dosenbach, N. U. F.</dc:creator>
<dc:creator>Snyder, A. Z.</dc:creator>
<dc:creator>Perlmutter, J. S.</dc:creator>
<dc:creator>Petersen, S. E.</dc:creator>
<dc:creator>Campbell, M. C.</dc:creator>
<dc:date>2019-11-09</dc:date>
<dc:identifier>doi:10.1101/837161</dc:identifier>
<dc:title><![CDATA[Removal of high frequency contamination from motion estimates in single-band fMRI saves data without biasing functional connectivity]]></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/10.1101/837294v1?rss=1">
<title>
<![CDATA[
B Cell Receptor-Responsive miR-141 and viral miR-BART9 promote Epstein-Barr Virus Reactivation Through FOXO3 Inhibition 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/837294v1?rss=1</link>
<description><![CDATA[
Antigen recognition by the B cell receptor (BCR) is a physiological trigger for reactivation of Epstein-Barr virus (EBV) and can be recapitulated in vitro by cross-linking of surface immunoglobulins. Previously, we identified a subset of EBV microRNAs (miRNAs) that attenuate BCR signal transduction and subsequently, dampen lytic replication in B cells. The roles of host miRNAs in virus reactivation are not completely understood. To investigate this process, we profiled the small RNAs in latently infected and reactivated Burkitts lymphoma cells, and identified several miRNAs, such as miR-141, that are induced upon BCR cross-linking. Notably, EBV encodes a viral miRNA, miR-BART9, with sequence homology to miR-141. To better understand the functions of these two miRNAs, we examined their molecular targets and experimentally validated multiple candidates commonly regulated by both miRNAs. Targets included transcriptional regulators of the EBV immediate early promoters and B cell transcription factors, leading us to hypothesize that these miRNAs modulate kinetics of the latent to lytic switch in B cells. Through functional assays, we identified roles for miR-141 and EBV miR-BART9 and one specific target, FOXO3, in lytic reactivation. Our data support a model whereby EBV exploits BCR-responsive miR-141 and further mimics activity of this miRNA family via a viral miRNA to promote productive virus replication.

ImportanceEBV is a human pathogen associated with several malignancies. A key aspect of lifelong virus persistence is the ability to switch between latent and lytic replication modes. Mechanisms governing latency and lytic reactivation are only partly understood, and how the EBV latent to lytic switch is post-transcriptionally regulated remains an outstanding question. This study sheds light on how miR-141 expression is regulated in Burkitts lymphoma cells, and identifies a role for FOXO3, a common target of both miR-141 and viral miR-BART9, in modulating EBV reactivation.
]]></description>
<dc:creator>Chen, Y.</dc:creator>
<dc:creator>Fachko, D. L.</dc:creator>
<dc:creator>Ivanov, N. S.</dc:creator>
<dc:creator>Skalsky, R. L.</dc:creator>
<dc:date>2019-11-10</dc:date>
<dc:identifier>doi:10.1101/837294</dc:identifier>
<dc:title><![CDATA[B Cell Receptor-Responsive miR-141 and viral miR-BART9 promote Epstein-Barr Virus Reactivation Through FOXO3 Inhibition]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-11-10</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/10.1101/848119v1?rss=1">
<title>
<![CDATA[
Pharmacological complementation remedies an inborn error of lipid metabolism 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/848119v1?rss=1</link>
<description><![CDATA[
X-linked adrenoleukodystrophy (X-ALD) is a rare, genetic disease in which increased very long chain fatty acids (VLCFAs) in the central nervous system (CNS) cause demyelination and axonal degeneration, leading to severe neurological deficits. Sobetirome, a potent thyroid hormone agonist, has been shown to lower VLCFA levels in the periphery and CNS. In this study, two pharmacological strategies for enhancing the effects of thyromimetics were tested in Abcd1 KO mice, a murine model that has the same inborn error in metabolism as X-ALD patients. First, a sobetirome prodrug (Sob-AM2) with increased CNS penetration lowered CNS VLCFAs more potently than sobetirome, and was better tolerated with lower peripheral exposure, but was unable to unable to break the efficacy threshold of CNS VLCFA lowering in Abcd1 KO mice. Second, co-administration of thyroid hormone with sobetirome enhanced VLCFA lowering in the periphery compared to sobetirome alone but did not produce greater lowering in the CNS. These data suggest that the extent of CNS VLCFA lowering in Abcd1 KO mice is limited by a mechanistic threshold related to slow turnover kinetics, potentially related to the lack of frank X-ALD disease in this model. However, Sob-AM2 has improved potency at correcting the lipid abnormality associated with X-ALD in the CNS with better tolerance than the parent drug sobetirome.
]]></description>
<dc:creator>Hartley, M. D.</dc:creator>
<dc:creator>Shokat, M. D.</dc:creator>
<dc:creator>DeBell, M. J.</dc:creator>
<dc:creator>Banerji, T.</dc:creator>
<dc:creator>Kirkemo, L. L.</dc:creator>
<dc:creator>Scanlan, T. S.</dc:creator>
<dc:date>2019-11-20</dc:date>
<dc:identifier>doi:10.1101/848119</dc:identifier>
<dc:title><![CDATA[Pharmacological complementation remedies an inborn error of lipid metabolism]]></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/10.1101/849083v1?rss=1">
<title>
<![CDATA[
Semaphorin 3A induces cytoskeletal paralysis in tumor-specific CD8+ T cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/849083v1?rss=1</link>
<description><![CDATA[
Semaphorin-3A (Sema3A) regulates tumor angiogenesis, but its role in modulating anti-tumor immunity is unclear. We demonstrate that Sema3A secreted within the tumor microenvironment (TME) suppresses tumor-specific CD8+ T cell function via Neuropilin-1 (NRP1), a receptor that is upregulated upon activation with T cells cognate antigen. Sema3A inhibits T cell migration, assembly of the immunological synapse, and tumor killing. It achieves these functional effects through hyper-activating the acto-myosin system in T cells leading to cellular paralysis. Finally, using a clear cell renal cell carcinoma patient cohort, we demonstrate that human tumor-specific CD8+ T cells express NRP1 and are trapped in Sema3A rich regions of tumors. Our study establishes Sema3A as a potent inhibitor of anti-tumor immunity.
]]></description>
<dc:creator>Barnkob, M. B.</dc:creator>
<dc:creator>Michaels, Y. S.</dc:creator>
<dc:creator>Andre, V.</dc:creator>
<dc:creator>Macklin, P. S.</dc:creator>
<dc:creator>Gileadi, U.</dc:creator>
<dc:creator>Valvo, S.</dc:creator>
<dc:creator>Rei, M.</dc:creator>
<dc:creator>Kulicke, C.</dc:creator>
<dc:creator>Chen, J.-L.</dc:creator>
<dc:creator>Jain, V.</dc:creator>
<dc:creator>Woodcock, V.</dc:creator>
<dc:creator>Colin-York, H.</dc:creator>
<dc:creator>Hadjinicolaou, A. V.</dc:creator>
<dc:creator>Kong, Y.</dc:creator>
<dc:creator>Mayya, V.</dc:creator>
<dc:creator>Bull, J. A.</dc:creator>
<dc:creator>Rijal, P.</dc:creator>
<dc:creator>Pugh, C. W.</dc:creator>
<dc:creator>Townsend, A.</dc:creator>
<dc:creator>Olsen, L. R.</dc:creator>
<dc:creator>Fritzsche, M.</dc:creator>
<dc:creator>Fulga, T. A.</dc:creator>
<dc:creator>Dustin, M. L.</dc:creator>
<dc:creator>Jones, E. Y.</dc:creator>
<dc:creator>Cerundolo, V.</dc:creator>
<dc:date>2019-11-20</dc:date>
<dc:identifier>doi:10.1101/849083</dc:identifier>
<dc:title><![CDATA[Semaphorin 3A induces cytoskeletal paralysis in tumor-specific CD8+ T cells]]></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/10.1101/850081v1?rss=1">
<title>
<![CDATA[
Head Stabilization During Standing in People with Persisting Symptoms after Mild Traumatic Brain Injury 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/850081v1?rss=1</link>
<description><![CDATA[
Increased postural sway is often observed in people with mild traumatic brain injury (mTBI), but our understanding of how individuals with mTBI control their head during stance is limited. The purpose of this study was to determine if people with mTBI exhibit increased sway at the head compared with healthy controls. People with persisting symptoms after mTBI (n = 59, 41 women) and control participants (n = 63, 38 women) stood quietly for one minute in four conditions: eyes open on a firm surface (EO-firm), eyes closed on a firm surface (EC-firm), eyes open on a foam pad (EO-foam), and eyes closed on foam (EC-foam). Inertial sensors at the head, sternum, and lumbar region collected tri-axial accelerations. Root-mean-square (RMS) accelerations in anteroposterior (AP) and mediolateral (ML) directions. Sway ratios between the head and sternum, head and lumbar, and sternum and lumbar region, were compared between groups. Temporal coupling of anti-phase motion between the upper and lower body angular accelerations was assessed with magnitude squared coherence and cross-spectral phase angles. People with mTBI demonstrated greater sway than controls across conditions and directions. During foam-surface conditions, the control group, but not the mTBI group, reduced ML sway at their head and trunk relative to their lumbar by increasing the expression of an anti-phase hip strategy within the frontal plane. These results are consistent with suggestions of inflexible or inappropriate postural control in people with mTBI.
]]></description>
<dc:creator>Fino, P. C.</dc:creator>
<dc:creator>Raffegeau, T. E.</dc:creator>
<dc:creator>Parrington, L.</dc:creator>
<dc:creator>Peterka, R. J.</dc:creator>
<dc:creator>King, L. A.</dc:creator>
<dc:date>2019-11-21</dc:date>
<dc:identifier>doi:10.1101/850081</dc:identifier>
<dc:title><![CDATA[Head Stabilization During Standing in People with Persisting Symptoms after Mild Traumatic Brain Injury]]></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/10.1101/852830v1?rss=1">
<title>
<![CDATA[
Divergent strategies for learning in males and females 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/852830v1?rss=1</link>
<description><![CDATA[
A frequent assumption in value-based decision-making tasks is that agents make decisions based on the feature dimension that reward probabilities vary on. However, in complex, multidimensional environments, stimuli can vary on multiple dimensions at once, meaning that the feature deserving the most credit for outcomes is not always obvious. As a result, individuals may vary in the strategies used to sample stimuli across dimensions, and these strategies may have an unrecognized influence on decision-making. Sex is a proxy for multiple genetic and endocrine influences that can influence decision-making strategies, including how environments are sampled. In this study, we examined the strategies adopted by female and male mice as they learned the value of stimuli that varied in both image and location in a visually-cued two-armed bandit, allowing two possible dimensions to learn about. Female mice acquired the correct image-value associations more quickly than male mice, and they used a fundamentally different strategy to do so. Female mice constrained their decision-space early in learning by preferentially sampling one location over which images varied. Conversely, male strategies were inconsistent, changing frequently and strongly influenced by the immediate experience of stochastic rewards. Individual strategies were related to sex-gated changes in neuronal activation in early learning. Together, we find that in mice, sex is linked with divergent strategies for sampling and learning about the world, revealing substantial unrecognized variability in the approaches implemented during value-based decision-making.
]]></description>
<dc:creator>Chen, C. S.</dc:creator>
<dc:creator>Ebitz, R. B.</dc:creator>
<dc:creator>Bindas, S.</dc:creator>
<dc:creator>Redish, A. D.</dc:creator>
<dc:creator>Hayden, B.</dc:creator>
<dc:creator>Grissom, N. M.</dc:creator>
<dc:date>2019-11-25</dc:date>
<dc:identifier>doi:10.1101/852830</dc:identifier>
<dc:title><![CDATA[Divergent strategies for learning in males and females]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-11-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/864082v1?rss=1">
<title>
<![CDATA[
Maternal Obesity and Western-style Diet Impair Fetal and Juvenile Offspring Skeletal Muscle Insulin-Stimulated Glucose Transport in Nonhuman Primates 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/864082v1?rss=1</link>
<description><![CDATA[
Infants born to mothers with obesity have a greater risk for childhood obesity and metabolic diseases; however, the underlying biological mechanisms remain poorly understood. We used a nonhuman primate model to investigate whether maternal obesity combined with a western-style diet (WSD) impairs offspring muscle insulin action. Briefly, adult females were fed a control (CON) or WSD prior to and during pregnancy and lactation. Offspring were weaned to a CON or WSD. Muscle glucose uptake and insulin signaling were measured ex vivo in fetal and juvenile offspring. In vivo signaling was evaluated before and after an intravenous insulin bolus just prior to weaning. We find that fetal muscle exposed to maternal WSD had reduced insulin-stimulated glucose uptake and impaired insulin signaling. In juvenile offspring, insulin-stimulated glucose uptake was similarly reduced by both maternal and post-weaning WSD. Analysis of insulin signaling activation revealed distinct changes between fetal and post-weaning WSD exposure. We conclude that maternal WSD leads to a persistent decrease in insulin-stimulated glucose uptake in juvenile offspring even in the absence of increased offspring adiposity or markers of systemic insulin resistance. Switching offspring to a healthy diet did not ameliorate the effects of maternal WSD suggesting earlier interventions may be necessary.
]]></description>
<dc:creator>Campodonico-Burnett, W.</dc:creator>
<dc:creator>Hetrick, B.</dc:creator>
<dc:creator>Wesolowski, S. R.</dc:creator>
<dc:creator>Schenk, S.</dc:creator>
<dc:creator>Takahashi, D. L.</dc:creator>
<dc:creator>Dean, T. A.</dc:creator>
<dc:creator>Sullivan, E. L.</dc:creator>
<dc:creator>Kievit, P.</dc:creator>
<dc:creator>Gannon, M.</dc:creator>
<dc:creator>Aagaard-Tillery, K.</dc:creator>
<dc:creator>Friedman, J. E.</dc:creator>
<dc:creator>McCurdy, C. E.</dc:creator>
<dc:date>2019-12-03</dc:date>
<dc:identifier>doi:10.1101/864082</dc:identifier>
<dc:title><![CDATA[Maternal Obesity and Western-style Diet Impair Fetal and Juvenile Offspring Skeletal Muscle Insulin-Stimulated Glucose Transport in Nonhuman Primates]]></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/10.1101/865493v1?rss=1">
<title>
<![CDATA[
Evolution of multifunctionality through a pleiotropic substitution in the innate immune protein S100A9 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/865493v1?rss=1</link>
<description><![CDATA[
Multifunctional proteins are evolutionary puzzles: how do proteins evolve to satisfy multiple functional constraints? S100A9 is one such multifunctional protein. It potently amplifies inflammation via Toll-like receptor 4 and is antimicrobial as part of a heterocomplex with S100A8. These two functions are seemingly regulated by proteolysis: S100A9 is readily degraded, while S100A8/S100A9 is resistant. We take an evolutionary biochemical approach to show that S100A9 evolved both functions and lost proteolytic resistance from a weakly proinflammatory, proteolytically resistant amniote ancestor. We identify a historical substitution that has pleiotropic effects on S100A9 proinflammatory activity and proteolytic resistance but has little effect on S100A8/S100A9 antimicrobial activity. We thus propose that mammals evolved S100A8/S100A9 antimicrobial and S100A9 proinflammatory activities concomitantly with a proteolytic "timer" to selectively regulate S100A9. This highlights how the same mutation can have pleiotropic effects on one functional state of a protein but not another, thus facilitating the evolution of multifunctionality.
]]></description>
<dc:creator>Harman, J. L.</dc:creator>
<dc:creator>Loes, A. N.</dc:creator>
<dc:creator>Warren, G. D.</dc:creator>
<dc:creator>Heaphy, M. C.</dc:creator>
<dc:creator>Lampi, K. J.</dc:creator>
<dc:creator>Harms, M. J.</dc:creator>
<dc:date>2019-12-05</dc:date>
<dc:identifier>doi:10.1101/865493</dc:identifier>
<dc:title><![CDATA[Evolution of multifunctionality through a pleiotropic substitution in the innate immune protein S100A9]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-12-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/865659v1?rss=1">
<title>
<![CDATA[
TrpML-mediated astrocyte microdomain Ca2+ transients regulate astrocyte-tracheal interactions in CNS 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/865659v1?rss=1</link>
<description><![CDATA[
Astrocytes exhibit spatially-restricted near-membrane microdomain Ca2+ transients in their fine processes. How these transients are generated and regulate brain function in vivo remains unclear. Here we show that Drosophila astrocytes exhibit spontaneous, activity-independent microdomain Ca2+ transients in their fine processes. Astrocyte microdomain Ca2+ transients are mediated by the TRP channel TrpML, stimulated by reactive oxygen species (ROS), and can be enhanced in frequency by the neurotransmitter tyramine via the TyrRII receptor. Interestingly, many astrocyte microdomain Ca2+ transients are closely associated with tracheal elements, which dynamically extend filopodia throughout the central nervous system (CNS) to deliver O2 and regulate gas exchange. Many astrocyte microdomain Ca2+ transients are spatio-temporally correlated with the initiation of tracheal filopodial retraction. Loss of TrpML leads to increased tracheal filopodial numbers, growth, and increased CNS ROS. We propose that local ROS production can activate astrocyte microdomain Ca2+ transients through TrpML, and that a subset of these microdomain transients promotes tracheal filopodial retraction and in turn modulate CNS gas exchange.
]]></description>
<dc:creator>Ma, Z.</dc:creator>
<dc:creator>Freeman, M. R.</dc:creator>
<dc:date>2019-12-08</dc:date>
<dc:identifier>doi:10.1101/865659</dc:identifier>
<dc:title><![CDATA[TrpML-mediated astrocyte microdomain Ca2+ transients regulate astrocyte-tracheal interactions in CNS]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-12-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/868299v1?rss=1">
<title>
<![CDATA[
Cocaine memory reactivation induces functional adaptations of fast-spiking interneurons in the rat medial prefrontal cortex 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/868299v1?rss=1</link>
<description><![CDATA[
Perineuronal nets (PNNs) are specialized extracellular matrix structures that ensheathe parvalbumin-containing fast-spiking interneurons (PV FSIs) and play a key role in neuroplasticity. We previously showed that PNNs within the prelimbic prefrontal cortex (PL PFC) are required for the maintenance of cocaine-associated memories following cocaine memory reactivation. However, how cocaine memory reactivation affects PNNs, PV, and corresponding changes in PV FSI function are unknown. In this study, we characterized the electrophysiological properties of PV FSIs and corresponding changes in PNN and PV intensity within the PL PFC prior to and after cocaine memory reactivation. Adult male Sprague-Dawley rats were trained to acquire cocaine-conditioned place preference (CPP) and, following cocaine-CPP memory reactivation (30 m, 2 h, and 24 h post-reactivation), we measured PNN intensity (determined by Wisteria floribunda agglutinin [WFA] staining) as well as PV intensity using immunohistochemistry. The intensity of PV staining was reduced at all time points following memory reactivation with no changes in WFA intensity. Using whole-cell electrophysiology we found a reduction in the number of action potentials at 30 m and 2 h that returned to control levels by 24 h. The attenuation in firing was accompanied by a presumed compensatory increase in excitatory synaptic transmission, which was corroborated by an increase in VGluT1 puncta apposing PV/PNN neurons. Collectively, our results indicate that cocaine memory reactivation decreases PV intensity, which may play a role in decreasing excitation of PV FSIs. Thus, the inhibitory tone onto pyramidal neurons may be decreased following memory reactivation, resulting in an increase in PFC output to promote cocaine-seeking behaviors.
]]></description>
<dc:creator>Jorgensen, E. T.</dc:creator>
<dc:creator>Gonzalez, A. E.</dc:creator>
<dc:creator>Harkness, J. H.</dc:creator>
<dc:creator>Hegarty, D. M.</dc:creator>
<dc:creator>Burchi, D. J.</dc:creator>
<dc:creator>Aadland, J. A.</dc:creator>
<dc:creator>Aicher, S. A.</dc:creator>
<dc:creator>Sorg, B. A.</dc:creator>
<dc:creator>Brown, T. E.</dc:creator>
<dc:date>2019-12-06</dc:date>
<dc:identifier>doi:10.1101/868299</dc:identifier>
<dc:title><![CDATA[Cocaine memory reactivation induces functional adaptations of fast-spiking interneurons in the rat medial prefrontal cortex]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-12-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/059204v1?rss=1">
<title>
<![CDATA[
Navigating the phenotype frontier: The Monarch Initiative 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/059204v1?rss=1</link>
<description><![CDATA[
Introduction Introduction References The principles of genetics apply across the entire tree of life. At the cellular level we share biological mechanisms with species from which we diverged millions, even billions of years ago. We can exploit this common ancestry to learn about health and disease, by analyzing DNA and protein sequences, but also through the observable outcomes of genetic differences, i.e. phenotypes.nnTo solve challenging disease problems we need to unify the heterogeneous data that relates genomics to disease traits. Most databases tend to focus either on a single data type across species, or on a single species across data types. Although each database may provide rich, high-quality information, none is unified and comprehensive across species, over biological scales, and throughout data types (Figure 1A).nnWithout ...
]]></description>
<dc:creator>Julie McMurry</dc:creator>
<dc:creator>Sebastian Kohler</dc:creator>
<dc:creator>James Balhoff</dc:creator>
<dc:creator>Charles Borromeo</dc:creator>
<dc:creator>Matthew Brush</dc:creator>
<dc:creator>Seth Carbon</dc:creator>
<dc:creator>Tom Conlin</dc:creator>
<dc:creator>Nathan Dunn</dc:creator>
<dc:creator>Mark Engelstad</dc:creator>
<dc:creator>Erin Foster</dc:creator>
<dc:creator>Jean-Philippe Gourdine</dc:creator>
<dc:creator>Julius Jacobsen</dc:creator>
<dc:creator>Daniel Keith</dc:creator>
<dc:creator>Bryan Laraway</dc:creator>
<dc:creator>Suzanna Lewis</dc:creator>
<dc:creator>Jeremy Nguyen Xuan</dc:creator>
<dc:creator>Kent Shefchek</dc:creator>
<dc:creator>Nicole Vasilevsky</dc:creator>
<dc:creator>Zhou Yuan</dc:creator>
<dc:creator>Nicole Washington</dc:creator>
<dc:creator>Harry Hochheiser</dc:creator>
<dc:creator>Christopher Mungall</dc:creator>
<dc:creator>Tudor Groza</dc:creator>
<dc:creator>Damian Smedley</dc:creator>
<dc:creator>Peter Robinson</dc:creator>
<dc:creator>Melissa Haendel</dc:creator>
<dc:creator></dc:creator>
<dc:date>2016-06-15</dc:date>
<dc:identifier>doi:10.1101/059204</dc:identifier>
<dc:title><![CDATA[Navigating the phenotype frontier: The Monarch Initiative]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2016-06-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/083428v1?rss=1">
<title>
<![CDATA[
Exonic somatic mutations contribute risk for autism spectrum disorder 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/083428v1?rss=1</link>
<description><![CDATA[
Genetic risk factors for autism spectrum disorder (ASD) have yet to be fully elucidated. Somatic mosaic mutations (SMMs) have been implicated in several neurodevelopmental disorders and overgrowth syndromes. Here, we systematically evaluate SMMs by leveraging whole-exome sequencing (WES) data on a large family-based ASD cohort, the Simons Simplex Collection (SSC). We find evidence that ~10% of previously published de novo mutations are potentially SMMs. When using a custom somatic calling pipeline, we recalled all SSC WES data. We validated high and low confidence mutation predictions for a subset of families with single molecule molecular inversion probes. With these validation data, we iteratively developed a high confidence calling approach integrating logistic regression modeling and additional heuristics and applied it to the full cohort. Surprisingly, we found evidence of significant synonymous SMM burden in probands, with mutations more likely to be close to splicing sites. Overall, we observe no strong evidence of missense SMM burden. However, we do observe nominally significant signal for missense SMMs in those families without germline mutations, which strengthens specifically in genes intolerant to mutations. In contrast to missense germline mutations, missense SMMs show potential enrichment for chromatin modifiers. We observe 7-10% of parental mosaics are transmitted germline to a child as occult de novo mutations, which has important implications for recurrence risk for families and potential subclinical ASD features. Finally, we find SMMs in previously implicated high-confidence ASD risk genes, including CHD2, CTNNB1, KMT2C, SYNGAP1, and RELN, further suggesting that this class of mutations contribute to population risk.
]]></description>
<dc:creator>Krupp, D. R.</dc:creator>
<dc:creator>Barnard, R. A.</dc:creator>
<dc:creator>Duffourd, Y.</dc:creator>
<dc:creator>Evans, S.</dc:creator>
<dc:creator>Bernier, R.</dc:creator>
<dc:creator>Riviere, J.-B.</dc:creator>
<dc:creator>Fombonne, E.</dc:creator>
<dc:creator>O'Roak, B. J.</dc:creator>
<dc:date>2016-10-26</dc:date>
<dc:identifier>doi:10.1101/083428</dc:identifier>
<dc:title><![CDATA[Exonic somatic mutations contribute risk for autism spectrum disorder]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2016-10-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/088682v1?rss=1">
<title>
<![CDATA[
In vivo imaging of coral tissue and skeleton with optical coherence tomography 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/088682v1?rss=1</link>
<description><![CDATA[
Optical coherence tomography (OCT) is a non-invasive three-dimensional imaging technique with micrometer resolution allowing microstructural characterization of tissues in vivo and in real time. We present the first application of OCT for in vivo imaging of tissue and skeleton structure of intact living corals spanning a variety of morphologies and tissue thickness. OCT visualized different coral tissue layers (e.g. endoderm vs ectoderm), special structures such as mesenterial filaments and skeletal cavities, as well as mucus release from living corals. We also developed a new approach for non-invasive imaging and quantification of chromatophores containing green fluorescent protein (GFP)-like host pigment granules in coral tissue. The chromatophore system is hyper-reflective and can thus be imaged with good optical contrast in OCT, enabling quantification of chromatophore size, distribution and abundance. Because of its rapid imaging speed, OCT can also be used to quantify coral tissue movement showing that maximal linear contraction velocity was ~120 m per second upon high light stimulation. Based on OCT imaging of tissue expansion and contraction, we made first estimates of dynamic changes in the coral tissue surface area, which varied by a factor of 2 between the contracted and expanded state of the coral Pocillopora damicornis. We conclude that OCT is an excellent novel tool for in vivo tomographic imaging of corals that can reveal tissue and skeleton organization as well as quantify dynamic changes in tissue structure and coral surface area non-invasively and at high spatio-temporal resolution.
]]></description>
<dc:creator>Wangpraseurt, D.</dc:creator>
<dc:creator>Wentzel, C.</dc:creator>
<dc:creator>Jacques, S. L.</dc:creator>
<dc:creator>Wagner, M.</dc:creator>
<dc:creator>Kuhl, M.</dc:creator>
<dc:date>2016-11-22</dc:date>
<dc:identifier>doi:10.1101/088682</dc:identifier>
<dc:title><![CDATA[In vivo imaging of coral tissue and skeleton with optical coherence tomography]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2016-11-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/104844v1?rss=1">
<title>
<![CDATA[
Comprehensive single cell transcriptional profiling of a multicellular organism by combinatorial indexing 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/104844v1?rss=1</link>
<description><![CDATA[
Conventional methods for profiling the molecular content of biological samples fail to resolve heterogeneity that is present at the level of single cells. In the past few years, single cell RNA sequencing has emerged as a powerful strategy for overcoming this challenge. However, its adoption has been limited by a paucity of methods that are at once simple to implement and cost effective to scale massively. Here, we describe a combinatorial indexing strategy to profile the transcriptomes of large numbers of single cells or single nuclei without requiring the physical isolation of each cell (Single cell Combinatorial Indexing RNA-seq or sci-RNA-seq). We show that sci-RNA-seq can be used to efficiently profile the transcriptomes of tens-of-thousands of single cells per experiment, and demonstrate that we can stratify cell types from these data. Key advantages of sci-RNA-seq over contemporary alternatives such as droplet-based single cell RNA-seq include sublinear cost scaling, a reliance on widely available reagents and equipment, the ability to concurrently process many samples within a single workflow, compatibility with methanol fixation of cells, cell capture based on DNA content rather than cell size, and the flexibility to profile either cells or nuclei. As a demonstration of sci-RNA-seq, we profile the transcriptomes of 42,035 single cells from C. elegans at the L2 stage, effectively 50-fold "shotgun cellular coverage" of the somatic cell composition of this organism at this stage. We identify 27 distinct cell types, including rare cell types such as the two distal tip cells of the developing gonad, estimate consensus expression profiles and define cell-type specific and selective genes. Given that C. elegans is the only organism with a fully mapped cellular lineage, these data represent a rich resource for future methods aimed at defining cell types and states. They will advance our understanding of developmental biology, and constitute a major step towards a comprehensive, single-cell molecular atlas of a whole animal.
]]></description>
<dc:creator>Cao, J.</dc:creator>
<dc:creator>Packer, J. S.</dc:creator>
<dc:creator>Ramani, V.</dc:creator>
<dc:creator>Cusanovich, D. A.</dc:creator>
<dc:creator>Huynh, C.</dc:creator>
<dc:creator>Daza, R.</dc:creator>
<dc:creator>Qiu, X.</dc:creator>
<dc:creator>Lee, C.</dc:creator>
<dc:creator>Furlan, S. N.</dc:creator>
<dc:creator>Steemers, F. J.</dc:creator>
<dc:creator>Adey, A.</dc:creator>
<dc:creator>Waterston, R. H.</dc:creator>
<dc:creator>Trapnell, C.</dc:creator>
<dc:creator>Shendure, J.</dc:creator>
<dc:date>2017-02-02</dc:date>
<dc:identifier>doi:10.1101/104844</dc:identifier>
<dc:title><![CDATA[Comprehensive single cell transcriptional profiling of a multicellular organism by combinatorial indexing]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-02-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/155952v1?rss=1">
<title>
<![CDATA[
Delineating the macroscale areal organization of the macaque cortex in vivo 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/155952v1?rss=1</link>
<description><![CDATA[
Complementing longstanding traditions centered around histology, fMRI approaches are rapidly maturing in delineating brain areal organization at the macroscale. The non-human primate (NHP) provides the opportunity to overcome critical barriers in translational research. Here, we establish the data and scanning conditions for achieving reproducible, stable and internally valid areal parcellations in individuals. We demonstrate that these functional boundaries serve as a functional fingerprint of the individual animals, and can be achieved under anesthesia or awake conditions (rest, naturalistic viewing), though differences between awake and anesthetized states precluded the detection of individual differences across states. Comparison of awake and anesthetized states suggested a more nuanced picture of changes in connectivity for higher order association areas, as well as visual and motor cortex. These results establish feasibility and data requirements for the generation of reproducible individual-specific parcellations in NHP, as well as provide insights into the impact of scan state and motivate efforts toward harmonizing protocols.
]]></description>
<dc:creator>Xu, T.</dc:creator>
<dc:creator>Falchier, A.</dc:creator>
<dc:creator>Sullivan, E.</dc:creator>
<dc:creator>Linn, G.</dc:creator>
<dc:creator>Ramirez, J.</dc:creator>
<dc:creator>Ross, D.</dc:creator>
<dc:creator>Feczko, E.</dc:creator>
<dc:creator>Opitz, A.</dc:creator>
<dc:creator>Bagley, J.</dc:creator>
<dc:creator>Sturgeon, D.</dc:creator>
<dc:creator>Earl, E.</dc:creator>
<dc:creator>Miranda-Domingue, O.</dc:creator>
<dc:creator>Perrone, A.</dc:creator>
<dc:creator>Craddock, C.</dc:creator>
<dc:creator>Schroeder, C.</dc:creator>
<dc:creator>Colcombe, S.</dc:creator>
<dc:creator>Fair, D.</dc:creator>
<dc:creator>Milham, M.</dc:creator>
<dc:date>2017-06-26</dc:date>
<dc:identifier>doi:10.1101/155952</dc:identifier>
<dc:title><![CDATA[Delineating the macroscale areal organization of the macaque cortex in vivo]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-06-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/162214v1?rss=1">
<title>
<![CDATA[
Common Garlic (Allium sativum) has Potent Anti-Bacillus anthracis Activity among Various Commonly Used Spices and Herbs 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/162214v1?rss=1</link>
<description><![CDATA[
Ethnopharmacological RelevanceGastrointestinal anthrax, a disease caused by Bacillus anthracis, remains an important but relatively neglected endemic disease of animals and humans in remote areas of the Indian subcontinent and some parts of Africa. Its initial symptoms include diarrhea and stomachache. In the current study, several common plants indicated for diarrhea, dysentery, stomachache or as stomachic as per traditional knowledge in the Indian subcontinent, i.e., Aegle marmelos (L.) Correa (Bael), Allium cepa L. (Onion), Allium sativum L. (Garlic), Azadirachta indica A. Juss. (Neem), Berberis asiatica Roxb. ex DC. (Daruharidra), Coriandrum sativum L. (Coriander), Curcuma longa L. (Turmeric), Cynodon dactylon (L.) Pers. (Bermuda grass), Mangifera indica L. (Mango), Morus indica L. (Black mulberry), Ocimum tenuiflorum L. (Ocimum sanctum L., Holy Basil), Ocimum gratissimum L. (Ram Tulsi), Psidium guajava L. (Guava), Zingiber officinale Roscoe (Ginger), were evaluated for their anti-Bacillus anthracis property. The usage of Azadirachta indica A. Juss. and Curcuma longa L. by Santals (India), and Allium sp. by biblical people to alleviate anthrax-like symptoms is well documented, but the usage of other plants is traditionally only indicated for different gastrointestinal disturbances/conditions.

Aim of the StudyEvaluate the above listed commonly available edible plants from the Indian subcontinent that are used in the traditional medicine to treat gastrointestinal diseases including those also indicated for anthrax-like symptoms for the presence of potent anti-B. anthracis activity in a form amenable to use by the general population in the endemic areas.

Materials and MethodsAqueous extracts made from fourteen plants indicated above were screened for their anti-B. anthracis activity using agar-well diffusion assay (AWDA) and broth microdilution methods. The Aqueous Garlic Extract (AGE) that displayed most potent anti-B. anthracis activity was assessed for its thermostability, stability under pH extremes encountered in the gastrointestinal tract, and potential antagonistic interaction with bile salts as well as the FDA-approved antibiotics used for anthrax control. The bioactive fractions from the AGE were isolated by TLC coupled bioautography followed by their characterization using GC-MS.

ResultsGarlic (Allium sativum L.) extract was identified as the most promising candidate with bactericidal activity against B. anthracis. It consistently inhibited the growth of B. anthracis in AWDA and decreased the viable colony-forming unit counts in liquid-broth cultures by 6-logs within 6-12 h. The AGE displayed acceptable thermostability (>80% anti-B. anthracis activity retained on incubation at 50{degrees}C for 12 h) and stability in gastric pH range (2-8). It did not antagonize the activity of FDA-approved antibiotics used for anthrax control. GC-MS analysis of the TLC separated bioactive fractions of AGE indicated the presence of previously unreported constituents such as phthalic acid derivatives, acid esters, phenyl group-containing compounds, steroids etc.

ConclusionThe Aqueous Garlic Extract (AGE) displayed potent anti-B. anthracis activity. It was better than that displayed by Azadirachta indica A. Juss. (Neem) and Mangifera indica L. while Curcuma longa L. (Turmeric) did not show any activity under the assay conditions used. Further work should be undertaken to explore the possible application of AGE in preventing anthrax incidences in endemic areas.



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]]></description>
<dc:creator>Kaur, R.</dc:creator>
<dc:creator>Tiwari, A.</dc:creator>
<dc:creator>Manish, M.</dc:creator>
<dc:creator>Maurya, I. K.</dc:creator>
<dc:creator>Bhatnagar, R.</dc:creator>
<dc:creator>Singh, S.</dc:creator>
<dc:date>2017-07-11</dc:date>
<dc:identifier>doi:10.1101/162214</dc:identifier>
<dc:title><![CDATA[Common Garlic (Allium sativum) has Potent Anti-Bacillus anthracis Activity among Various Commonly Used Spices and Herbs]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-07-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/195701v1?rss=1">
<title>
<![CDATA[
Ocular and uteroplacental pathology in macaque congenital Zika virus infection 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/195701v1?rss=1</link>
<description><![CDATA[
Congenital Zika virus (ZIKV) infection impacts fetal development and pregnancy outcomes. We infected a pregnant rhesus macaque with a Puerto Rican ZIKV isolate in the first trimester. The pregnancy was complicated by preterm premature rupture of membranes (PPROM) and fetal demise 49 days post infection (gestational day 95). Significant pathology at the maternal-fetal interface included acute chorioamnionitis, placental infarcts, and leukocytoclastic vasculitis of the myometrial radial arteries. ZIKV RNA was disseminated throughout the fetus tissues and maternal immune system at necropsy, as assessed by quantitative RT-PCR for viral RNA. Replicating ZIKV was identified in fetal tissues, maternal lymph node, and maternal spleen by fluorescent in situ hybridization for viral replication intermediates. Fetal ocular pathology included a choroidal coloboma, suspected anterior segment dysgenesis, and a dysplastic retina. This is the first report of ocular pathology and prolonged viral replication in both maternal and fetal tissues following congenital ZIKV infection in rhesus macaques. PPROM followed by fetal demise and severe pathology of the visual system have not been described in macaque congenital infection previously; further nonhuman primate studies are needed to determine if an increased risk for PPROM is associated with congenital Zika virus infection.nnAuthor summaryA ZIKV infection during pregnancy is associated with malformations in fetal development including, but not limited to, ocular and brain anomalies, such as microcephaly, and stillbirth. The development of an accurate pregnancy model to study the effects of ZIKV will provide insight into vertical transmission, ZIKV tissue distribution, and fetal injury and malformations. Non-human primates closely resemble human in terms of the reproductive system, immunity, placentation and pregnancy. Our study demonstrates that the rhesus macaque is a compelling model in which to study ZIKV during pregnancy due to similar outcomes between the human and rhesus macaque. These similarities include prolonged viremia, vertical transmission, adverse pregnancy outcomes and fetal pathology, including defects in the visual system.
]]></description>
<dc:creator>Mohr, E. L.</dc:creator>
<dc:creator>Block, L. N.</dc:creator>
<dc:creator>Newman, C. M.</dc:creator>
<dc:creator>Stewart, L. M.</dc:creator>
<dc:creator>Koenig, M.</dc:creator>
<dc:creator>Semler, M.</dc:creator>
<dc:creator>Breitbach, M. E.</dc:creator>
<dc:creator>Teixeira, L. B. C.</dc:creator>
<dc:creator>Zeng, X.</dc:creator>
<dc:creator>Weiler, A. M.</dc:creator>
<dc:creator>Barry, G. L.</dc:creator>
<dc:creator>Thoong, T. H.</dc:creator>
<dc:creator>Wiepz, G. J.</dc:creator>
<dc:creator>Dudley, D. M.</dc:creator>
<dc:creator>Simmons, H. A.</dc:creator>
<dc:creator>Mejia, A.</dc:creator>
<dc:creator>Morgan, T. K.</dc:creator>
<dc:creator>Salamat, M. S.</dc:creator>
<dc:creator>Kohn, S.</dc:creator>
<dc:creator>Antony, K. M.</dc:creator>
<dc:creator>Aliota, M. T.</dc:creator>
<dc:creator>Mohns, M. S.</dc:creator>
<dc:creator>Hayes, J. M.</dc:creator>
<dc:creator>Schultz-Darken, N.</dc:creator>
<dc:creator>Schotzko, M. L.</dc:creator>
<dc:creator>Peterson, E.</dc:creator>
<dc:creator>Capuano, S.</dc:creator>
<dc:creator>Osorio, J. E.</dc:creator>
<dc:creator>O'Connor, S. L.</dc:creator>
<dc:creator>Friedrich, T. C.</dc:creator>
<dc:creator>O'Connor, D. H.</dc:creator>
<dc:creator>Golos, T. G.</dc:creator>
<dc:date>2017-09-30</dc:date>
<dc:identifier>doi:10.1101/195701</dc:identifier>
<dc:title><![CDATA[Ocular and uteroplacental pathology in macaque congenital Zika virus infection]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-09-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/196634v1?rss=1">
<title>
<![CDATA[
Mapping Cortical Brain Asymmetry in 17,141 Healthy Individuals Worldwide via the ENIGMA Consortium 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/196634v1?rss=1</link>
<description><![CDATA[
Hemispheric asymmetry is a cardinal feature of human brain organization. Altered brain asymmetry has also been linked to some cognitive and neuropsychiatric disorders. Here the ENIGMA consortium presents the largest ever analysis of cerebral cortical asymmetry and its variability across individuals. Cortical thickness and surface area were assessed in MRI scans of 17,141 healthy individuals from 99 datasets worldwide. Results revealed widespread asymmetries at both hemispheric and regional levels, with a generally thicker cortex but smaller surface area in the left hemisphere relative to the right. Regionally, asymmetries of cortical thickness and/or surface area were found in the inferior frontal gyrus, transverse temporal gyrus, parahippocampal gyrus, and entorhinal cortex. These regions are involved in lateralized functions, including language and visuospatial processing. In addition to population-level asymmetries, variability in brain asymmetry was related to sex, age, and brain size (indexed by intracranial volume). Interestingly, we did not find significant associations between asymmetries and handedness. Finally, with two independent pedigree datasets (N = 1,443 and 1,113, respectively), we found several asymmetries showing modest but highly reliable heritability. The structural asymmetries identified, and their variabilities and heritability provide a reference resource for future studies on the genetic basis of brain asymmetry and altered laterality in cognitive, neurological, and psychiatric disorders.nnSignificance StatementLeft-right asymmetry is a key feature of the human brain's structure and function. It remains unclear which cortical regions are asymmetrical on average in the population, and how biological factors such as age, sex and genetic variation affect these asymmetries. Here we describe by far the largest ever study of cerebral cortical brain asymmetry, based on data from 17,141 participants. We found a global anterior-posterior 'torque' pattern in cortical thickness, together with various regional asymmetries at the population level, which have not been previously described, as well as effects of age, sex, and heritability estimates. From these data, we have created an on-line resource that will serve future studies of human brain anatomy in health and disease.
]]></description>
<dc:creator>Kong, X.-Z.</dc:creator>
<dc:creator>Mathias, S.</dc:creator>
<dc:creator>Guadalupe, T.</dc:creator>
<dc:creator>Abe, C.</dc:creator>
<dc:creator>Agartz, I.</dc:creator>
<dc:creator>Akudjedu, T. N.</dc:creator>
<dc:creator>Aleman, A.</dc:creator>
<dc:creator>Alhusaini, S.</dc:creator>
<dc:creator>Allen, N. B.</dc:creator>
<dc:creator>Ames, D.</dc:creator>
<dc:creator>Andreassen, O. A.</dc:creator>
<dc:creator>Armstrong, N. J.</dc:creator>
<dc:creator>Bergo, F.</dc:creator>
<dc:creator>Bastin, M. E.</dc:creator>
<dc:creator>Batalla, A.</dc:creator>
<dc:creator>Bauer, J.</dc:creator>
<dc:creator>Baune, B.</dc:creator>
<dc:creator>Baur, R.</dc:creator>
<dc:creator>Biederman, J.</dc:creator>
<dc:creator>Blaine, S. K.</dc:creator>
<dc:creator>Boedhoe, P.</dc:creator>
<dc:creator>Boen, E.</dc:creator>
<dc:creator>Bose, A.</dc:creator>
<dc:creator>Bralten, J.</dc:creator>
<dc:creator>Brandeis, D.</dc:creator>
<dc:creator>Brem, S.</dc:creator>
<dc:creator>Brodaty, H.</dc:creator>
<dc:creator>Bröhl, H.</dc:creator>
<dc:creator>Buitelaar, J.</dc:creator>
<dc:creator>Bürger, C.</dc:creator>
<dc:creator>Bülow, R.</dc:creator>
<dc:creator>Calhoun, V.</dc:creator>
<dc:creator>Calvo, A.</dc:creator>
<dc:creator>Canales-Rodriguez, E. J.</dc:creator>
<dc:creator>Canive, J. M.</dc:creator>
<dc:creator>Cannon, D. M.</dc:creator>
<dc:creator>Caparelli, E. C.</dc:creator>
<dc:creator>Castellanos, F. X.</dc:creator>
<dc:creator>Cavalleri, G. L.</dc:creator>
<dc:creator>Cendes, F.</dc:creator>
<dc:creator>Chaim-Avancini, T. M.</dc:creator>
<dc:creator>Chant</dc:creator>
<dc:date>2017-10-01</dc:date>
<dc:identifier>doi:10.1101/196634</dc:identifier>
<dc:title><![CDATA[Mapping Cortical Brain Asymmetry in 17,141 Healthy Individuals Worldwide via the ENIGMA Consortium]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-10-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.01.12.903708v1?rss=1">
<title>
<![CDATA[
Shotgun Proteomics of non-depleted and depleted hemolymph of human Schistosomiasis snail vector; Biomphalaria glabrata, revealed vital immune related proteins 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.01.12.903708v1?rss=1</link>
<description><![CDATA[
BackgroundBiomphalaria glabrata is an important host in the transmission of human schistosomiasis in the Caribbean and South America. There are serious setbacks in the current control measures, hence need to explore alternatives.

ObjectiveTo unveil Biomphalaria glabrata hemolymph proteome for possible detailed knowledge of its immunity in host-pathogen relationship.

MethodsIn this study we performed a shotgun proteomic and bioinformatic analyses of the non-depleted and depleted [0.5 and 0.75% Trifluoroacetic acid (TFA) depletion] hemolymph of B. glabrata (LE strain).

ResultsOur analysis was able to mine a total of 148 proteins from the hemolymph. The highest number of protein (148) was obtained from the 0.5% TFA-depleted sample, followed by 62 proteins from the 0.75% TFA-depleted sample and only 59 from non-depleted hemolymph.

ConclusionMost of these proteins are novel and some of them may perform an important role in the immunity of this snail against invading pathogens.
]]></description>
<dc:creator>Otarigho, B.</dc:creator>
<dc:date>2020-01-13</dc:date>
<dc:identifier>doi:10.1101/2020.01.12.903708</dc:identifier>
<dc:title><![CDATA[Shotgun Proteomics of non-depleted and depleted hemolymph of human Schistosomiasis snail vector; Biomphalaria glabrata, revealed vital immune related proteins]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-01-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.04.29.065136v1?rss=1">
<title>
<![CDATA[
Specificity of Gβ and γ subunits to SNARE complex both at rest and after α2aadrenergic receptor stimulation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.04.29.065136v1?rss=1</link>
<description><![CDATA[
Though much is known about the various physiological functions of each GPCR and the specificity of G subunits, the specificity of G{beta}{gamma} activated by a given GPCR and activating each effector in vivo is not known. Previously, we identified different G{beta} and G{gamma} subunits interacting specifically with 2a-adrenergic receptors (2aAR). In this study, we examined its in vivo specificity to the soluble NSF attachment proteins (SNARE) complex in adrenergic (auto-2aAR) and non-adrenergic (hetero-2aAR) neurons. We applied a quantitative targeted multiple reaction monitoring proteomic analysis of G{beta} and G{gamma} subunits bound to the SNARE complex, and found only a subset of G{beta} and G{gamma} bound. Without stimulation of auto-2aAR, G{beta}1 and G{gamma}3 interacted with the SNARE complex. When auto-2aAR were activated, G{beta}1, G{beta}2, and G{gamma}3 were found. Further understanding of in vivo G{beta}{gamma} specificity to its effectors provides new insights into the multiplicity of genes for G{beta} and G{gamma}.

SummarySpecific G{beta}{gamma} dimers interact with the SNARE complex following presynaptic 2aAR activation in both adrenergic and non-adrenergic neurons.
]]></description>
<dc:creator>Yim, Y. Y.</dc:creator>
<dc:creator>McDonald, W. H.</dc:creator>
<dc:creator>Betke, K. M.</dc:creator>
<dc:creator>Kaya, A.</dc:creator>
<dc:creator>Hyde, K.</dc:creator>
<dc:creator>Gilsbach, R.</dc:creator>
<dc:creator>Hein, L.</dc:creator>
<dc:creator>Hamm, H. E.</dc:creator>
<dc:date>2020-05-01</dc:date>
<dc:identifier>doi:10.1101/2020.04.29.065136</dc:identifier>
<dc:title><![CDATA[Specificity of Gβ and γ subunits to SNARE complex both at rest and after α2aadrenergic receptor stimulation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-05-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.05.02.074575v1?rss=1">
<title>
<![CDATA[
Autoantibody landscape of advanced prostate cancer 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.05.02.074575v1?rss=1</link>
<description><![CDATA[
Although the importance of T-cell immune responses is well appreciated in cancer, autoantibody responses are less well-characterized. Nevertheless, autoantibody responses are of great interest, as they may be concordant with T-cell responses to cancer antigens or predictive of response to cancer immunotherapies. We performed serum epitope repertoire analysis (SERA) on a total of 1,229 serum samples obtained from a cohort of 72 men with metastatic castration-resistant prostate cancer (mCRPC) and 1,157 healthy control patients to characterize the autoantibody landscape of mCRPC. Using whole-genome sequencing results from paired solid-tumor metastasis biopsies and germline specimens, we identified tumor-specific epitopes in 29 mutant and 11 non-mutant proteins. Autoantibody enrichments for the top candidate autoantigen (NY-ESO-1) were validated using ELISA performed on the prostate cancer cohort and an independent cohort of 106 patients with melanoma. Our study recovers antigens of known importance and identifies novel tumor-specific epitopes of translational interest in advanced prostate cancer.

Statement of significanceAutoantibodies have been shown to inform treatment response and candidate drug targets in various cancers. We present the first large-scale profiling of autoantibodies in advanced prostate cancer, utilizing a new next-generation sequencing-based approach to antibody profiling to reveal novel cancer-specific antigens and epitopes.

Disclosure of Potential Conflicts of InterestJJA reports receiving consulting income from Janssen Biotech and Merck and honoraria from Astellas for speakers fees. MR reports receiving commercial research support from Novartis, Johnson & Johnson, Merck, Astellas, and Medivation, and is a consultant/advisory board member for Constellation Pharmaceuticals, Amgen, Ambrx, Johnson & Johnson, and Bayer. A.R. has received honoraria from consulting with Amgen, Bristol-Myers Squibb, Chugai, Dynavax, Genentech, Merck, Nektar, Novartis, Roche and Sanofi, is or has been a member of the scientific advisory board and holds stock in Advaxis, Arcus Biosciences, Bioncotech Therapeutics, Compugen, CytomX, Five Prime, RAPT, ImaginAb, Isoplexis, Kite-Gilead, Lutris Pharma, Merus, PACT Pharma, Rgenix and Tango Therapeutics. FYF serves on the advisory board for Dendreon, EMD Serono, Janssen Oncology, Ferring, Sanofi, Blue Earth Diagnostics, Celgene, consults for Bayer, Medivation/Astellas, Genetech, and Nutcracker Therapeutics, has honoraria from Clovis Oncology, and is a founder and has an ownership stake in PFS Genomics. SGZ and FYF have patent applications with Decipher Biosciences. SGZ and FYF have a patent application licensed to PFS Genomics. SGZ and FYF have patent applications with Celgene. WAH, RW, KK, PSD, and JCS have ownership of stocks or shares at Serimmune, paid employment at Serimmune, board membership at Serimmune, and patent applications on behalf of Serimmune.
]]></description>
<dc:creator>Chen, W. S.</dc:creator>
<dc:creator>Haynes, W. A.</dc:creator>
<dc:creator>Waitz, R.</dc:creator>
<dc:creator>Kamath, K.</dc:creator>
<dc:creator>Vega-Crespo, A.</dc:creator>
<dc:creator>Shrestha, R.</dc:creator>
<dc:creator>Foye, A.</dc:creator>
<dc:creator>Baselga Carretero, I.</dc:creator>
<dc:creator>Zhang, M.</dc:creator>
<dc:creator>Zhao, S. G.</dc:creator>
<dc:creator>Sjostrom, M.</dc:creator>
<dc:creator>Quigley, D. A.</dc:creator>
<dc:creator>Chou, J.</dc:creator>
<dc:creator>Beer, T.</dc:creator>
<dc:creator>Rettig, M.</dc:creator>
<dc:creator>Gleave, M.</dc:creator>
<dc:creator>Evans, C.</dc:creator>
<dc:creator>Lara, P.</dc:creator>
<dc:creator>Chi, K. N.</dc:creator>
<dc:creator>Reiter, R. E.</dc:creator>
<dc:creator>Alumkal, J. J.</dc:creator>
<dc:creator>Aggarwal, R.</dc:creator>
<dc:creator>Small, E. J.</dc:creator>
<dc:creator>Daugherty, P. S.</dc:creator>
<dc:creator>Ribas, A.</dc:creator>
<dc:creator>Oh, D. Y.</dc:creator>
<dc:creator>Shon, J. C.</dc:creator>
<dc:creator>Feng, F. Y.</dc:creator>
<dc:date>2020-05-03</dc:date>
<dc:identifier>doi:10.1101/2020.05.02.074575</dc:identifier>
<dc:title><![CDATA[Autoantibody landscape of advanced prostate cancer]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-05-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.05.17.100982v1?rss=1">
<title>
<![CDATA[
Rod signals are routed through specific Off cone bipolar cells in primate retina 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.05.17.100982v1?rss=1</link>
<description><![CDATA[
Adapting between scotopic and photopic illumination involves switching the routing of retinal signals between rod and cone-dominated circuits. In the daytime, cone signals pass through parallel On and Off cone bipolar cells, that are sensitive to increments and decrements in luminance, respectively. At night, rod signals are routed into these cone-pathways via a key glycinergic interneuron, the AII amacrine cell (AII-AC). In primates, it is not known whether AII-ACs contact all Off-bipolar cell types indiscriminately, or whether their outputs are biased towards specific Off-bipolar cell types. Here, we show that the rod-driven glycinergic output of AII-ACs is strongly biased towards a subset of macaque Off-cone bipolar cells. The Off-bipolar types that receive this glycinergic input have sustained physiological properties and include the Off-midget bipolar cells, which provide excitatory input to the Off-midget ganglion cells (parvocellular pathway). The kinetics of the glycinergic events are consistent with the involvement of the 1 glycine receptor subunit. Taken together with results in mouse retina, our findings point towards a conserved motif whereby rod signals are preferentially routed into sustained Off signaling pathways.

Significance StatementVisual signals pass through different retinal neurons depending on the prevailing level of illumination. Under night-time light levels, signals from rods pass through the AII amacrine cell, an inhibitory interneuron that routes rod signals into On and Off bipolar cells to detect increments and decrements in light intensity, respectively. Here, we show in primate retina that the output of AII amacrine cells is strongly biased towards specific Off bipolar cell types, which suggests that rod signals reach the brain via specific neural channels. Our results further our understanding of how visual signals are routed through visual circuits during night-time vision.
]]></description>
<dc:creator>McLaughlin, A. J.</dc:creator>
<dc:creator>Percival, K. A.</dc:creator>
<dc:creator>Gayet-Primo, J.</dc:creator>
<dc:creator>Puthussery, T.</dc:creator>
<dc:date>2020-05-19</dc:date>
<dc:identifier>doi:10.1101/2020.05.17.100982</dc:identifier>
<dc:title><![CDATA[Rod signals are routed through specific Off cone bipolar cells in primate retina]]></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/10.1101/2020.06.19.162214v1?rss=1">
<title>
<![CDATA[
FREQUENT GENE CONVERSION IN HUMAN EMBRYOS INDUCED BY DOUBLE STRAND BREAKS 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.06.19.162214v1?rss=1</link>
<description><![CDATA[
Applications of genome editing ultimately depend on DNA repair triggered by targeted double-strand breaks (DSBs). However, repair mechanisms in human cells remain poorly understood and vary across different cell types. Here we report that DSBs selectively induced on a mutant allele in heterozygous human embryos are repaired by gene conversion using an intact wildtype homolog as a template in up to 40% of targeted embryos. We also show that targeting of homozygous loci facilitates an interplay of non-homologous end joining (NHEJ) and gene conversion and results in embryos which carry identical indel mutations on both loci. Additionally, conversion tracks may expand bidirectionally well beyond the target region leading to an extensive loss of heterozygosity (LOH). Our study demonstrates that gene conversion and NHEJ are two major DNA DSB repair mechanisms in preimplantation human embryos. While gene conversion could be applicable for gene correction, extensive LOH presents a serious safety concern.
]]></description>
<dc:creator>Mitalipov, S.</dc:creator>
<dc:date>2020-06-20</dc:date>
<dc:identifier>doi:10.1101/2020.06.19.162214</dc:identifier>
<dc:title><![CDATA[FREQUENT GENE CONVERSION IN HUMAN EMBRYOS INDUCED BY DOUBLE STRAND BREAKS]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-06-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.07.15.204073v1?rss=1">
<title>
<![CDATA[
The ITS region provides a reliable DNA barcode for identifying reishi/lingzhi (Ganoderma) from herbal supplements 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.07.15.204073v1?rss=1</link>
<description><![CDATA[
The dietary supplement industry is a growing enterprise, valued at over $100 billion by 2025 yet, a recent study revealed that up to 60% of herbal supplements may have substituted ingredients not listed on their labels, some with harmful contaminants. Substituted ingredients make rigorous quality control testing a necessary aspect in the production of supplements. Traditionally, species have been verified morphologically or biochemically, but this is not possible for all species if the identifying characteristics are lost in the processing of the material. One approach to validating plant and fungal ingredients in herbal supplements is through DNA barcoding complemented with a molecular phylogenetic analysis. This method provides an efficient, objective, rigorous and repeatable method for species identification. We employed a molecular phylogenetic analysis for species authentication of the commonly used fungal supplement, reishi (Ganoderma lingzhi), by amplifying and sequencing the nuclear ribosomal internal transcribed spacer regions (ITS) with genus-specific primers. PCR of six powdered samples and one dried sample sold as G. lucidum representing independent suppliers produced single, strong amplification products in the expected size-range for Ganoderma. Both best-hit BLAST and molecular phylogenetic analyses using a reference panel assembled from Genbank clearly identified the predominant fungal DNA was G. lingzhi in all seven herbal supplements. We detected variation in ITS among our samples, but all samples still fall within a large clade of G. lingzhi. ITS is a successful and cost-effective method for DNA-based species authentication that could be used in the herbal supplement industry for this and other fungal and plant species that are otherwise difficult to identify.
]]></description>
<dc:creator>Whittall, J. B.</dc:creator>
<dc:creator>McFerrin, J.</dc:creator>
<dc:creator>Creswell, M.</dc:creator>
<dc:creator>Gunnels, T.</dc:creator>
<dc:date>2020-07-15</dc:date>
<dc:identifier>doi:10.1101/2020.07.15.204073</dc:identifier>
<dc:title><![CDATA[The ITS region provides a reliable DNA barcode for identifying reishi/lingzhi (Ganoderma) from herbal supplements]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-07-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.09.25.313098v1?rss=1">
<title>
<![CDATA[
Rcn3 is Involved in Postnatal Tendon Development by Regulating Collagen Modification and Fibrillogenesis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.09.25.313098v1?rss=1</link>
<description><![CDATA[
Tendon plays a critical role in the joint movement by transmitting force from muscle to bone. This transmission of force is facilitated by its specialized structure, which consists of highly aligned extracellular matrix consisting predominantly of type I collagen. Tenocytes, fibroblast-like tendon cells residing between the parallel collagen fibers, regulate this specialized tendon matrix. Despite the importance of collagen structure and tenocyte function, the biological mechanisms regulating fibrillogenesis and tenocyte maturation are not well understood. Here we examine the function of Reticulocalbin 3 (Rcn3) in collagen fibrillogenesis and tenocyte maturation during postnatal tendon development using a genetic mouse model. Loss of Rcn3 in tendon caused decreased tendon thickness, abnormal tendon cell maturation, and decreased mechanical properties. Interestingly, Rcn3 deficient mice exhibited a smaller collagen fibril distribution and over-hydroxylation in C-telopeptide cross-linking lysine from 1(1) chain. Additionally, the proline 3-hydroxylation sites in type I collagen were also over-hydroxylated in Rcn3 deficient mice. Our data collectively suggest that Rcn3 is a pivotal regulator of collagen fibrillogenesis and tenocyte maturation during postnatal tendon development.
]]></description>
<dc:creator>Park, N. R.</dc:creator>
<dc:creator>Shetye, S.</dc:creator>
<dc:creator>Keene, D. R.</dc:creator>
<dc:creator>Tufa, S.</dc:creator>
<dc:creator>Hudson, D. M.</dc:creator>
<dc:creator>Archer, M.</dc:creator>
<dc:creator>Soslowsky, L. J.</dc:creator>
<dc:creator>Dyment, N. A.</dc:creator>
<dc:creator>Joeng, K. S.</dc:creator>
<dc:date>2020-09-25</dc:date>
<dc:identifier>doi:10.1101/2020.09.25.313098</dc:identifier>
<dc:title><![CDATA[Rcn3 is Involved in Postnatal Tendon Development by Regulating Collagen Modification and Fibrillogenesis]]></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/10.1101/2020.10.26.356014v1?rss=1">
<title>
<![CDATA[
COVID-19 Disease Map, a computational knowledge repository of SARS-CoV-2 virus-host interaction mechanisms 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.10.26.356014v1?rss=1</link>
<description><![CDATA[
We describe a large-scale community effort to build an open-access, interoperable, and computable repository of COVID-19 molecular mechanisms - the COVID-19 Disease Map. We discuss the tools, platforms, and guidelines necessary for the distributed development of its contents by a multi-faceted community of biocurators, domain experts, bioinformaticians, and computational biologists. We highlight the role of relevant databases and text mining approaches in enrichment and validation of the curated mechanisms. We describe the contents of the Map and their relevance to the molecular pathophysiology of COVID-19 and the analytical and computational modelling approaches that can be applied for mechanistic data interpretation and predictions. We conclude by demonstrating concrete applications of our work through several use cases and highlight new testable hypotheses.
]]></description>
<dc:creator>Ostaszewski, M.</dc:creator>
<dc:creator>Niarakis, A.</dc:creator>
<dc:creator>Mazein, A.</dc:creator>
<dc:creator>Kuperstein, I.</dc:creator>
<dc:creator>Phair, R.</dc:creator>
<dc:creator>Orta-Resendiz, A.</dc:creator>
<dc:creator>Singh, V.</dc:creator>
<dc:creator>Aghamiri, S. S.</dc:creator>
<dc:creator>Acencio, M. L.</dc:creator>
<dc:creator>Glaab, E.</dc:creator>
<dc:creator>Ruepp, A.</dc:creator>
<dc:creator>Fobo, G.</dc:creator>
<dc:creator>Montrone, C.</dc:creator>
<dc:creator>Brauner, B.</dc:creator>
<dc:creator>Frishman, G.</dc:creator>
<dc:creator>Monraz Gomez, L. C.</dc:creator>
<dc:creator>Somers, J.</dc:creator>
<dc:creator>Hoch, M.</dc:creator>
<dc:creator>Gupta, S. K.</dc:creator>
<dc:creator>Scheel, J.</dc:creator>
<dc:creator>Borlinghaus, H.</dc:creator>
<dc:creator>Czauderna, T.</dc:creator>
<dc:creator>Schreiber, F.</dc:creator>
<dc:creator>Montagud, A.</dc:creator>
<dc:creator>Ponce de Leon, M.</dc:creator>
<dc:creator>Funahashi, A.</dc:creator>
<dc:creator>Hiki, Y.</dc:creator>
<dc:creator>Hiroi, N.</dc:creator>
<dc:creator>Yamada, T. G.</dc:creator>
<dc:creator>Drager, A.</dc:creator>
<dc:creator>Renz, A.</dc:creator>
<dc:creator>Naveez, M.</dc:creator>
<dc:creator>Bocskei, Z.</dc:creator>
<dc:creator>Messina, F.</dc:creator>
<dc:creator>Bornigen, D.</dc:creator>
<dc:creator>Fergusson, L.</dc:creator>
<dc:creator>Conti, M.</dc:creator>
<dc:creator>Rameil, M.</dc:creator>
<dc:creator>Nakonecnij, V.</dc:creator>
<dc:creator>Vanhoefer, J.</dc:creator>
<dc:creator>Schmiester, L.</dc:creator>
<dc:creator>Wang, M.</dc:creator>
<dc:creator>Ackerman, E. E.</dc:creator>
<dc:creator>Shoemake</dc:creator>
<dc:date>2020-10-28</dc:date>
<dc:identifier>doi:10.1101/2020.10.26.356014</dc:identifier>
<dc:title><![CDATA[COVID-19 Disease Map, a computational knowledge repository of SARS-CoV-2 virus-host interaction mechanisms]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-10-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.11.10.375279v1?rss=1">
<title>
<![CDATA[
Benchmarking DNA isolation kits used in analyses of the urinary microbiome 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.11.10.375279v1?rss=1</link>
<description><![CDATA[
The urinary microbiome has been increasingly characterized using next-generation sequencing. However, many of the technical methods have not yet been specifically optimized for urine. We sought to compare the performance of several DNA isolation kits used in urinary microbiome studies. A total of 11 voided urine samples and one buffer control were divided into 5 equal aliquots and processed in parallel using five commercial DNA isolation kits. DNA was quantified and the V4 segment of the 16S rRNA gene was sequenced. Data were processed to identify the microbial composition and to assess alpha and beta diversity of the samples. Tested DNA isolation kits result in significantly different DNA yields from urine samples but non-significant differences in the number of reads recovered, alpha, or beta diversity. DNA extracted with the Qiagen Biostic Bacteremia and DNeasy Blood & Tissue kits showed the fewest technical issues in downstream analyses, with the DNeasy Blood & Tissue kit also demonstrating the highest DNA yield. The Promega kit recovered fewer Gram positive bacteria compared to other kits. The Promega and DNeasy PowerSoil kits also appear to have some important biases towards over-representing certain Gram negative bacteria of biologic relevance within the urinary microbiome.
]]></description>
<dc:creator>Karstens, L.</dc:creator>
<dc:creator>Siddiqui, N. Y.</dc:creator>
<dc:creator>Zaza, T.</dc:creator>
<dc:creator>Barstad, A.</dc:creator>
<dc:creator>Amundsen, C. L.</dc:creator>
<dc:creator>Sysoeva, T. A.</dc:creator>
<dc:date>2020-11-10</dc:date>
<dc:identifier>doi:10.1101/2020.11.10.375279</dc:identifier>
<dc:title><![CDATA[Benchmarking DNA isolation kits used in analyses of the urinary microbiome]]></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/10.1101/2020.11.18.388876v1?rss=1">
<title>
<![CDATA[
Cas12a-Capture: a novel, low-cost, and scalable method for targeted sequencing 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.11.18.388876v1?rss=1</link>
<description><![CDATA[
Targeted sequencing remains a valuable technique for clinical and research applications. However, many existing technologies suffer from pervasive GC sequence content bias, high input DNA requirements, and high cost for custom panels. We have developed Cas12a-Capture, a low-cost and highly scalable method for targeted sequencing. The method utilizes preprogramed guide RNAs to direct CRISPR-Cas12a cleavage of double stranded DNA in vitro and then takes advantage of the resulting four to five nucleotide overhangs for selective ligation with a custom sequencing adapter. Addition of a second sequencing adapter and enrichment for ligation products generates a targeted sequence library. We first performed a pilot experiment with 7,176 guides targeting 3.5 megabases of DNA. Using these data, we modeled the sequence determinants of Cas12a-Capture efficiency, then designed an optimized set of 11,438 guides targeting 3.0 megabases. The optimized guide set achieves an average 64-fold enrichment of targeted regions with minimal GC bias. Cas12a-Capture variant calls had strong concordance with Illumina Platinum Genome calls, especially for SNVs, which could be improved by applying basic variant quality heuristics. We believe Cas12a-Capture has a wide variety of potential clinical and research applications and is amendable for selective enrichment for any double stranded DNA template or genome.
]]></description>
<dc:creator>Mighell, T. L.</dc:creator>
<dc:creator>Nishida, A.</dc:creator>
<dc:creator>O'Connell, B. L.</dc:creator>
<dc:creator>Miller, C. V.</dc:creator>
<dc:creator>Grindstaff, S.</dc:creator>
<dc:creator>Thornton, C. A.</dc:creator>
<dc:creator>Adey, A. C.</dc:creator>
<dc:creator>Doherty, D.</dc:creator>
<dc:creator>O'Roak, B. J.</dc:creator>
<dc:date>2020-11-20</dc:date>
<dc:identifier>doi:10.1101/2020.11.18.388876</dc:identifier>
<dc:title><![CDATA[Cas12a-Capture: a novel, low-cost, and scalable method for targeted sequencing]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-11-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2020.12.20.423638v1?rss=1">
<title>
<![CDATA[
Thyroid hormone and thyromimetics inhibit myelin and axonal degeneration and oligodendrocyte loss in EAE 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2020.12.20.423638v1?rss=1</link>
<description><![CDATA[
We have previously demonstrated that thyromimetics stimulate oligodendrocyte precursor cell differentiation and promote remyelination in murine demyelination models. We investigated whether a thyroid receptor-beta selective thyromimetic, sobetirome (Sob), and its CNS-targeted prodrug, Sob-AM2, could prevent myelin and axonal degeneration in experimental autoimmune encephalomyelitis (EAE). Compared to controls, EAE mice receiving triiodothyronine (T3, 0.4mg/kg), Sob (5mg/kg) or Sob-AM2 (5mg/kg) had reduced clinical disease and, within the spinal cord, less tissue damage, more normally myelinated axons, fewer degenerating axons and more oligodendrocytes. T3 and Sob also protected cultured oligodendrocytes against cell death. Thyromimetics thus might protect against oligodendrocyte death, demyelination and axonal degeneration as well as stimulate remyelination in multiple sclerosis.

HighlightsO_LIThyroid hormone, the thyromimetic Sob and its CNS penetrating prodrug, Sob-AM2, reduce disease severity, reduce myelin and axonal degeneration and protect oligodendrocytes in EAE.
C_LIO_LIThe benefits of Sob and Sob-AM2 may be via direct protective effects on oligodendrocytes and reduction in activity of microglia/macrophages.
C_LI
]]></description>
<dc:creator>Chaudhary, P.</dc:creator>
<dc:creator>Marracci, G. H.</dc:creator>
<dc:creator>Calkins, E.</dc:creator>
<dc:creator>Pocius, E.</dc:creator>
<dc:creator>Bensen, A. L.</dc:creator>
<dc:creator>Scanlan, T. S.</dc:creator>
<dc:creator>Emery, B.</dc:creator>
<dc:creator>Bourdette, D. N.</dc:creator>
<dc:date>2020-12-21</dc:date>
<dc:identifier>doi:10.1101/2020.12.20.423638</dc:identifier>
<dc:title><![CDATA[Thyroid hormone and thyromimetics inhibit myelin and axonal degeneration and oligodendrocyte loss in EAE]]></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/10.1101/2021.05.14.444205v1?rss=1">
<title>
<![CDATA[
Common Mechanism of SARS-CoV and SARS-CoV-2 Pathogenesis across Species 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.05.14.444205v1?rss=1</link>
<description><![CDATA[
Sarbecovirus (CoV) infections, including Severe Acute Respiratory CoV (SARS-CoV) and SARS-CoV-2, are considerable human threats. Human GWAS studies have recently identified loci associated with variation in SARS-CoV-2 susceptibility. However, genetically tractable models that reproduce human CoV disease outcomes are needed to mechanistically evaluate genetic determinants of CoV susceptibility. We used the Collaborative Cross (CC) and human GWAS datasets to elucidate host susceptibility loci that regulate CoV infections and to identify host quantitative trait loci that modulate severe CoV and pan-CoV disease outcomes including a major disease regulating loci including CCR9. CCR9 ablation resulted in enhanced titer, weight loss, respiratory dysfunction, mortality, and inflammation, providing mechanistic support in mitigating protection from severe SARS-CoV-2 pathogenesis across species. This study represents a comprehensive analysis of susceptibility loci for an entire genus of human pathogens conducted, identifies a large collection of susceptibility loci and candidate genes that regulate multiple aspects type-specific and cross-CoV pathogenesis, and also validates the paradigm of using the CC platform to identify common cross-species susceptibility loci and genes for newly emerging and pre-epidemic viruses.
]]></description>
<dc:creator>Schafer, A.</dc:creator>
<dc:creator>Gralinski, L. E.</dc:creator>
<dc:creator>Leist, S. R.</dc:creator>
<dc:creator>Winkler, E. S.</dc:creator>
<dc:creator>Hampton, B. K.</dc:creator>
<dc:creator>Mooney, M. A.</dc:creator>
<dc:creator>Jensen, K. L.</dc:creator>
<dc:creator>Graham, R. L.</dc:creator>
<dc:creator>Agnihothram, S.</dc:creator>
<dc:creator>Jeng, S.</dc:creator>
<dc:creator>Chamberlin, S.</dc:creator>
<dc:creator>Bell, T. A.</dc:creator>
<dc:creator>Scobey, D. T.</dc:creator>
<dc:creator>VanBlargan, L. A.</dc:creator>
<dc:creator>Thackray, L. B.</dc:creator>
<dc:creator>Hock, P.</dc:creator>
<dc:creator>Miller, D. R.</dc:creator>
<dc:creator>Shaw, G. D.</dc:creator>
<dc:creator>Pardo Manuel de Villena, F.</dc:creator>
<dc:creator>McWeeney, S. K.</dc:creator>
<dc:creator>Montgomery, S. A.</dc:creator>
<dc:creator>Diamond, M. S.</dc:creator>
<dc:creator>Heise, M. T.</dc:creator>
<dc:creator>Menachery, V. D.</dc:creator>
<dc:creator>Ferris, M. T.</dc:creator>
<dc:creator>Baric, R. S.</dc:creator>
<dc:date>2021-05-14</dc:date>
<dc:identifier>doi:10.1101/2021.05.14.444205</dc:identifier>
<dc:title><![CDATA[Common Mechanism of SARS-CoV and SARS-CoV-2 Pathogenesis across Species]]></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/10.1101/2021.05.17.444458v1?rss=1">
<title>
<![CDATA[
Alcohol Use Disrupts Age-Appropriate Cortical Thinning in Adolescence: A Data Driven Approach 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.05.17.444458v1?rss=1</link>
<description><![CDATA[
ObjectiveCortical thickness changes dramatically during development and is influenced by adolescent drinking. However, previous findings have been inconsistent and limited by region-of-interest approaches that are underpowered because they do not conform to the underlying heterogeneity from the effects of alcohol.

MethodsAdolescents (n=657; 12-22 years at baseline) from the National Consortium on Alcohol and Neurodevelopment in Adolescence (NCANDA) who endorsed little to no alcohol use at baseline were assessed with structural MRI and followed longitudinally at four yearly intervals. Seven unique spatially covarying patterns of cortical thickness were obtained from the baseline scans by applying a novel data-driven method called non-negative matrix factorization (NMF). The cortical thickness maps of all participants longitudinal scans were projected onto vertex-level cortical patterns to obtain participant-specific coefficients for each pattern. Linear mixed-effects models were fit to each pattern to investigate longitudinal effects of alcohol consumption on cortical thickness.

ResultsIn most NMF-derived cortical thickness patterns, the longitudinal rate of decline in no/low drinkers was similar for all age cohorts, among moderate drinkers the decline was faster in the younger cohort and slower in the older cohort, among heavy drinkers the decline was fastest in the younger cohort and slowest in the older cohort (FDR corrected p-values < 0.01).

ConclusionsThe NMF method can delineate spatially coordinated patterns of cortical thickness at the vertex level that are unconstrained by anatomical features. Age-appropriate cortical thinning is more rapid in younger adolescent drinkers and slower in older adolescent drinkers.
]]></description>
<dc:creator>Sun, D.</dc:creator>
<dc:creator>Adduru, V. R.</dc:creator>
<dc:creator>Phillips, R. D.</dc:creator>
<dc:creator>Bouchard, H. C.</dc:creator>
<dc:creator>Sotiras, A.</dc:creator>
<dc:creator>Michael, A. M.</dc:creator>
<dc:creator>Baker, F. C.</dc:creator>
<dc:creator>Tapert, S. F.</dc:creator>
<dc:creator>Brown, S. A.</dc:creator>
<dc:creator>Clark, D. B.</dc:creator>
<dc:creator>Goldston, D.</dc:creator>
<dc:creator>Nooner, K. B.</dc:creator>
<dc:creator>Nagel, B. J.</dc:creator>
<dc:creator>Thompson, W. K.</dc:creator>
<dc:creator>De Bellis, M.</dc:creator>
<dc:creator>Morey, R. A.</dc:creator>
<dc:date>2021-05-17</dc:date>
<dc:identifier>doi:10.1101/2021.05.17.444458</dc:identifier>
<dc:title><![CDATA[Alcohol Use Disrupts Age-Appropriate Cortical Thinning in Adolescence: A Data Driven Approach]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-05-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.05.21.445138v1?rss=1">
<title>
<![CDATA[
recount3: summaries and queries for large-scale RNA-seq expression and splicing 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.05.21.445138v1?rss=1</link>
<description><![CDATA[
We present recount3, a resource consisting of over 750,000 publicly available human and mouse RNA sequencing (RNA-seq) samples uniformly processed by our new Monorail analysis pipeline. To facilitate access to the data, we provide the recount3 and snapcount R/Bioconductor packages as well as complementary web resources. Using these tools, data can be downloaded as study-level summaries or queried for specific exon-exon junctions, genes, samples, or other features. Monorail can be used to process local and/or private data, allowing results to be directly compared to any study in recount3. Taken together, our tools help biologists maximize the utility of publicly available RNA-seq data, especially to improve their understanding of newly collected data. recount3 is available from http://rna.recount.bio.
]]></description>
<dc:creator>Wilks, C.</dc:creator>
<dc:creator>Zheng, S. C.</dc:creator>
<dc:creator>Chen, F. Y.</dc:creator>
<dc:creator>Charles, R.</dc:creator>
<dc:creator>Solomon, B.</dc:creator>
<dc:creator>Ling, J. P.</dc:creator>
<dc:creator>Imada, E. L.</dc:creator>
<dc:creator>Zhang, D.</dc:creator>
<dc:creator>Joseph, L.</dc:creator>
<dc:creator>Leek, J. T.</dc:creator>
<dc:creator>Jaffe, A. E.</dc:creator>
<dc:creator>Nellore, A.</dc:creator>
<dc:creator>Collado-Torres, L.</dc:creator>
<dc:creator>Hansen, K. D.</dc:creator>
<dc:creator>Langmead, B.</dc:creator>
<dc:date>2021-05-23</dc:date>
<dc:identifier>doi:10.1101/2021.05.21.445138</dc:identifier>
<dc:title><![CDATA[recount3: summaries and queries for large-scale RNA-seq expression and splicing]]></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/10.1101/2021.06.01.446587v1?rss=1">
<title>
<![CDATA[
OBO Foundry in 2021: Operationalizing Open Data Principles to Evaluate Ontologies 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.06.01.446587v1?rss=1</link>
<description><![CDATA[
Biological ontologies are used to organize, curate, and interpret the vast quantities of data arising from biological experiments. While this works well when using a single ontology, integrating multiple ontologies can be problematic, as they are developed independently, which can lead to incompatibilities. The Open Biological and Biomedical Ontologies (OBO) Foundry was created to address this by facilitating the development, harmonization, application, and sharing of ontologies, guided by a set of overarching principles. One challenge in reaching these goals was that the OBO principles were not originally encoded in a precise fashion, and interpretation was subjective. Here we show how we have addressed this by formally encoding the OBO principles as operational rules and implementing a suite of automated validation checks and a dashboard for objectively evaluating each ontologys compliance with each principle. This entailed a substantial effort to curate metadata across all ontologies and to coordinate with individual stakeholders. We have applied these checks across the full OBO suite of ontologies, revealing areas where individual ontologies require changes to conform to our principles. Our work demonstrates how a sizable federated community can be organized and evaluated on objective criteria that help improve overall quality and interoperability, which is vital for the sustenance of the OBO project and towards the overall goals of making data FAIR.
]]></description>
<dc:creator>Jackson, R. C.</dc:creator>
<dc:creator>Matentzoglu, N.</dc:creator>
<dc:creator>Overton, J. A.</dc:creator>
<dc:creator>Vita, R.</dc:creator>
<dc:creator>Balhoff, J. P.</dc:creator>
<dc:creator>Buttigieg, P. L.</dc:creator>
<dc:creator>Carbon, S.</dc:creator>
<dc:creator>Courtot, M.</dc:creator>
<dc:creator>Diehl, A. D.</dc:creator>
<dc:creator>Dooley, D.</dc:creator>
<dc:creator>Duncan, W.</dc:creator>
<dc:creator>Harris, N. L.</dc:creator>
<dc:creator>Haendel, M. A.</dc:creator>
<dc:creator>Lewis, S. E.</dc:creator>
<dc:creator>Natale, D. A.</dc:creator>
<dc:creator>Osumi-Sutherland, D.</dc:creator>
<dc:creator>Ruttenberg, A.</dc:creator>
<dc:creator>Schriml, L. M.</dc:creator>
<dc:creator>Smith, B.</dc:creator>
<dc:creator>Stoeckert, C. J.</dc:creator>
<dc:creator>Vasilevsky, N. A.</dc:creator>
<dc:creator>Walls, R. L.</dc:creator>
<dc:creator>Zheng, J.</dc:creator>
<dc:creator>Mungall, C. J.</dc:creator>
<dc:creator>Peters, B.</dc:creator>
<dc:date>2021-06-02</dc:date>
<dc:identifier>doi:10.1101/2021.06.01.446587</dc:identifier>
<dc:title><![CDATA[OBO Foundry in 2021: Operationalizing Open Data Principles to Evaluate Ontologies]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-06-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.06.04.447154v1?rss=1">
<title>
<![CDATA[
Utilizing proteomics and phosphoproteomics to predict ex vivo drug sensitivity across genetically diverse AML patients 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.06.04.447154v1?rss=1</link>
<description><![CDATA[
Acute Myeloid Leukemia (AML) affects 20,000 patients in the US annually with a five-year survival rate of approximately 25%. One reason for the low survival rate is the high prevalence of clonal evolution that gives rise to heterogeneous sub-populations of leukemia. This genetic heterogeneity is difficult to treat using conventional therapies that are generally based on the detection of a single driving mutation. Thus, the use of molecular signatures, consisting of multiple functionally related transcripts or proteins, in making treatment decisions may overcome this hurdle and provide a more effective way to inform drug treatment protocols. Toward this end, the Beat AML research program prospectively collected genomic and transcriptomic data from over 1000 AML patients and carried out ex vivo drug sensitivity assays to identify signatures that could predict patient-specific drug responses. The Clinical Proteomic Tumor Analysis Consortium is in the process of extending this cohort to collect proteomic and phosphoproteomic measurements from a subset of these patient samples to evaluate the hypothesis that proteomic signatures can robustly predict drug response in AML patients. We sought to examine this hypothesis on a sub-cohort of 38 patient samples from Beat AML with proteomic and drug response data and evaluate our ability to identify proteomic signatures that predict drug response with high accuracy. For this initial analysis we built predictive models of patient drug responses across 26 drugs of interest using the proteomics and phosphproteomics data. We found that proteomics-derived signatures provide an accurate and robust signature of drug response in the AML ex vivo samples, as well as related cell lines, with better performance than those signatures derived from mutations or mRNA expression. Furthermore, we found that in specific drug-resistant cell lines, the proteins in our prognostic signatures represented dysregulated signaling pathways compared to parental cell lines, confirming the role of the proteins in the signatures in drug resistance. In conclusion, this pilot study demonstrates strong promise for proteomics-based patient stratification to predict drug sensitivity in AML.
]]></description>
<dc:creator>Gosline, S. J.</dc:creator>
<dc:creator>Tognon, C.</dc:creator>
<dc:creator>Nestor, M.</dc:creator>
<dc:creator>Joshi, S.</dc:creator>
<dc:creator>Modak, R.</dc:creator>
<dc:creator>Damnernsawad, A.</dc:creator>
<dc:creator>Moon, J.</dc:creator>
<dc:creator>Hansen, J. R.</dc:creator>
<dc:creator>Hutchinson-Bunch, C.</dc:creator>
<dc:creator>Gritsenko, M. A.</dc:creator>
<dc:creator>Weitz, K. K.</dc:creator>
<dc:creator>Traer, E.</dc:creator>
<dc:creator>Drucker, B.</dc:creator>
<dc:creator>Agarwal, A.</dc:creator>
<dc:creator>Piehowski, P.</dc:creator>
<dc:creator>McDermott, J. E.</dc:creator>
<dc:creator>Rodland, K.</dc:creator>
<dc:date>2021-06-06</dc:date>
<dc:identifier>doi:10.1101/2021.06.04.447154</dc:identifier>
<dc:title><![CDATA[Utilizing proteomics and phosphoproteomics to predict ex vivo drug sensitivity across genetically diverse AML patients]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-06-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.06.08.447375v1?rss=1">
<title>
<![CDATA[
Pulsation changes link to impaired glymphatic function in a mouse model of vascular cognitive impairment 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.06.08.447375v1?rss=1</link>
<description><![CDATA[
Large vessel disease and carotid stenosis are key mechanisms contributing to vascular cognitive impairment (VCI) and dementia. Our previous work, and that of others, using rodent models, demonstrated that bilateral common carotid stenosis (BCAS) leads to cognitive impairment via gradual deterioration of the glial-vascular unit and accumulation of amyloid-{beta} (A{beta}) protein. Since brain-wide drainage pathways (glymphatic) for waste clearance, including A{beta} removal, have been implicated in the pathophysiology of VCI via glial mechanisms, we hypothesized that glymphatic function would be impaired in a BCAS model and exacerbated in the presence of A{beta}. Male wild- type and Tg-SwDI (model of microvascular amyloid) mice were subjected to BCAS or sham surgery which led to a reduction in cerebral perfusion and impaired spatial learning and memory. After 3 months survival, glymphatic function was evaluated by cerebrospinal fluid (CSF) fluorescent tracer influx. We demonstrated that BCAS caused a marked regional reduction of CSF tracer influx in the dorsolateral cortex and CA1-DG molecular layer. In parallel to these changes increased reactive astrogliosis was observed post-BCAS. To further investigate the mechanisms that may lead to these changes, we measured the pulsation of cortical vessels. BCAS impaired vascular pulsation in pial arteries in WT and Tg-SwDI mice. Since our findings show that BCAS may influence VCI by impaired glymphatic drainage and reduced vascular pulsation we propose that these additional targets need to be considered when treating VCI.
]]></description>
<dc:creator>Li, M.</dc:creator>
<dc:creator>Kitamura, A.</dc:creator>
<dc:creator>Beverley, J.</dc:creator>
<dc:creator>Koudelka, J.</dc:creator>
<dc:creator>Duncombe, J.</dc:creator>
<dc:creator>Platt, B.</dc:creator>
<dc:creator>Wiegand, U. K.</dc:creator>
<dc:creator>Carare, R. O.</dc:creator>
<dc:creator>Kalaria, R. N.</dc:creator>
<dc:creator>Iliff, J. J.</dc:creator>
<dc:creator>Horsburgh, K.</dc:creator>
<dc:date>2021-06-09</dc:date>
<dc:identifier>doi:10.1101/2021.06.08.447375</dc:identifier>
<dc:title><![CDATA[Pulsation changes link to impaired glymphatic function in a mouse model of vascular cognitive impairment]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-06-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.06.08.447553v1?rss=1">
<title>
<![CDATA[
Predictors of cognitive impairment in primary age-related tauopathy: an autopsy study 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.06.08.447553v1?rss=1</link>
<description><![CDATA[
Primary age-related tauopathy (PART) is a form of Alzheimer-type neurofibrillary degeneration occurring in the absence of amyloid-beta (A{beta}) plaques. While PART shares some features with Alzheimer disease (AD), such as progressive accumulation of neurofibrillary tangle pathology in the medial temporal lobe and other brain regions, it does not progress extensively to neocortical regions. Given this restricted pathoanatomical pattern and variable symptomatology, there is a need to reexamine and improve upon how PART is neuropathologically assessed and staged. We performed a retrospective autopsy study in a collection (n=174) of post-mortem PART brains and used logistic regression to determine the extent to which a set of clinical and neuropathological features predict cognitive impairment. We compared Braak staging, which focuses on hierarchical neuroanatomical progression of AD tau and A{beta} pathology, with quantitative assessments of neurofibrillary burden using computer-derived positive pixel counts on digitized whole slide images of sections stained immunohistochemically with antibodies targeting abnormal hyperphosphorylated tau (p-tau) in the entorhinal region and hippocampus. We also assessed other factors affecting cognition, including aging-related tau astrogliopathy (ARTAG) and atrophy. We found no association between Braak stage and cognitive impairment when controlling for age (p=0.76). In contrast, p-tau burden was significantly correlated with cognitive impairment even when adjusting for age (p=0.03). The strongest correlate of cognitive impairment was cerebrovascular disease, a well-known risk factor (p<0.0001), but other features including ARTAG (p=0.03) and hippocampal atrophy (p=0.04) were also associated. In contrast, sex, APOE, psychiatric illness, education, argyrophilic grains, and incidental Lewy bodies were not. These findings support the hypothesis that comorbid pathologies contribute to cognitive impairment in subjects with PART. Quantitative approaches beyond Braak staging are critical for advancing our understanding of the extent to which age-related tauopathy changes impact cognitive function.
]]></description>
<dc:creator>Iida, M. A.</dc:creator>
<dc:creator>Farrell, K.</dc:creator>
<dc:creator>Walker, J. M.</dc:creator>
<dc:creator>Richardson, T. E.</dc:creator>
<dc:creator>Marx, G.</dc:creator>
<dc:creator>Bryce, C. H.</dc:creator>
<dc:creator>Purohit, D.</dc:creator>
<dc:creator>Ayalon, G.</dc:creator>
<dc:creator>Beach, T. G.</dc:creator>
<dc:creator>Bigio, E. H.</dc:creator>
<dc:creator>Cortes, E.</dc:creator>
<dc:creator>Gearing, M.</dc:creator>
<dc:creator>Haroutunian, V.</dc:creator>
<dc:creator>McMillan, C. T.</dc:creator>
<dc:creator>Lee, E. B.</dc:creator>
<dc:creator>Dickson, D.</dc:creator>
<dc:creator>McKee, A. C.</dc:creator>
<dc:creator>Stein, T. D.</dc:creator>
<dc:creator>Trojanowski, J. Q.</dc:creator>
<dc:creator>Woltjer, R. L.</dc:creator>
<dc:creator>Kovacs, G. G.</dc:creator>
<dc:creator>Kofler, J. K.</dc:creator>
<dc:creator>Kaye, J.</dc:creator>
<dc:creator>White, C. L.</dc:creator>
<dc:creator>Crary, J. F.</dc:creator>
<dc:date>2021-06-09</dc:date>
<dc:identifier>doi:10.1101/2021.06.08.447553</dc:identifier>
<dc:title><![CDATA[Predictors of cognitive impairment in primary age-related tauopathy: an autopsy study]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-06-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.07.22.451363v1?rss=1">
<title>
<![CDATA[
A community-based approach to image analysis of cells, tissues and tumors 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.07.22.451363v1?rss=1</link>
<description><![CDATA[
Emerging multiplexed imaging platforms provide an unprecedented view of an increasing number of molecular markers at subcellular resolution and the dynamic evolution of tumor cellular composition. As such, they are capable of elucidating cell-to-cell interactions within the tumor microenvironment that impact clinical outcome and therapeutic response. However, the rapid development of these platforms has far outpaced the computational methods for processing and analyzing the data they generate. While being technologically disparate, all imaging assays share many computational requirements for post-collection data processing. We convened a workshop to characterize these shared computational challenges and a follow-up hackathon to implement solutions for a selected subset of them. Here, we delineate these areas that reflect major axes of research within the field, including image registration, segmentation of cells and subcellular structures, and identification of cell types from their morphology. We further describe the logistical organization of these events, believing our lessons learned can aid others in uniting the imaging community around self-identified topics of mutual interest, in designing and implementing operational procedures to address those topics and in mitigating issues inherent in image analysis (e.g., sharing exemplar images of large datasets and disseminating baseline solutions to hackathon challenges through open-source code repositories).
]]></description>
<dc:creator>CSBC/PS-ON Image Analysis Working Group,</dc:creator>
<dc:creator>Vizcarra, J. C.</dc:creator>
<dc:creator>Burlingame, E. A.</dc:creator>
<dc:creator>Hug, C. B.</dc:creator>
<dc:creator>Goltsev, Y.</dc:creator>
<dc:creator>White, B. S.</dc:creator>
<dc:creator>Tyson, D. R.</dc:creator>
<dc:creator>Sokolov, A.</dc:creator>
<dc:date>2021-07-25</dc:date>
<dc:identifier>doi:10.1101/2021.07.22.451363</dc:identifier>
<dc:title><![CDATA[A community-based approach to image analysis of cells, tissues and tumors]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-07-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.09.05.458975v1?rss=1">
<title>
<![CDATA[
Metformin in nucleus accumbens core reduces cue-induced cocaine seeking in male and female rats 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.09.05.458975v1?rss=1</link>
<description><![CDATA[
This study investigated the potential therapeutic effects of the FDA-approved drug metformin on cue-induced reinstatement of cocaine seeking. Metformin (dimethyl-biguanide) is a first-line treatment for type II diabetes that, among other mechanisms, is involved in the activation of adenosine monophosphate activated protein kinase (AMPK). Cocaine self-administration and extinction is associated with decreased levels of phosphorylated AMPK within the nucleus accumbens core (NAcore). Previously it was shown that increasing AMPK activity in the NAcore decreased cue-induced reinstatement of cocaine seeking. Decreasing AMPK activity produced the opposite effect. The goal of the present study was to determine if metformin in the NAcore reduces cue-induced cocaine seeking in adult male and female Sprague Dawley rats. Rats were trained to self-administer cocaine followed by extinction prior to cue-induced reinstatement trials. Metformin microinjected in the NAcore attenuated cue-induced reinstatement in male and female rats. Importantly, metformins effects on cocaine seeking were not due to a general depression of spontaneous locomotor activity. In female rats, metformins effects did generalize to a reduction in cue-induced reinstatement of sucrose seeking. These data support a potential role for metformin as a pharmacotherapy for cocaine use disorder, but warrant caution given the potential for metformins effects to generalize to a natural reward in female rats.
]]></description>
<dc:creator>Chan, A.</dc:creator>
<dc:creator>Willard, A.</dc:creator>
<dc:creator>Mulloy, S.</dc:creator>
<dc:creator>Ibrahim, N.</dc:creator>
<dc:creator>Sciaccotta, A.</dc:creator>
<dc:creator>Schonfeld, M.</dc:creator>
<dc:creator>Spencer, S.</dc:creator>
<dc:date>2021-09-06</dc:date>
<dc:identifier>doi:10.1101/2021.09.05.458975</dc:identifier>
<dc:title><![CDATA[Metformin in nucleus accumbens core reduces cue-induced cocaine seeking in male and female rats]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-09-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.09.08.459549v1?rss=1">
<title>
<![CDATA[
OpenMonkeyChallenge: Dataset and Benchmark Challenges for Pose Tracking of Non-human Primates 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.09.08.459549v1?rss=1</link>
<description><![CDATA[
The ability to automatically track non-human primates as they move through the world is important for several subfields in biology and biomedicine. Inspired by the recent success of computer vision models enabled by benchmark challenges (e.g., object detection), we propose a new benchmark challenge called OpenMonkeyChallenge that facilitates collective community efforts through an annual competition to build generalizable non- human primate pose tracking models. To host the benchmark challenge, we provide a new public dataset consisting of 111,529 annotated (17 body landmarks) photographs of non-human primates in naturalistic contexts obtained from various sources including the internet, three National Primate Research Centers, and the Minnesota Zoo. Such annotated datasets will be used for the training and testing datasets to develop generalizable models with standardized evaluation metrics. We demonstrate the effectiveness of our dataset quantitatively by comparing it with existing datasets based on seven state-of-the-art pose tracking models.
]]></description>
<dc:creator>Yao, Y.</dc:creator>
<dc:creator>Abhiraj Mohan, A.</dc:creator>
<dc:creator>Bliss-Moreau, E.</dc:creator>
<dc:creator>Coleman, K.</dc:creator>
<dc:creator>Freeman, S. M.</dc:creator>
<dc:creator>Machado, C. J.</dc:creator>
<dc:creator>Raper, J.</dc:creator>
<dc:creator>Zimmermann, J.</dc:creator>
<dc:creator>Hayden, B. Y.</dc:creator>
<dc:creator>Park, H. S.</dc:creator>
<dc:date>2021-09-10</dc:date>
<dc:identifier>doi:10.1101/2021.09.08.459549</dc:identifier>
<dc:title><![CDATA[OpenMonkeyChallenge: Dataset and Benchmark Challenges for Pose Tracking of Non-human Primates]]></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/10.1101/2021.11.12.468428v1?rss=1">
<title>
<![CDATA[
#COVIDisAirborne: AI-Enabled Multiscale Computational Microscopy of Delta SARS-CoV-2 in a Respiratory Aerosol 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.11.12.468428v1?rss=1</link>
<description><![CDATA[
We seek to completely revise current models of airborne transmission of respiratory viruses by providing never-before-seen atomic-level views of the SARS-CoV-2 virus within a respiratory aerosol. Our work dramatically extends the capabilities of multiscale computational microscopy to address the significant gaps that exist in current experimental methods, which are limited in their ability to interrogate aerosols at the atomic/molecular level and thus ob-scure our understanding of airborne transmission. We demonstrate how our integrated data-driven platform provides a new way of exploring the composition, structure, and dynamics of aerosols and aerosolized viruses, while driving simulation method development along several important axes. We present a series of initial scientific discoveries for the SARS-CoV-2 Delta variant, noting that the full scientific impact of this work has yet to be realized.

ACM Reference FormatAbigail Dommer1{dagger}, Lorenzo Casalino1{dagger}, Fiona Kearns1{dagger}, Mia Rosenfeld1, Nicholas Wauer1, Surl-Hee Ahn1, John Russo,2 Sofia Oliveira3, Clare Morris1, AnthonyBogetti4, AndaTrifan5,6, Alexander Brace5,7, TerraSztain1,8, Austin Clyde5,7, Heng Ma5, Chakra Chennubhotla4, Hyungro Lee9, Matteo Turilli9, Syma Khalid10, Teresa Tamayo-Mendoza11, Matthew Welborn11, Anders Christensen11, Daniel G. A. Smith11, Zhuoran Qiao12, Sai Krishna Sirumalla11, Michael OConnor11, Frederick Manby11, Anima Anandkumar12,13, David Hardy6, James Phillips6, Abraham Stern13, Josh Romero13, David Clark13, Mitchell Dorrell14, Tom Maiden14, Lei Huang15, John McCalpin15, Christo- pherWoods3, Alan Gray13, MattWilliams3, Bryan Barker16, HarindaRajapaksha16, Richard Pitts16, Tom Gibbs13, John Stone6, Daniel Zuckerman2*, Adrian Mulholland3*, Thomas MillerIII11,12*, ShantenuJha9*, Arvind Ramanathan5*, Lillian Chong4*, Rommie Amaro1*. 2021. #COVIDisAirborne: AI-Enabled Multiscale Computational Microscopy ofDeltaSARS-CoV-2 in a Respiratory Aerosol. In Supercomputing  21: International Conference for High Perfor-mance Computing, Networking, Storage, and Analysis. ACM, New York, NY, USA, 14 pages. https://doi.org/finalDOI
]]></description>
<dc:creator>Dommer, A.</dc:creator>
<dc:creator>Casalino, L.</dc:creator>
<dc:creator>Kearns, F.</dc:creator>
<dc:creator>Rosenfeld, M.</dc:creator>
<dc:creator>Wauer, N.</dc:creator>
<dc:creator>Ahn, S.-H.</dc:creator>
<dc:creator>Russo, J.</dc:creator>
<dc:creator>Oliveira, S.</dc:creator>
<dc:creator>Morris, C.</dc:creator>
<dc:creator>Bogetti, A.</dc:creator>
<dc:creator>Trifan, A.</dc:creator>
<dc:creator>Brace, A.</dc:creator>
<dc:creator>Sztain, T.</dc:creator>
<dc:creator>Clyde, A.</dc:creator>
<dc:creator>Ma, H.</dc:creator>
<dc:creator>Chennubhotla, C.</dc:creator>
<dc:creator>Lee, H.</dc:creator>
<dc:creator>Turilli, M.</dc:creator>
<dc:creator>Khalid, S.</dc:creator>
<dc:creator>Tamayo-Mendoza, T.</dc:creator>
<dc:creator>Welborn, M.</dc:creator>
<dc:creator>Christiansen, A.</dc:creator>
<dc:creator>Smith, D. G. A.</dc:creator>
<dc:creator>Qiao, Z.</dc:creator>
<dc:creator>Sirumalla, S. K.</dc:creator>
<dc:creator>O'Connor, M.</dc:creator>
<dc:creator>Manby, F.</dc:creator>
<dc:creator>Anandkumar, A.</dc:creator>
<dc:creator>Hardy, D.</dc:creator>
<dc:creator>Phillips, J.</dc:creator>
<dc:creator>Stern, A.</dc:creator>
<dc:creator>Romero, J.</dc:creator>
<dc:creator>Clark, D.</dc:creator>
<dc:creator>Dorrell, M.</dc:creator>
<dc:creator>Maiden, T.</dc:creator>
<dc:creator>Huang, L.</dc:creator>
<dc:creator>McCalpin, J.</dc:creator>
<dc:creator>Woods, C.</dc:creator>
<dc:creator>Gray, A.</dc:creator>
<dc:creator>Williams, M.</dc:creator>
<dc:creator>Barker, B.</dc:creator>
<dc:creator>Rajapaksha, H.</dc:creator>
<dc:creator>Pitts, R.</dc:creator>
<dc:creator>Gibbs, T.</dc:creator>
<dc:creator>Stone, J.</dc:creator>
<dc:creator>Zuckerman, D.</dc:creator>
<dc:creator>Muholland,</dc:creator>
<dc:date>2021-11-15</dc:date>
<dc:identifier>doi:10.1101/2021.11.12.468428</dc:identifier>
<dc:title><![CDATA[#COVIDisAirborne: AI-Enabled Multiscale Computational Microscopy of Delta SARS-CoV-2 in a Respiratory Aerosol]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-11-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2021.12.17.473201v1?rss=1">
<title>
<![CDATA[
Overlapping stimulons arising in response to divergent stresses in Escherichia coli 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2021.12.17.473201v1?rss=1</link>
<description><![CDATA[
Cellular responses to stress can cause a similar change in some facets of fitness even if the stresses are different. Lactose as a sole carbon source for Escherichia coli is an established example: too little causes starvation while excessive lactose import causes toxicity as a side-effect. In an E. coli strain that is robust to osmotic and ionic differences in growth media, B REL606, the rate of antibiotic-tolerant persister formation is elevated in both starvation-inducing and toxicity-inducing concentrations of lactose in comparison to less stressful intermediate concentrations. Such similarities between starvation and toxification raise the question of how much the global stress response stimulon differs between them. We hypothesized that a common stress response is conserved between the two conditions, but that a previously shown threshold driving growth rate heterogeneity in a lactose-toxifying medium would reveal that the growing fraction of cells in that medium to be missing key stress responses that curb growth. To test this, we performed RNA-seq in three representative conditions for differential expression analysis. In comparison to nominally unstressed cultures, both stress conditions showed global shifts in gene expression, with informative similarities and differences. Functional analysis of pathways, gene ontology terms, and clusters of orthogonal groups revealed signatures of overflow metabolism, membrane component shifts, and altered cytosolic and periplasmic contents in toxified cultures. Starving cultures showed an increased tendency toward stringent response-like regulatory signatures. Along with other emerging evidence, our results show multiple possible pathways to stress responses, persistence, and possibly other phenotypes. These results suggest a set of overlapping responses that drives emergence of stress-tolerant phenotypes in diverse conditions.
]]></description>
<dc:creator>Wang, H.</dc:creator>
<dc:creator>McElfresh, G.</dc:creator>
<dc:creator>Wijesuriya, N.</dc:creator>
<dc:creator>Podgorny, A.</dc:creator>
<dc:creator>Hecht, A. D.</dc:creator>
<dc:creator>Ray, J. C. J.</dc:creator>
<dc:date>2021-12-18</dc:date>
<dc:identifier>doi:10.1101/2021.12.17.473201</dc:identifier>
<dc:title><![CDATA[Overlapping stimulons arising in response to divergent stresses in Escherichia coli]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-12-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.02.08.479578v1?rss=1">
<title>
<![CDATA[
Functional, transcriptional, and microbial shifts associated with healthy pulmonary aging: insights from rhesus macaques 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.02.08.479578v1?rss=1</link>
<description><![CDATA[
Older individuals are at increased risk of developing severe respiratory infections due to age-related changes in the immunological, microbial, and functional landscape of the lung. However, our understanding of the impact of age on the respiratory tract remains limited as samples from healthy humans are challenging to obtain and confounding variables such as smoking and environmental pollutant exposure make it difficult to assess the true impact of aging. On the other hand, studies in rodent models are biased by their specific pathogen free status. In this study, we utilize a rhesus macaque model of healthy aging to examine the functional, immunological, and microbial consequences of aging in the lung. Pulmonary function testing in this large (n=34 adult, n=49 aged) cross-sectional study established age and sex differences similar to humans supporting the translational accuracy of this model. Additionally, an increased abundance of myeloid cells (alveolar and infiltrating macrophages) and a concomitant decrease in T-cells were also observed in aged animals. Single cell RNA sequencing indicated a transcriptional shift in the pulmonary CD8+ T-cell population from GRZMB expressing cells to IFN expressing cells, while frequency of IL-1B expressing alveolar macrophages was significantly reduced. Interestingly, the lung microbiome of many animals was dominated by a single microbe, Tropheryma spp., the prevalence of which decreased with age. These data provide a comprehensive picture of the functional, microbial and immunological changes of the lung in healthy macaque aging and provide insight into the increased prevalence and severity of respiratory disease in the elderly.
]]></description>
<dc:creator>Rhoades, N. S.</dc:creator>
<dc:creator>Davies, M.</dc:creator>
<dc:creator>Lewis, S. A.</dc:creator>
<dc:creator>Cinco, I. R.</dc:creator>
<dc:creator>Kohama, S. G.</dc:creator>
<dc:creator>Bermudez, L. E.</dc:creator>
<dc:creator>Winthorp, K. L.</dc:creator>
<dc:creator>Fuss, C.</dc:creator>
<dc:creator>Mattison, J. A.</dc:creator>
<dc:creator>Spindel, E. R.</dc:creator>
<dc:creator>Messaoudi, I.</dc:creator>
<dc:date>2022-02-10</dc:date>
<dc:identifier>doi:10.1101/2022.02.08.479578</dc:identifier>
<dc:title><![CDATA[Functional, transcriptional, and microbial shifts associated with healthy pulmonary aging: insights from rhesus macaques]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-02-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.05.10.491275v1?rss=1">
<title>
<![CDATA[
HIV rapidly targets a diverse pool of CD4+ T cells to establish productive and latent infections 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.05.10.491275v1?rss=1</link>
<description><![CDATA[
Upon infection, HIV disseminates throughout the human body within 1-2 weeks. However, its early cellular targets remain poorly characterized. We analyzed productively and latently infected cells in blood and lymphoid tissue from individuals in acute infection. The phenotype of productively infected cells rapidly evolved with time and differed between blood and lymph nodes. The TCR repertoire of productively infected cells was heavily biased, with preferential infection of previously expanded/disseminated cells, but composed almost exclusively of unique clonotypes, indicating that they were the product of independent infection events. Latent genetically intact proviruses were already archived early in infection. Hence, productive infection is initially established in a pool of phenotypically and clonotypically distinct T cells in blood and lymph nodes and latently infected cells are generated simultaneously.

One-Sentence SummaryHIV initially infects phenotypically and clonotypically distinct T cells and establishes a latent reservoir concomitantly.
]]></description>
<dc:creator>Gantner, P.</dc:creator>
<dc:creator>Buranapraditkun, S.</dc:creator>
<dc:creator>Pagliuzza, A.</dc:creator>
<dc:creator>Dufour, C.</dc:creator>
<dc:creator>Pardons, M.</dc:creator>
<dc:creator>Mitchell, J. L.</dc:creator>
<dc:creator>Kroon, E.</dc:creator>
<dc:creator>Sacdalan, C.</dc:creator>
<dc:creator>Tulmethakaan, N.</dc:creator>
<dc:creator>Pinyakorn, S.</dc:creator>
<dc:creator>Robb, M. L.</dc:creator>
<dc:creator>Phanuphak, N.</dc:creator>
<dc:creator>Ananworanich, J.</dc:creator>
<dc:creator>Hsu, D.</dc:creator>
<dc:creator>Vasan, S.</dc:creator>
<dc:creator>Trautmann, L.</dc:creator>
<dc:creator>Fromentin, R.</dc:creator>
<dc:creator>Chomont, N.</dc:creator>
<dc:date>2022-05-10</dc:date>
<dc:identifier>doi:10.1101/2022.05.10.491275</dc:identifier>
<dc:title><![CDATA[HIV rapidly targets a diverse pool of CD4+ T cells to establish productive and latent infections]]></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/10.1101/2022.06.20.496874v1?rss=1">
<title>
<![CDATA[
Adenosine Reduces Sinoatrial Node Cell AP Firing Rate by uncoupling its Membrane and Calcium Clocks 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.06.20.496874v1?rss=1</link>
<description><![CDATA[
AO_SCPLOWBSTRACTC_SCPLOWThe spontaneous action potential (AP) firing rate of sinoatrial nodal cells (SANC) is regulated by a system of intracellular Ca2+ and membrane ion current clocks driven by Ca2+-calmodulin-activated adenylyl cyclase-protein kinase A (PKA) signaling. The mean AP cycle length (APCL) and APCL variability inform on the effectiveness of clock coupling. Endogenous ATP metabolite adenosine (ado) binds to adenosine receptors that couple to Gi protein-coupled receptors, reducing spontaneous AP firing rate via G{beta}{gamma} signaling that activates an membrane-clock outward current, IKACh. Ado also inhibits adenylyl cyclase activity via Gi signaling, impacting cAMP-mediated PKA-dependent protein phosphorylation and intracellular Ca2+ cycling. We hypothesize that in addition to IKAdo activation, ado signaling impacts Ca2+ via Gi signaling and that both effects reduce AP firing rate by reducing the effectiveness of the Ca2+ and membrane clock coupling. To this end, we measured Ca2+ and membrane potential characteristics in enzymatically isolated single rabbit SANC. 10 {micro}M ado substantially increased both the mean APCL (on average by 43%, n=10) and AP beat-to-beat variability from 5.1{+/-}1.7% to 7.2{+/-}2.0% (n=10) measured via membrane potential and 5.0{+/-}2.2 to 10.6{+/-}5.9 (n=40) measured via Ca2+ (assessed as the coefficient of variability, CV=SD/mean). These effects were mediated by hyperpolarization of the maximum diastolic membrane potential (membrane clock effect) and suppression of diastolic spontaneous, local Ca2+ releases (LCRs) (Ca2+ clock effect): as LCR size distributions shifted from larger to smaller values, the time of LCR occurrence during diastolic depolarization (LCR period) became prolonged, and the ensemble LCR Ca2+ signal became reduced. The tight linear relationship of coupling between LCR period to the APCL in the presence of ado "drifted" upward and leftward, i.e. for a given LCR period, APCL was prolonged, becoming non-linear indicating clock uncoupling. An extreme case of uncoupling occurred at higher ado concentrations (>100 {micro}M): small stochastic LCRs of the Ca2+ clock failed to self-organize and synchronize to the membrane clock, thus creating a failed attempt to generate an AP resulting in arrhythmia and cessation of AP firing. Thus, the effects of ado to activate G{beta}{gamma} and IKACh, Ado and to activate Gi, suppressing adenylyl cyclase activity, both contribute to the ado-induced increase in the mean APCL and APCL variability by reducing the fidelity of clock coupling and AP firing rate.
]]></description>
<dc:creator>Wirth, A. N.</dc:creator>
<dc:creator>Tsutsui, K.</dc:creator>
<dc:creator>Maltsev, V. A.</dc:creator>
<dc:creator>Lakatta, E. G.</dc:creator>
<dc:date>2022-06-20</dc:date>
<dc:identifier>doi:10.1101/2022.06.20.496874</dc:identifier>
<dc:title><![CDATA[Adenosine Reduces Sinoatrial Node Cell AP Firing Rate by uncoupling its Membrane and Calcium Clocks]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-06-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.06.30.498290v1?rss=1">
<title>
<![CDATA[
Meta-analysis of metagenomes via machine learning and assembly graphs reveals strain switches in Crohn's disease 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.06.30.498290v1?rss=1</link>
<description><![CDATA[
Microbial strains have closely related genomes but may have different phenotypes in the same environment. Shotgun metagenomic sequencing can capture the genomes of all strains present in a community but strain-resolved analysis from shotgun sequencing alone remains difficult. We developed an approach to identify and interrogate strain-level differences in groups of metagenomes. We use this approach to perform a meta-analysis of stool microbiomes from individuals with and without inflammatory bowel disease (IBD; Crohns disease, ulcerative colitis; n = 605), a disease for which there are not specific microbial biomarkers but some evidence that microbial strain variation may stratify by disease state. We first developed a machine learning classifier based on compressed representations of complete metagenomes (FracMinHash sketches) and identified genomes that correlate with IBD subtype. To rescue variation that may not have been present in the genomes, we then used assembly graph genome queries to recover strain variation for correlated genomes. Lastly, we developed a novel differential abundance framework that works directly on the assembly graph to uncover all sequence variants correlated with IBD. We refer to this approach as dominating set differential abundance analysis and have implemented it in the spacegraphcats software package. Using this approach, we identified five bacterial strains that are associated with Crohns disease. Our method captures variation within the entire sequencing data set, allowing for discovery of previously hidden disease associations.
]]></description>
<dc:creator>Reiter, T. E.</dc:creator>
<dc:creator>Irber, L. C.</dc:creator>
<dc:creator>Gingrich, A. A.</dc:creator>
<dc:creator>Haynes, D.</dc:creator>
<dc:creator>Pierce-Ward, N. T.</dc:creator>
<dc:creator>Brooks, P. T.</dc:creator>
<dc:creator>Mizutani, Y.</dc:creator>
<dc:creator>Moritz, D.</dc:creator>
<dc:creator>Reidl, F.</dc:creator>
<dc:creator>Willis, A. D.</dc:creator>
<dc:creator>Sullivan, B. D.</dc:creator>
<dc:creator>Brown, C. T.</dc:creator>
<dc:date>2022-07-05</dc:date>
<dc:identifier>doi:10.1101/2022.06.30.498290</dc:identifier>
<dc:title><![CDATA[Meta-analysis of metagenomes via machine learning and assembly graphs reveals strain switches in Crohn's disease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-07-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2022.08.02.502565v1?rss=1">
<title>
<![CDATA[
An adhesive signaling axis regulates the establishment of the cortical glial scaffold. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.08.02.502565v1?rss=1</link>
<description><![CDATA[
The mature mammalian cortex is composed of six architecturally and functionally distinct layers. Two key steps in the assembly of this layered structure are the initial establishment of the glial scaffold and the subsequent migration of post-mitotic neurons to their final position. These processes involve the precise and timely regulation of adhesion and detachment of neural cells from their substrates. Although much is known about the roles of adhesive substrates during neuronal migration and the formation of the glial scaffold, less is understood about how these signals are interpreted and integrated within these neural cells. Here, we provide in vivo evidence that Cas proteins, a family of cytoplasmic adaptors, serve a functional and redundant role during cortical lamination. Cas triple conditional knock-out (Cas TcKO) mice display severe cortical phenotypes that feature cobblestone malformations. Molecular epistasis and genetic experiments suggest that Cas proteins act downstream of transmembrane Dystroglycan and {beta}1-Integrin in a radial glial cell-autonomous manner. Overall, these data establish a new and essential role for Cas adaptor proteins during the formation of cortical circuits and reveal a signaling axis controlling cortical scaffold formation.
]]></description>
<dc:creator>Wong, W.</dc:creator>
<dc:creator>Estep, J. A.</dc:creator>
<dc:creator>Ubina, T.</dc:creator>
<dc:creator>Jahncke, J. N.</dc:creator>
<dc:creator>Wright, K. M.</dc:creator>
<dc:creator>Riccomagno, M. M.</dc:creator>
<dc:date>2022-08-05</dc:date>
<dc:identifier>doi:10.1101/2022.08.02.502565</dc:identifier>
<dc:title><![CDATA[An adhesive signaling axis regulates the establishment of the cortical glial scaffold.]]></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/10.1101/2022.10.11.511633v1?rss=1">
<title>
<![CDATA[
Physics-based Deep Learning for Imaging Neuronal Activity via Two-photon and Light Field Microscopy 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.10.11.511633v1?rss=1</link>
<description><![CDATA[
Light Field Microscopy (LFM) is an imaging technique that offers the opportunity to study fast dynamics in biological systems due to its rapid 3D imaging rate. In particular, it is attractive to analyze neuronal activity in the brain. Unlike scanning-based imaging methods, LFM simultaneously encodes the spatial and angular information of light in a single snapshot. However, LFM is limited by a trade-off between spatial and angular resolution and is affected by scattering at deep layers in the brain tissue. In contrast, two-photon (2P) microscopy is a point-scanning 3D imaging technique that achieves higher spatial resolution, deeper tissue penetration, and reduced scattering effects. However, point-scanning acquisition limits the imaging speed in 2P microscopy and cannot be used to simultaneously monitor the activity of a large population of neurons. This work introduces a physics-driven deep neural network to image neuronal activity in scattering volume tissues using LFM. The architecture of the network is obtained by unfolding the ISTA algorithm and is based on the observation that the neurons in the tissue are sparse. The deep-network architecture is also based on a novel imaging system modeling that uses a linear convolutional neural network and fits the physics of the acquisition process. To achieve the high-quality reconstruction of neuronal activity in 3D brain tissues from temporal sequences of light field (LF) images, we train the network in a semi-supervised manner using generative adversarial networks (GANs). We use the TdTomato indicator to obtain static structural information of the tissue with the microscope operating in 2P scanning modality, representing the target reconstruction quality. We also use additional functional data in LF modality with GCaMP indicators to train the network. Our approach is tested under adverse conditions: limited training data, background noise, and scattering samples. We experimentally show that our method performs better than model-based reconstruction strategies and typical artificial neural networks for imaging neuronal activity in mammalian brain tissue, considering reconstruction quality, generalization to functional imaging, and reconstruction speed.
]]></description>
<dc:creator>Verinaz-Jadan, H.</dc:creator>
<dc:creator>Howe, C. L.</dc:creator>
<dc:creator>Song, P.</dc:creator>
<dc:creator>Lesept, F.</dc:creator>
<dc:creator>Kittler, J.</dc:creator>
<dc:creator>Foust, A. J.</dc:creator>
<dc:creator>Dragotti, P. L.</dc:creator>
<dc:date>2022-10-13</dc:date>
<dc:identifier>doi:10.1101/2022.10.11.511633</dc:identifier>
<dc:title><![CDATA[Physics-based Deep Learning for Imaging Neuronal Activity via Two-photon and Light Field Microscopy]]></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/10.1101/2022.10.17.512460v1?rss=1">
<title>
<![CDATA[
A Machine-learning-based Method to Detect Degradation of Motor Control Stability with Applications to Diagnosis of Presymptomatic Parkinson's Disease 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2022.10.17.512460v1?rss=1</link>
<description><![CDATA[
Parkinsons disease (PD), a neuro-degenerative disorder, is often detected by onset of its motor symptoms such as rest tremor. Unfortunately, motor symptoms appear only when approximately 40%-60% of the dopaminergic neurons in the substantia nigra are lost. In most cases, by the time PD is clinically diagnosed, the disease may already have started 4 to 6 years beforehand. So there is a need for developing a test for detecting PD before the onset of the motor symptoms. This phase of PD is referred to as Presymptomatic PD (PPD). The motor symptoms of Parkinsons Disease are manifestations of instability in the sensorimotor system that develops gradually due to the neuro-degenerative process. In this paper, based on the above insight, we propose a new method that can potentially be used to detect degradation of motor control stability which can be employed for the detection of PPD. The proposed method tracks the tendency of a feedback control system to transition to an unstable state, and uses machine learning algorithm for its robust detection. This method is explored using simulations of a simple pendulum with PID controller as a conceptual representation for both healthy and PPD individuals. We also propose an example task with physiological measurements that can be used with this method and potentially be employed in a clinical setting. We present representative data collected through such a task, thereby demonstrating the feasibility to generate data for the proposed method.

Author summaryParkinsons disease (PD) is a neuro-degenerative disorder that develops and progresses over several years. Currently, one is able to diagnose PD only after the appearance of motor symptoms (symptoms in movements of body parts), which unfortunately may be 4 to 6 years after the neuro-degeneration may have started. It has been shown that there are benefits to diagnosing PD at early stages, motivating the need to explore tools for diagnosing PD in the pre-symptomatic stage referred to as Presymptomatic Parkinsons disease (PPD). In this paper, a novel approach is explored that utilises the insight that the motor symptoms in PD may be seen as an instability in the feedback-control system that controls movements of body parts (sensory-motor loop). The proposed method uses a series of simple movement tasks performed by an individual in a clinic as the input to detect any gradual degradation of movement control that is leading to an instability, but before the instability and consequently the symptoms are manifested. This method is tested through extensive simulations and a potential experimental realisation with preliminary data. While a full-fledged validation will be undertaken as part of future work, initial results show promise and feasibility of further data collection.
]]></description>
<dc:creator>Shah, V. V.</dc:creator>
<dc:creator>Jadav, S.</dc:creator>
<dc:creator>Goyal, S.</dc:creator>
<dc:creator>Palanthandalam-Madapusi, H. J.</dc:creator>
<dc:date>2022-10-20</dc:date>
<dc:identifier>doi:10.1101/2022.10.17.512460</dc:identifier>
<dc:title><![CDATA[A Machine-learning-based Method to Detect Degradation of Motor Control Stability with Applications to Diagnosis of Presymptomatic Parkinson's Disease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-10-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.01.02.522522v1?rss=1">
<title>
<![CDATA[
Systematic assessment of the contribution of structural variants to inherited retinal diseases 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.01.02.522522v1?rss=1</link>
<description><![CDATA[
Despite increasing success in determining genetic diagnosis for patients with inherited retinal diseases (IRDs), mutations in about 30% of the IRD cases remain unclear or unsettled after targeted gene panel or whole exome sequencing. In this study, we aimed to investigate the contributions of structural variants (SVs) to settling the molecular diagnosis of IRD with whole-genome sequencing (WGS). A cohort of 755 IRD patients whose pathogenic mutations remain undefined was subjected to WGS. Four SV calling algorithms including include MANTA, DELLY, LUMPY, and CNVnator were used to detect SVs throughout the genome. All SVs identified by any one of these four algorithms were included for further analysis. AnnotSV was used to annotate these SVs. SVs that overlap with known IRD-associated genes were examined with sequencing coverage, junction reads, and discordant read pairs. PCR followed by Sanger sequencing was used to further confirm the SVs and identify the breakpoints. Segregation of the candidate pathogenic alleles with the disease was performed when possible. In total, sixteen candidate pathogenic SVs were identified in sixteen families, including deletions and inversions, representing 2.1% of patients with previously unsolved IRDs. Autosomal dominant, autosomal recessive, and X-linked inheritance of disease-causing SVs were observed in 12 different genes. Among these, SVs in CLN3, EYS, PRPF31 were found in multiple families. Our study suggests that the contribution of SVs detected by short-read WGS is about 0.25% of our IRD patient cohort and is significantly lower than that of single nucleotide changes and small insertions and deletions.
]]></description>
<dc:creator>Wen, S.</dc:creator>
<dc:creator>Wang, M.</dc:creator>
<dc:creator>Qian, X.</dc:creator>
<dc:creator>Li, Y.</dc:creator>
<dc:creator>Wang, K.</dc:creator>
<dc:creator>Choi, J.</dc:creator>
<dc:creator>Pennesi, M. E.</dc:creator>
<dc:creator>Yang, P.</dc:creator>
<dc:creator>Marra, M.</dc:creator>
<dc:creator>Koenekoop, R. K.</dc:creator>
<dc:creator>Lopez, I.</dc:creator>
<dc:creator>Matynia, A.</dc:creator>
<dc:creator>Gorin, M.</dc:creator>
<dc:creator>Sui, R.</dc:creator>
<dc:creator>Yao, F.</dc:creator>
<dc:creator>Goetz, K.</dc:creator>
<dc:creator>Porto, F. B. O.</dc:creator>
<dc:creator>Chen, R.</dc:creator>
<dc:date>2023-01-03</dc:date>
<dc:identifier>doi:10.1101/2023.01.02.522522</dc:identifier>
<dc:title><![CDATA[Systematic assessment of the contribution of structural variants to inherited retinal diseases]]></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/10.1101/2023.02.14.528487v1?rss=1">
<title>
<![CDATA[
CXCR6 promotes dermal CD8+ T cell survival and transition to long-term tissue residence 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.02.14.528487v1?rss=1</link>
<description><![CDATA[
Tissue resident memory T cells (TRM) provide protection against local re-infection, and yet the interstitial signals necessary for their formation and persistence remain incompletely understood. Here we show that antigen-dependent induction of the chemokine receptor, CXCR6, is a conserved adaptation to peripheral tissue infiltration that promotes TRM formation after viral infection. Deficient TRM formation in the absence of CXCR6 was not explained by canonical trafficking as CXCR6 was not required for tissue entry, was dispensable for the early accumulation of antigen-specific CD8+ T cells in skin, and did not restrain their exit. Further, single cell sequencing indicated that Cxcr6-/- CD8+ T cells were competent to acquire a transcriptional program of residence and TRM that formed were equally functional compared to their WT counterparts when reactivated greater than 100 days post primary infection. The reduced numbers observed at memory time points, where instead found to associate with impaired redox homeostasis and antioxidant capacity during the transition from effector to memory states. As such, Cxcr6-/- CD8+ T cells exhibited increased rates of apoptosis in the dermis relative to controls, which led to reduced numbers of TRM in the epidermis at memory. CXCR6 therefore promotes the metabolic adaptation of T cells as they engage antigen in tissue to increase the probability of memory differentiation and long-term residence.

One Sentence SummaryCXCR6 promotes mechanisms of cellular adaptation to tissue that support local survival and the transition to tissue residence.
]]></description>
<dc:creator>Heim, T. A.</dc:creator>
<dc:creator>Lin, Z.</dc:creator>
<dc:creator>Steele, M. M.</dc:creator>
<dc:creator>Mudianto, T.</dc:creator>
<dc:creator>Lund, A. W.</dc:creator>
<dc:date>2023-02-15</dc:date>
<dc:identifier>doi:10.1101/2023.02.14.528487</dc:identifier>
<dc:title><![CDATA[CXCR6 promotes dermal CD8+ T cell survival and transition to long-term tissue residence]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-02-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.02.23.529781v1?rss=1">
<title>
<![CDATA[
The dual role of TonB genes in turnerbactin uptake and carbohydrate utilization in the shipworm symbiont Teredinibacter turnerae 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.02.23.529781v1?rss=1</link>
<description><![CDATA[
Teredinibacter turnerae is an intracellular bacterial symbiont that resides in the gills of shipworms, wood-eating bivalve mollusks. This bacterium produces a catechol siderophore, turnerbactin, required for the survival of this bacterium under iron limiting conditions. The turnerbactin biosynthetic genes are contained in one of the secondary metabolite clusters conserved among T. turnerae strains. However, Fe(III)-turnerbactin uptake mechanisms are largely unknown. Here, we show that the first gene of the cluster, fttA a homologue of Fe(III)-siderophore TonB-dependent outer membrane receptor (TBDR) genes is indispensable for iron uptake via the endogenous siderophore, turnerbactin, as well as by an exogenous siderophore, amphi-enterobactin, ubiquitously produced by marine vibrios. Furthermore, three TonB clusters containing four tonB genes were identified, and two of these genes, tonB1b and tonB2, functioned not only for iron transport but also for carbohydrate utilization when cellulose was a sole carbon source. Gene expression analysis revealed that none of the tonB genes and other genes in those clusters were clearly regulated by iron concentration while turnerbactin biosynthesis and uptake genes were up-regulated under iron limiting conditions, highlighting the importance of tonB genes even in iron rich conditions, possibly for utilization of carbohydrates derived from cellulose.
]]></description>
<dc:creator>Naka, H.</dc:creator>
<dc:creator>Haygood, M. G.</dc:creator>
<dc:date>2023-02-24</dc:date>
<dc:identifier>doi:10.1101/2023.02.23.529781</dc:identifier>
<dc:title><![CDATA[The dual role of TonB genes in turnerbactin uptake and carbohydrate utilization in the shipworm symbiont Teredinibacter turnerae]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-02-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.03.22.533696v1?rss=1">
<title>
<![CDATA[
BIBSNet: A Deep Learning Baby Image Brain Segmentation Network for MRI Scans 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.03.22.533696v1?rss=1</link>
<description><![CDATA[
ObjectivesBrain segmentation of infant magnetic resonance (MR) images is vitally important for studying typical and atypical brain development. The infant brain undergoes many changes throughout the first years of postnatal life, making tissue segmentation difficult for most existing algorithms. Here we introduce a deep neural network BIBSNet (Baby and Infant Brain Segmentation Neural Network), an open-source, community-driven model for robust and generalizable brain segmentation leveraging data augmentation and a large sample size of manually annotated images.

Experimental DesignIncluded in model training and testing were MR brain images from 90 participants with an age range of 0-8 months (median age 4.6 months). Using the BOBs repository of manually annotated real images along with synthetic segmentation images produced using SynthSeg, the model was trained using a 10-fold procedure. Model performance of segmentations was assessed by comparing BIBSNet, joint label fusion (JLF) inferred segmentation to ground truth segmentations using Dice Similarity Coefficient (DSC). Additionally, MR data along with the FreeSurfer compatible segmentations were processed with the DCAN labs infant-ABCD-BIDS processing pipeline from ground truth, JLF, and BIBSNet to further assess model performance on derivative data, including cortical thickness, resting state connectivity and brain region volumes.

Principal ObservationsBIBSNet segmentations outperforms JLF across all regions based on DSC comparisons. Additionally, with processed derived metrics, BIBSNet segmentations outperforms JLF segmentations across nearly all metrics.

ConclusionsBIBSNet segmentation shows marked improvement over JLF across all age groups analyzed. The BIBSNet model is 600x faster compared to JLF, produces FreeSurfer-compatible segmentation labels, and can be easily included in other processing pipelines. BIBSNet provides a viable alternative for segmenting the brain in the earliest stages of development.
]]></description>
<dc:creator>Hendrickson, T. J.</dc:creator>
<dc:creator>Reiners, P.</dc:creator>
<dc:creator>Moore, L. A.</dc:creator>
<dc:creator>Perrone, A. J.</dc:creator>
<dc:creator>Alexopoulos, D.</dc:creator>
<dc:creator>Lee, E. G.</dc:creator>
<dc:creator>Styner, M.</dc:creator>
<dc:creator>Kardan, O.</dc:creator>
<dc:creator>Chamberlain, T. A.</dc:creator>
<dc:creator>Mummaneni, A.</dc:creator>
<dc:creator>Caldas, H. A.</dc:creator>
<dc:creator>Bower, B.</dc:creator>
<dc:creator>Stoyell, S.</dc:creator>
<dc:creator>Martin, T.</dc:creator>
<dc:creator>Sung, S.</dc:creator>
<dc:creator>Fair, E.</dc:creator>
<dc:creator>Uriarte-Lopez, J.</dc:creator>
<dc:creator>Rueter, A. R.</dc:creator>
<dc:creator>Rosenberg, M. D.</dc:creator>
<dc:creator>Smyser, C. D.</dc:creator>
<dc:creator>Elison, J. T.</dc:creator>
<dc:creator>Graham, A.</dc:creator>
<dc:creator>Fair, D. A.</dc:creator>
<dc:creator>Feczko, E.</dc:creator>
<dc:date>2023-03-24</dc:date>
<dc:identifier>doi:10.1101/2023.03.22.533696</dc:identifier>
<dc:title><![CDATA[BIBSNet: A Deep Learning Baby Image Brain Segmentation Network for MRI Scans]]></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/10.1101/212860v1?rss=1">
<title>
<![CDATA[
An automated statistical technique for counting distinct multiple sclerosis lesions can recover aspects of lesion history and provide relevant disease information 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/212860v1?rss=1</link>
<description><![CDATA[
BackgroundLesion load is a common biomarker in multiple sclerosis, yet it has historically shown modest associations with clinical outcomes. Lesion count, which encapsulates the natural history of lesion formation and is thought to provide complementary information, is difficult to assess in patients with confluent (i.e. spatially overlapping) lesions. We introduce a statistical technique for cross-sectionally counting pathologically distinct lesions.nnMethodsMRI is used to assess the probability of lesion at each location. The texture of this map is quantified using a novel technique, and clusters resembling the center of a lesion are counted.nnResultsValidity was demonstrated by comparing the proposed count to a gold-standard count in 60 subjects observed longitudinally. The counts were highly correlated (r = .97, p < .001) and not significantly different (t59 = -0.83, p > .40). Reliability was determined using 14 scans of a clinically stable subject acquired at 7 sites, and variability of lesion count was equivalent to that of lesion load. Accounting for lesion load and age, lesion count was negatively associated (t58 = -2.73, p < .01) with the Expanded Disability Status Scale (EDSS). Average lesion size had a higher association with EDSS (r =.35, p < .01) than lesion load (r = .10, p > .40) or lesion count (r = -.12, p > .30) alone.nnConclusionThese findings demonstrate that it is possible to recover important aspects of the natural history of lesion formation without longitudinal data, and suggest that lesion size provides complementary information about disease.nnGrant SupportThe project described was supported in part by the NIH grants R01 NS085211, R21 NS093349, and R01 NS094456 from the National Institute of Neurological Disorders and Stroke (NINDS). The study was also supported by the Intramural Research Program of NINDS and the Race to Erase MS Foundation. The content is solely the responsibility of the authors and does not necessarily represent the official views of the funding agencies.
]]></description>
<dc:creator>Dworkin, J. D.</dc:creator>
<dc:creator>Linn, K. A.</dc:creator>
<dc:creator>Oguz, I.</dc:creator>
<dc:creator>Fleishman, G. M.</dc:creator>
<dc:creator>Bakshi, R.</dc:creator>
<dc:creator>Nair, G.</dc:creator>
<dc:creator>Calabresi, P. A.</dc:creator>
<dc:creator>Henry, R. G.</dc:creator>
<dc:creator>Oh, J.</dc:creator>
<dc:creator>Papinutto, N.</dc:creator>
<dc:creator>Pelletier, D.</dc:creator>
<dc:creator>Rooney, W.</dc:creator>
<dc:creator>Stern, W.</dc:creator>
<dc:creator>Sicotte, N. L.</dc:creator>
<dc:creator>Reich, D. S.</dc:creator>
<dc:creator>Shinohara, R. T.</dc:creator>
<dc:date>2017-11-01</dc:date>
<dc:identifier>doi:10.1101/212860</dc:identifier>
<dc:title><![CDATA[An automated statistical technique for counting distinct multiple sclerosis lesions can recover aspects of lesion history and provide relevant disease information]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-11-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/224733v1?rss=1">
<title>
<![CDATA[
Combining accurate tumour genome simulation with crowd sourcing to benchmark somatic structural variant detection 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/224733v1?rss=1</link>
<description><![CDATA[
BackgroundThe phenotypes of cancer cells are driven in part by somatic structural variants. Structural variants can initiate tumors, enhance their aggressiveness and provide unique therapeutic opportunities. Whole-genome sequencing of tumors can allow exhaustive identification of the specific structural variants present in an individual cancer, facilitating both clinical diagnostics and the discovery of novel mutagenic mechanisms. A plethora of somatic structural variant detection algorithms have been created to enable these discoveries, however there are no systematic benchmarks of them. Rigorous performance evaluation of somatic structural variant detection methods has been challenged by the lack of gold-standards, extensive resource requirements and difficulties arising from the need to share personal genomic information.nnResultsTo facilitate structural variant detection algorithm evaluations, we create a robust simulation framework for somatic structural variants by extending the BAMSurgeon algorithm. We then organize and enable a crowd-sourced benchmarking within the ICGC-TCGA DREAM Somatic Mutation Calling Challenge (SMC-DNA). We report here the results of structural variant benchmarking on three different tumors, comprising 204 submissions from 15 teams. In addition to ranking methods, we identify characteristic error-profiles of individual algorithms and general trends across them. Surprisingly, we find that ensembles of analysis pipelines do not always outperform the best individual method, indicating a need for new ways to aggregate somatic structural variant detection approaches.nnConclusionsThe synthetic tumors and somatic structural variant detection leaderboards remain available as a community benchmarking resource, and BAMSurgeon is available at https://github.com/adamewing/bamsurgeon.
]]></description>
<dc:creator>Lee, A. Y.-W.</dc:creator>
<dc:creator>Ewing, A. D.</dc:creator>
<dc:creator>Ellrott, K.</dc:creator>
<dc:creator>Hu, Y.</dc:creator>
<dc:creator>Houlahan, K. E.</dc:creator>
<dc:creator>Bare, J. C.</dc:creator>
<dc:creator>Espiritu, S. M. G.</dc:creator>
<dc:creator>Huang, V.</dc:creator>
<dc:creator>Dang, K.</dc:creator>
<dc:creator>Chong, Z.</dc:creator>
<dc:creator>Caloian, C.</dc:creator>
<dc:creator>Yamaguchi, T. N.</dc:creator>
<dc:creator>ICGC-TCGA DREAM Somatic Mutation Calling Challenge,</dc:creator>
<dc:creator>Kellen, M. R.</dc:creator>
<dc:creator>Chen, K.</dc:creator>
<dc:creator>Norman, T. C.</dc:creator>
<dc:creator>Friend, S. H.</dc:creator>
<dc:creator>Guinney, J.</dc:creator>
<dc:creator>Stolovitzky, G.</dc:creator>
<dc:creator>Haussler, D.</dc:creator>
<dc:creator>Margolin, A. A.</dc:creator>
<dc:creator>Stuart, J. M.</dc:creator>
<dc:creator>Boutros, P. C.</dc:creator>
<dc:date>2017-11-25</dc:date>
<dc:identifier>doi:10.1101/224733</dc:identifier>
<dc:title><![CDATA[Combining accurate tumour genome simulation with crowd sourcing to benchmark somatic structural variant detection]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-11-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/232611v1?rss=1">
<title>
<![CDATA[
Teaching data science fundamentals through realistic synthetic clinical cardiovascular data 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/232611v1?rss=1</link>
<description><![CDATA[
ObjectiveOur goal was to create a synthetic dataset and curricular materials to assist in teaching fundamentals of translational data science.nnMaterials and MethodsA literature review was conducted to extract current cardiovascular risk score logic, data elements, and population characteristics. Then, clinical data elements in the models were pulled from clinical data and transformed to the Observational Medical Outcomes Partnership (OMOP) common data model; genetic data elements were added based on population rates. A hybrid Bayesian network was used to create synthetic data from the logical elements of the risk scores and the underlying population frequencies of the clinical data.nnResultsA synthetic dataset of 446,000 patients was created. A two-day curriculum was created based on this synthetic data with exploratory data analysis and machine learning components. The curriculum was offered on two separate occasions; the two groups of learners were given the curriculum and data, and results were tallied, summarized, and compared. Students ability to complete the challenge was mixed; more experienced students achieved a range of 70%-85% in balanced accuracy, but many others did not perform better than the baseline model.nnDiscussionOverall, students enjoyed the course and dataset, but some struggled to consistently apply machine learning techniques. The curriculum, data set, techniques for generation, and results are available for others to use for their own training.nnConclusionA realistic synthetic data with clinical and genetic components helps students learn issues in cardiovascular risk scoring, practice data science skills, and compete in a challenge to improve identification of risk.
]]></description>
<dc:creator>Laderas, T.</dc:creator>
<dc:creator>Vasilevsky, N.</dc:creator>
<dc:creator>Pederson, B.</dc:creator>
<dc:creator>Haendel, M.</dc:creator>
<dc:creator>McWeeney, S.</dc:creator>
<dc:creator>Dorr, D.</dc:creator>
<dc:date>2017-12-12</dc:date>
<dc:identifier>doi:10.1101/232611</dc:identifier>
<dc:title><![CDATA[Teaching data science fundamentals through realistic synthetic clinical cardiovascular data]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-12-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/254839v1?rss=1">
<title>
<![CDATA[
Valection: Design Optimization for Validation and Verification Studies 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/254839v1?rss=1</link>
<description><![CDATA[
BackgroundPlatform-specific error profiles necessitate confirmatory studies where predictions made on data generated using one technology are additionally verified by processing the same samples on an orthogonal technology. In disciplines that rely heavily on high-throughput data generation, such as genomics, reducing the impact of false positive and false negative rates in results is a top priority. However, verifying all predictions can be costly and redundant, and testing a subset of findings is often used to estimate the true error profile. To determine how to create subsets of predictions for validation that maximize inference of global error profiles, we developed Valection, a software program that implements multiple strategies for the selection of verification candidates.nnResultsTo evaluate these selection strategies, we obtained 261 sets of somatic mutation calls from a single-nucleotide variant caller benchmarking challenge where 21 teams competed on whole-genome sequencing datasets of three computationally-simulated tumours. By using synthetic data, we had complete ground truth of the tumours mutations and, therefore, we were able to accurately determine how estimates from the selected subset of verification candidates compared to the complete prediction set. We found that selection strategy performance depends on several verification study characteristics. In particular the verification budget of the experiment (i.e. how many candidates can be selected) is shown to influence estimates.nnConclusionsThe Valection framework is flexible, allowing for the implementation of additional selection algorithms in the future. Its applicability extends to any discipline that relies on experimental verification and will benefit from the optimization of verification candidate selection.
]]></description>
<dc:creator>Cooper, C. I.</dc:creator>
<dc:creator>Yao, D.</dc:creator>
<dc:creator>Sendorek, D. H.</dc:creator>
<dc:creator>Yamaguchi, T. N.</dc:creator>
<dc:creator>P'ng, C.</dc:creator>
<dc:creator>Houlahan, K. E.</dc:creator>
<dc:creator>Caloian, C.</dc:creator>
<dc:creator>Fraser, M.</dc:creator>
<dc:creator>SMC-DNA Challenge Participants,</dc:creator>
<dc:creator>Ellrott, K.</dc:creator>
<dc:creator>Margolin, A. A.</dc:creator>
<dc:creator>Bristow, R. G.</dc:creator>
<dc:creator>Stuart, J. M.</dc:creator>
<dc:creator>Boutros, P. C.</dc:creator>
<dc:date>2018-01-28</dc:date>
<dc:identifier>doi:10.1101/254839</dc:identifier>
<dc:title><![CDATA[Valection: Design Optimization for Validation and Verification Studies]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-01-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/255679v1?rss=1">
<title>
<![CDATA[
Individual differences in functional brain connectivity predict temporal discounting preference in the transition to adolescence 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/255679v1?rss=1</link>
<description><![CDATA[
The transition from childhood to adolescence is marked by distinct changes in behavior, including how one values waiting for a large reward compared to receiving an immediate, yet smaller, reward. While previous research has emphasized the relationship between this preference and age, it is also proposed that this behavior is related to circuitry between valuation and cognitive control systems. In this study, we examined how age and intrinsic functional connectivity strength within and between these neural systems relate to changes in discounting behavior across the transition into adolescence. We used mixed-effects modeling and linear regression to assess the contributions of age and connectivity strength in predicting discounting behavior. First, we identified relevant connections in a longitudinal sample of 64 individuals who completed MRI scans and behavioral assessments 2-3 times across ages 7-15 years (137 scans). We then repeated the analysis in a separate, cross-sectional, sample of 84 individuals (7-13 years). Both samples showed an age-related increase in preference for waiting for larger rewards. Connectivity strength within and between valuation and cognitive control systems accounted for further variance not explained by age. These results suggest that individual differences in functional neural organization can account for behavioral changes typically associated with age.
]]></description>
<dc:creator>Anandakumar, J.</dc:creator>
<dc:creator>Mills, K.</dc:creator>
<dc:creator>Earl, E.</dc:creator>
<dc:creator>Irwin, L.</dc:creator>
<dc:creator>Miranda-Dominguez, O.</dc:creator>
<dc:creator>Demeter, D. V.</dc:creator>
<dc:creator>Walton-Weston, A.</dc:creator>
<dc:creator>Karalunas, S.</dc:creator>
<dc:creator>Nigg, J.</dc:creator>
<dc:creator>Fair, D. A.</dc:creator>
<dc:date>2018-01-29</dc:date>
<dc:identifier>doi:10.1101/255679</dc:identifier>
<dc:title><![CDATA[Individual differences in functional brain connectivity predict temporal discounting preference in the transition to adolescence]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-01-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/285171v1?rss=1">
<title>
<![CDATA[
Small effect size leads to reproducibility failure in resting-state fMRI studies 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/285171v1?rss=1</link>
<description><![CDATA[
Thousands of papers using resting-state functional magnetic resonance imaging (RS-fMRI) have been published on brain disorders. Results in each paper may have survived correction for multiple comparison. However, since there have been no robust results from large scale meta-analysis, we do not know how many of published results are truly positives. The present meta-analytic work included 60 original studies, with 57 studies (4 datasets, 2266 participants) that used a between-group design and 3 studies (1 dataset, 107 participants) that employed a within-group design. To evaluate the effect size of brain disorders, a very large neuroimaging dataset ranging from neurological to psychiatric isorders together with healthy individuals have been analyzed. Parkinsons disease off levodopa (PD-off) included 687 participants from 15 studies. PD on levodopa (PD-on) included 261 participants from 9 studies. Autism spectrum disorder (ASD) included 958 participants from 27 studies. The meta-analyses of a metric named amplitude of low frequency fluctuation (ALFF) showed that the effect size (Hedges g) was 0.19 - 0.39 for the 4 datasets using between-group design and 0.46 for the dataset using within-group design. The effect size of PD-off, PD-on and ASD were 0.23, 0.39, and 0.19, respectively. Using the meta-analysis results as the robust results, the between-group design results of each study showed high false negative rates (median 99%), high false discovery rates (median 86%), and low accuracy (median 1%), regardless of whether stringent or liberal multiple comparison correction was used. The findings were similar for 4 RS-fMRI metrics including ALFF, regional homogeneity, and degree centrality, as well as for another widely used RS-fMRI metric namely seed-based functional connectivity. These observations suggest that multiple comparison correction does not control for false discoveries across multiple studies when the effect sizes are relatively small. Meta-analysis on un-thresholded t-maps is critical for the recovery of ground truth. We recommend that to achieve high reproducibility through meta-analysis, the neuroimaging research field should share raw data or, at minimum, provide un-thresholded statistical images.
]]></description>
<dc:creator>Jia, X.-Z.</dc:creator>
<dc:creator>Zhao, N.</dc:creator>
<dc:creator>Barton, B.</dc:creator>
<dc:creator>Burciu, R.</dc:creator>
<dc:creator>Carriere, N.</dc:creator>
<dc:creator>Cerasa, A.</dc:creator>
<dc:creator>Chen, B.-Y.</dc:creator>
<dc:creator>Chen, J.</dc:creator>
<dc:creator>Coombes, S.</dc:creator>
<dc:creator>Defebvre, L.</dc:creator>
<dc:creator>Delmaire, C.</dc:creator>
<dc:creator>Dujardin, K.</dc:creator>
<dc:creator>Esposito, F.</dc:creator>
<dc:creator>Fan, G.-G.</dc:creator>
<dc:creator>Federica, D. N.</dc:creator>
<dc:creator>Feng, Y.-X.</dc:creator>
<dc:creator>Fling, B. W.</dc:creator>
<dc:creator>Garg, S.</dc:creator>
<dc:creator>Gilat, M.</dc:creator>
<dc:creator>Gorges, M.</dc:creator>
<dc:creator>Ho, S.-L.</dc:creator>
<dc:creator>Horak, F. B.</dc:creator>
<dc:creator>Hu, X.</dc:creator>
<dc:creator>Hu, X.-F.</dc:creator>
<dc:creator>Huang, B.</dc:creator>
<dc:creator>Huang, P.-Y.</dc:creator>
<dc:creator>Jia, Z.-J.</dc:creator>
<dc:creator>Jones, C.</dc:creator>
<dc:creator>Kassubek, J.</dc:creator>
<dc:creator>Krajcovicova, L.</dc:creator>
<dc:creator>Kurani, A.</dc:creator>
<dc:creator>Li, J.</dc:creator>
<dc:creator>Li, Q.</dc:creator>
<dc:creator>Liu, A.-P.</dc:creator>
<dc:creator>Liu, B.</dc:creator>
<dc:creator>Liu, H.</dc:creator>
<dc:creator>Liu, W.-G.</dc:creator>
<dc:creator>Lopes, R.</dc:creator>
<dc:creator>Lou, Y.-T.</dc:creator>
<dc:creator>Luo, W.</dc:creator>
<dc:creator>Madhyastha, T.</dc:creator>
<dc:creator>Mao, N.-N.</dc:creator>
<dc:creator>McAlonan, G.</dc:creator>
<dc:creator>McKeown, M. J.</dc:creator>
<dc:creator>Pang, S. Y.</dc:creator>
<dc:creator>Quattrone, A.</dc:creator>
<dc:creator>Rektorova, I.</dc:creator>
<dc:creator>Sarica, A.</dc:creator>
<dc:creator>Shang, H</dc:creator>
<dc:date>2018-03-20</dc:date>
<dc:identifier>doi:10.1101/285171</dc:identifier>
<dc:title><![CDATA[Small effect size leads to reproducibility failure in resting-state fMRI studies]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-03-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/286138v1?rss=1">
<title>
<![CDATA[
Divergent gene expression among phytoplankton taxa in response to upwelling 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/286138v1?rss=1</link>
<description><![CDATA[
Frequent blooms of phytoplankton occur in coastal upwelling zones creating hotspots of biological productivity in the ocean. As cold, nutrient-rich water is brought up to sunlit layers from depth, phytoplankton are also transported upwards to seed surface blooms that are often dominated by diatoms. The physiological response of phytoplankton to this process, commonly referred to as shift-up, is characterized by rapid growth rates and increases in nitrate assimilation. To examine the molecular underpinnings behind this phenomenon, metatranscriptomics was applied to a simulated upwelling experiment using natural phytoplankton communities from the California Upwelling Zone. An increase in diatom growth following five days of incubation was attributed to the genera Chaetoceros and Pseudo-nitzschia. Here we show that certain bloom-forming diatoms exhibit a distinct transcriptional response that coordinates shift-up where diatoms exhibited the greatest transcriptional change following upwelling; however, comparison of coexpressed genes exposed overrepresentation of distinct sets within each of the dominant phytoplankton groups. The analysis revealed that diatoms frontload genes involved in nitrogen assimilation likely in order to outcompete other groups for available nitrogen during upwelling events. We speculate that the evolutionary success of diatoms may be due, in part, to this proactive response to frequently encountered changes in their environment.
]]></description>
<dc:creator>Lampe, R. H.</dc:creator>
<dc:creator>Cohen, N. R.</dc:creator>
<dc:creator>Ellis, K. A.</dc:creator>
<dc:creator>Bruland, K. W.</dc:creator>
<dc:creator>Maldonado, M. T.</dc:creator>
<dc:creator>Peterson, T. D.</dc:creator>
<dc:creator>Till, C. P.</dc:creator>
<dc:creator>Brzezinski, M. A.</dc:creator>
<dc:creator>Bargu, S.</dc:creator>
<dc:creator>Thamatrakoln, K.</dc:creator>
<dc:creator>Kuzminov, F. I.</dc:creator>
<dc:creator>Twining, B. S.</dc:creator>
<dc:creator>Marchetti, A.</dc:creator>
<dc:date>2018-03-23</dc:date>
<dc:identifier>doi:10.1101/286138</dc:identifier>
<dc:title><![CDATA[Divergent gene expression among phytoplankton taxa in response to upwelling]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-03-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/304717v1?rss=1">
<title>
<![CDATA[
Sorting Nexin 27 (SNX27): A Novel Regulator of Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) Trafficking 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/304717v1?rss=1</link>
<description><![CDATA[
The underlying defect in cystic fibrosis is mutation of the cystic fibrosis transmembrane conductance regulator (CFTR), a cAMP-activated chloride channel expressed at the apical surface of lung epithelia. In addition to its export and maintenance at the cell surface, CFTR regulation involves repeated cycles of transport through the endosomal trafficking system, including endocytosis and recycling. Many of the known disease mutations cause CFTR intracellular trafficking defects that result in failure of ion channel delivery to the apical plasma membrane. Corrective maneuvers directed at improving transport to the plasma membrane are thwarted by rapid internalization and degradation of the mutant CFTR proteins. The molecular mechanisms involved in these processes are not completely understood but may involve protein-protein interactions with the C-terminal type I PDZ-binding motif of CFTR. Using a proteomic approach, we identify sorting nexin 27 (SNX27) as a novel CFTR binding partner in human airway epithelial Calu-3 cells. SNX27 and CFTR interact directly, with the SNX27 PDZ domain being both necessary and sufficient for this interaction. SNX27 co-localizes with internalized CFTR at sub-apical endosomal sites in polarized Calu-3 cells, and either knockdown of the endogenous SNX27, or over-expression of a dominant-negative SNX27 mutant, resulted in significant decreases in cell surface CFTR levels. CFTR internalization was not affected by SNX27 knockdown, but defects were observed in the recycling arm of CFTR trafficking through the endosomal system. Furthermore, knockdown of SNX27 in Calu-3 cells resulted in significant decreases in CFTR protein levels, consistent with degradation of the internalized pool. These data identify SNX27 as a physiologically significant regulator of CFTR trafficking and homeostasis in epithelial cells.
]]></description>
<dc:creator>McDermott, M. I.</dc:creator>
<dc:creator>Thelin, W. R.</dc:creator>
<dc:creator>Chen, Y.</dc:creator>
<dc:creator>Lyons, P. T.</dc:creator>
<dc:creator>Reilly, G.</dc:creator>
<dc:creator>Gentzsch, M.</dc:creator>
<dc:creator>Lei, C.</dc:creator>
<dc:creator>Hong, W.</dc:creator>
<dc:creator>Stutts, M. J.</dc:creator>
<dc:creator>Playford, M. P.</dc:creator>
<dc:creator>Bankaitis, V. A.</dc:creator>
<dc:date>2018-04-19</dc:date>
<dc:identifier>doi:10.1101/304717</dc:identifier>
<dc:title><![CDATA[Sorting Nexin 27 (SNX27): A Novel Regulator of Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) Trafficking]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-04-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/334979v1?rss=1">
<title>
<![CDATA[
Transmembrane Protein 135 (TMEM135) is a Liver X Receptor Target Gene that Mediates an Auxiliary Peroxisome Matrix Protein Import Pathway 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/334979v1?rss=1</link>
<description><![CDATA[
The liver x receptors (LXRs) are key regulators of systemic lipid metabolism. We determined whether transmembrane protein 135 (TMEM135) is an LXR target gene and its physiologic function. An LXR agonist increased TMEM135 mRNA and protein in human hepatocyte and macrophage cell lines, which was prevented by LXR knockdown. The human TMEM135 promoter contains an LXR response element that bound the LXRs via EMSA and ChIP, and mediated LXR-induced transcription in reporter assays. Knockdown of TMEM135 in HepG2 cells caused triglyceride accumulation despite reduced lipogenic gene expression, indicating a potential role in {beta}-oxidation. To determine physiologic importance, TMEM135 was knocked-down via siRNA in livers of fed and fasted C57BL/6 mice. Fasting increased hepatic fatty acid and NADH concentrations in control mice, consistent with increased fatty acid uptake and {beta}-oxidation. However, in fasted TMEM135 knockdown mice, there was a further significant increase in hepatic fatty acid concentrations and a significant decrease in NADH, indicating an impairment in {beta}-oxidation by peroxisomes and/or mitochondria. Conversely, hepatic ketones tended to increase in fasted TMEM135 knockdown compared to control mice, and because ketogenesis is exclusively dependent on mitochondrial {beta}-oxidation, this indicates peroxisomal {beta}-oxidation was impaired in knockdown mice. Localization studies demonstrated that TMEM135 co-localized with peroxisomes but not mitochondria. Mechanistically, proteomic and Western blot analyses indicated that TMEM135 regulates concentrations of matrix enzymes within peroxisomes. In conclusion, TMEM135 is a novel LXR target gene in humans that mediates peroxisomal metabolism, and thus TMEM135 may be a therapeutic target for metabolic disorders associated with peroxisome dysfunction.
]]></description>
<dc:creator>Renquist, B. J.</dc:creator>
<dc:creator>Madanayake, T. W.</dc:creator>
<dc:creator>Ghimire, S.</dc:creator>
<dc:creator>Geisler, C. E.</dc:creator>
<dc:creator>Xu, Y.</dc:creator>
<dc:creator>Bogan, R. L.</dc:creator>
<dc:date>2018-05-30</dc:date>
<dc:identifier>doi:10.1101/334979</dc:identifier>
<dc:title><![CDATA[Transmembrane Protein 135 (TMEM135) is a Liver X Receptor Target Gene that Mediates an Auxiliary Peroxisome Matrix Protein Import Pathway]]></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/10.1101/357095v1?rss=1">
<title>
<![CDATA[
Amyloid β oligomers constrict human capillaries in Alzheimer’s disease via signalling to pericytes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/357095v1?rss=1</link>
<description><![CDATA[
Vascular compromise occurs early in Alzheimers disease (AD) and other dementias1-3. Amyloid {beta} (A{beta}) reduces cerebral blood flow4-6 and, as most of the cerebral vasculature resistance is in capillaries7, A{beta} might mainly act on contractile pericytes on capillary walls8-10. Employing human tissue to establish disease-relevance, and rodent experiments to define mechanism, we now show that A{beta} constricts brain capillaries at pericyte locations in human subjects with cognitive decline. Applying soluble A{beta}1-42 oligomers to live human cortical tissue constricted capillaries. Using rat cortical slices, this was shown to reflect A{beta} evoking capillary pericyte contraction, with an EC50 of 4.7 nM, via the generation of reactive oxygen species and activation of endothelin ET-A receptors. In freshly-fixed diagnostic biopsies from human patients investigated for cognitive decline, mean capillary diameters were less in subjects showing A{beta} deposition than in subjects without A{beta} deposition. For patients with A{beta} deposition, the capillary diameter was 31% less at pericyte somata than away from somata, predicting a halving of blood flow. Constriction of capillaries by A{beta} will contribute to the energy lack1-3 occurring in AD, which promotes further A{beta} generation11,12. This mechanism reconciles the amyloid hypothesis13-15 with the earliest events in AD being vascular1.
]]></description>
<dc:creator>Nortley, R.</dc:creator>
<dc:creator>Mishra, A.</dc:creator>
<dc:creator>Jaunmuktane, Z.</dc:creator>
<dc:creator>Kyrargyri, V.</dc:creator>
<dc:creator>Madry, C.</dc:creator>
<dc:creator>Gong, H.</dc:creator>
<dc:creator>Richard-Loendt, A.</dc:creator>
<dc:creator>Brandner, S.</dc:creator>
<dc:creator>Sethi, H.</dc:creator>
<dc:creator>Attwell, D.</dc:creator>
<dc:date>2018-06-27</dc:date>
<dc:identifier>doi:10.1101/357095</dc:identifier>
<dc:title><![CDATA[Amyloid β oligomers constrict human capillaries in Alzheimer’s disease via signalling to pericytes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-06-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/418772v1?rss=1">
<title>
<![CDATA[
Stochastic Simulation to Visualize Gene Expression and Error Correction in Living Cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/418772v1?rss=1</link>
<description><![CDATA[
Stochastic simulation can make the molecular processes of cellular control more vivid than the traditional differential-equation approach by generating typical system histories instead of just statistical measures such as the mean and variance of a population. Simple simulations are now easy for students to construct from scratch, that is, without recourse to black-box packages. In some cases, their results can also be compared directly to single-molecule experimental data. After introducing the stochastic simulation algorithm, this article gives two case studies, involving gene expression and error correction, respectively. Code samples and resulting animations showing results are given in the online supplement.
]]></description>
<dc:creator>Chen, K. Y.</dc:creator>
<dc:creator>Zuckerman, D. M.</dc:creator>
<dc:creator>Nelson, P. C.</dc:creator>
<dc:date>2018-09-15</dc:date>
<dc:identifier>doi:10.1101/418772</dc:identifier>
<dc:title><![CDATA[Stochastic Simulation to Visualize Gene Expression and Error Correction in Living Cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-09-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/448142v1?rss=1">
<title>
<![CDATA[
Comprehensive human cell-type methylation atlas reveals origins of circulating cell-free DNA in health and disease 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/448142v1?rss=1</link>
<description><![CDATA[
Methylation patterns of circulating cell-free DNA (cfDNA) contain rich information about recent cell death events in the body. Here, we present an approach for unbiased determination of the tissue origins of cfDNA, using a reference methylation atlas of 25 human tissues and cell types. The method is validated using in silico simulations as well as in vitro mixes of DNA from different tissue sources at known proportions. We show that plasma cfDNA of healthy donors originates from white blood cells (55%), erythrocyte progenitors (30%), vascular endothelial cells (10%) and hepatocytes (1%). Deconvolution of cfDNA from patients reveals tissue contributions that agree with clinical findings in sepsis, islet transplantation, cancer of the colon, lung, breast and prostate, and cancer of unknown primary. We propose a procedure which can be easily adapted to study the cellular contributors to cfDNA in many settings, opening a broad window into healthy and pathologic human tissue dynamics.
]]></description>
<dc:creator>Moss, J.</dc:creator>
<dc:creator>Magenheim, J.</dc:creator>
<dc:creator>Neiman, D.</dc:creator>
<dc:creator>Zemmour, H.</dc:creator>
<dc:creator>Loyfer, N.</dc:creator>
<dc:creator>Korach, A.</dc:creator>
<dc:creator>Samet, Y.</dc:creator>
<dc:creator>Maoz, M.</dc:creator>
<dc:creator>Druid, H.</dc:creator>
<dc:creator>Arner, P.</dc:creator>
<dc:creator>Fu, K.-Y.</dc:creator>
<dc:creator>Kiss, E.</dc:creator>
<dc:creator>Spalding, K. L.</dc:creator>
<dc:creator>Landesberg, G.</dc:creator>
<dc:creator>Zick, A.</dc:creator>
<dc:creator>Grinshpun, A.</dc:creator>
<dc:creator>Shapiro, A. J.</dc:creator>
<dc:creator>Grompe, M.</dc:creator>
<dc:creator>Dreazan Wittenberg, A.</dc:creator>
<dc:creator>Glaser, B.</dc:creator>
<dc:creator>Shemer, R.</dc:creator>
<dc:creator>Kaplan, T.</dc:creator>
<dc:creator>Dor, Y.</dc:creator>
<dc:date>2018-10-20</dc:date>
<dc:identifier>doi:10.1101/448142</dc:identifier>
<dc:title><![CDATA[Comprehensive human cell-type methylation atlas reveals origins of circulating cell-free DNA in health and disease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-10-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/496513v1?rss=1">
<title>
<![CDATA[
Electron cryo-tomography of vestibular hair-cell stereocilia 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/496513v1?rss=1</link>
<description><![CDATA[
High-resolution imaging of hair-cell stereocilia of the inner ear has contributed substantially to our understanding of auditory and vestibular function. To provide three-dimensional views of the structure of stereocilia cytoskeleton and membranes, we developed a method for rapidly freezing unfixed stereocilia on electron microscopy grids, which allowed subsequent 3D imaging by electron cryo-tomography. Structures of stereocilia tips, shafts, and tapers were revealed, demonstrating that the actin paracrystal was not perfectly ordered. This sample-preparation and imaging procedure will allow for examination of structural features of stereocilia in a near-native state.
]]></description>
<dc:creator>Metlagel, Z.</dc:creator>
<dc:creator>Krey, J. F.</dc:creator>
<dc:creator>Song, J.</dc:creator>
<dc:creator>Swift, M. F.</dc:creator>
<dc:creator>Tivol, W. J.</dc:creator>
<dc:creator>Dumont, R. A.</dc:creator>
<dc:creator>Thai, J.</dc:creator>
<dc:creator>Chang, A.</dc:creator>
<dc:creator>Seifikar, H.</dc:creator>
<dc:creator>Volkmann, N.</dc:creator>
<dc:creator>Hanein, D.</dc:creator>
<dc:creator>Barr-Gillespie, P. G.</dc:creator>
<dc:creator>Auer, M.</dc:creator>
<dc:date>2018-12-13</dc:date>
<dc:identifier>doi:10.1101/496513</dc:identifier>
<dc:title><![CDATA[Electron cryo-tomography of vestibular hair-cell stereocilia]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-12-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/517813v1?rss=1">
<title>
<![CDATA[
HLA alleles associated with risk of ankylosing spondylitis and rheumatoid arthritis influence the gut microbiome 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/517813v1?rss=1</link>
<description><![CDATA[
ObjectivesHLA alleles affect susceptibility to more than 100 diseases, but the mechanisms to account for these genotype-disease associations are largely unknown. HLA-alleles strongly influence predisposition to ankylosing spondylitis (AS) and rheumatoid arthritis (RA). Both AS and RA patients have discrete intestinal and faecal microbiome signatures. Whether these changes are cause or consequence of the diseases themselves is unclear. To distinguish these possibilities, we examine the effect of HLA-B27 and HLA-DRB1 RA-risk alleles on the composition of the intestinal microbiome in healthy individuals.nnMethods568 samples from 6 intestinal sites were collected from 107 otherwise healthy unrelated subjects and stool samples from 696 twin pairs from the TwinsUK cohort. Microbiome profiling was performed using sequencing of the 16S rRNA bacterial marker gene. All patients were genotyped using the Illumina CoreExome SNP microarray, and HLA genotypes were imputed from these data.nnResultsAssociation was observed between HLA-B27 genotype, and RA-risk HLA-DRB1 alleles, and overall microbial composition (P=0.0002 and P=0.00001 respectively). These associations were replicated in the TwinsUK cohort stool samples (P=0.023 and P=0.033 respectively).nnConclusionsThis study shows that the changes in intestinal microbiome composition seen in AS and RA are at least partially due to effects of HLA-B27 and -DRB1 on the gut microbiome. These findings support the hypothesis that HLA alleles operate to cause or increase the risk of these diseases through interaction with the intestinal microbiome, and suggest that therapies targeting the microbiome may be effective in their prevention and/or treatment.
]]></description>
<dc:creator>Asquith, M.</dc:creator>
<dc:creator>Sternes, P. R.</dc:creator>
<dc:creator>Costello, M.-E.</dc:creator>
<dc:creator>Karstens, L.</dc:creator>
<dc:creator>Diamond, S.</dc:creator>
<dc:creator>Martin, T. M.</dc:creator>
<dc:creator>Spector, T. D.</dc:creator>
<dc:creator>le Cao, K.-A.</dc:creator>
<dc:creator>Rosenbaum, J. T.</dc:creator>
<dc:creator>Brown, M. A.</dc:creator>
<dc:date>2019-01-11</dc:date>
<dc:identifier>doi:10.1101/517813</dc:identifier>
<dc:title><![CDATA[HLA alleles associated with risk of ankylosing spondylitis and rheumatoid arthritis influence the gut microbiome]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-01-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/519231v1?rss=1">
<title>
<![CDATA[
Semantic Integration of Clinical Laboratory Tests from Electronic Health Records for Deep Phenotyping and Biomarker Discovery 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/519231v1?rss=1</link>
<description><![CDATA[
Electronic Health Record (EHR) systems typically define laboratory test results using the Laboratory Observation Identifier Names and Codes (LOINC) and can transmit them using Fast Healthcare Interoperability Resource (FHIR) standards. LOINC has not yet been semantically integrated with computational resources for phenotype analysis. Here, we provide a method for mapping LOINC-encoded laboratory test results transmitted in FHIR standards to the Human Phenotype Ontology (HPO) terms. We annotated the medical implications of 2421 commonly used laboratory tests with HPO terms. Using these annotations, a software assesses laboratory test results and converts each into an HPO term. We validated our approach with EHR data from 15,681 patients with respiratory complaints and identified known biomarkers for asthma. Finally, we provide a freely available SMART on FHIR application that can be used within EHR systems. Our approach allows reusing readily available laboratory tests in EHR for deep phenotyping and using the hierarchical structure of HPO for association studies with medical outcomes and genomics.nnOne Sentence SummaryWe present an approach to semantically integrating LOINC-encoded laboratory data with the Human Phenotype Ontology and show that the integrated LOINC data can be used to identify biomarkers for asthma from electronic health record data.
]]></description>
<dc:creator>Zhang, X. A.</dc:creator>
<dc:creator>Yates, A.</dc:creator>
<dc:creator>Vasilevsky, N.</dc:creator>
<dc:creator>Gourdine, J.</dc:creator>
<dc:creator>Carmody, L. C.</dc:creator>
<dc:creator>Danis, D.</dc:creator>
<dc:creator>Joachimiak, M. P.</dc:creator>
<dc:creator>Ravanmehr, V.</dc:creator>
<dc:creator>Pfaff, E. R.</dc:creator>
<dc:creator>Champion, J.</dc:creator>
<dc:creator>Robasky, K.</dc:creator>
<dc:creator>Xu, H.</dc:creator>
<dc:creator>Fecho, K.</dc:creator>
<dc:creator>Walton, N. A.</dc:creator>
<dc:creator>Zhu, R.</dc:creator>
<dc:creator>Ramsdill, J.</dc:creator>
<dc:creator>Mungall, C.</dc:creator>
<dc:creator>Kohler, S.</dc:creator>
<dc:creator>Haendel, M. A.</dc:creator>
<dc:creator>McDonald, C.</dc:creator>
<dc:creator>Vreeman, D. J.</dc:creator>
<dc:creator>Peden, D. B.</dc:creator>
<dc:creator>Chute, C. G.</dc:creator>
<dc:creator>Robinson, P. N.</dc:creator>
<dc:date>2019-01-13</dc:date>
<dc:identifier>doi:10.1101/519231</dc:identifier>
<dc:title><![CDATA[Semantic Integration of Clinical Laboratory Tests from Electronic Health Records for Deep Phenotyping and Biomarker Discovery]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-01-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/524934v1?rss=1">
<title>
<![CDATA[
Feeling left out or just surprised? Neural correlates of social exclusion and expectancy violations in adolescence 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/524934v1?rss=1</link>
<description><![CDATA[
Social belonging and affiliation are important human drives that impact decision-making and health outcomes. While neural responses to social exclusion are increasingly well-characterized, studies rarely identify whether these responses reflect the affective distress or expectancy violation associated with exclusion. The present study compares neural responses to exclusion and over-inclusion, a similarly unexpected social occurrence that violates fair play expectations, but does not involve rejection, with a focus on implications for models of dorsal anterior cingulate cortex (dACC) function. In an fMRI adaptation of the Cyberball paradigm with human adolescents 11.1-17.7 years of age (N=69), we employed parametric modulators to examine scaling of neural signal with cumulative exclusion and inclusion events. The use of parametric modulators overcomes arbitrary definitions of condition onsets and offsets imposed on fluid, continuous gameplay. In support of the notion that the dACC plays a role in processing affective distress, this region exhibited greater signal with cumulative exclusion than cumulative inclusion events. However, this difference was partly driven by diminished signal accompanying cumulative inclusion, a finding that does not conform to either affective distress or expectancy violation model predictions. Additionally, we found that signal in the insula and dorsomedial prefrontal cortex scaled uniquely with social exclusion and over-inclusion, respectively, while conjunction analyses revealed that the rostromedial prefrontal cortex and left intraparietal sulcus responded similarly to both conditions. These findings shed light on which neural regions may exhibit patterns of differential sensitivity to exclusion or over-inclusion, as well as those that are more broadly engaged by both types of social interaction.
]]></description>
<dc:creator>Cheng, T.</dc:creator>
<dc:creator>Vijayakumar, N.</dc:creator>
<dc:creator>Flournoy, J. C.</dc:creator>
<dc:creator>Op de Macks, Z.</dc:creator>
<dc:creator>Peake, S. J.</dc:creator>
<dc:creator>Flannery, J. E.</dc:creator>
<dc:creator>Mobasser, A.</dc:creator>
<dc:creator>Alberti, S. L.</dc:creator>
<dc:creator>Fisher, P. A.</dc:creator>
<dc:creator>Pfeifer, J. H.</dc:creator>
<dc:date>2019-01-20</dc:date>
<dc:identifier>doi:10.1101/524934</dc:identifier>
<dc:title><![CDATA[Feeling left out or just surprised? Neural correlates of social exclusion and expectancy violations in adolescence]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-01-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/570655v1?rss=1">
<title>
<![CDATA[
Altered structural brain asymmetry in autism spectrum disorder: large-scale analysis via the ENIGMA Consortium 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/570655v1?rss=1</link>
<description><![CDATA[
BackgroundLeft-right asymmetry is an important organizing feature of the healthy brain. Various studies have reported altered structural brain asymmetry in autism spectrum disorder (ASD). However, findings have been inconsistent, likely due to limited sample sizes and low statistical power.nnMethodsWe investigated 1,774 subjects with ASD and 1,809 controls, from 54 datasets, for differences in the asymmetry of thickness and surface area of 34 cerebral cortical regions. We also examined global hemispheric measures of cortical thickness and area asymmetry, and volumetric asymmetries of subcortical structures. Data were obtained via the ASD Working Group of the ENIGMA (Enhancing NeuroImaging Genetics through Meta-Analysis) consortium. T1-weighted MRI data were processed with a single protocol using FreeSurfer and the Desikan-Killiany atlas.nnResultsASD was significantly associated with reduced leftward asymmetry of total hemispheric average cortical thickness, compared to controls. Eight regional thickness asymmetries, distributed over the cortex, also showed significant associations with diagnosis after correction for multiple comparisons, for which asymmetry was again generally lower in ASD versus controls. In addition, the medial orbitofrontal surface area was less rightward asymmetric in ASD than controls, and the putamen volume was more leftward asymmetric in ASD than controls. The largest effect size had Cohens d = 0.15. Most effects did not depend on age, sex, IQ, or disorder severity.nnConclusionAltered lateralized neurodevelopment is suggested in ASD, affecting widespread cortical regions with diverse functions. Large-scale analysis was necessary to reliably detect, and accurately describe, subtle alterations of structural brain asymmetry in this disorder.
]]></description>
<dc:creator>Postema, M. C.</dc:creator>
<dc:creator>van Rooij, D.</dc:creator>
<dc:creator>Anagnostou, E.</dc:creator>
<dc:creator>Arango, C.</dc:creator>
<dc:creator>Auzias, G.</dc:creator>
<dc:creator>Behrmann, M.</dc:creator>
<dc:creator>Filho, G. B.</dc:creator>
<dc:creator>Calderoni, S.</dc:creator>
<dc:creator>Calvo, R.</dc:creator>
<dc:creator>Daly, E.</dc:creator>
<dc:creator>Deruelle, C.</dc:creator>
<dc:creator>Di Martino, A.</dc:creator>
<dc:creator>Dinstein, I.</dc:creator>
<dc:creator>Duran, F. L. S.</dc:creator>
<dc:creator>Durston, S.</dc:creator>
<dc:creator>Ecker, C.</dc:creator>
<dc:creator>Ehrlich, S.</dc:creator>
<dc:creator>Fair, D.</dc:creator>
<dc:creator>Fedor, J.</dc:creator>
<dc:creator>Feng, X.</dc:creator>
<dc:creator>Fitzgerald, J.</dc:creator>
<dc:creator>Floris, D. L.</dc:creator>
<dc:creator>Freitag, C. M.</dc:creator>
<dc:creator>Gallagher, L.</dc:creator>
<dc:creator>Glahn, D. C.</dc:creator>
<dc:creator>Gori, I.</dc:creator>
<dc:creator>Haar, S.</dc:creator>
<dc:creator>Hoekstra, L.</dc:creator>
<dc:creator>Jahansad, N.</dc:creator>
<dc:creator>Jalbrzikowski, M.</dc:creator>
<dc:creator>Janssen, J.</dc:creator>
<dc:creator>King, J. A.</dc:creator>
<dc:creator>Zaro, L. L.</dc:creator>
<dc:creator>Lerch, J. P.</dc:creator>
<dc:creator>Luna, B.</dc:creator>
<dc:creator>Martinho, M. M.</dc:creator>
<dc:creator>McGrath, J.</dc:creator>
<dc:creator>Medland, S. E.</dc:creator>
<dc:creator>Muratori, F.</dc:creator>
<dc:creator>Murphy, C. M.</dc:creator>
<dc:creator>Murphy, D. G. M.</dc:creator>
<dc:creator>O'Hearn, K.</dc:creator>
<dc:creator>Oranje, B.</dc:creator>
<dc:creator>Parellada,</dc:creator>
<dc:date>2019-03-09</dc:date>
<dc:identifier>doi:10.1101/570655</dc:identifier>
<dc:title><![CDATA[Altered structural brain asymmetry in autism spectrum disorder: large-scale analysis via the ENIGMA Consortium]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-03-09</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/10.1101/633941v1?rss=1">
<title>
<![CDATA[
Somatodendritic HCN channels in hippocampal OLM cells revealed by a convergence of computational models and experiments 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/633941v1?rss=1</link>
<description><![CDATA[
Determining details of spatially extended neurons is a challenge that needs to be overcome. The oriens-lacunosum/moleculare (OLM) interneuron has been implicated as a critical controller of hippocampal memory making it essential to understand how its biophysical properties contribute to function. We previously used computational models to show that OLM cells exhibit theta spiking resonance frequencies that depend on their dendrites having hyperpolarization-activated cation channels (h-channels). However, whether OLM cells have dendritic h-channels is unknown. We performed a set of whole-cell recordings of OLM cells from mouse hippocampus and constructed multi-compartment models using morphological and electrophysiological parameters extracted from the same cell. The models matched experiments only when dendritic h-channels were present. Immunohistochemical localization of the HCN2 subunit confirmed dendritic expression. These models can be used to obtain insight into hippocampal function. Our work shows that a tight integration of model and experiment tackles the challenge of characterizing spatially extended neurons.
]]></description>
<dc:creator>Sekulic, V.</dc:creator>
<dc:creator>Lawrence, J.</dc:creator>
<dc:creator>Guet-McCreight, A.</dc:creator>
<dc:creator>Yi, F.</dc:creator>
<dc:creator>Garrett, T.</dc:creator>
<dc:creator>Wang, R.</dc:creator>
<dc:creator>Liu, X.</dc:creator>
<dc:creator>Lopez, Y.</dc:creator>
<dc:creator>Solis-Wheeler, M.</dc:creator>
<dc:creator>Skinner, F. K.</dc:creator>
<dc:date>2019-05-13</dc:date>
<dc:identifier>doi:10.1101/633941</dc:identifier>
<dc:title><![CDATA[Somatodendritic HCN channels in hippocampal OLM cells revealed by a convergence of computational models and experiments]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-05-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/647321v1?rss=1">
<title>
<![CDATA[
Pseudomonas isolates degrade and form biofilms on polyethylene terephthalate (PET) plastic 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/647321v1?rss=1</link>
<description><![CDATA[
Bioaugmentation is a possible remediation strategy for the massive amounts of plastic waste in our oceans and landfills. For this study, soil samples were collected from petroleum polluted locations in the Houston, Texas area to isolate microorganisms capable of plastic degradation. Bacteria were propagated and screened for lipase activity, which has been associated with the bacterial degradation of some plastics to date. We identified three lipase-positive Pseudomonas species, and Bacillus cereus as part of two consortia, which we predict enhances biofilm formation and plastic degradation. Lipase-positive consortia bacteria were incubated alongside blank and E.coli controls with UV-irradiated polyethylene terephthalate (PET), high-density polyethylene (HDPE), or low-density polyethylene (LDPE) as sole sources of carbon. Surface degradation of PET plastic was quantified by changes in molecular vibrations by infrared spectroscopy. The bacteria formed biofilms on PET, observed by scanning electron microscopy, and induced molecular changes on the plastic surface, indicating the initial stages of plastic degradation. We also found molecular evidence that one of the Pseudomonas isolates degrades LDPE. To date, lipase positive Pseudomonas spp. degradation of PET has not been well described, and this work highlights the potential for using consortia of common soil bacteria to degrade plastic waste.
]]></description>
<dc:creator>Vague, M.</dc:creator>
<dc:creator>Chan, G.</dc:creator>
<dc:creator>Roberts, C.</dc:creator>
<dc:creator>Swartz, N.</dc:creator>
<dc:creator>Mellies, J.</dc:creator>
<dc:date>2019-05-24</dc:date>
<dc:identifier>doi:10.1101/647321</dc:identifier>
<dc:title><![CDATA[Pseudomonas isolates degrade and form biofilms on polyethylene terephthalate (PET) plastic]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-05-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/692616v1?rss=1">
<title>
<![CDATA[
Cross-species Functional Alignment Reveals Evolutionary Hierarchy Within the Connectome 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/692616v1?rss=1</link>
<description><![CDATA[
Evolution provides an important window into how cortical organization shapes function and vice versa. The complex mosaic of changes in brain morphology and functional organization that have shaped the mammalian cortex during evolution, complicates attempts to chart cortical differences across species. It limits our ability to fully appreciate how evolution has shaped our brain, especially in systems associated with unique human cognitive capabilities that lack anatomical homologues in other species. Here, we demonstrate a function-based method for cross-species cortical alignment that leverages recent advances in understanding cortical organization and that enables the quantification of homologous regions across species, even when their location is decoupled from anatomical landmarks. Critically, our method establishes that cross-species similarity in cortical organization decreases with geodesic distance from unimodal systems, and culminates in the most pronounced changes in posterior regions of the default network (angular gyrus, posterior cingulate and middle temporal cortices). Our findings suggest that the establishment of the default network, as the apex of a cognitive hierarchy, as is seen in humans, is a relatively recent evolutionary adaptation. They also highlight functional changes in regions such as the posterior cingulate cortex and angular gyrus as key influences on uniquely human features of cognition.
]]></description>
<dc:creator>Xu, T.</dc:creator>
<dc:creator>Nenning, K.-H.</dc:creator>
<dc:creator>Schwartz, E.</dc:creator>
<dc:creator>Hong, S.-J.</dc:creator>
<dc:creator>Vogelstein, J. T.</dc:creator>
<dc:creator>Fair, D. A.</dc:creator>
<dc:creator>Schroeder, C. E.</dc:creator>
<dc:creator>Margulies, D. S.</dc:creator>
<dc:creator>Smallwood, J.</dc:creator>
<dc:creator>Milham, M. P.</dc:creator>
<dc:creator>Langs, G.</dc:creator>
<dc:date>2019-07-04</dc:date>
<dc:identifier>doi:10.1101/692616</dc:identifier>
<dc:title><![CDATA[Cross-species Functional Alignment Reveals Evolutionary Hierarchy Within the Connectome]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-07-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/871608v1?rss=1">
<title>
<![CDATA[
Effective buprenorphine use and tapering strategies: Endorsements and insights by people in recovery from opioid use disorder on a Reddit forum 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/871608v1?rss=1</link>
<description><![CDATA[
Opioid use disorder (OUD) is a public health emergency in the United States. Over 47,000 overdose-related deaths in 2017 involved opioids. Medication-assisted treatment (MAT), in particular, buprenorphine and buprenorphine combination products such as Suboxone(R), is the most effective, evidence-based treatment for OUD. However, there are a limited number of conclusive scientific studies that provide guidance to medical professionals about strategies for using buprenorphine to achieve stable recovery. In this study, we used data-driven natural language processing methods to mine a total of 16,146 posts about buprenorphine from 1933 unique users on the anonymous social network Reddit. Analysis of a sample of these posts showed that 74% of the posts described users personal experiences and that the top three topics included advice on using Suboxone(R) (55.0%), Suboxone(R) dosage information (35.5%) and information about Suboxone(R) tapering (32.0%). Based on two models, one that incorporated  upvoting by other members and one that did not, we found that Reddit users reported more successful recovery with longer tapering schedules, particularly from 2.0 mg to 0.0 mg (median: 93 days; mean: 95 days), as compared to shorter tapering schedules investigated in past clinical trials. Diarrhea, insomnia, restlessness, and fatigue were commonly reported adverse events. Physical exercise, clonidine, and Imodium(R) were frequently reported to help during the recovery process. Due to the difficulties of conducting longer-term clinical trials involving patients with OUD, clinicians should consider other information sources including peer discussions from the abundant, real-time information available on Reddit.

Significance StatementOpioid use disorder (OUD) is a national crisis in the United States and buprenorphine is one of the most effective evidence-based treatments. However, few studies have explored successful strategies for using and tapering buprenorphine to achieve stable recovery, particularly due to the difficulties of conducting long-term studies involving patients with OUD. In this study, we show that discussions on the anonymous social network Reddit may be leveraged, via automatic text mining methods, to discover successful buprenorphine use and tapering strategies. We discovered that longer tapering schedules, compared to those investigated in past clinical trials, may lead to (self-reported) sustained recovery. Furthermore, Reddit posts also provide key information regarding buprenorphine withdrawal, cravings, adjunct medications for withdrawal symptoms and relapse prevention strategies.
]]></description>
<dc:creator>Graves, R. L.</dc:creator>
<dc:creator>Sarker, A.</dc:creator>
<dc:creator>Al-Garadi, M. A.</dc:creator>
<dc:creator>Yang, Y.-c.</dc:creator>
<dc:creator>Love, J. S.</dc:creator>
<dc:creator>O'Connor, K.</dc:creator>
<dc:creator>Gonzalez-Hernandez, G.</dc:creator>
<dc:creator>Perrone, J.</dc:creator>
<dc:date>2019-12-11</dc:date>
<dc:identifier>doi:10.1101/871608</dc:identifier>
<dc:title><![CDATA[Effective buprenorphine use and tapering strategies: Endorsements and insights by people in recovery from opioid use disorder on a Reddit forum]]></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/10.1101/2023.05.15.539065v1?rss=1">
<title>
<![CDATA[
Comprehensively defining cellular specializations for initiating parallel auditory pathways in the mouse cochlear nucleus 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.05.15.539065v1?rss=1</link>
<description><![CDATA[
The cochlear nuclear complex (CN), the starting point for all central auditory processing, comprises a suite of neuronal cell types that are highly specialized for neural coding of acoustic signals, yet molecular logic governing cellular specializations remains unknown. By combining single-nucleus RNA sequencing and Patch-seq analysis, we reveal a set of transcriptionally distinct cell populations encompassing all previously observed types and discover multiple new subtypes with anatomical and physiological identity. The resulting comprehensive cell-type taxonomy reconciles anatomical position, morphological, physiological, and molecular criteria, enabling the determination of the molecular basis of the remarkable cellular phenotypes in the CN. In particular, CN cell-type identity is encoded in a transcriptional architecture that orchestrates functionally congruent expression across a small set of gene families to customize projection patterns, input-output synaptic communication, and biophysical features required for encoding distinct aspects of acoustic signals. This high-resolution account of cellular heterogeneity from the molecular to the circuit level illustrates molecular logic for cellular specializations and enables genetic dissection of auditory processing and hearing disorders with unprecedented specificity.
]]></description>
<dc:creator>Jing, J.</dc:creator>
<dc:creator>Hu, M.</dc:creator>
<dc:creator>Ngodup, T.</dc:creator>
<dc:creator>Ma, Q.</dc:creator>
<dc:creator>Lau, S. N. N.</dc:creator>
<dc:creator>Ljungberg, C.</dc:creator>
<dc:creator>McGinley, M. J.</dc:creator>
<dc:creator>Trussell, L. O.</dc:creator>
<dc:creator>Jiang, X.</dc:creator>
<dc:date>2023-05-15</dc:date>
<dc:identifier>doi:10.1101/2023.05.15.539065</dc:identifier>
<dc:title><![CDATA[Comprehensively defining cellular specializations for initiating parallel auditory pathways in the mouse cochlear nucleus]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-05-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.05.17.541057v1?rss=1">
<title>
<![CDATA[
Recovery of a learned behavior despite partial restoration of neuronal dynamics after chronic inactivation of inhibitory neurons 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.05.17.541057v1?rss=1</link>
<description><![CDATA[
Maintaining motor behaviors throughout life is crucial for an individuals survival and reproductive success. The neuronal mechanisms that preserve behavior are poorly understood. To address this question, we focused on the zebra finch, a bird that produces a highly stereotypical song after learning it as a juvenile. Using cell-specific viral vectors, we chronically silenced inhibitory neurons in the pre-motor song nucleus called the high vocal center (HVC), which caused drastic song degradation. However, after producing severely degraded vocalizations for around 2 months, the song rapidly improved, and animals could sing songs that highly resembled the original. In adult birds, single-cell RNA sequencing of HVC revealed that silencing interneurons elevated markers for microglia and increased expression of the Major Histocompatibility Complex I (MHC I), mirroring changes observed in juveniles during song learning. Interestingly, adults could restore their songs despite lesioning the lateral magnocellular nucleus of the anterior neostriatum (LMAN), a brain nucleus crucial for juvenile song learning. This suggests that while molecular mechanisms may overlap, adults utilize different neuronal mechanisms for song recovery. Chronic and acute electrophysiological recordings within HVC and its downstream target, the robust nucleus of the archistriatum (RA), revealed that neuronal activity in the circuit permanently altered with higher spontaneous firing in RA and lower in HVC compared to control even after the song had fully recovered. Together, our findings show that a complex learned behavior can recover despite extended periods of perturbed behavior and permanently altered neuronal dynamics. These results show that loss of inhibitory tone can be compensated for by recovery mechanisms partly local to the perturbed nucleus and do not require circuits necessary for learning.
]]></description>
<dc:creator>Torok, Z.</dc:creator>
<dc:creator>Luebbert, L.</dc:creator>
<dc:creator>Feldman, J.</dc:creator>
<dc:creator>Duffy, A.</dc:creator>
<dc:creator>Nevue, A.</dc:creator>
<dc:creator>Wongso, S.</dc:creator>
<dc:creator>Mello, C.</dc:creator>
<dc:creator>Fairhall, A.</dc:creator>
<dc:creator>Pachter, L.</dc:creator>
<dc:creator>Gonzalez, W.</dc:creator>
<dc:creator>Lois, C.</dc:creator>
<dc:date>2023-05-18</dc:date>
<dc:identifier>doi:10.1101/2023.05.17.541057</dc:identifier>
<dc:title><![CDATA[Recovery of a learned behavior despite partial restoration of neuronal dynamics after chronic inactivation of inhibitory neurons]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-05-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.05.18.540717v1?rss=1">
<title>
<![CDATA[
Human Herpesvirus 8 ORF57 protein is able to reduce TDP-43 pathology: Network analysis Identifies Interacting Pathways 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.05.18.540717v1?rss=1</link>
<description><![CDATA[
Aggregation of TAR DNA-binding protein 43kDa (TDP-43) is thought to drive the pathophysiology of ALS and some Frontotemporal dementias. TDP-43 is normally a nuclear protein that in neurons translocates to the cytoplasm and forms insoluble aggregates upon activation of the integrated stress response (ISR). Viruses evolved to control the ISR. In the case of Herpesvirus 8, the protein ORF57 acts to bind protein kinase R, inhibit phosphorylation of eIF2 and reduce activation of the ISR. We hypothesized that ORF57 might also possess the ability to inhibit aggregation of TDP-43. ORF57 was expressed in the neuronal SH-SY5Y line and its effects on TDP-43 aggregation characterized. We report that ORF57 inhibits TDP-43 aggregation by 55% and elicits a 2.45-fold increase in the rate of dispersion of existing TDP-43 granules. These changes were associated with a 50% decrease in cell death. Proteomic studies were carried out to identify the protein interaction network of ORF57. We observed that ORF57 directly binds to TDP-43 as well as interacts with many components of the ISR, including elements of the proteostasis machinery known to reduce TDP-43 aggregation. We propose that viral proteins designed to inhibit a chronic ISR can be engineered to remove aggregated proteins and dampen a chronic ISR.
]]></description>
<dc:creator>Webber, C. J.</dc:creator>
<dc:creator>Murphy, C. N.</dc:creator>
<dc:creator>Rondon-Ortiz, A. N.</dc:creator>
<dc:creator>van der Spek, S. J.</dc:creator>
<dc:creator>Kelly, E. X.</dc:creator>
<dc:creator>Lampl, N. M.</dc:creator>
<dc:creator>Chiesa, G.</dc:creator>
<dc:creator>Khalil, A. S.</dc:creator>
<dc:creator>Emili, A.</dc:creator>
<dc:creator>Wolozin, B.</dc:creator>
<dc:date>2023-05-18</dc:date>
<dc:identifier>doi:10.1101/2023.05.18.540717</dc:identifier>
<dc:title><![CDATA[Human Herpesvirus 8 ORF57 protein is able to reduce TDP-43 pathology: Network analysis Identifies Interacting Pathways]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-05-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.05.18.541362v1?rss=1">
<title>
<![CDATA[
Transcriptional profiling of mucus production and modification in rhesus macaque endocervical cells under hormonal regulation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.05.18.541362v1?rss=1</link>
<description><![CDATA[
ObjectiveEndocervical mucus production is a key regulator of fertility throughout the menstrual cycle. With cycle-dependent variability in mucus quality and quantity, cervical mucus can either facilitate or block sperm ascension into the upper female reproductive tract. This study seeks to identify genes involved in the hormonal regulation of mucus production, modification, and regulation through profiling the transcriptome of endocervical cells from the non-human primate, the Rhesus Macaque (Macaca mulatta).

DesignExperimental.

SettingTranslational science laboratory.

InterventionWe treated differentiated primary endocervical cultures with estradiol (E2) and progesterone (P4) to mimic peri-ovulatory and luteal-phase hormonal changes. Using RNA-sequencing, we identified differential expression of gene pathways and mucus producing and modifying genes in cells treated with E2 compared to hormone-free conditions and E2 compared to E2-primed cells treated with P4.

Main Outcome MeasuresWe pursued differential gene expression analysis on RNA-sequenced cells. Sequence validation was done using qPCR.

ResultsOur study identified 158 genes that show significant differential expression in E2-only conditions compared to hormone-free control, and 250 genes that show significant differential expression in P4-treated conditions compared to E2-only conditions. From this list, we found hormone-induced changes in transcriptional profiles for genes across several classes of mucus production, including ion channels and enzymes involved in post-translational mucin modification that have not previously been described as hormonally regulated.

ConclusionOur study is the first to use an in vitro culture system to create an epithelial-cell specific transcriptome of the endocervix. As a result, our study identifies new genes and pathways that are altered by sex-steroids in cervical mucus production.
]]></description>
<dc:creator>Rapp, K.</dc:creator>
<dc:creator>Wei, S.</dc:creator>
<dc:creator>Roberts, M.</dc:creator>
<dc:creator>Yao, S.</dc:creator>
<dc:creator>Fei, S.</dc:creator>
<dc:creator>Gao, L.</dc:creator>
<dc:creator>Ray, K.</dc:creator>
<dc:creator>Wang, A.</dc:creator>
<dc:creator>Godiah, R.</dc:creator>
<dc:creator>Han, L.</dc:creator>
<dc:date>2023-05-18</dc:date>
<dc:identifier>doi:10.1101/2023.05.18.541362</dc:identifier>
<dc:title><![CDATA[Transcriptional profiling of mucus production and modification in rhesus macaque endocervical cells under hormonal regulation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-05-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.05.17.541249v1?rss=1">
<title>
<![CDATA[
Computational modeling of methylation impact of AML drivers reveals new pathways and refines AML risk-stratification 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.05.17.541249v1?rss=1</link>
<description><![CDATA[
Decades before its clinical onset, epigenetic changes start to accumulate in the progenitor cells of Acute Myelogenous Leukemia (AML). Delineating these changes can improve risk-stratification for patients and shed insights into AML etiology, dynamics and mechanisms. Towards this goal, we extracted "epigenetic signatures" through two parallel machine learning approaches: a supervised regression model using frequently mutated genes as labels and an unsupervised topic modeling approach to factorize covarying epigenetic changes into a small number of "topics". First, we created regression models for DNMT3A and TET2, the two most frequently mutated epigenetic drivers in AML. Our model differentiated wild-type vs. mutant genotypes based on their downstream epigenetic impacts with very high accuracy: AUROC 0.9 and 0.8, respectively. Methylation loci frequently selected by the models recapitulated known downstream pathways and identified several novel recurrent targets. Second, we used topic modeling to systematically factorize the high dimensional methylation profiles to a latent space of 15 topics. We annotated identified topics with biological and clinical features such as mutation status, prior malignancy and ELN criteria. Topic modeling successfully deconvoluted the combined effects of multiple upstream epigenetic drivers into individual topics including relatively infrequent cytogenetic events, improving the methylation-based subtyping of AML. Furthermore, they revealed complimentary and synergistic interactions between drivers, grouped them based on the similarity of their downstream methylation impact and linked them to prognostic criteria. Our models identify new signatures and methylation pathways, refine risk-stratification and inform detection and drug response studies for AML patients.

KEY POINTSO_LISupervised and unsupervised models reveal new methylation pathways of AML driver events and validate previously known associations.
C_LIO_LIIndividual DNMT3A and TET2 signatures are accurate and robust, despite the complex genetic and epigenetic make-up of samples at diagnosis.
C_LIO_LIUnsupervised topic modeling factorizes covarying methylation changes and isolates methylation signatures caused by rare mutations.
C_LIO_LITopic modeling reveals a group of mutations with similar downstream methylation impacts and mapped to adverse-risk class by ELN.
C_LIO_LITopic modeling uncovers methylation signatures of infrequent cytogenetic events, significantly improving methylation-based subtyping.
C_LIO_LIOur models can be leveraged to build predictive models for AML-risk.
C_LIO_LIOur models show that cytogenetic events, such as t(15;17) have widespread trans downstream methylation impacts.
C_LI
]]></description>
<dc:creator>Gurun, B.</dc:creator>
<dc:creator>Tyner, J. W.</dc:creator>
<dc:creator>Demir, E.</dc:creator>
<dc:creator>Druker, B. J.</dc:creator>
<dc:creator>Spellman, P. T.</dc:creator>
<dc:date>2023-05-21</dc:date>
<dc:identifier>doi:10.1101/2023.05.17.541249</dc:identifier>
<dc:title><![CDATA[Computational modeling of methylation impact of AML drivers reveals new pathways and refines AML risk-stratification]]></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/10.1101/2023.05.22.540988v1?rss=1">
<title>
<![CDATA[
Muscle-specific MEF2D alpha 2 isoform regulates mice exercise capacity, muscle ketolysis, and systemic ketone body availability in mice 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.05.22.540988v1?rss=1</link>
<description><![CDATA[
Ketone bodies are an alternate fuel source generated by the liver in response to low carbohydrate availability in neonates and after starvation and exhausting exercise in adulthood. The postnatal alternative splicing generates a highly conserved muscle-specific MEF2D2 protein isoform of the transcription factor MEF2D. Here, we discovered that compared to WT mice, MEF2D2 exon knockout (Eko) mice displayed reduced running capacity and muscle expression of all three ketolytic genes, BDH1, OXCT1, and ACAT1. Consistent with reduced muscle utilization of ketone bodies, MEF2D2 Eko mice also showed increased ketone body levels in a tolerance test, after exercise, and upon feeding a ketogenic diet. Lastly, using mitochondria isolated from skeletal muscle, we showed reduced ketone body utilization and respiration in Eko compared to WT mice. Thus, we identified a new role of MEF2D2 protein isoform in regulating skeletal muscle ketone body oxidation, exercise capacity, and its effect on systemic ketone body levels.
]]></description>
<dc:creator>Kumar, S.</dc:creator>
<dc:creator>Xiangnan, G.</dc:creator>
<dc:creator>Mis, B.</dc:creator>
<dc:creator>Iqbal, H.</dc:creator>
<dc:creator>Kumar, S.</dc:creator>
<dc:creator>Besler, J.</dc:creator>
<dc:creator>Xia, Z.</dc:creator>
<dc:creator>Singh, R. K.</dc:creator>
<dc:date>2023-05-22</dc:date>
<dc:identifier>doi:10.1101/2023.05.22.540988</dc:identifier>
<dc:title><![CDATA[Muscle-specific MEF2D alpha 2 isoform regulates mice exercise capacity, muscle ketolysis, and systemic ketone body availability in mice]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-05-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.05.23.541932v1?rss=1">
<title>
<![CDATA[
Accelerated prime-and-trap vaccine regimen in mice using repRNA-based CSP malaria vaccine 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.05.23.541932v1?rss=1</link>
<description><![CDATA[
Malaria, caused by Plasmodium parasites, remains one of the most devastating infectious diseases worldwide, despite control efforts that have lowered morbidity and mortality. The only P. falciparum vaccine candidates to show field efficacy are those targeting the asymptomatic pre-erythrocytic (PE) stages of infection. The subunit (SU) RTS,S/AS01 vaccine, the only licensed malaria vaccine to date, is only modestly effective against clinical malaria. Both RTS,S/AS01 and the SU R21 vaccine candidate target the PE sporozoite (spz) circumsporozoite (CS) protein. These candidates elicit high-titer antibodies that provide short-term protection from disease, but do not induce the liver-resident memory CD8+ T cells (Trm) that confer strong PE immunity and long-term protection. In contrast, whole-organism (WO) vaccines, employing for example radiation-attenuated spz (RAS), elicit both high antibody titers and Trm, and have achieved high levels of sterilizing protection. However, they require multiple intravenous (IV) doses, which must be administered at intervals of several weeks, complicating mass administration in the field. Moreover, the quantities of spz required present production difficulties. To reduce reliance on WO while maintaining protection via both antibodies and Trm responses, we have developed an accelerated vaccination regimen that combines two distinct agents in a prime-and-trap strategy. While the priming dose is a self-replicating RNA encoding P. yoelii CS protein, delivered via an advanced cationic nanocarrier (LIONTM), the trapping dose consists of WO RAS. This accelerated regime confers sterile protection in the P. yoelii mouse model of malaria. Our approach presents a clear path to late-stage preclinical and clinical testing of dose-sparing, same-day regimens that can confer sterilizing protection against malaria.
]]></description>
<dc:creator>MacMillen, Z.</dc:creator>
<dc:creator>Hatzakis, K.</dc:creator>
<dc:creator>Simpson, A.</dc:creator>
<dc:creator>Shears, M. J.</dc:creator>
<dc:creator>Watson, F.</dc:creator>
<dc:creator>Erasmus, J.</dc:creator>
<dc:creator>Khandhar, A. P.</dc:creator>
<dc:creator>Wilder, B. K.</dc:creator>
<dc:creator>Murphy, S. C.</dc:creator>
<dc:creator>Reed, S. G.</dc:creator>
<dc:creator>Davie, J. W.</dc:creator>
<dc:creator>Avril, M.</dc:creator>
<dc:date>2023-05-23</dc:date>
<dc:identifier>doi:10.1101/2023.05.23.541932</dc:identifier>
<dc:title><![CDATA[Accelerated prime-and-trap vaccine regimen in mice using repRNA-based CSP malaria vaccine]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-05-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.05.28.542637v1?rss=1">
<title>
<![CDATA[
Adaptor protein complex 2 in the orbitofrontal cortexpredicts alcohol use disorder 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.05.28.542637v1?rss=1</link>
<description><![CDATA[
Alcohol use disorder (AUD) is a life-threatening disease characterized by compulsive drinking, cognitive deficits, and social impairment that continue despite negative consequences. The inability of individuals with AUD to regulate drinking may involve functional deficits in cortical areas that normally balance actions that have aspects of both reward and risk. Among these, the orbitofrontal cortex (OFC) is critically involved in goal-directed behavior and is thought to maintain a representation of reward value that guides decision making. In the present study, we analyzed post-mortem OFC brain samples collected from age- and sex-matched control subjects and those with AUD using proteomics, bioinformatics, machine learning, and reverse genetics approaches. Of the 4,500+ total unique proteins identified in the proteomics screen, there were 47 proteins that differed significantly by sex that were enriched in processes regulating extracellular matrix and axonal structure. Gene ontology enrichment analysis revealed that proteins differentially expressed in AUD cases were involved in synaptic and mitochondrial function, as well as transmembrane transporter activity. Alcohol-sensitive OFC proteins also mapped to abnormal social behaviors and social interactions. Machine learning analysis of the post-mortem OFC proteome revealed dysregulation of presynaptic (e.g., AP2A1) and mitochondrial proteins that predicted the occurrence and severity of AUD. Using a reverse genetics approach to validate a target protein, we found that prefrontal Ap2a1 expression significantly correlated with voluntary alcohol drinking in male and female genetically diverse mouse strains. Moreover, recombinant inbred strains that inherited the C57BL/6J allele at the Ap2a1 interval consumed higher amounts of alcohol than those that inherited the DBA/2J allele. Together, these findings highlight the impact of excessive alcohol consumption on the human OFC proteome and identify important cross-species cortical mechanisms and proteins that control drinking in individuals with AUD.
]]></description>
<dc:creator>Mulholland, P. J.</dc:creator>
<dc:creator>Berto, S.</dc:creator>
<dc:creator>Wilmarth, P.</dc:creator>
<dc:creator>McMahan, C.</dc:creator>
<dc:creator>Ball, L.</dc:creator>
<dc:creator>Woodward, J.</dc:creator>
<dc:date>2023-05-30</dc:date>
<dc:identifier>doi:10.1101/2023.05.28.542637</dc:identifier>
<dc:title><![CDATA[Adaptor protein complex 2 in the orbitofrontal cortexpredicts alcohol use disorder]]></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/10.1101/2023.05.30.542954v1?rss=1">
<title>
<![CDATA[
N-glycans show distinct spatial distribution in mouse brain 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.05.30.542954v1?rss=1</link>
<description><![CDATA[
Protein N-linked glycosylation is a ubiquitous modification in the secretory pathway that plays a critical role in the development and function of the brain. N-glycans have a distinct composition and undergo tight regulation in the brain, but the spatial distribution of these structures remains relatively unexplored. Here, we systematically employed carbohydrate binding lectins with differing specificities to various classes of N-glycans and appropriate controls to identify multiple regions of the mouse brain. Lectins binding high-mannose-type N-glycans, the most abundant class of brain N-glycans, showed diffuse staining with some punctate structures observed on high magnification. Lectins binding specific motifs of complex N-glycans, including fucose and bisecting GlcNAc, showed more partitioned labeling, including to the synapse-rich molecular layer of the cerebellum. Understanding the distribution of N-glycans across the brain will aid future studies of these critical protein modifications in development and disease of the brain.
]]></description>
<dc:creator>Noel, M.</dc:creator>
<dc:creator>Cummings, R. D.</dc:creator>
<dc:creator>Mealer, R. G.</dc:creator>
<dc:date>2023-05-31</dc:date>
<dc:identifier>doi:10.1101/2023.05.30.542954</dc:identifier>
<dc:title><![CDATA[N-glycans show distinct spatial distribution in mouse brain]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-05-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.05.31.543132v1?rss=1">
<title>
<![CDATA[
Ribosomal protein control of hematopoietic stem cell transformation through direct, non-canonical regulation of metabolism 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.05.31.543132v1?rss=1</link>
<description><![CDATA[
We report here that expression of the ribosomal protein, RPL22, is frequently reduced in human myelodysplastic syndrome (MDS) and acute myelogenous leukemia (AML); and, reduced RPL22 expression is associated with worse outcomes. Mice null for Rpl22 display characteristics of an MDS-like syndrome and develop leukemia at an accelerated rate. Rpl22-deficient mice also display enhanced hematopoietic stem cell (HSC) self-renewal and obstructed differentiation potential, which arises not from reduced protein synthesis but from altered metabolism including increased fatty acid oxidation (FAO) and a striking induction of the stemness factor Lin28b in the resulting leukemia. Lin28b promotes a substantial increase in lipid content, upon which the survival of Rpl22-deficient leukemias depends. Altogether, these findings reveal that Rpl22 insufficiency enhances the leukemia potential of HSC through regulation of FAO and promotes leukemogenesis through Lin28b promotion of lipid synthesis.

HighlightsO_LIRPL22 insufficiency is observed in MDS/AML and is associated with reduced survival
C_LIO_LIRpl22-deficiency produces an MDS-like syndrome and facilitates leukemogenesis
C_LIO_LIRpl22-deficiency does not impair global protein synthesis by HSC
C_LIO_LIRpl22 controls leukemia survival through control of lipid synthesis
C_LI

eTOC: Rpl22 controls the function and transformation potential of hematopoietic stem cells through regulation of lipid metabolism.
]]></description>
<dc:creator>Harris, B.</dc:creator>
<dc:creator>Singh, D. K.</dc:creator>
<dc:creator>Verma, M.</dc:creator>
<dc:creator>Fahl, S. P.</dc:creator>
<dc:creator>Rhodes, M.</dc:creator>
<dc:creator>Sprinkle, S. R.</dc:creator>
<dc:creator>Wang, M.</dc:creator>
<dc:creator>Zhang, Y.</dc:creator>
<dc:creator>Perrigoue, J.</dc:creator>
<dc:creator>Kessel, R.</dc:creator>
<dc:creator>Peri, S.</dc:creator>
<dc:creator>West, J.</dc:creator>
<dc:creator>Giricz, O.</dc:creator>
<dc:creator>Boultwood, J.</dc:creator>
<dc:creator>Pellagatti, A.</dc:creator>
<dc:creator>Ramesh, K.</dc:creator>
<dc:creator>Montangna, C.</dc:creator>
<dc:creator>Pradhan, K.</dc:creator>
<dc:creator>Tyner, J.</dc:creator>
<dc:creator>Kennedy, B. K.</dc:creator>
<dc:creator>Holinstat, M.</dc:creator>
<dc:creator>Steidl, U.</dc:creator>
<dc:creator>Sykes, S.</dc:creator>
<dc:creator>Verma, A.</dc:creator>
<dc:creator>Wiest, D. L.</dc:creator>
<dc:date>2023-06-01</dc:date>
<dc:identifier>doi:10.1101/2023.05.31.543132</dc:identifier>
<dc:title><![CDATA[Ribosomal protein control of hematopoietic stem cell transformation through direct, non-canonical regulation of metabolism]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-06-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.06.02.543485v1?rss=1">
<title>
<![CDATA[
Small regulatory RNAs are mediators of the Streptococcus mutans SloR regulon 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.06.02.543485v1?rss=1</link>
<description><![CDATA[
Dental caries is among the most prevalent chronic infectious diseases worldwide. Streptococcus mutans, the chief causative agent of caries, uses a 25 kDa manganese dependent SloR protein to coordinate the uptake of essential manganese with the transcription of its virulence attributes. Small non-coding RNAs (sRNAs) can either enhance or repress gene expression and reports in the literature ascribe an emerging role for sRNAs in the environmental stress response. Herein, we identify 18-50 nt sRNAs as mediators of the S. mutans SloR and manganese regulons. Specifically, the results of sRNA-seq revealed 56 sRNAs in S. mutans that were differentially transcribed in the SloR-proficient UA159 and SloR-deficient GMS584 strains, and 109 sRNAs that were differentially expressed in UA159 cells grown in the presence of low versus high manganese. We describe SmsR1532 and SmsR1785 as SloR- and/or manganese-responsive sRNAs that are processed from large transcripts, and that bind SloR directly in their promoter regions. The predicted targets of these sRNAs include regulators of metal ion transport, growth management via a toxin-antitoxin operon, and oxidative stress tolerance. These findings support a role for sRNAs in coordinating intracellular metal ion homeostasis with virulence gene control in an important oral cariogen.

IMPORTANCESmall regulatory RNAs (sRNAs) are critical mediators of environmental signaling, particularly in bacterial cells under stress, but their role in Streptococcus mutans is poorly understood. S. mutans, the principal causative agent of dental caries, uses a 25 kDa manganese-dependent protein, called SloR, to coordinate the regulated uptake of essential metal ions with the transcription of its virulence genes. In the present study, we identified and characterize sRNAs that are both SloR- and manganese-responsive. Taken together, this research can elucidate the details of regulatory networks that engage sRNAs in an important oral pathogen, and that can enable the development of an effective anti-caries therapeutic.
]]></description>
<dc:creator>Drummond, I. Y.</dc:creator>
<dc:creator>DePaolo, A.</dc:creator>
<dc:creator>Krieger, M.</dc:creator>
<dc:creator>Driscoll, H. E.</dc:creator>
<dc:creator>Eckstrom, K.</dc:creator>
<dc:creator>Spatafora, G. A.</dc:creator>
<dc:date>2023-06-03</dc:date>
<dc:identifier>doi:10.1101/2023.06.02.543485</dc:identifier>
<dc:title><![CDATA[Small regulatory RNAs are mediators of the Streptococcus mutans SloR regulon]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-06-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.06.05.543335v1?rss=1">
<title>
<![CDATA[
Illuminating Dark Proteins using Reactome Pathways 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.06.05.543335v1?rss=1</link>
<description><![CDATA[
Limited knowledge about a substantial portion of protein coding genes, known as "dark" proteins, hinders our understanding of their functions and potential therapeutic applications. To address this, we leveraged Reactome, the most comprehensive, open source, open-access pathway knowledgebase, to contextualize dark proteins within biological pathways. By integrating multiple resources and employing a random forest classifier trained on 106 protein/gene pairwise features, we predicted functional interactions between dark proteins and Reactome-annotated proteins. We then developed three scores to measure the interactions between dark proteins and Reactome pathways, utilizing enrichment analysis and fuzzy logic simulations. Correlation analysis of these scores with an independent single-cell RNA sequencing dataset provided supporting evidence for this approach. Furthermore, systematic natural language processing (NLP) analysis of over 22 million PubMed abstracts and manual checking of the literature associated with 20 randomly selected dark proteins reinforced the predicted interactions between proteins and pathways. To enhance the visualization and exploration of dark proteins within Reactome pathways, we developed the Reactome IDG portal, deployed at https://idg.reactome.org, a web application featuring tissue-specific protein and gene expression overlay, as well as drug interactions. Our integrated computational approach, together with the user-friendly web platform, offers a valuable resource for uncovering potential biological functions and therapeutic implications of dark proteins.
]]></description>
<dc:creator>Brunson, T.</dc:creator>
<dc:creator>Sanati, N.</dc:creator>
<dc:creator>Matthews, L.</dc:creator>
<dc:creator>Haw, R.</dc:creator>
<dc:creator>Beavers, D.</dc:creator>
<dc:creator>Shorser, S.</dc:creator>
<dc:creator>Sevilla, C.</dc:creator>
<dc:creator>Viteri, G.</dc:creator>
<dc:creator>Conley, P.</dc:creator>
<dc:creator>Rothfels, K.</dc:creator>
<dc:creator>Hermjakob, H.</dc:creator>
<dc:creator>Stein, L.</dc:creator>
<dc:creator>D'Eustachio, P.</dc:creator>
<dc:creator>Wu, G.</dc:creator>
<dc:date>2023-06-05</dc:date>
<dc:identifier>doi:10.1101/2023.06.05.543335</dc:identifier>
<dc:title><![CDATA[Illuminating Dark Proteins using Reactome Pathways]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-06-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.06.05.543783v1?rss=1">
<title>
<![CDATA[
Bacterial mimicry of eukaryotic HECT ubiquitin ligation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.06.05.543783v1?rss=1</link>
<description><![CDATA[
HECT E3 ubiquitin (Ub) ligases direct their modified substrates toward a range of cellular fates dictated by the specific form of monomeric or polymeric Ub (polyUb) signal that is attached. How polyUb specificity is achieved has been a longstanding mystery, despite extensive study ranging from yeast to human. Two outlying examples of bacterial "HECT-like" (bHECT) E3 ligases have been reported in the human pathogens Enterohemorrhagic Escherichia coli and Salmonella Typhimurium, but what parallels can be drawn to eukaryotic HECT (eHECT) mechanism and specificity had not been explored. Here, we expanded the bHECT family and identified catalytically active, bona fide examples in both human and plant pathogens. By determining structures for three bHECT complexes in their primed, Ub-loaded states, we resolved key details of the full bHECT Ub ligation mechanism. One structure provided the first glimpse of a HECT E3 ligase in the act of ligating polyUb, yielding a means to rewire the polyUb specificity of both bHECT and eHECT ligases. Through studying this evolutionarily distinct bHECT family, we have not only gained insight into the function of key bacterial virulence factors but also revealed fundamental principles underlying HECT-type Ub ligation.
]]></description>
<dc:creator>Franklin, T. G.</dc:creator>
<dc:creator>Brzovic, P. S.</dc:creator>
<dc:creator>Pruneda, J. N.</dc:creator>
<dc:date>2023-06-05</dc:date>
<dc:identifier>doi:10.1101/2023.06.05.543783</dc:identifier>
<dc:title><![CDATA[Bacterial mimicry of eukaryotic HECT ubiquitin ligation]]></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/10.1101/2023.06.06.543939v1?rss=1">
<title>
<![CDATA[
Single cell RNA-seq analysis of spinal locomotor circuitry in larval zebrafish 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.06.06.543939v1?rss=1</link>
<description><![CDATA[
Identification of the neuronal types that form the specialized circuits controlling distinct behaviors has benefited greatly from the simplicity offered by zebrafish. Electrophysiological studies have shown that additional to connectivity, understanding of circuitry requires identification of functional specializations among individual circuit components, such as those that regulate levels of transmitter release and neuronal excitability. In this study we use single cell RNA sequencing (scRNAseq) to identify the molecular bases for functional distinctions between motoneuron types that are causal to their differential roles in swimming. The primary motoneuron (PMn) in particular, expresses high levels of a unique combination of voltage-dependent ion channel types and synaptic proteins termed functional  cassettes. The ion channel types are specialized for promoting high frequency firing of action potentials and augmented transmitter release at the neuromuscular junction, both contributing to greater power generation. Our transcriptional profiling of spinal neurons further assigns expression of this cassette to specific interneuron types also involved in the central circuitry controlling high speed swimming and escape behaviors. Our analysis highlights the utility of scRNAseq in functional characterization of neuronal circuitry, in addition to providing a gene expression resource for studying cell type diversity.
]]></description>
<dc:creator>Kelly, J. J.</dc:creator>
<dc:creator>Wen, H.</dc:creator>
<dc:creator>Brehm, P.</dc:creator>
<dc:date>2023-06-07</dc:date>
<dc:identifier>doi:10.1101/2023.06.06.543939</dc:identifier>
<dc:title><![CDATA[Single cell RNA-seq analysis of spinal locomotor circuitry in larval zebrafish]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-06-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.06.13.544801v1?rss=1">
<title>
<![CDATA[
Structural basis for flagellin induced NAIP5 activation. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.06.13.544801v1?rss=1</link>
<description><![CDATA[
The NAIP/NLRC4 inflammasome is activated when NAIP binds to a gram-negative bacterial ligand. Initially, NAIP exists in an inactive state with a wide-open conformation. Upon ligand binding, the winged helix domain (WHD) of NAIP is activated and forms steric clash with NLRC4 to open it up. However, how ligand binding induces the conformational change of NAIP is less clear. To understand this process, we investigated the dynamics of the ligand binding region of inactive NAIP5 and solved the cryo-EM structure of NAIP5 in complex with its specific ligand, FliC from flagellin, at 2.93 [A] resolution. The structure revealed a "trap and lock" mechanism in FliC recognition, whereby FliC-D0C is first trapped by the hydrophobic pocket of NAIP5, then locked in the binding site by the insertion domain (ID) and C-terminal tail (CTT) of NAIP5. The FliC-D0N domain further inserts into the loop of ID to stabilize the complex. According to this mechanism, FliC activates NAIP5 by bringing multiple flexible domains together, particularly the ID, HD2, and LRR domains, to form the active conformation and support the WHD loop in triggering NLRC4 activation.
]]></description>
<dc:creator>Zhang, L.</dc:creator>
<dc:creator>Paidimuddala, B.</dc:creator>
<dc:creator>Cao, J.</dc:creator>
<dc:date>2023-06-13</dc:date>
<dc:identifier>doi:10.1101/2023.06.13.544801</dc:identifier>
<dc:title><![CDATA[Structural basis for flagellin induced NAIP5 activation.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-06-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.06.14.544866v1?rss=1">
<title>
<![CDATA[
Sensory context for coding of natural sounds in auditory cortex 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.06.14.544866v1?rss=1</link>
<description><![CDATA[
Accurate sound perception can require integrating information over hundreds of milliseconds or even seconds. Spectro-temporal models of sound coding by single neurons in auditory cortex indicate that the majority of sound-evoked activity can be attributed to stimuli with a few tens of milliseconds. It remains uncertain how the auditory system integrates information about sensory context on a longer timescale. Here we characterized long-lasting contextual effects in auditory cortex (AC) using a diverse set of natural sound stimuli. We measured context effects as the difference in a neurons response to a single probe sound following two different context sounds. Many AC neurons showed context effects lasting longer than the temporal window of a traditional spectro-temporal receptive field. The duration and magnitude of context effects varied substantially across neurons and stimuli. This diversity of context effects formed a sparse code across the neural population that encoded a wider range of contexts than any constituent neuron. Encoding model analysis indicates that context effects can be explained by activity in the local neural population, suggesting that recurrent local circuits support a long-lasting representation of sensory context in auditory cortex.
]]></description>
<dc:creator>Lopez Espejo, M.</dc:creator>
<dc:creator>David, S. V.</dc:creator>
<dc:date>2023-06-14</dc:date>
<dc:identifier>doi:10.1101/2023.06.14.544866</dc:identifier>
<dc:title><![CDATA[Sensory context for coding of natural sounds in auditory cortex]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-06-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.06.21.545873v1?rss=1">
<title>
<![CDATA[
Calcineurin inhibition enhances Caenorhabditis elegans lifespan by defecation defects-mediated calorie restriction and nuclear hormone signaling 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.06.21.545873v1?rss=1</link>
<description><![CDATA[
Calcineurin is a highly conserved calcium/calmodulin-dependent serine/threonine protein phosphatase with diverse functions. Inhibition of calcineurin is known to enhance the lifespan of Caenorhabditis elegans through multiple signaling pathways. Aiming to study the role of calcineurin in regulating innate immunity, we discover that calcineurin is required for the rhythmic defecation motor program (DMP) in C. elegans. Calcineurin inhibition leads to defects in the DMP, resulting in intestinal bloating, rapid colonization of the gut by bacteria, and increased susceptibility to bacterial infection. We demonstrate that intestinal bloating caused by calcineurin inhibition mimics the effects of calorie restriction, resulting in enhanced lifespan. The TFEB ortholog, HLH-30, is required for lifespan extension mediated by calcineurin inhibition. Finally, we show that the nuclear hormone receptor, NHR-8, is upregulated by calcineurin inhibition and is necessary for the increased lifespan. Our studies uncover a role for calcineurin in the C. elegans DMP and provide a new mechanism for calcineurin inhibition-mediated longevity extension.
]]></description>
<dc:creator>Das, P.</dc:creator>
<dc:creator>Aballay, A.</dc:creator>
<dc:creator>Singh, J.</dc:creator>
<dc:date>2023-06-24</dc:date>
<dc:identifier>doi:10.1101/2023.06.21.545873</dc:identifier>
<dc:title><![CDATA[Calcineurin inhibition enhances Caenorhabditis elegans lifespan by defecation defects-mediated calorie restriction and nuclear hormone signaling]]></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/10.1101/2023.06.23.546323v1?rss=1">
<title>
<![CDATA[
The Na+ leak channel NALCN controls spontaneous activity and mediates synaptic modulation by α2-adrenergic receptors in auditory neurons 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.06.23.546323v1?rss=1</link>
<description><![CDATA[
Cartwheel interneurons of the dorsal cochlear nucleus (DCN) potently suppress multisensory signals that converge with primary auditory afferent input, and thus regulate auditory processing. Noradrenergic fibers from locus coeruleus project to the DCN, and 2-adrenergic receptors inhibit spontaneous spike activity but simultaneously enhance synaptic strength in cartwheel cells, a dual effect leading to enhanced signal- to-noise for inhibition. However, the ionic mechanism of this striking modulation is unknown. We generated a glycinergic neuron-specific knockout of the Na+ leak channel NALCN, and found that its presence was required for spontaneous firing in cartwheel cells. Activation of 2-adrenergic receptors inhibited both NALCN and spike generation, and this modulation was absent in the NALCN knockout. Moreover, 2-dependent enhancement of synaptic strength was also absent in the knockout. GABAB receptors mediated inhibition through NALCN as well, acting on the same population of channels as 2 receptors, suggesting close apposition of both receptor subtypes with NALCN. Thus, multiple neuromodulatory systems determine the impact of synaptic inhibition by suppressing the excitatory leak channel, NALCN.
]]></description>
<dc:creator>Ngodup, T.</dc:creator>
<dc:creator>Irie, T.</dc:creator>
<dc:creator>Elkins, S.</dc:creator>
<dc:creator>Trussell, L. O.</dc:creator>
<dc:date>2023-06-25</dc:date>
<dc:identifier>doi:10.1101/2023.06.23.546323</dc:identifier>
<dc:title><![CDATA[The Na+ leak channel NALCN controls spontaneous activity and mediates synaptic modulation by α2-adrenergic receptors in auditory neurons]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-06-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.06.26.546619v1?rss=1">
<title>
<![CDATA[
Bats possess the anatomical substrate for a laryngeal motor cortex 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.06.26.546619v1?rss=1</link>
<description><![CDATA[
Cortical neurons that make direct connections to motor neurons in the brainstem and spinal cord are specialized for fine motor control and learning [1, 2]. Imitative vocal learning, the basis for human speech, requires the precise control of the larynx muscles [3]. While much knowledge on vocal learning systems has been gained from studying songbirds [4], an accessible laboratory model for mammalian vocal learning is highly desirable. Evidence indicative of complex vocal repertoires and dialects suggests that bats are vocal learners [5, 6], however the circuitry that underlies vocal control and learning in bats is largely unknown. A key feature of vocal learning animals is a direct cortical projection to the brainstem motor neurons that innervate the vocal organ [7]. A recent study [8] described a direct connection from the primary motor cortex to medullary nucleus ambiguus in the Egyptian fruit bat (Rousettus aegyptiacus). Here we show that a distantly related bat, Sebas short-tailed bat (Carollia perspicillata) also possesses a direct projection from the primary motor cortex to nucleus ambiguus. Our results, in combination with Wirthlin et al. [8], suggest that multiple bat lineages possess the anatomical substrate for cortical control of vocal output. We propose that bats would be an informative mammalian model for vocal learning studies to better understand the genetics and circuitry involved in human vocal communication.
]]></description>
<dc:creator>Nevue, A. A.</dc:creator>
<dc:creator>Mello, C. V.</dc:creator>
<dc:creator>Portfors, C. V.</dc:creator>
<dc:date>2023-06-26</dc:date>
<dc:identifier>doi:10.1101/2023.06.26.546619</dc:identifier>
<dc:title><![CDATA[Bats possess the anatomical substrate for a laryngeal motor cortex]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-06-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.06.29.547009v1?rss=1">
<title>
<![CDATA[
Task-specific invariant representation in auditory cortex 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.06.29.547009v1?rss=1</link>
<description><![CDATA[
Categorical sensory representations are critical for many behaviors, including speech perception. In the auditory system, categorical information is thought to arise hierarchically, becoming increasingly prominent in higher order cortical regions. The neural mechanisms that support this robust and flexible computation remain poorly understood. Here, we studied sound representations in primary and non-primary auditory cortex while animals engaged in a challenging sound discrimination task. Population-level decoding of simultaneously recorded single neurons revealed that task engagement caused categorical sound representations to emerge in non-primary auditory cortex. In primary auditory cortex, task engagement caused a general enhancement of sound decoding that was not specific to task-relevant categories. These findings are consistent with mixed selectivity models of neural disentanglement, in which early sensory regions build an overcomplete representation of the world and allow neurons in downstream brain regions to flexibly and selectively read out behaviorally relevant, categorical information.
]]></description>
<dc:creator>Heller, C. R.</dc:creator>
<dc:creator>Hamersky, G. R.</dc:creator>
<dc:creator>David, S. V.</dc:creator>
<dc:date>2023-06-29</dc:date>
<dc:identifier>doi:10.1101/2023.06.29.547009</dc:identifier>
<dc:title><![CDATA[Task-specific invariant representation in auditory cortex]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-06-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.06.29.547125v1?rss=1">
<title>
<![CDATA[
Tuberculosis susceptibility in genetically diverse mice reveals functional diversity of neutrophils 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.06.29.547125v1?rss=1</link>
<description><![CDATA[
Tuberculosis (TB) is a heterogenous disease in humans with individuals exhibiting a wide range of susceptibility. This heterogeneity is not captured by standard laboratory mouse lines. We used a new collection of 19 wild-derived inbred mouse lines collected from diverse geographic sites to identify novel phenotypes during Mycobacterium tuberculosis (Mtb) infection. Wild derived mice have heterogenous immune responses to infection that result in differential ability to control disease at early timepoints. Correlation analysis with multiple parameters including sex, weight, and cellular immune responses in the lungs revealed that enhanced control of infection is associated with increased numbers of CD4 T cells, CD8 T cells and B cells. Surprisingly, we did not observe strong correlations between IFN-{gamma} production and control of infection. Although in most lines high neutrophils were associated with susceptibility, we identified a mouse line that harbors high neutrophils numbers yet controls infection. Using single-cell RNA sequencing, we identified a novel neutrophil signature associated with failure to control infection.
]]></description>
<dc:creator>Ravesloot-Chavez, M. M.</dc:creator>
<dc:creator>Van Dis, E.</dc:creator>
<dc:creator>Fox, D.</dc:creator>
<dc:creator>Nguyenla, X. H.</dc:creator>
<dc:creator>Rawal, S. L.</dc:creator>
<dc:creator>Ballinger, M. A.</dc:creator>
<dc:creator>Thomas, H.</dc:creator>
<dc:creator>Kotov, D. I.</dc:creator>
<dc:creator>Vance, R.</dc:creator>
<dc:creator>Nachman, M.</dc:creator>
<dc:creator>Stanley, S.</dc:creator>
<dc:date>2023-06-30</dc:date>
<dc:identifier>doi:10.1101/2023.06.29.547125</dc:identifier>
<dc:title><![CDATA[Tuberculosis susceptibility in genetically diverse mice reveals functional diversity of neutrophils]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-06-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.06.30.547175v1?rss=1">
<title>
<![CDATA[
Three-dimensional genome architecture persists in a 52,000-year-old woolly mammoth skin sample 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.06.30.547175v1?rss=1</link>
<description><![CDATA[
Ancient DNA (aDNA) sequencing analysis typically involves alignment to a modern reference genome assembly from a related species. Since aDNA molecules are fragmentary, these alignments yield information about small-scale differences, but provide no information about larger features such as the chromosome structure of ancient species. We report the genome assembly of a female Late Pleistocene woolly mammoth (Mammuthus primigenius) with twenty-eight chromosome-length scaffolds, generated using mammoth skin preserved in permafrost for roughly 52,000 years. We began by creating a modified Hi-C protocol, dubbed PaleoHi-C, optimized for ancient samples, and using it to map chromatin contacts in a woolly mammoth. Next, we developed "reference-assisted 3D genome assembly," which begins with a reference genome assembly from a related species, and uses Hi-C and DNA-Seq data from a target species to split, order, orient, and correct sequences on the basis of their 3D proximity, yielding accurate chromosome-length scaffolds for the target species. By means of this reference-assisted 3D genome assembly, PaleoHi-C data reveals the 3D architecture of a woolly mammoth genome, including chromosome territories, compartments, domains, and loops. The active (A) and inactive (B) genome compartments in mammoth skin more closely resemble those observed in Asian elephant skin than the compartmentalization patterns seen in other Asian elephant tissues. Differences in compartmentalization between these skin samples reveal sequences whose transcription was potentially altered in mammoth. We observe a tetradic structure for the inactive X chromosome in mammoth, distinct from the bipartite architecture seen in human and mouse. Generating chromosome-length genome assemblies for two other elephantids (Asian and African elephant), we find that the overall karyotype, and this tetradic Xi structure, are conserved throughout the clade. These results illustrate that cell-type specific epigenetic information can be preserved in ancient samples, in the form of DNA geometry, and that it may be feasible to perform de novo genome assembly of some extinct species.
]]></description>
<dc:creator>Sandoval-Velasco, M.</dc:creator>
<dc:creator>Dudchenko, O.</dc:creator>
<dc:creator>Antonio Rodriguez, J.</dc:creator>
<dc:creator>Perez Estrada, C.</dc:creator>
<dc:creator>Dehasque, M.</dc:creator>
<dc:creator>Fontsere, C.</dc:creator>
<dc:creator>Mak, S. S. T.</dc:creator>
<dc:creator>Plotnikov, V.</dc:creator>
<dc:creator>Khan, R.</dc:creator>
<dc:creator>Weisz, D.</dc:creator>
<dc:creator>Contessoto, V. G.</dc:creator>
<dc:creator>Oliveira Junior, A. B.</dc:creator>
<dc:creator>Kalluchi, A.</dc:creator>
<dc:creator>Omer, A. D.</dc:creator>
<dc:creator>Batra, S. S.</dc:creator>
<dc:creator>Shamim, M. S.</dc:creator>
<dc:creator>Durand, N. C.</dc:creator>
<dc:creator>O'Connell, B.</dc:creator>
<dc:creator>Roca, A. L.</dc:creator>
<dc:creator>Gnirke, A.</dc:creator>
<dc:creator>Garcia-Trevino, I.</dc:creator>
<dc:creator>Coke, R.</dc:creator>
<dc:creator>Flanagan, J. P.</dc:creator>
<dc:creator>Pletch, K.</dc:creator>
<dc:creator>Ruiz-Herrera, A.</dc:creator>
<dc:creator>Lander, E. S.</dc:creator>
<dc:creator>Rowley, M. J.</dc:creator>
<dc:creator>Onuchic, J. N.</dc:creator>
<dc:creator>Dalen, L.</dc:creator>
<dc:creator>Marti-Renom, M. A.</dc:creator>
<dc:creator>Gilbert, M. T. P.</dc:creator>
<dc:creator>Aiden, E. L.</dc:creator>
<dc:date>2023-06-30</dc:date>
<dc:identifier>doi:10.1101/2023.06.30.547175</dc:identifier>
<dc:title><![CDATA[Three-dimensional genome architecture persists in a 52,000-year-old woolly mammoth skin sample]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-06-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.06.30.547249v1?rss=1">
<title>
<![CDATA[
Gestational and postnatal age associations for striatal tissue iron deposition in early infancy 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.06.30.547249v1?rss=1</link>
<description><![CDATA[
Striatal development is crucial for later motor, cognitive, and reward behavior, but age-related change in striatal physiology during the neonatal period remains understudied. An MRI-based measure of tissue iron deposition, T2*, is a non-invasive way to probe striatal physiology neonatally, linked to dopaminergic processing and cognition in children and adults. Striatal subregions have distinct functions that may come online at different time periods in early life. To identify if there are critical periods before or after birth, we measured if striatal iron accrued with gestational age at birth [range=34.57-41.85 weeks] or postnatal age at scan [range=5-64 days], using MRI to probe the T2* signal in N=83 neonates in three striatal subregions. We found iron increased with postnatal age in the pallidum and putamen but not the caudate. No significant relationship between iron and gestational age was observed. Using a subset of infants scanned at preschool age (N=26), we show distributions of iron shift between timepoints. In infants, the pallidum had the least iron of the three regions but had the most by preschool age. Together, this provides evidence of distinct change for striatal subregions, a possible differentiation between motor and cognitive systems, identifying a mechanism that may impact future trajectories.

HighlightsO_LINeonatal striatal tissue iron can be measured using the T2* signal from rsfMRI
C_LIO_LInT2* changed with postnatal age in the pallidum and putamen but not in the caudate
C_LIO_LInT2* did not change with gestational age in any of the three regions
C_LIO_LIPatterns of iron deposition (nT2*) among regions shift from infancy to preschool
C_LI
]]></description>
<dc:creator>Cabral, L.</dc:creator>
<dc:creator>Calabro, F.</dc:creator>
<dc:creator>Rasmussen, J.</dc:creator>
<dc:creator>Foran, W.</dc:creator>
<dc:creator>Moore, L. A.</dc:creator>
<dc:creator>Graham, A.</dc:creator>
<dc:creator>O'Connor, T. G.</dc:creator>
<dc:creator>Wadhwa, P. D.</dc:creator>
<dc:creator>Entringer, S.</dc:creator>
<dc:creator>Fair, D.</dc:creator>
<dc:creator>Buss, C.</dc:creator>
<dc:creator>Panigrahy, A.</dc:creator>
<dc:creator>Luna, B.</dc:creator>
<dc:date>2023-06-30</dc:date>
<dc:identifier>doi:10.1101/2023.06.30.547249</dc:identifier>
<dc:title><![CDATA[Gestational and postnatal age associations for striatal tissue iron deposition in early infancy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-06-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.06.30.547228v1?rss=1">
<title>
<![CDATA[
Environmental enrichment promotes adaptive responding during tests of behavioral regulation in male heterogeneous stock rats 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.06.30.547228v1?rss=1</link>
<description><![CDATA[
Organisms must regulate their behavior flexibly in the face of environmental challenges. Failure can lead to a host of maladaptive behavioral traits associated with a range of neuropsychiatric disorders, including attention deficit hyperactivity disorder, autism, and substance use disorders. This maladaptive dysregulation of behavior is influenced by genetic and environmental factors. For example, environmental enrichment produces beneficial neurobehavioral effects in animal models of such disorders. The present study determined the effects of environmental enrichment on a range of measures related to behavioral regulation using a large cohort of male, outbred heterogeneous stock (HS) rats as subjects to mimic the genetic variability found in the human population. Subjects were reared from late adolescence onwards either in pairs in standard housing with minimal enrichment (n=200) or in groups of 16 in a highly enriched environment consisting of a large multi-level cage filled with toys, running wheels, and shelters (n=64). Rats were subjected to a battery of tests, including: (i) locomotor response to novelty, (iI) light reinforcement, (iii) social reinforcement, (iv) reaction time, (v) a patch-depletion foraging test, (vi) Pavlovian conditioned approach, (vii) conditioned reinforcement, and (viii) cocaine conditioned cue preference. Results indicated that rats housed in the enriched environment were able to filter out irrelevant stimuli more effectively and thereby regulate their behavior more efficiently than standard-housing rats. The dramatic impact of environmental enrichment suggests that behavioral studies using standard housing conditions may not generalize to more complex environments that may be more ethologically relevant.
]]></description>
<dc:creator>Ishiwari, K.</dc:creator>
<dc:creator>King, C. P.</dc:creator>
<dc:creator>Martin, C. D.</dc:creator>
<dc:creator>Tripi, J. A.</dc:creator>
<dc:creator>George, A. M.</dc:creator>
<dc:creator>Lamparelli, A. C.</dc:creator>
<dc:creator>Chitre, A.</dc:creator>
<dc:creator>Polesskaya, O.</dc:creator>
<dc:creator>Richards, J. B.</dc:creator>
<dc:creator>Solberg Woods, L. C.</dc:creator>
<dc:creator>Gancarz, A.</dc:creator>
<dc:creator>Palmer, A. A.</dc:creator>
<dc:creator>Dietz, D. M.</dc:creator>
<dc:creator>Mitchell, S. H.</dc:creator>
<dc:creator>Meyer, P. J.</dc:creator>
<dc:date>2023-07-03</dc:date>
<dc:identifier>doi:10.1101/2023.06.30.547228</dc:identifier>
<dc:title><![CDATA[Environmental enrichment promotes adaptive responding during tests of behavioral regulation in male heterogeneous stock rats]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-07-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.07.01.547283v1?rss=1">
<title>
<![CDATA[
KRAS-mediated upregulation of CIP2A promotes suppression of PP2A-B56α to initiate pancreatic cancer development 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.07.01.547283v1?rss=1</link>
<description><![CDATA[
Oncogenic mutations in KRAS are present in approximately 95% of patients diagnosed with pancreatic ductal adenocarcinoma (PDAC) and are considered the initiating event of pancreatic intraepithelial neoplasia (PanIN) precursor lesions. While it is well established that KRAS mutations drive the activation of oncogenic kinase cascades during pancreatic oncogenesis, the effects of oncogenic KRAS signaling on regulation of phosphatases during this process is not fully appreciated. Protein Phosphatase 2A (PP2A) has been implicated in suppressing KRAS-driven cellular transformation. However, low PP2A activity is observed in PDAC cells compared to non-transformed cells, suggesting that suppression of PP2A activity is an important step in the overall development of PDAC. In the current study, we demonstrate that KRASG12D induces the expression of both an endogenous inhibitor of PP2A activity, Cancerous Inhibitor of PP2A (CIP2A), and the PP2A substrate, c-MYC. Consistent with these findings, KRASG12D sequestered the specific PP2A subunit responsible for c-MYC degradation, B56, away from the active PP2A holoenzyme in a CIP2A-dependent manner. During PDAC initiation in vivo, knockout of B56 promoted KRASG12D tumorigenesis by accelerating acinar-to-ductal metaplasia (ADM) and the formation of PanIN lesions. The process of ADM was attenuated ex vivo in response to pharmacological re-activation of PP2A utilizing direct small molecule activators of PP2A (SMAPs). Together, our results suggest that suppression of PP2A-B56 through KRAS signaling can promote the MYC-driven initiation of pancreatic tumorigenesis.
]]></description>
<dc:creator>Tinsley, S. L.</dc:creator>
<dc:creator>Shelley, R. A.</dc:creator>
<dc:creator>Mall, G. K.</dc:creator>
<dc:creator>Chianis, E. R. D.</dc:creator>
<dc:creator>Dhiman, A.</dc:creator>
<dc:creator>Baral, G.</dc:creator>
<dc:creator>Kothandaraman, H.</dc:creator>
<dc:creator>Thoma, M. C.</dc:creator>
<dc:creator>Daniel, C.</dc:creator>
<dc:creator>Lanman, N. A.</dc:creator>
<dc:creator>Pasca di Magliano, M.</dc:creator>
<dc:creator>Narla, G.</dc:creator>
<dc:creator>Solorio, L.</dc:creator>
<dc:creator>Dykhuizen, E.</dc:creator>
<dc:creator>Sears, R. C.</dc:creator>
<dc:creator>Allen-Petersen, B.</dc:creator>
<dc:date>2023-07-03</dc:date>
<dc:identifier>doi:10.1101/2023.07.01.547283</dc:identifier>
<dc:title><![CDATA[KRAS-mediated upregulation of CIP2A promotes suppression of PP2A-B56α to initiate pancreatic cancer development]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-07-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.07.03.547507v1?rss=1">
<title>
<![CDATA[
Deep immunophenotyping reveals circulating activated lymphocytes in individuals at risk for rheumatoid arthritis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.07.03.547507v1?rss=1</link>
<description><![CDATA[
Rheumatoid arthritis (RA) is a systemic autoimmune disease with currently no universally highly effective prevention strategies. Identifying pathogenic immune phenotypes in  At-Risk populations prior to clinical disease onset is crucial to establishing effective prevention strategies. Here, we applied mass cytometry to deeply characterize the immunophenotypes in blood from At-Risk individuals identified through the presence of serum antibodies to citrullinated protein antigens (ACPA) and/or first-degree relative (FDR) status (n=52), as compared to established RA (n=67), and healthy controls (n=48). We identified significant cell expansions in At-Risk individuals compared with controls, including CCR2+CD4+ T cells, T peripheral helper (Tph) cells, type 1 T helper cells, and CXCR5+CD8+ T cells. We also found that CD15+ classical monocytes were specifically expanded in ACPA-negative FDRs, and an activated PAX5low naive B cell population was expanded in ACPA-positive FDRs. Further, we developed an "RA immunophenotype score" classification method based on the degree of enrichment of cell states relevant to established RA patients. This score significantly distinguished At-Risk individuals from controls. In all, we systematically identified activated lymphocyte phenotypes in At-Risk individuals, along with immunophenotypic differences among both ACPA+ and ACPA-FDR At-Risk subpopulations. Our classification model provides a promising approach for understanding RA pathogenesis with the goal to further improve prevention strategies and identify novel therapeutic targets.
]]></description>
<dc:creator>Inamo, J.</dc:creator>
<dc:creator>Keegan, J.</dc:creator>
<dc:creator>Griffith, A.</dc:creator>
<dc:creator>Ghosh, T.</dc:creator>
<dc:creator>Horisberger, A.</dc:creator>
<dc:creator>Howard, K.</dc:creator>
<dc:creator>Pulford, J.</dc:creator>
<dc:creator>Murzin, E.</dc:creator>
<dc:creator>Hancock, B.</dc:creator>
<dc:creator>Jonsson, A. H.</dc:creator>
<dc:creator>Seifert, J.</dc:creator>
<dc:creator>Feser, M. L.</dc:creator>
<dc:creator>Norris, J. M.</dc:creator>
<dc:creator>Cao, Y.</dc:creator>
<dc:creator>Apruzzese, W.</dc:creator>
<dc:creator>Bridges, S. L.</dc:creator>
<dc:creator>Bykerk, V.</dc:creator>
<dc:creator>Goodman, S.</dc:creator>
<dc:creator>Donlin, L.</dc:creator>
<dc:creator>Firestein, G. S.</dc:creator>
<dc:creator>Perlman, H.</dc:creator>
<dc:creator>Bathon, J. M.</dc:creator>
<dc:creator>Hughes, L. B.</dc:creator>
<dc:creator>Tabechian, D.</dc:creator>
<dc:creator>Filer, A.</dc:creator>
<dc:creator>Pitzalis, C.</dc:creator>
<dc:creator>H. Anolik, J.</dc:creator>
<dc:creator>Moreland, L.</dc:creator>
<dc:creator>Guthridge, J. M.</dc:creator>
<dc:creator>James, J. A.</dc:creator>
<dc:creator>Brenner, M. B.</dc:creator>
<dc:creator>Raychaudhuri, S.</dc:creator>
<dc:creator>Sparks, J. A.</dc:creator>
<dc:creator>The Accelerating Medicines Partnership RA/SLE Network,</dc:creator>
<dc:creator>Holers, V. M.</dc:creator>
<dc:creator>Deane, K. D.</dc:creator>
<dc:creator>Lederer, J. A.</dc:creator>
<dc:creator>Rao, D. A.</dc:creator>
<dc:creator>Zhang, F.</dc:creator>
<dc:date>2023-07-04</dc:date>
<dc:identifier>doi:10.1101/2023.07.03.547507</dc:identifier>
<dc:title><![CDATA[Deep immunophenotyping reveals circulating activated lymphocytes in individuals at risk for rheumatoid arthritis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-07-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.07.09.548280v1?rss=1">
<title>
<![CDATA[
DISCOVERY AND VALIDATION OF GENES DRIVING DRUG-INTAKE AND RELATED BEHAVIORAL TRAITS IN MICE 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.07.09.548280v1?rss=1</link>
<description><![CDATA[
Substance use disorders (SUDs) are heritable disorders characterized by compulsive drug use, but the biological mechanisms driving addiction remain largely unknown. Genetic correlations reveal that predisposing drug-naive phenotypes, including anxiety, depression, novelty preference, and sensation seeking, are predictive of drug-use phenotypes, implicating shared genetic mechanisms. Because of this relationship, high-throughput behavioral screening of predictive phenotypes in knockout (KO) mice allows efficient discovery of genes likely to be involved in drug use. We used this strategy in two rounds of screening in which we identified 33 drug-use candidate genes and ultimately validated the perturbation of 22 of these genes as causal drivers of substance intake. In our initial round of screening, we employed the two-bottle-choice paradigms to assess alcohol, methamphetamine, and nicotine intake. We identified 19 KO strains that were extreme responders on at least one predictive phenotype. Thirteen of the 19 gene deletions (68%) significantly affected alcohol use three methamphetamine use, and two both. In the second round of screening, we employed a multivariate approach to identify outliers and performed validation using methamphetamine two-bottle choice and ethanol drinking-in-the-dark protocols. We identified 15 KO strains that were extreme responders across the predisposing drug-naive phenotypes. Eight of the 15 gene deletions (53%) significantly affected intake or preference for three alcohol, eight methamphetamine or three both (3). We observed multiple relations between predisposing behaviors and drug intake, revealing many distinct biobehavioral processes underlying these relationships. The set of mouse models identified in this study can be used to characterize these addiction-related processes further.
]]></description>
<dc:creator>Roy, T.</dc:creator>
<dc:creator>Bubier, J. A.</dc:creator>
<dc:creator>Dickson, P. E.</dc:creator>
<dc:creator>Wilcox, T.</dc:creator>
<dc:creator>Ndukum, J.</dc:creator>
<dc:creator>Clark, J. W.</dc:creator>
<dc:creator>Rizzo, S. J. S.</dc:creator>
<dc:creator>Crabbe, J. C.</dc:creator>
<dc:creator>Denegre, J.</dc:creator>
<dc:creator>Svenson, K. L.</dc:creator>
<dc:creator>Braun, R. E.</dc:creator>
<dc:creator>Kumar, V.</dc:creator>
<dc:creator>Murray, S. A.</dc:creator>
<dc:creator>White, J. K.</dc:creator>
<dc:creator>Philip, V.</dc:creator>
<dc:creator>Chesler, E.</dc:creator>
<dc:date>2023-07-10</dc:date>
<dc:identifier>doi:10.1101/2023.07.09.548280</dc:identifier>
<dc:title><![CDATA[DISCOVERY AND VALIDATION OF GENES DRIVING DRUG-INTAKE AND RELATED BEHAVIORAL TRAITS IN MICE]]></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/10.1101/2023.07.12.548718v1?rss=1">
<title>
<![CDATA[
sciMET-cap: High-throughput single-cell methylation analysis with a reduced sequencing burden 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.07.12.548718v1?rss=1</link>
<description><![CDATA[
DNA methylation is a key component of the mammalian epigenome, playing a regulatory role in development, disease, and other processes. Robust, high-throughput single-cell DNA methylation assays are now possible (sciMET); however, the genome-wide nature of DNA methylation results in a high sequencing burden per cell. Here, we leverage target enrichment with sciMET to capture sufficient information per cell for cell type assignment using substantially fewer sequence reads (sciMET-cap). Sufficient off-target coverage further enables the production of near-complete methylomes for individual cell types. We characterize sciMET-cap on human PBMCs and brain (middle frontal gyrus).
]]></description>
<dc:creator>Acharya, S.</dc:creator>
<dc:creator>Nichols, R. V.</dc:creator>
<dc:creator>Rylaarsdam, L. E.</dc:creator>
<dc:creator>O'Connell, B. L.</dc:creator>
<dc:creator>Braun, T. P.</dc:creator>
<dc:creator>Adey, A. C.</dc:creator>
<dc:date>2023-07-14</dc:date>
<dc:identifier>doi:10.1101/2023.07.12.548718</dc:identifier>
<dc:title><![CDATA[sciMET-cap: High-throughput single-cell methylation analysis with a reduced sequencing burden]]></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/10.1101/2023.07.16.549116v1?rss=1">
<title>
<![CDATA[
Cell networks in the mouse liver during partial hepatectomy 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.07.16.549116v1?rss=1</link>
<description><![CDATA[
In solid tissues homeostasis and regeneration after injury involve a complex interplay between many different cell types. The mammalian liver harbors numerous epithelial and non-epithelial cells and little is known about the global signaling networks that govern their interactions. To better understand the hepatic cell network, we isolated and purified 10 different cell populations from normal and regenerative mouse livers. Their transcriptomes were analyzed by bulk RNA-seq and a computational platform was used to analyze the cell-cell and ligand-receptor interactions among the 10 populations. Over 50,000 potential cell-cell interactions were found in both the ground state and after partial hepatectomy. Importantly, about half of these differed between the two states, indicating massive changes in the cell network during regeneration. Our study provides the first comprehensive database of potential cell-cell interactions in mammalian liver cell homeostasis and regeneration. With the help of this prediction model, we identified and validated two previously unknown signaling interactions involved in accelerating and delaying liver regeneration. Overall, we provide a novel platform for investigating autocrine/paracrine pathways in tissue regeneration, which can be adapted to other complex multicellular systems.

HighlightsA platform predicting cell-cell interactions in liver regeneration was established

This platform identified the BMP4 pathway antagonist Fstl1 as a stimulator of hepatocyte proliferation

This platform also discovered the role of Wnt pathway inhibitor Sfrp1 delaying liver regeneration
]]></description>
<dc:creator>Li, B.</dc:creator>
<dc:creator>Rodrigo-Torres, D.</dc:creator>
<dc:creator>Pelz, C.</dc:creator>
<dc:creator>Innes, B. T.</dc:creator>
<dc:creator>Canaday, P.</dc:creator>
<dc:creator>Chai, S.</dc:creator>
<dc:creator>Zandstra, P.</dc:creator>
<dc:creator>Bader, G.</dc:creator>
<dc:creator>Grompe, M.</dc:creator>
<dc:date>2023-07-18</dc:date>
<dc:identifier>doi:10.1101/2023.07.16.549116</dc:identifier>
<dc:title><![CDATA[Cell networks in the mouse liver during partial hepatectomy]]></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/10.1101/2023.07.19.549735v1?rss=1">
<title>
<![CDATA[
The Brain Electroencephalogram Microdisplay for Precision Neurosurgery 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.07.19.549735v1?rss=1</link>
<description><![CDATA[
Brain surgeries are among the most delicate clinical procedures and must be performed with the most technologically robust and advanced tools. When such surgical procedures are performed in functionally critical regions of the brain, functional mapping is applied as a standard practice that involves direct coordinated interactions between the neurosurgeon and the clinical neurology electrophysiology team. However, information flow during these interactions is commonly verbal as well as time consuming which in turn increases the duration and cost of the surgery, possibly compromising the patient outcomes. Additionally, the grids that measure brain activity and identify the boundaries of pathological versus functional brain regions suffer from low resolution (3-10 mm contact to contact spacing) with limited conformity to the brain surface. Here, we introduce a brain intracranial electroencephalogram microdisplay (Brain-iEEG-microdisplay) which conforms to the brain to measure the brain activity and display changes in near real-time (40 Hz refresh rate) on the surface of the brain in the surgical field. We used scalable engineered gallium nitride (GaN) substrates with 6" diameter to fabricate, encapsulate, and release free-standing arrays of up to 2048 GaN light emitting diodes (LEDs) in polyimide substrates. We then laminated the LED arrays on the back of micro-electrocorticography (ECoG) platinum nanorod grids (PtNRGrids) and developed hardware and software to perform near real-time intracranial EEG analysis and activation of light patterns that correspond to specific cortical activities. Using the Brain-iEEG-microdisplay, we precisely ideFSntified and displayed important cortical landmarks and pharmacologically induced pathological activities. In the rat model, we identified and displayed individual cortical columns corresponding to individual whiskers and the near real-time evolution of epileptic discharges. In the pig animal model, we demonstrated near real-time mapping and display of cortical functional boundaries using somatosensory evoked potentials (SSEP) and display of responses to direct electrical stimulation (DES) from the surface or within the brain tissue. Using a dual-color Brain-iEEG-microdisplay, we demonstrated co-registration of the functional cortical boundaries with one color and displayed the evolution of electrical potentials associated with epileptiform activity with another color. The Brain-iEEG-microdisplay holds the promise of increasing the efficiency of diagnosis and possibly surgical treatment, thereby reducing the cost and improving patient outcomes which would mark a major advancement in neurosurgery. These advances can also be translated to broader applications in neuro-oncology and neurophysiology.

One Sentence SummaryA brain intracranial electroencephalogram microdisplay (Brain-iEEG-microdisplay) measures and displays real-time brain activity in the surgical field.
]]></description>
<dc:creator>Tchoe, Y.</dc:creator>
<dc:creator>Wu, T.</dc:creator>
<dc:creator>U, H. S.</dc:creator>
<dc:creator>Roth, D. M.</dc:creator>
<dc:creator>Kim, D.</dc:creator>
<dc:creator>Lee, J.</dc:creator>
<dc:creator>Cleary, D. R.</dc:creator>
<dc:creator>Pizarro, P.</dc:creator>
<dc:creator>Tonsfeldt, K. J.</dc:creator>
<dc:creator>Lee, K.</dc:creator>
<dc:creator>Chen, P. C.</dc:creator>
<dc:creator>Bourhis, A. M.</dc:creator>
<dc:creator>Galton, I.</dc:creator>
<dc:creator>Coughlin, B.</dc:creator>
<dc:creator>Yang, J. C.</dc:creator>
<dc:creator>Paulk, A. C.</dc:creator>
<dc:creator>Halgren, E.</dc:creator>
<dc:creator>Cash, S. S.</dc:creator>
<dc:creator>Dayeh, S. A.</dc:creator>
<dc:date>2023-07-21</dc:date>
<dc:identifier>doi:10.1101/2023.07.19.549735</dc:identifier>
<dc:title><![CDATA[The Brain Electroencephalogram Microdisplay for Precision Neurosurgery]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-07-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.07.21.548450v1?rss=1">
<title>
<![CDATA[
Addressing persistent challenges in digital image analysis of cancerous tissues 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.07.21.548450v1?rss=1</link>
<description><![CDATA[
The National Cancer Institute (NCI) supports many research programs and consortia, many of which use imaging as a major modality for characterizing cancerous tissue. A trans-consortia Image Analysis Working Group (IAWG) was established in 2019 with a mission to disseminate imaging-related work and foster collaborations. In 2022, the IAWG held a virtual hackathon focused on addressing challenges of analyzing high dimensional datasets from fixed cancerous tissues. Standard image processing techniques have automated feature extraction, but the next generation of imaging data requires more advanced methods to fully utilize the available information. In this perspective, we discuss current limitations of the automated analysis of multiplexed tissue images, the first steps toward deeper understanding of these limitations, what possible solutions have been developed, any new or refined approaches that were developed during the Image Analysis Hackathon 2022, and where further effort is required. The outstanding problems addressed in the hackathon fell into three main themes: 1) challenges to cell type classification and assessment, 2) translation and visual representation of spatial aspects of high dimensional data, and 3) scaling digital image analyses to large (multi-TB) datasets. We describe the rationale for each specific challenge and the progress made toward addressing it during the hackathon. We also suggest areas that would benefit from more focus and offer insight into broader challenges that the community will need to address as new technologies are developed and integrated into the broad range of image-based modalities and analytical resources already in use within the cancer research community.
]]></description>
<dc:creator>Prabhakaran, S.</dc:creator>
<dc:creator>Yapp, C.</dc:creator>
<dc:creator>Baker, G. J.</dc:creator>
<dc:creator>Beyer, J.</dc:creator>
<dc:creator>Chang, Y. H.</dc:creator>
<dc:creator>Creason, A. L.</dc:creator>
<dc:creator>Krueger, R.</dc:creator>
<dc:creator>Muhlich, J.</dc:creator>
<dc:creator>Patterson, N. H.</dc:creator>
<dc:creator>Sidak, K.</dc:creator>
<dc:creator>Sudar, D.</dc:creator>
<dc:creator>Taylor, A. J.</dc:creator>
<dc:creator>Ternes, L.</dc:creator>
<dc:creator>Troidl, J.</dc:creator>
<dc:creator>Yubin, X.</dc:creator>
<dc:creator>Sokolov, A.</dc:creator>
<dc:creator>Tyson, D. R.</dc:creator>
<dc:creator>Participants of the Cell Imaging Hackathon 2022,</dc:creator>
<dc:date>2023-07-24</dc:date>
<dc:identifier>doi:10.1101/2023.07.21.548450</dc:identifier>
<dc:title><![CDATA[Addressing persistent challenges in digital image analysis of cancerous tissues]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-07-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.07.25.550547v1?rss=1">
<title>
<![CDATA[
Systematic analysis of the sphingomyelin synthase family in C. elegans 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.07.25.550547v1?rss=1</link>
<description><![CDATA[
Sphingomyelin (SM) is a major component of mammalian cell membranes and particularly abundant in the myelin sheath that surrounds nerve fibers. Its production is catalyzed by SM synthases SMS1 and SMS2, which interconvert phosphatidylcholine and ceramide to diacylglycerol and SM in the Golgi and at the plasma membrane, respectively. As the lipids participating in this reaction fulfill both structural and signaling functions, SMS enzymes have considerable potential to influence diverse important cellular processes. The nematode Caenorhabditis elegans is an attractive model for studying both animal development and human disease. The organism contains five SMS homologues but none of these have been characterized in any detail. Here, we carried out the first systematic analysis of SMS family members in C. elegans. Using heterologous expression systems, genetic ablation, metabolic labeling and lipidome analyses, we show that C. elegans harbors at least three distinct SM synthases and one ceramide phosphoethanolamine (CPE) synthase. Moreover, C. elegans SMS family members have partially overlapping but also unique subcellular distributions and together occupy all principal compartments of the secretory pathway. Our findings shed light on crucial aspects of sphingolipid metabolism in a valuable animal model and opens avenues for exploring the role of SM and its metabolic intermediates in organismal development.
]]></description>
<dc:creator>Guzman, G. D.</dc:creator>
<dc:creator>Farley, S. E.</dc:creator>
<dc:creator>Kyle, J. E.</dc:creator>
<dc:creator>Bramer, L.</dc:creator>
<dc:creator>Hoeltzl, S.</dc:creator>
<dc:creator>van den Dikkenberg, J.</dc:creator>
<dc:creator>Holthuis, J. C. M.</dc:creator>
<dc:creator>Tafesse, F.</dc:creator>
<dc:date>2023-07-25</dc:date>
<dc:identifier>doi:10.1101/2023.07.25.550547</dc:identifier>
<dc:title><![CDATA[Systematic analysis of the sphingomyelin synthase family in C. elegans]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-07-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.07.25.550459v1?rss=1">
<title>
<![CDATA[
Elevated peripheral and nervous system inflammation is associated with decreased short-chain fatty acid levels in Zika-virus infected macaques 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.07.25.550459v1?rss=1</link>
<description><![CDATA[
Zika virus (ZIKV) infection of central nervous system (CNS) tissue is associated with CNS inflammation, which contributes to ZIKV pathology. Similarly, ZIKV infection has been associated with increased vaginal and rectal mucosal inflammation. As mucosal dysfunction may contribute to elevated systemic inflammation, ZIKV-induced mucosal alterations could potentiate CNS disruptions, leading to ZIKV pathogenesis. However, the potential link between mucosal dysfunction, CNS inflammation and the underlying mechanisms causing these disruptions in ZIKV infection has not been well described. Here, we assessed plasma and CSF indicators of inflammation, including neopterin, tryptophan, kynurenine and serotonin by liquid chromatography tandem mass spectrometry. We observed significant increases in neopterin formation, tryptophan catabolism and serotonin levels in the plasma and CSF of ZIKV-infected pigtail macaques (PTM), rhesus macaques (RM) and in the plasma of ZIKV-infected humans. We next examined whether ZIKV infection resulted in microbial translocation across mucosal surfaces by evaluating plasma and cerebrospinal fluid (CSF) levels of soluble CD14 (sCD14) and lipopolysaccharide-binding protein (LBP) by enzyme-linked immunosorbent assay (ELISA). Increased sCD14 was observed in the CSF of PTM and rhesus macaque (RM), while increased LBP was observed in pigtail macaque (PTM) plasma. Finally, to examine whether ZIKV-induced microbial dysbiosis could underlie increased microbial translocation and inflammation, we characterized intestinal microbial communities by 16s rRNA gene sequencing and microbial functional changes by quantifying short-chain fatty acid (SCFA) concentrations by gas chromatography mass spectrometry. We observed that although ZIKV infection of PTM did not result in significant taxonomic shifts in microbial communities, there were significant reductions in SCFA levels. Loss of microbial function in ZIKV infection could cause decreased intestinal integrity, thereby contributing to elevated microbial translocation and systemic and CNS inflammation, providing a possible mechanism underlying ZIKV pathogenesis. Further, this may represent a mechanism underlying inflammation and pathogenesis in other diseases.

Author SummaryZika virus (ZIKV) can be transmitted to humans via the bite of an infected mosquito or between humans during sexual intercourse, typically resulting in mild symptoms, which has been linked to elevated inflammation in the CNS and the development of more serious conditions, including severe neurological syndromes. Previous studies have observed that ZIKV infection is associated with increased mucosal dysfunction, including elevated inflammation in rectal and vaginal mucosal tissue. However, the mechanism of ZIKV-induced mucosal dysfunction may contribute to systemic and CNS inflammation has not been previously investigated. Here, we used the non-human primate (NHP) model and clinical specimens from ZIKV-infected humans to examine markers of systemic and CNS inflammation and microbial translocation. We observed elevated markers indicative of microbial translocation and inflammation in the CNS of ZIKV-infected macaques and humans. A potential association with mucosal dysfunction in ZIKV infection is shifts in microbial dysbiosis. We also observed that there were no significant overall taxonomic shifts in microbial communities, but a reduction of bacterial-derived short-chain fatty acid (SCFA) levels. Finally, we observed that the decrease in SCFA levels significantly negatively correlated with the elevated peripheral and CNS inflammatory markers, suggesting a link between ZIKV-driven disease pathology and microbial function. Taken together, our study provides new insight into a previously unconsidered mechanism underlying ZIKV pathogenesis.
]]></description>
<dc:creator>Miller, C.</dc:creator>
<dc:creator>Manuzak, J.</dc:creator>
<dc:creator>Gustin, A.</dc:creator>
<dc:creator>Basting, C.</dc:creator>
<dc:creator>Cheu, R.</dc:creator>
<dc:creator>Schroeder, T.</dc:creator>
<dc:creator>Velez, A.</dc:creator>
<dc:creator>Driscoll, C.</dc:creator>
<dc:creator>Tisoncik-Go, J.</dc:creator>
<dc:creator>Schifanella, L.</dc:creator>
<dc:creator>Hensley-McBain, T.</dc:creator>
<dc:creator>Evandy, C.</dc:creator>
<dc:creator>Smith, E.</dc:creator>
<dc:creator>Bratt, D.</dc:creator>
<dc:creator>Smedley, J.</dc:creator>
<dc:creator>O'Connor, M.</dc:creator>
<dc:creator>Fuller, D.</dc:creator>
<dc:creator>Barouch, D.</dc:creator>
<dc:creator>Gale, M.</dc:creator>
<dc:creator>Klatt, N. R.</dc:creator>
<dc:date>2023-07-26</dc:date>
<dc:identifier>doi:10.1101/2023.07.25.550459</dc:identifier>
<dc:title><![CDATA[Elevated peripheral and nervous system inflammation is associated with decreased short-chain fatty acid levels in Zika-virus infected macaques]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-07-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.07.27.550887v1?rss=1">
<title>
<![CDATA[
Calcium-sensitive subthreshold oscillations and electrical coupling in principal cells of mouse dorsal cochlear nucleus 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.07.27.550887v1?rss=1</link>
<description><![CDATA[
In higher sensory brain regions, slow oscillations (0.5-5 Hz) associated with quiet wakefulness and attention modulate multisensory integration, predictive coding, and perception. Although often assumed to originate via thalamocortical mechanisms, the extent to which sub-cortical sensory pathways are independently capable of slow oscillatory activity is unclear. We find that in the first station for auditory processing, the cochlear nucleus, fusiform cells from juvenile mice (of either sex) generate robust 1-2 Hz oscillations in membrane potential and exhibit electrical resonance. Such oscillations were absent prior to the onset of hearing, intrinsically generated by hyperpolarization-activated cyclic nucleotide-gated (HCN) and persistent Na+ conductances (NaP) interacting with passive membrane properties, and reflected the intrinsic resonance properties of fusiform cells. Cx36-containing gap junctions facilitated oscillation strength and promoted pairwise synchrony of oscillations between neighboring neurons. The strength of oscillations were strikingly sensitive to external Ca2+, disappearing at concentrations > 1.7 mM, due in part to the shunting effect of small-conductance calcium-activated potassium (SK) channels. This effect explains their apparent absence in previous in vitro studies of cochlear nucleus which routinely employed high-Ca2+ extracellular solution. In contrast, oscillations were amplified in reduced Ca2+ solutions, due to relief of suppression by Ca2+ of Na+ channel gating. Our results thus reveal mechanisms for synchronous oscillatory activity in auditory brainstem, suggesting that slow oscillations, and by extension their perceptual effects, may originate at the earliest stages of sensory processing.

SIGNIFICANCE STATEMENT (120 words max.)Many studies show that electrical activity in higher brain regions is regulated by brain oscillations. Here we show that such oscillatory activity can arise even in the first levels of auditory processing in the cochlear nucleus of the brainstem (fusiform cells), and is generated not by neural networks but by the biophysical properties of individual neurons. Oscillations are highly sensitive to external Ca2+ due to interplay of multiple ionic conductances. Gap junctions between cells allows for amplification and synchrony of such activity. Oscillations are absent in pre-hearing neurons, suggesting that sound activity might be important for their emergence. We propose that such early-level oscillations may serve to enhance signaling associated with particular environmental stimuli.
]]></description>
<dc:creator>Hong, H.</dc:creator>
<dc:creator>Moore, L. A.</dc:creator>
<dc:creator>Apostolides, P. F.</dc:creator>
<dc:creator>Trussell, L. O.</dc:creator>
<dc:date>2023-07-28</dc:date>
<dc:identifier>doi:10.1101/2023.07.27.550887</dc:identifier>
<dc:title><![CDATA[Calcium-sensitive subthreshold oscillations and electrical coupling in principal cells of mouse dorsal cochlear nucleus]]></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/10.1101/2023.07.27.550570v1?rss=1">
<title>
<![CDATA[
Neuronal NPR-15 controls the interplay between molecular and behavioral immune responses through the amphid sensory neuron-intestinal axis in C. elegans 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.07.27.550570v1?rss=1</link>
<description><![CDATA[
The survival of hosts during infections relies on their ability to mount effective molecular and behavioral immune responses. Despite extensive research on these defense strategies in various species, including the model organism Caenorhabditis elegans, the neural mechanisms underlying their interaction remain poorly understood. Previous studies have highlighted the role of neural G protein-coupled receptors (GPCRs) in regulating both immunity and pathogen avoidance, which is particularly dependent on aerotaxis. To address this knowledge gap, we conducted a screen of mutants in neuropeptide receptor family genes. We found that loss-of-function mutations in npr-15 activated immunity while suppressing pathogen avoidance behavior. Through further analysis, NPR-15 was found to regulate immunity by modulating the activity of key transcription factors, namely GATA/ELT-2 and TFEB/HLH-30. Surprisingly, the lack of pathogen avoidance of npr-15 mutant animals was not influenced by oxygen levels. Moreover, our studies revealed that the amphid sensory neuron ASJ is involved in mediating the immune and behavioral responses orchestrated by NPR-15. Additionally, NPR-15 was found to regulate avoidance behavior via the TRPM gene, GON-2, which may sense the intestinal distension caused by bacterial colonization to elicit pathogen avoidance. Our study contributes to a broader understanding of host defense strategies and mechanisms underlining the interaction between molecular and behavioral immune responses.
]]></description>
<dc:creator>Otarigho, B.</dc:creator>
<dc:creator>Butts, A. F.</dc:creator>
<dc:creator>Aballay, A.</dc:creator>
<dc:date>2023-07-29</dc:date>
<dc:identifier>doi:10.1101/2023.07.27.550570</dc:identifier>
<dc:title><![CDATA[Neuronal NPR-15 controls the interplay between molecular and behavioral immune responses through the amphid sensory neuron-intestinal axis in C. elegans]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-07-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.07.28.548760v1?rss=1">
<title>
<![CDATA[
Enhancing gene transfer to renal tubules and podocytes by context-dependent selection of AAV capsids 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.07.28.548760v1?rss=1</link>
<description><![CDATA[
Despite recent remarkable advancements in adeno-associated virus (AAV) vector technologies, effective gene delivery to the kidney remains a significant challenge. Here we show that AAV vector transduction in proximal tubules and podocytes, the crucial targets for renal gene therapy, can be enhanced remarkably through a meticulous selection of both AAV capsids and route of administration, tailored to the condition of the kidney. In this study, we performed a side-by-side comparison of 47 AAV capsids using AAV Barcode-Seq and identified six AAV capsids, including AAV-KP1, that exhibit remarkable enhancement of renal transduction in mice when delivered locally via the renal vein or the renal pelvis. Individual capsid validation analyses revealed that local delivery of AAV-KP1, but not AAV9, enables remarkably enhanced proximal tubule transduction while minimizing off-target liver transduction. In a mouse model of chronic kidney disease, intravenous administration of AAV9, not AAV-KP1, showed efficient renal tubule and podocyte transduction, which was not observed in the control wild-type mice. We also provide evidence that these contrasting observations between AAV-KP1 and AAV9 are attributed to their distinct pharmacokinetic profiles. Thus, this study highlights the importance of context-dependent capsid selection and engineering for successful renal gene therapy.
]]></description>
<dc:creator>Furusho, T.</dc:creator>
<dc:creator>Adachi, K.</dc:creator>
<dc:creator>Galbraith-Liss, M.</dc:creator>
<dc:creator>Sairavi, A.</dc:creator>
<dc:creator>Das, R.</dc:creator>
<dc:creator>Nakai, H.</dc:creator>
<dc:date>2023-07-29</dc:date>
<dc:identifier>doi:10.1101/2023.07.28.548760</dc:identifier>
<dc:title><![CDATA[Enhancing gene transfer to renal tubules and podocytes by context-dependent selection of AAV capsids]]></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/10.1101/2023.08.01.551546v1?rss=1">
<title>
<![CDATA[
Structure of an open KATP channel reveals tandem PIP2 binding sites mediating the Kir6.2 and SUR1 regulatory interface 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.08.01.551546v1?rss=1</link>
<description><![CDATA[
ATP-sensitive potassium (KATP) channels, composed of four pore-lining Kir6.2 subunits and four regulatory sulfonylurea receptor 1 (SUR1) subunits, control insulin secretion in pancreatic {beta}-cells. KATP channel opening is stimulated by PIP2 and inhibited by ATP. Mutations that increase channel opening by PIP2 reduce ATP inhibition and cause neonatal diabetes. Although considerable evidence has implicated a role for PIP2 in KATP channel function, previously solved open-channel structures have lacked bound PIP2, and mechanisms by which PIP2 regulates KATP channels remain unresolved. Here, we report cryoEM structure of a KATP channel harboring the neonatal diabetes mutation Kir6.2-Q52R, bound to natural C18:0/C20:4 long-chain PI(4,5)P2 in open conformation. The structure reveals two adjacent PIP2 molecules between SUR1 and Kir6.2. The first PIP2 binding site is conserved among PIP2-gated Kir channels. The second site forms uniquely in KATP at the interface of Kir6.2 and SUR1. Functional studies demonstrate both binding sites determine channel activity. Kir6.2 pore opening is associated with a twist of the Kir6.2 cytoplasmic domain and a rotation of the N-terminal transmembrane domain of SUR1, which widens the inhibitory ATP binding pocket to disfavor ATP binding. The open conformation is particularly stabilized by the Kir6.2-Q52R residue through cation-{pi} bonding with SUR1 - W51. Together, these results uncover the cooperation between SUR1 and Kir6.2 in PIP2 binding and gating, explain the antagonistic regulation of KATP channels by PIP2 and ATP, and provide the mechanism by which Kir6.2-Q52R stabilizes an open channel to cause neonatal diabetes.
]]></description>
<dc:creator>Driggers, C. M.</dc:creator>
<dc:creator>Kuo, Y.-Y.</dc:creator>
<dc:creator>Zhu, P.</dc:creator>
<dc:creator>ElSheikh, A.</dc:creator>
<dc:creator>Shyng, S.-L.</dc:creator>
<dc:date>2023-08-01</dc:date>
<dc:identifier>doi:10.1101/2023.08.01.551546</dc:identifier>
<dc:title><![CDATA[Structure of an open KATP channel reveals tandem PIP2 binding sites mediating the Kir6.2 and SUR1 regulatory interface]]></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/10.1101/2023.08.01.551556v1?rss=1">
<title>
<![CDATA[
Specific configurations of electrical synapses filter sensory information to drive choices in behavior 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.08.01.551556v1?rss=1</link>
<description><![CDATA[
Synaptic configurations in precisely wired circuits underpin how sensory information is processed by the nervous system, and the emerging animal behavior. This is best understood for chemical synapses, but far less is known about how electrical synaptic configurations modulate, in vivo and in specific neurons, sensory information processing and context-specific behaviors. We discovered that INX-1, a gap junction protein that forms electrical synapses, is required to deploy context-specific behavioral strategies during C. elegans thermotaxis behavior. INX-1 couples two bilaterally symmetric interneurons, and this configuration is required for the integration of sensory information during migration of animals across temperature gradients. In inx-1 mutants, uncoupled interneurons display increased excitability and responses to subthreshold temperature stimuli, resulting in abnormally longer run durations and context-irrelevant tracking of isotherms. Our study uncovers a conserved configuration of electrical synapses that, by increasing neuronal capacitance, enables differential processing of sensory information and the deployment of context-specific behavioral strategies.

One-Sentence SummaryCoupling of interneurons by electrical synapses reduces membrane resistance and filters sensory inputs to guide sensory-dependent behavioral choices.
]]></description>
<dc:creator>Almoril-Porras, A.</dc:creator>
<dc:creator>Calvo, A.</dc:creator>
<dc:creator>Niu, L.-G.</dc:creator>
<dc:creator>Beagan, J.</dc:creator>
<dc:creator>Hawk, J. D.</dc:creator>
<dc:creator>Aljobeh, A.</dc:creator>
<dc:creator>Wisdom, E.</dc:creator>
<dc:creator>Ren, I.</dc:creator>
<dc:creator>Diaz-Garcia, M.</dc:creator>
<dc:creator>Wang, Z.-W.</dc:creator>
<dc:creator>Colon-Ramos, D.</dc:creator>
<dc:date>2023-08-03</dc:date>
<dc:identifier>doi:10.1101/2023.08.01.551556</dc:identifier>
<dc:title><![CDATA[Specific configurations of electrical synapses filter sensory information to drive choices in behavior]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-08-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.08.03.551861v1?rss=1">
<title>
<![CDATA[
Gbx2 controls amacrine cell dendrite stratification through Robo1/2 receptors 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.08.03.551861v1?rss=1</link>
<description><![CDATA[
Within the neuronal classes of the retina, amacrine cells (ACs) exhibit the greatest neuronal diversity in morphology and function. We show that the selective expression of the transcription factor Gbx2 is required for cell fate specification and dendritic stratification of an individual AC subtype in the mouse retina. We identify Robo1 and Robo2 as downstream effectors that when deleted, phenocopy the dendritic misprojections seen in Gbx2 mutants. Slit1 and Slit2, the ligands of Robo receptors, are localized to the OFF layers of the inner plexiform layer where we observe the dendritic misprojections in both Gbx2 and Robo1/2 mutants. We show that Robo receptors also are required for the proper dendritic stratification of additional AC subtypes, such as Vglut3+ ACs. These results show both that Gbx2 functions as a terminal selector in a single AC subtype and identify Slit-Robo signaling as a developmental mechanism for ON-OFF pathway segregation in the retina.
]]></description>
<dc:creator>Kerstein, P. C.</dc:creator>
<dc:creator>Santana Agreda, Y.</dc:creator>
<dc:creator>Curran, B. M.</dc:creator>
<dc:creator>Ma, L.</dc:creator>
<dc:creator>Wright, K. M.</dc:creator>
<dc:date>2023-08-04</dc:date>
<dc:identifier>doi:10.1101/2023.08.03.551861</dc:identifier>
<dc:title><![CDATA[Gbx2 controls amacrine cell dendrite stratification through Robo1/2 receptors]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-08-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.08.04.552039v1?rss=1">
<title>
<![CDATA[
Intrinsic control of DRG sensory neuron diversification by Pten 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.08.04.552039v1?rss=1</link>
<description><![CDATA[
Autism spectrum disorder (ASD) is a complex neurodevelopmental disorder characterized by deficits in social interactions, repetitive behaviors, and hyper- or hyposensitivity to sensory stimuli. The mechanisms underlying the emergence of sensory features in ASD are not fully understood, but recent studies in rodent models highlight that these may result from differences in primary sensory neurons themselves. We examined sensory behaviors in a Pten haploinsufficient mouse model (PtenHet) for syndromic ASD and identified elevated responses to mechanical stimuli and a higher threshold to thermal responses. Transcriptomic and in vivo anatomical analysis identified alterations in subtype-specific markers of primary somatosensory neurons in PtenHet dorsal root ganglia (DRG). These defects emerge early during DRG development and involve dysregulation of multiple signaling pathways downstream of Pten. Finally, we show that mice harboring an ASD-associated mutation (PtenY69H) also show altered expression of somatosensory neuron subtype-specific markers. Together, these results show that precise levels of Pten are required for proper somatosensory development and provide insight into the molecular and cellular basis of sensory abnormalities in a model for syndromic ASD.
]]></description>
<dc:creator>Fernandez, A.</dc:creator>
<dc:creator>Sarn, N.</dc:creator>
<dc:creator>Eng, C.</dc:creator>
<dc:creator>Wright, K. M.</dc:creator>
<dc:date>2023-08-06</dc:date>
<dc:identifier>doi:10.1101/2023.08.04.552039</dc:identifier>
<dc:title><![CDATA[Intrinsic control of DRG sensory neuron diversification by Pten]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-08-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.08.07.552340v1?rss=1">
<title>
<![CDATA[
A Novel Burn / Synovectomy Mouse Model for Temporomandibular Joint Osteoarthritis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.08.07.552340v1?rss=1</link>
<description><![CDATA[
Temporomandibular Disorders (TMDs) is present in 33% of the U.S. population. Currently available animal models do not faithfully simulate the native disease progression of TMJ OA. The initiation of TMJ OA requires both local trauma and systemic inflammation. In this study, we present a novel mouse model which reproduces these two conditions. This is achieved by a procedure involving both synovectomy (local trauma) and a distant burn injury (systemic inflammation). Its efficacy at inducing TMJ OA was assessed with histomorphology and radiographic evaluation at 1,3, and 9 weeks after the procedure. We found that burn/synovectomy mice demonstrated significantly more degenerative hard and soft tissue changes in TMJ than uninjured control mice or synovotomy mice. The observed histology in burn/synoectomy mice mimicked native TMJ OA disease progression in a faithful manner. This animal model is invaluable in future research of the mechanism and risk factors of TMJ OA.
]]></description>
<dc:creator>Hsu, G. C.-Y.</dc:creator>
<dc:date>2023-08-08</dc:date>
<dc:identifier>doi:10.1101/2023.08.07.552340</dc:identifier>
<dc:title><![CDATA[A Novel Burn / Synovectomy Mouse Model for Temporomandibular Joint Osteoarthritis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-08-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.08.10.552874v1?rss=1">
<title>
<![CDATA[
Thyroid hormone rewires cortical circuits to coordinate body-wide metabolism and exploratory drive 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.08.10.552874v1?rss=1</link>
<description><![CDATA[
Animals adapt to varying environmental conditions by modifying the function of their internal organs, including the brain. To be adaptive, alterations in behavior must be coordinated with the functional state of organs throughout the body. Here we find that thyroid hormone-- a prominent regulator of metabolism in many peripheral organs-- activates cell-type specific transcriptional programs in anterior regions of cortex of adult mice via direct activation of thyroid hormone receptors. These programs are enriched for axon-guidance genes in glutamatergic projection neurons, synaptic regulators across both astrocytes and neurons, and pro-myelination factors in oligodendrocytes, suggesting widespread remodeling of cortical circuits. Indeed, whole-cell electrophysiology recordings revealed that thyroid hormone induces local transcriptional programs that rewire cortical neural circuits via pre-synaptic mechanisms, resulting in increased excitatory drive with a concomitant sensitization of recruited inhibition. We find that thyroid hormone bidirectionally regulates innate exploratory behaviors and that the transcriptionally mediated circuit changes in anterior cortex causally promote exploratory decision-making. Thus, thyroid hormone acts directly on adult cerebral cortex to coordinate exploratory behaviors with whole-body metabolic state.
]]></description>
<dc:creator>Hochbaum, D. R.</dc:creator>
<dc:creator>Dubinsky, A. C.</dc:creator>
<dc:creator>Farnsworth, H. C.</dc:creator>
<dc:creator>Hulshof, L.</dc:creator>
<dc:creator>Kleinberg, G.</dc:creator>
<dc:creator>Urke, A.</dc:creator>
<dc:creator>Wang, W.</dc:creator>
<dc:creator>Hakim, R.</dc:creator>
<dc:creator>Robertson, K.</dc:creator>
<dc:creator>Park, C.</dc:creator>
<dc:creator>Solberg, A.</dc:creator>
<dc:creator>Yang, Y.</dc:creator>
<dc:creator>Baynard, C.</dc:creator>
<dc:creator>Nadaf, N. M.</dc:creator>
<dc:creator>Beron, C.</dc:creator>
<dc:creator>Girasole, A.</dc:creator>
<dc:creator>Chantranupong, L.</dc:creator>
<dc:creator>Cortopassi, M.</dc:creator>
<dc:creator>Prouty, S.</dc:creator>
<dc:creator>Geistlinger, L.</dc:creator>
<dc:creator>Banks, A.</dc:creator>
<dc:creator>Scanlan, T.</dc:creator>
<dc:creator>Greenberg, M.</dc:creator>
<dc:creator>Boulting, G. L.</dc:creator>
<dc:creator>Macosko, E.</dc:creator>
<dc:creator>Sabatini, B. L.</dc:creator>
<dc:date>2023-08-10</dc:date>
<dc:identifier>doi:10.1101/2023.08.10.552874</dc:identifier>
<dc:title><![CDATA[Thyroid hormone rewires cortical circuits to coordinate body-wide metabolism and exploratory drive]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-08-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.08.11.553011v1?rss=1">
<title>
<![CDATA[
Cyclin B3 is a dominant fast-acting cyclin that drives rapid early embryonic mitoses 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.08.11.553011v1?rss=1</link>
<description><![CDATA[
In many species, early embryonic mitoses proceed at a very rapid pace, but how this pace is achieved is not understood. Here we show that in the early C. elegans embryo, cyclin B3 is the dominant driver of rapid embryonic mitoses. Metazoans typically have three cyclin B isoforms that associate with and activate Cdk1 kinase to orchestrate mitotic events: the related cyclins B1 and B2 and the more divergent cyclin B3. We show that whereas embryos expressing cyclins B1 and B2 support slow mitosis (NEBD to Anaphase [~] 600s), the presence of cyclin B3 dominantly drives the [~]3-fold faster mitosis observed in wildtype embryos. CYB-1/2-driven mitosis is longer than CYB-3-driven mitosis primarily because the progression of mitotic events itself is slower, rather than delayed anaphase onset due to activation of the spindle checkpoint or inhibitory phosphorylation of the anaphase activator CDC-20. Addition of cyclin B1 to cyclin B3-only mitosis introduces an [~]60s delay between the completion of chromosome alignment and anaphase onset, which likely ensures segregation fidelity; this delay is mediated by inhibitory phosphorylation on CDC-20. Thus, the dominance of cyclin B3 in driving mitotic events, coupled to introduction of a short cyclin B1-dependent delay in anaphase onset, sets the rapid pace and ensures fidelity of mitoses in the early C. elegans embryo.
]]></description>
<dc:creator>Lara-Gonzalez, P.</dc:creator>
<dc:creator>Variyar, S.</dc:creator>
<dc:creator>Budrewicz, J.</dc:creator>
<dc:creator>Schlientz, A.</dc:creator>
<dc:creator>Varshney, N.</dc:creator>
<dc:creator>Bellaart, A.</dc:creator>
<dc:creator>Moghareh, S.</dc:creator>
<dc:creator>Nguyen, A. C. N.</dc:creator>
<dc:creator>Oegema, K.</dc:creator>
<dc:creator>Desai, A.</dc:creator>
<dc:date>2023-08-12</dc:date>
<dc:identifier>doi:10.1101/2023.08.11.553011</dc:identifier>
<dc:title><![CDATA[Cyclin B3 is a dominant fast-acting cyclin that drives rapid early embryonic mitoses]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-08-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.08.10.552735v1?rss=1">
<title>
<![CDATA[
Cryopreservation of cerebrospinal fluid cells preserves transcriptomics integrity for single-cell analysis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.08.10.552735v1?rss=1</link>
<description><![CDATA[
Cerebrospinal fluid (CSF) matrix biomarkers have become increasingly valuable surrogate markers of neuropsychiatric diseases in research and clinical practice. In contrast, CSF cells have been rarely investigated due to their relative scarcity and fragility, and lack of common collection and cryopreservation protocols, with limited exceptions for neurooncology and primary immune-based diseases like multiple sclerosis. the advent of a microfluidics-based multi-omics approaches to studying individual cells has allowed for the study of cellular phenotyping, intracellular dynamics, and intercellular relationships that provide multidimensionality unable to be obtained through acellular fluid-phase analyses. challenges to cell-based research include site-to-site differences in handling, storage, and thawing methods, which can lead to inaccuracy and inter-assay variability. In the present study, we performed single-cell RNA sequencing (10x Genomics) on fresh or previously cryopreserved human CSF samples from three alternative cryopreservation methods: Fetal Bovine Serum with Dimethyl sulfoxide (FBS/DMSO), FBS/DMSO after a DNase step (a step often included in epigenetic studies), and cryopreservation using commercially available Recovery(C) media. In comparing relative differences between fresh and cryopreserved samples, we found little effect of the cryopreservation method on being able to resolve donor-linked cell type proportions, markers of cellular stress, and overall gene expression at the single-cell level, whereas donor-specific differences were readily discernable. We further demonstrate the compatibility of fresh and cryopreserved CSF immune cell sequencing using biologically relevant sexually dimorphic gene expression differences by donor. Our findings support the utility and interchangeability of FBS/DMSO and Recovery cryopreservation with fresh sample analysis, providing a methodological grounding that will enable researchers to further expand our understanding of the CSF immune cell contributions to neurological and psychiatric disease.
]]></description>
<dc:creator>Kodali, M. C.</dc:creator>
<dc:creator>Antone, J.</dc:creator>
<dc:creator>Alsop, E.</dc:creator>
<dc:creator>Jayakuymar, R.</dc:creator>
<dc:creator>Parikh, K.</dc:creator>
<dc:creator>Sanchez-Molina, P.</dc:creator>
<dc:creator>Ajami, B.</dc:creator>
<dc:creator>Arnold, S. E.</dc:creator>
<dc:creator>Van Keuren-Jensen, K.</dc:creator>
<dc:creator>Das, S.</dc:creator>
<dc:creator>Weinberg, M. S.</dc:creator>
<dc:date>2023-08-14</dc:date>
<dc:identifier>doi:10.1101/2023.08.10.552735</dc:identifier>
<dc:title><![CDATA[Cryopreservation of cerebrospinal fluid cells preserves transcriptomics integrity for single-cell analysis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-08-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.08.14.553195v1?rss=1">
<title>
<![CDATA[
TIGIT drives the immunosuppressive environment by downregulation of metalloproteinases MMP2 and MMP14 in perihilar cholangiocarcinoma. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.08.14.553195v1?rss=1</link>
<description><![CDATA[
BackgroundCheckpoint blockade in cholangiocarcinoma (CCA) is promising; however, little is known about the response to treatment in CCA subtypes. In this study, we investigated the spatial immune environment in combination with checkpoint expression in perihilar CCA (pCCA).

Materials & MethodsThe levels of checkpoint molecules (PD-1, PD-L1, PD-L2, LAG-3, ICOS, TIGIT, TIM-3, and CTLA-4), macrophages (CD68), and T cells (CD4 and CD8) were assessed by multiplex immunofluorescence (mIF) in 50 patients. We investigated the transcriptomic profile using the NanoString Cancer Progression Panel, and validation was performed by mIF on tissue sections from 24 patients.

ResultsThe expression of checkpoint molecules TIGIT, CTLA-4, and LAG-3 alone and in combination with other checkpoint molecules was more abundant in the Central Tumor (CT) and Invasive Margin (IM) than in peritumoral tissue (PT) (CD4 and CD8 TIGIT p<0.0001 for both CD4 and CD8 CTLA-4, p<0.0001 and p < 0.001, respectively, and CD8 LAG-3 p < 0.05). MMP2 and MMP14 were differentially expressed in patients with high TIGIT expression.

ConclusionThe immune environment in pCCA is characterized by the expression of multiple checkpoints, demonstrating the complexity of ICI treatment. High TIGIT expression drives an immunosuppressive environment by modulating the extracellular matrix. Future clinical trials in pCCA could consider TIGIT as a therapeutically relevant target for (combination) treatment.
]]></description>
<dc:creator>Heij, L.</dc:creator>
<dc:creator>Reichel, K.</dc:creator>
<dc:creator>de Koning, W.</dc:creator>
<dc:creator>Bednarsch, J.</dc:creator>
<dc:creator>Tan, X.</dc:creator>
<dc:creator>Campello Deierl, J.</dc:creator>
<dc:creator>Clahsen-van Groningen, M.</dc:creator>
<dc:creator>Al-Masri, T.</dc:creator>
<dc:creator>van Dam, R.</dc:creator>
<dc:creator>Garcia Vallejo, J.</dc:creator>
<dc:creator>Ulmer, F.</dc:creator>
<dc:creator>Lang, S.</dc:creator>
<dc:creator>Luedde, T.</dc:creator>
<dc:creator>Rocha, F.</dc:creator>
<dc:creator>Dahl, E.</dc:creator>
<dc:creator>Jonigk, D.</dc:creator>
<dc:creator>Kuehnel, M.</dc:creator>
<dc:creator>Sivakumar, S.</dc:creator>
<dc:creator>Neumann, U.</dc:creator>
<dc:date>2023-08-15</dc:date>
<dc:identifier>doi:10.1101/2023.08.14.553195</dc:identifier>
<dc:title><![CDATA[TIGIT drives the immunosuppressive environment by downregulation of metalloproteinases MMP2 and MMP14 in perihilar cholangiocarcinoma.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-08-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.08.16.553572v1?rss=1">
<title>
<![CDATA[
Long-term combination therapy with Metformin and Oxymetholone in a Fanconi Anemia mouse model 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.08.16.553572v1?rss=1</link>
<description><![CDATA[
Fanconi Anemia (FA) is a disease caused by defective DNA repair which manifests as bone marrow failure, cancer predisposition, and developmental defects. Mice containing inactivating mutations in one or more genes in the FA pathway partially mimic the human disease. We previously reported that monotherapy with either metformin (MET) or oxymetholone (OXM) improved peripheral blood (PB) counts and the number and functionality of bone marrow (BM) hematopoietic stem progenitor cells (HSPCs) number in Fancd2-/- mice. To evaluate whether the combination treatment of these drugs has a synergistic effect to prevent bone marrow failure in FA, we treated cohorts of Fancd2-/- mice and wild-type controls with either MET alone, OXM alone, MET+OXM or placebo diet. Both male and female mice were treated from age 3 weeks to 18 months. The OXM treated animals showed modest improvements in blood parameters including platelet count (p=0.01) and hemoglobin levels (p<0.05). In addition, the percentage of quiescent HSC (LSK) was significantly increased (p=0.001) by long-term treatment with MET alone. However, the absolute number of progenitors, measured by LSK frequency or CFU-S, was not significantly altered by MET therapy. The combination of metformin and oxymetholone did not result in a significant synergistic effect on any parameter. Male animals on MET+OXM or MET alone were significantly leaner than controls at 18 months, regardless of genotype. Gene expression analysis of liver tissue from these animals showed that some of the expression changes caused by Fancd2 deletion were partially normalized by metformin treatment. Importantly, no adverse effects of the individual or combination therapies were observed, despite the long-term administration.

Highlights- Long-term coadministration of metformin in combination oxymetholone is well tolerated by Fancd2-/- mice.
- HSC quiescence in mutant mice was enhanced by treatment with metformin alone.
- Metformin treatment caused a partial normalization of gene expression in the livers of mutant mice.
]]></description>
<dc:creator>Grompe, M.</dc:creator>
<dc:creator>Dorrell, C. S.</dc:creator>
<dc:creator>Peters, A. M.</dc:creator>
<dc:creator>Zhang, Q.</dc:creator>
<dc:creator>Balaji, N.</dc:creator>
<dc:creator>Mochizuki-Kashio, M.</dc:creator>
<dc:creator>Major, A.</dc:creator>
<dc:creator>Finegold, M. J.</dc:creator>
<dc:creator>Liu, C.</dc:creator>
<dc:creator>Lu, K.</dc:creator>
<dc:date>2023-08-16</dc:date>
<dc:identifier>doi:10.1101/2023.08.16.553572</dc:identifier>
<dc:title><![CDATA[Long-term combination therapy with Metformin and Oxymetholone in a Fanconi Anemia mouse model]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-08-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.08.15.553435v1?rss=1">
<title>
<![CDATA[
The α-crystallin chaperones undergo a quasi-ordered co-aggregation process in response to saturating client interaction 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.08.15.553435v1?rss=1</link>
<description><![CDATA[
Small heat shock proteins (sHSPs) are ATP-independent chaperones vital to cellular proteostasis, preventing protein aggregation events linked to various human diseases including cataract. The -crystallins, A-crystallin (Ac) and B-crystallin (Bc), represent archetypal sHSPs that exhibit complex polydispersed oligomeric assemblies and rapid subunit exchange dynamics. Yet, our understanding of how this plasticity contributes to chaperone function remains poorly understood. This study investigates structural changes in Ac and Bc during client sequestration under varying degree of chaperone saturation. Using biochemical and biophysical analyses combined with single-particle electron microscopy (EM), we examined Ac and Bc in their apo-states and at various stages of client-induced co-aggregation, using lysozyme as a model client. Quantitative single-particle analysis unveiled a continuous spectrum of oligomeric states formed during the co-aggregation process, marked by significant client-triggered expansion and quasi-ordered elongation of the sHSP scaffold. These structural modifications culminated in an apparent amorphous collapse of chaperone-client complexes, resulting in the creation of co-aggregates capable of scattering visible light. Intriguingly, these co-aggregates maintain internal morphological features of highly elongated sHSP scaffolding with striking resemblance to polymeric -crystallin species isolated from aged lens tissue. This mechanism appears consistent across both Ac and Bc, albeit with varying degrees of susceptibility to client-induced co-aggregation. Importantly, our findings suggest that client-induced co-aggregation follows a distinctive mechanistic and quasi-ordered trajectory, distinct from a purely amorphous process. These insights reshape our understanding of the physiological and pathophysiological co-aggregation processes of sHSPs, carrying potential implications for a pathway toward cataract formation.
]]></description>
<dc:creator>Miller, A. P.</dc:creator>
<dc:creator>O'Neill, S. E.</dc:creator>
<dc:creator>Lampi, K. J.</dc:creator>
<dc:creator>Reichow, S. L.</dc:creator>
<dc:date>2023-08-17</dc:date>
<dc:identifier>doi:10.1101/2023.08.15.553435</dc:identifier>
<dc:title><![CDATA[The α-crystallin chaperones undergo a quasi-ordered co-aggregation process in response to saturating client interaction]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-08-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.08.16.553618v1?rss=1">
<title>
<![CDATA[
Structure of C. elegans TMC-2 complex suggests roles of lipid-mediated subunit contacts in mechanosensory transduction 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.08.16.553618v1?rss=1</link>
<description><![CDATA[
Mechanotransduction is the process by which a mechanical force, such as touch, is converted into an electrical signal. Transmembrane channel-like (TMC) proteins are an evolutionarily-conserved family of ion channels whose function has been linked to a variety of mechanosensory processes, including hearing and balance sensation in vertebrates and locomotion in Drosophila. The molecular features that tune homologous TMC ion channel complexes to diverse mechanical stimuli are unknown. Caenorhabditis elegans express two TMC homologs, TMC-1 and TMC-2, both of which are the likely pore-forming subunits of mechanosensitive ion channels but differ in their expression pattern and functional role in the worm. Here we present the single particle cryo-electron microscopy structure of the native TMC-2 complex isolated from C. elegans. The complex is composed of two copies each of the pore-forming TMC-2 subunit, the calcium and integrin binding protein CALM-1 and the transmembrane inner ear protein TMIE. Comparison of the TMC-2 complex to the recently published cryo-EM structure of the C. elegans TMC-1 complex reveals differences in subunit composition and highlights conserved protein-lipid interactions, as well as other structural features, that together suggest a mechanism for TMC-mediated mechanosensory transduction.

Significance StatementOne mechanism by which organisms sense their environment is through the perception of mechanical stimuli such as sound, touch, and vibration. Transmembrane channel-like (TMC) proteins are ion channels whose function has been linked to a variety of mechanosensitive processes, including hearing and balance in vertebrates and touch sensation in worms. The molecular mechanisms by which TMCs respond to mechanical stimuli are unknown. Here we present the structure of the TMC-2 complex isolated from worms. Comparison of the TMC-2 complex to the recently solved structure of the worm TMC-1 complex highlights common structural features that are likely important for sensing mechanical stimuli yet also illuminates key differences that may explain the distinct functional roles of TMC-1 and TMC-2 in the worm.
]]></description>
<dc:creator>Clark, S.</dc:creator>
<dc:creator>Jeong, H.</dc:creator>
<dc:creator>Posert, R.</dc:creator>
<dc:creator>Goehring, A.</dc:creator>
<dc:creator>Gouaux, E.</dc:creator>
<dc:date>2023-08-17</dc:date>
<dc:identifier>doi:10.1101/2023.08.16.553618</dc:identifier>
<dc:title><![CDATA[Structure of C. elegans TMC-2 complex suggests roles of lipid-mediated subunit contacts in mechanosensory transduction]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-08-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.08.16.553641v1?rss=1">
<title>
<![CDATA[
ESAT-6 undergoes self-association at phagosomal pH and an ESAT-6 specific nanobody restricts M. tuberculosis growth in macrophages 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.08.16.553641v1?rss=1</link>
<description><![CDATA[
Mycobacterium tuberculosis (Mtb) is known to survive within macrophages by compromising the integrity of the phagosomal compartment in which it resides. This activity primarily relies on the ESX-1 secretion system, predominantly involving the protein duo ESAT-6 and CFP-10. CFP-10 likely acts as a chaperone, while ESAT-6 likely disrupts phagosomal membrane stability via a largely unknown mechanism. we employ a series of biochemical analyses, protein modeling techniques, and a novel ESAT-6-specific nanobody to gain insight into the ESAT-6s mode of action. First, we measure the binding kinetics of the tight 1:1 complex formed by ESAT-6 and CFP-10 at neutral pH. Subsequently, we demonstrate a rapid self-association of ESAT-6 into large complexes under acidic conditions, leading to the identification of a stable tetrameric ESAT-6 species. Using molecular dynamics simulations, we pinpoint the most probable interaction interface. Furthermore, we show that cytoplasmic expression of an anti-ESAT-6 nanobody blocks Mtb replication, thereby underlining the pivotal role of ESAT-6 in intracellular survival. Together, these data suggest that ESAT-6 acts by a pH dependent mechanism to establish two-way communication between the cytoplasm and the Mtb-containing phagosome.
]]></description>
<dc:creator>Bates, T. A.</dc:creator>
<dc:creator>Trank-Greene, M.</dc:creator>
<dc:creator>Nguyenla, X.</dc:creator>
<dc:creator>Anastas, A.</dc:creator>
<dc:creator>Merutka, I. R.</dc:creator>
<dc:creator>Dixon, S. D.</dc:creator>
<dc:creator>Shumate, A.</dc:creator>
<dc:creator>Groncki, A. R.</dc:creator>
<dc:creator>Parson, M. A.</dc:creator>
<dc:creator>Barklis, E.</dc:creator>
<dc:creator>Burke, J. E.</dc:creator>
<dc:creator>Shinde, U.</dc:creator>
<dc:creator>Ploegh, H. L.</dc:creator>
<dc:creator>Tafesse, F. G.</dc:creator>
<dc:date>2023-08-17</dc:date>
<dc:identifier>doi:10.1101/2023.08.16.553641</dc:identifier>
<dc:title><![CDATA[ESAT-6 undergoes self-association at phagosomal pH and an ESAT-6 specific nanobody restricts M. tuberculosis growth in macrophages]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-08-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.08.17.553761v1?rss=1">
<title>
<![CDATA[
Complementary roles of orbitofrontal and prelimbic cortices in adaption of reward motivated actions to learned anxiety. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.08.17.553761v1?rss=1</link>
<description><![CDATA[
BackgroundAnxiety is a common symptom of several mental health disorders and adversely affects motivated behaviors. Anxiety can emerge from associating risk of future harm while engaged in goal-guided actions. Using a recently developed behavioral paradigm to model this aspect of anxiety, we investigated the role of two cortical subregions, the prelimbic medial frontal cortex (PL) and lateral orbitofrontal cortex (lOFC), which have been implicated in anxiety and outcome expectation, in flexible representation of actions associated with harm risk.

MethodsA seek-take reward-guided instrumental task design was used to train animals to associate the seek action with a variable risk of punishment. After learning, animals underwent extinction training for this association. Fiber photometry was used to measure and compare neuronal activity in PL and lOFC during learning and extinction.

ResultsAnimals increased action suppression in response to punishment contingencies. This increase dissipated after extinction training. These behavioral changes were associated with region specific changes in neuronal activity. PL neuronal activity preferentially adapted to threat of punishment whereas lOFC activity adapted to safe aspects of the task. Moreover, correlated activity between these regions was suppressed during actions associated with harm risk suggesting that these regions may guide behavior independently under anxiety.

ConclusionsThese findings suggest the PL and lOFC serve distinct but complementary roles in the representation of learned anxiety. This dissociation may provide a mechanism for how overlapping cortical systems are implicated in reward-guided action execution during anxiety.
]]></description>
<dc:creator>Jacobs, D. S.</dc:creator>
<dc:creator>Bogachuk, A. P.</dc:creator>
<dc:creator>Moghaddam, B.</dc:creator>
<dc:date>2023-08-17</dc:date>
<dc:identifier>doi:10.1101/2023.08.17.553761</dc:identifier>
<dc:title><![CDATA[Complementary roles of orbitofrontal and prelimbic cortices in adaption of reward motivated actions to learned anxiety.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-08-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.08.17.553732v1?rss=1">
<title>
<![CDATA[
Enlarged Perivascular Spaces are Associated with White Matter Injury, Brain Atrophy, Cognitive Decline and Markers of Inflammation in an Autosomal Dominant Vascular Neurodegenerative Disease (CADASIL) 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.08.17.553732v1?rss=1</link>
<description><![CDATA[
Background and ObjectivesEnlarged perivascular spaces (ePVS) have been previously reported in Cerebral Autosomal Dominant Arteriopathy with Subcortical Infarcts and Leucoencephalopathy (CADASIL), but their significance and pathophysiology remains unclear. We investigated associations of ePVS with classical imaging measures, cognitive measures and plasma proteins to better understand what ePVS represents in CADASIL and whether radiographic measures of ePVS would be of value in future therapeutic discovery studies for CADASIL.

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

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

DiscussionBased on associations uncovered between ePVS volumes and cognitive functions, imaging and plasma proteins, we conclude that ePVS volumes capture pathologies contributing to chronic brain dysfunction and degeneration in CADASIL, with relevance to future clinical trials for novel therapeutic discoveries to prevent decline and injury in individuals carrying NOTCH3 mutations.
]]></description>
<dc:creator>Karvelas, N.</dc:creator>
<dc:creator>Oh, B.</dc:creator>
<dc:creator>Wang, E.</dc:creator>
<dc:creator>Cobigo, Y.</dc:creator>
<dc:creator>Tsuei, T.</dc:creator>
<dc:creator>Fitzsimons, S.</dc:creator>
<dc:creator>Ehrenberg, A.</dc:creator>
<dc:creator>Geschwind, M.</dc:creator>
<dc:creator>Schwartz, D.</dc:creator>
<dc:creator>Kramer, J.</dc:creator>
<dc:creator>Ferguson, A. R.</dc:creator>
<dc:creator>Miller, B. L.</dc:creator>
<dc:creator>Silbert, L.</dc:creator>
<dc:creator>Rosen, H.</dc:creator>
<dc:creator>Elahi, F. M.</dc:creator>
<dc:date>2023-08-19</dc:date>
<dc:identifier>doi:10.1101/2023.08.17.553732</dc:identifier>
<dc:title><![CDATA[Enlarged Perivascular Spaces are Associated with White Matter Injury, Brain Atrophy, Cognitive Decline and Markers of Inflammation in an Autosomal Dominant Vascular Neurodegenerative Disease (CADASIL)]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-08-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.08.18.553854v1?rss=1">
<title>
<![CDATA[
Multiplex imaging of localized prostate tumors reveals changes in mast cell type composition and spatial organization of AR-positive cells in the tumor microenvironment 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.08.18.553854v1?rss=1</link>
<description><![CDATA[
Mapping spatial interactions of cancer, immune and stromal cells present novel opportunities for patient stratification and for advancing immunotherapy. While single-cell studies revealed significant molecular heterogeneity in prostate tumors, there is currently no understanding of how immune cell heterogeneity impacts spatial coordination between tumor and stromal cells in localized tumors. Here, we used cyclic immunofluorescent imaging on whole-tissue sections to uncover novel spatial associations between cancer and stromal cells in low- and high-grade prostate tumors and tumor-adjacent normal tissues. Our results provide a spatial map of 699,461 single-cells that show epigenetic and molecular differences in distinct clinical grades. We report unique populations of mast cells that differentially express CD44, CD90 and Granzyme B (GZMB) and demonstrate GZMB+ mast cells are spatially associated with M2 macrophages in prostate tumors. Finally, we uncover recurrent neighborhoods that are primarily driven by androgen receptor positive (AR+) stromal cells and identify transcriptional networks active in AR+ prostate stroma.
]]></description>
<dc:creator>Eksi, S. E.</dc:creator>
<dc:creator>Ak, C.</dc:creator>
<dc:creator>Sayar, Z.</dc:creator>
<dc:creator>Thibault, G.</dc:creator>
<dc:creator>Burlingame, E.</dc:creator>
<dc:creator>Eng, J.</dc:creator>
<dc:creator>Chitsazan, A.</dc:creator>
<dc:creator>Adey, A.</dc:creator>
<dc:creator>Boniface, C.</dc:creator>
<dc:creator>Spellman, P. T.</dc:creator>
<dc:creator>Thomas, G. V.</dc:creator>
<dc:creator>Kopp, R.</dc:creator>
<dc:creator>Demir, E.</dc:creator>
<dc:creator>Chang, Y. H.</dc:creator>
<dc:creator>Stavrinides, V.</dc:creator>
<dc:date>2023-08-21</dc:date>
<dc:identifier>doi:10.1101/2023.08.18.553854</dc:identifier>
<dc:title><![CDATA[Multiplex imaging of localized prostate tumors reveals changes in mast cell type composition and spatial organization of AR-positive cells in the tumor microenvironment]]></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/10.1101/2023.08.24.554559v1?rss=1">
<title>
<![CDATA[
Improved dual-color GRAB sensors for monitoring dopaminergic activity in vivo 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.08.24.554559v1?rss=1</link>
<description><![CDATA[
Dopamine (DA) plays multiple roles in a wide range of physiological and pathological processes via a vast network of dopaminergic projections. To fully dissect the spatiotemporal dynamics of DA release in both dense and sparsely innervated brain regions, we developed a series of green and red fluorescent GPCR activation-based DA (GRABDA) sensors using a variety of DA receptor subtypes. These sensors have high sensitivity, selectivity, and signal-to-noise properties with subsecond response kinetics and the ability to detect a wide range of DA concentrations. We then used these sensors in freely moving mice to measure both optogenetically evoked and behaviorally relevant DA release while measuring neurochemical signaling in the nucleus accumbens, amygdala, and cortex. Using these sensors, we also detected spatially resolved heterogeneous cortical DA release in mice performing various behaviors. These next-generation GRABDA sensors provide a robust set of tools for imaging dopaminergic activity under a variety of physiological and pathological conditions.
]]></description>
<dc:creator>Zhuo, Y.</dc:creator>
<dc:creator>Luo, B.</dc:creator>
<dc:creator>Yi, X.</dc:creator>
<dc:creator>Dong, H.</dc:creator>
<dc:creator>Wan, J.</dc:creator>
<dc:creator>Cai, R.</dc:creator>
<dc:creator>Williams, J. T.</dc:creator>
<dc:creator>Qian, T.</dc:creator>
<dc:creator>Campbell, M. G.</dc:creator>
<dc:creator>Miao, X.</dc:creator>
<dc:creator>Li, B.</dc:creator>
<dc:creator>Wei, Y.</dc:creator>
<dc:creator>Li, G.</dc:creator>
<dc:creator>Wang, H.</dc:creator>
<dc:creator>Zheng, Y.</dc:creator>
<dc:creator>Watabe-Uchida, M.</dc:creator>
<dc:creator>Li, Y.</dc:creator>
<dc:date>2023-08-25</dc:date>
<dc:identifier>doi:10.1101/2023.08.24.554559</dc:identifier>
<dc:title><![CDATA[Improved dual-color GRAB sensors for monitoring dopaminergic activity in vivo]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-08-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.08.24.554733v1?rss=1">
<title>
<![CDATA[
Quantitative image analysis pipeline for detecting circulating hybrid cells in immunofluorescence images with human-level accuracy 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.08.24.554733v1?rss=1</link>
<description><![CDATA[
Circulating hybrid cells (CHCs) are a newly discovered, tumor-derived cell population identified in the peripheral blood of cancer patients and are thought to contribute to tumor metastasis. However, identifying CHCs by immunofluorescence (IF) imaging of patient peripheral blood mononuclear cells (PBMCs) is a time-consuming and subjective process that currently relies on manual annotation by laboratory technicians. Additionally, while IF is relatively easy to apply to tissue sections, its application on PBMC smears presents challenges due to the presence of biological and technical artifacts. To address these challenges, we present a robust image analysis pipeline to automate the detection and analyses of CHCs in IF images. The pipeline incorporates quality control to optimize specimen preparation protocols and remove unwanted artifacts, leverages a {beta}-variational autoencoder (VAE) to learn meaningful latent representations of single-cell images and employs a support vector machine (SVM) classifier to achieve human-level CHC detection. We created a rigorously labeled IF CHC dataset including 9 patients and 2 disease sites with the assistance of 10 annotators to evaluate the pipeline. We examined annotator variation and bias in CHC detection and then provided guidelines to optimize the accuracy of CHC annotation. We found that all annotators agreed on CHC identification for only 65% of the cells in the dataset and had a tendency to underestimate CHC counts for regions of interest (ROI) containing relatively large amounts of cells (>50,000) when using conventional enumeration methods. On the other hand, our proposed approach is unbiased to ROI size. The SVM classifier trained on the {beta}-VAE encodings achieved an F1 score of 0.80, matching the average performance of annotators. Our pipeline enables researchers to explore the role of CHCs in cancer progression and assess their potential as a clinical biomarker for metastasis. Further, we demonstrate that the pipeline can identify discrete cellular phenotypes among PBMCs, highlighting its utility beyond CHCs.
]]></description>
<dc:creator>Anderson, A.</dc:creator>
<dc:creator>Theison, H.</dc:creator>
<dc:creator>Baik, J.</dc:creator>
<dc:creator>Gibbs, S.</dc:creator>
<dc:creator>Wong, M. H.</dc:creator>
<dc:creator>Chang, Y. H.</dc:creator>
<dc:date>2023-08-26</dc:date>
<dc:identifier>doi:10.1101/2023.08.24.554733</dc:identifier>
<dc:title><![CDATA[Quantitative image analysis pipeline for detecting circulating hybrid cells in immunofluorescence images with human-level accuracy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-08-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.08.25.554774v1?rss=1">
<title>
<![CDATA[
Local and dynamic regulation of neuronal glycolysis in vivo 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.08.25.554774v1?rss=1</link>
<description><![CDATA[
Energy metabolism supports neuronal function. While it is well established that changes in energy metabolism underpin brain plasticity and function, less is known about how individual neurons modulate their metabolic states to meet varying energy demands. This is because most approaches used to examine metabolism in living organisms lack the resolution to visualize energy metabolism within individual circuits, cells, or subcellular regions. Here we adapted a biosensor for glycolysis, HYlight, for use in C. elegans to image dynamic changes in glycolysis within individual neurons and in vivo. We determined that neurons perform glycolysis cell-autonomously, and modulate glycolytic states upon energy stress. By examining glycolysis in specific neurons, we documented a neuronal energy landscape comprising three general observations: 1) glycolytic states in neurons are diverse across individual cell types; 2) for a given condition, glycolytic states within individual neurons are reproducible across animals; and 3) for varying conditions of energy stress, glycolytic states are plastic and adapt to energy demands. Through genetic analyses, we uncovered roles for regulatory enzymes and mitochondrial localization in the cellular and subcellular dynamic regulation of glycolysis. Our study demonstrates the use of a single-cell glycolytic biosensor to examine how energy metabolism is distributed across cells and coupled to dynamic states of neuronal function, and uncovers new relationships between neuronal identities and metabolic landscapes in vivo.

Significance statementWhile it is generally accepted that energy metabolism underpins neuronal function, how it is distributed and dynamically regulated in different tissues of the brain to meet varying energy demands is not well understood. Here we utilized a fluorescent biosensor, HYlight, to observe glycolytic metabolism at cellular and subcellular scales in vivo. By leveraging both the stereotyped identities of individual neurons in C. elegans, and genetic tools for manipulating glycolytic metabolism, we determined that neurons perform and dynamically regulate glycolysis to match changing cellular demands for energy. Our findings support a model whereby glycolytic states should be considered distinct and related to individual neuron identities in vivo, and introduce new questions about the interconnected nature of metabolism and neuronal function.
]]></description>
<dc:creator>Wolfe, A. D.</dc:creator>
<dc:creator>Koberstein, J. N.</dc:creator>
<dc:creator>Smith, C. B.</dc:creator>
<dc:creator>Stewart, M. L.</dc:creator>
<dc:creator>Hammarlund, M.</dc:creator>
<dc:creator>Hyman, A. A.</dc:creator>
<dc:creator>Stork, P. J. S.</dc:creator>
<dc:creator>Goodman, R. H.</dc:creator>
<dc:creator>Colon-Ramos, D. A.</dc:creator>
<dc:date>2023-08-26</dc:date>
<dc:identifier>doi:10.1101/2023.08.25.554774</dc:identifier>
<dc:title><![CDATA[Local and dynamic regulation of neuronal glycolysis in vivo]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-08-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.08.26.554902v1?rss=1">
<title>
<![CDATA[
Functional subtypes of rodent melanopsin ganglion cells switch roles between night and day illumination. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.08.26.554902v1?rss=1</link>
<description><![CDATA[
Intrinsically photosensitive retinal ganglion cells (ipRGCs), contain the photopigment melanopsin, and influence both image and non-image forming behaviors. Despite being categorized into multiple types (M1-M6), physiological variability within these types suggests our current understanding of ipRGCs is incomplete. We used multi-electrode array (MEA) recordings and unbiased cluster analysis under synaptic blockade to identify 8 functional clusters of ipRGCs, each with distinct photosensitivity and response timing. We used Cre mice to drive the expression of channelrhodopsin in SON-ipRGCs, enabling the localization of distinct ipRGCs in the dorsal retina. Additionally, we conducted a retrospective unbiased cluster analysis of ipRGC photoresponses to light stimuli across scotopic, mesopic, and photopic intensities, aimed at activating both rod and cone inputs to ipRGCs. Our results revealed shared and distinct synaptic inputs to the identified functional clusters, demonstrating that ipRGCs encode visual information with high fidelity at low light intensities, but poorly at photopic light intensities, when melanopsin activation is highest. Collectively, our findings support a framework with at least 8 functional subtypes of ipRGCs, each encoding luminance with distinct spike outputs, highlighting the inherent functional diversity and complexity of ipRGCs and suggesting a reevaluation of their contributions to retinal function and visual perception under varying light conditions.
]]></description>
<dc:creator>Berry, M. H.</dc:creator>
<dc:creator>Leffler, J.</dc:creator>
<dc:creator>Allen, C. N.</dc:creator>
<dc:creator>Sivyer, B.</dc:creator>
<dc:date>2023-08-27</dc:date>
<dc:identifier>doi:10.1101/2023.08.26.554902</dc:identifier>
<dc:title><![CDATA[Functional subtypes of rodent melanopsin ganglion cells switch roles between night and day illumination.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-08-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.08.26.554953v1?rss=1">
<title>
<![CDATA[
Single-cell analysis of Rohon-Beard neurons implicates Fgf signaling in axon maintenance and cell survival 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.08.26.554953v1?rss=1</link>
<description><![CDATA[
Peripheral sensory neurons are a critical part of the nervous system that transmit a multitude of sensory stimuli to the central nervous system. During larval and juvenile stages in zebrafish, this function is mediated by Rohon-Beard somatosensory neurons (RBs). RBs are optically accessible and amenable to experimental manipulation, making them a powerful system for mechanistic investigation of sensory neurons. Previous studies provided evidence that RBs fall into multiple subclasses; however, the number and molecular make up of these potential RB subtypes have not been well defined. Using a single-cell RNA sequencing (scRNA-seq) approach, we demonstrate that larval RBs in zebrafish fall into three, largely non-overlapping classes of neurons. We also show that RBs are molecularly distinct from trigeminal neurons in zebrafish. Cross-species transcriptional analysis indicates that one RB subclass is similar to a mammalian group of A-fiber sensory neurons. Another RB subclass is predicted to sense multiple modalities, including mechanical stimulation and chemical irritants. We leveraged our scRNA-seq data to determine that the fibroblast growth factor (Fgf) pathway is active in RBs. Pharmacological and genetic inhibition of this pathway led to defects in axon maintenance and RB cell death. Moreover, this can be phenocopied by treatment with dovitinib, an FDA-approved Fgf inhibitor with a common side effect of peripheral neuropathy. Importantly, dovitinib-mediated axon loss can be suppressed by loss of Sarm1, a positive regulator of neuronal cell death and axonal injury. This offers a molecular target for future clinical intervention to fight neurotoxic effects of this drug.
]]></description>
<dc:creator>nechiporuk, a.</dc:creator>
<dc:creator>Tuttle, A. M.</dc:creator>
<dc:creator>Royer, L. J.</dc:creator>
<dc:creator>Kelly, J. J.</dc:creator>
<dc:creator>Wen, H.</dc:creator>
<dc:creator>Calistri, N.</dc:creator>
<dc:creator>Heiser, L. M.</dc:creator>
<dc:date>2023-08-28</dc:date>
<dc:identifier>doi:10.1101/2023.08.26.554953</dc:identifier>
<dc:title><![CDATA[Single-cell analysis of Rohon-Beard neurons implicates Fgf signaling in axon maintenance and cell survival]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-08-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.08.27.554228v1?rss=1">
<title>
<![CDATA[
Complete correction of murine phenylketonuria by selection-enhanced hepatocyte transplantation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.08.27.554228v1?rss=1</link>
<description><![CDATA[
Hepatocyte transplantation for genetic liver diseases has several potential advantages over gene therapy. However, low efficiency of cell engraftment has limited its clinical implementation. This problem could be overcome by selectively expanding transplanted donor cells until they replace enough of the liver mass to achieve therapeutic benefit. We previously described a gene therapy method to selectively expand hepatocytes deficient in cytochrome p450 reductase (Cypor) using acetaminophen (APAP). Because Cypor is required for the transformation of APAP to a hepatotoxic metabolite, Cypor deficient cells are protected from toxicity and are able to expand following APAP-induced liver injury. Here, we apply this selection system to correct a mouse model of phenylketonuria (PKU) by cell transplantation. Hepatocytes from a wildtype donor animal were edited in vitro to create Cypor deficiency and then transplanted into PKU animals. Following selection with APAP, blood phenylalanine concentrations were fully normalized and remained stable following APAP withdrawal. Cypor-deficient hepatocytes expanded from <1% to [~]14% in corrected animals, and they showed no abnormalities in blood chemistries, liver histology, or drug metabolism. We conclude that APAP-mediated selection of transplanted hepatocytes is a potential therapeutic for PKU with long-term efficacy and a favorable safety profile.
]]></description>
<dc:creator>Vonada, A.</dc:creator>
<dc:creator>Wakefield, L.</dc:creator>
<dc:creator>Martinez, M.</dc:creator>
<dc:creator>Harding, C. O.</dc:creator>
<dc:creator>Grompe, M.</dc:creator>
<dc:creator>Tiyaboonchai, A.</dc:creator>
<dc:date>2023-08-28</dc:date>
<dc:identifier>doi:10.1101/2023.08.27.554228</dc:identifier>
<dc:title><![CDATA[Complete correction of murine phenylketonuria by selection-enhanced hepatocyte transplantation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-08-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.08.30.555301v1?rss=1">
<title>
<![CDATA[
Modulation of riboflavin biosynthesis and utilization in mycobacteria 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.08.30.555301v1?rss=1</link>
<description><![CDATA[
Riboflavin (vitamin B2) is the precursor of the flavin coenzymes, FAD and FMN, which play a central role in cellular redox metabolism. While humans must obtain riboflavin from dietary sources, certain microbes, including Mycobacterium tuberculosis (Mtb), can biosynthesize riboflavin de novo. Riboflavin precursors have also been implicated in the activation of mucosal-associated invariant T (MAIT) cells which recognize metabolites derived from the riboflavin biosynthesis pathway complexed to the MHC-I-like molecule, MR1. To investigate the biosynthesis and function of riboflavin and its pathway intermediates in mycobacterial metabolism, physiology and MAIT cell recognition, we constructed conditional knockdowns (hypomorphs) in riboflavin biosynthesis and utilization genes in Mycobacterium smegmatis (Msm) and Mtb by inducible CRISPR interference. Using this comprehensive panel of hypomorphs, we analyzed the impact of gene silencing on viability, on the transcription of (other) riboflavin pathway genes, on the levels of the pathway proteins and on riboflavin itself. Our results revealed that (i) despite lacking a canonical transporter, both Msm and Mtb assimilate exogenous riboflavin when supplied at high concentration; (ii) there is functional redundancy in lumazine synthase activity in Msm; (iii) silencing of ribA2 or ribF is profoundly bactericidal in Mtb; and (iv) in Msm, ribA2 silencing results in concomitant knockdown of other pathway genes coupled with RibA2 and riboflavin depletion and is also bactericidal. In addition to their use in genetic validation of potential drug targets for tuberculosis, this collection of hypomorphs provides a useful resource for investigating the role of pathway intermediates in MAIT cell recognition of mycobacteria.

IMPORTANCEThe pathway for biosynthesis and utilization of riboflavin, precursor of the essential coenzymes, FMN and FAD, is of particular interest in the flavin-rich pathogen, Mycobacterium tuberculosis (Mtb), for two important reasons: (i) the pathway includes potential tuberculosis (TB) drug targets; and (ii) intermediates from the riboflavin biosynthesis pathway provide ligands for mucosal associated invariant T (MAIT) cells, which have been implicated in TB pathogenesis. However, the riboflavin pathway is poorly understood in mycobacteria, which lack canonical mechanisms to transport this vitamin and to regulate flavin coenzyme homeostasis. By conditionally disrupting each step of the pathway and assessing the impact on mycobacterial viability and on the levels of the pathway proteins as well as riboflavin, our work provides genetic validation of the riboflavin pathway as a target for TB drug discovery and offers a resource for further exploring the association between riboflavin biosynthesis, MAIT cell activation and TB infection and disease.
]]></description>
<dc:creator>Chengalroyen, M. D.</dc:creator>
<dc:creator>Mehaffy, C.</dc:creator>
<dc:creator>Lucas, M.</dc:creator>
<dc:creator>Raphela, M. L.</dc:creator>
<dc:creator>Warner, D. F.</dc:creator>
<dc:creator>Lewinsohn, D.</dc:creator>
<dc:creator>Lewinsohn, D.</dc:creator>
<dc:creator>Dobos, K.</dc:creator>
<dc:creator>Mizrahi, V.</dc:creator>
<dc:date>2023-08-30</dc:date>
<dc:identifier>doi:10.1101/2023.08.30.555301</dc:identifier>
<dc:title><![CDATA[Modulation of riboflavin biosynthesis and utilization in mycobacteria]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-08-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.08.29.555369v1?rss=1">
<title>
<![CDATA[
Lymphatic vessel transit seeds precursors to cytotoxic resident memory T cells in skin draining lymph nodes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.08.29.555369v1?rss=1</link>
<description><![CDATA[
Resident memory T cells (TRM) provide rapid, localized protection in peripheral tissues to pathogens and cancer. While TRM are also found in lymph nodes (LN), how they develop during primary infection and their functional significance remains largely unknown. Here, we track the anatomical distribution of anti-viral CD8+ T cells as they simultaneously seed skin and LN TRM using a model of skin infection with restricted antigen distribution. We find exquisite localization of LN TRM to the draining LN of infected skin. LN TRM formation depends on lymphatic transport and specifically egress of effector CD8+ T cells that appear poised for residence as early as 12 days post infection. Effector CD8+ T cell transit through skin is necessary and sufficient to populate LN TRM in draining LNs, a process reinforced by antigen encounter in skin. Importantly, we demonstrate that LN TRM are sufficient to provide protection against pathogenic rechallenge. These data support a model whereby a subset of tissue infiltrating CD8+ T cells egress during viral clearance, and establish regional protection in the draining lymphatic basin as a mechanism to prevent pathogen spread.

One Sentence SummaryT cell egress out of virally infected skin via afferent lymphatic vessels seeds CD8+ resident memory T cells in the draining lymph node.
]]></description>
<dc:creator>Heim, T.</dc:creator>
<dc:creator>Schultz, A. C.</dc:creator>
<dc:creator>Delclaux, I.</dc:creator>
<dc:creator>Cristaldi, V.</dc:creator>
<dc:creator>Churchill, M. J.</dc:creator>
<dc:creator>Lund, A. W.</dc:creator>
<dc:date>2023-08-31</dc:date>
<dc:identifier>doi:10.1101/2023.08.29.555369</dc:identifier>
<dc:title><![CDATA[Lymphatic vessel transit seeds precursors to cytotoxic resident memory T cells in skin draining lymph nodes]]></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/10.1101/2023.08.31.555803v1?rss=1">
<title>
<![CDATA[
A Linkable, Polycarbonate Gut Microbiome-Distal Tumor Chip Platform for Interrogating Cancer Promoting Mechanisms 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.08.31.555803v1?rss=1</link>
<description><![CDATA[
Gut microbiome composition has been tied to diseases ranging from arthritis to cancer to depression. However, mechanisms of action are poorly understood, limiting development of relevant therapeutics. Organ-on-chip platforms, which model minimal functional units of tissues and can tightly control communication between them, are ideal platforms to study these relationships. Many gut microbiome models have been published to date but devices are typically fabricated using oxygen permeable PDMS, requiring interventions to support anaerobic bacteria. To address this challenge, a novel platform was developed where the chips were fabricated entirely from gas-impermeable polycarbonate without tapes or gaskets. These chips replicated polarized villus-like structures of the native tissue. Further, they enabled co-cultures of commensal anaerobic bacteria Blautia coccoides on the surface of gut epithelia for two days within a standard incubator. Another complication of PDMS devices is high ad-/absorption, limiting applications in high-resolution microscopy and biomolecule interaction studies. For future communication studies between gut microbiota and distal tumors, an additional polycarbonate chip design was developed to support hydrogel-embedded tissue culture. These chips enable high-resolution microscopy with all relevant processing done on-chip. Designed for facile linking, this platform will make a variety of mechanistic studies possible.
]]></description>
<dc:creator>Brasino, D. S.</dc:creator>
<dc:creator>Speese, S. D.</dc:creator>
<dc:creator>Schilling, K.</dc:creator>
<dc:creator>Schutt, C. E.</dc:creator>
<dc:creator>Barton, M. C.</dc:creator>
<dc:date>2023-09-03</dc:date>
<dc:identifier>doi:10.1101/2023.08.31.555803</dc:identifier>
<dc:title><![CDATA[A Linkable, Polycarbonate Gut Microbiome-Distal Tumor Chip Platform for Interrogating Cancer Promoting Mechanisms]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-09-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.09.03.556067v1?rss=1">
<title>
<![CDATA[
Human milk extracellular vesicles survive neonatal digestion in vivo to be absorbed by neonatal enteroids 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.09.03.556067v1?rss=1</link>
<description><![CDATA[
Human milk contains extracellular vesicles (HMEVs). Pre-clinical models suggest that HMEVs may enhance intestinal function and limit inflammation; however, it is unknown if HMEVs or their cargo survive neonatal human digestion. This limits the ability to leverage HMEV cargo as additives to infant nutrition or as therapeutics. This study aimed to develop an EV isolation pipeline from small volumes of human milk and neonatal intestinal contents after milk feeding (digesta) to address the hypothesis that HMEVs survive in vivo neonatal digestion to be taken up intestinal epithelial cells (IECs). Digesta was collected from nasoduodenal sampling tubes or ostomies. EVs were isolated from raw and pasteurized human milk and digesta by density-gradient ultracentrifugation following two-step skimming, acid precipitation of caseins, and multi-step filtration. EVs were validated by electron microscopy, western blotting, nanoparticle tracking analysis, resistive pulse sensing, and super-resolution microscopy. EV uptake was tested in human neonatal enteroids. HMEVs and digesta EVs (dEVs) show typical EV morphology and are enriched in CD81 and CD9, but depleted of {beta}-casein and lactalbumin. HMEV and some dEV fractions contain mammary gland-derived protein BTN1A1. Neonatal human enteroids rapidly take up dEVs in part via clathrin-mediated endocytosis. Our data suggest that EVs can be isolated from digestive fluid and that these dEVs can be absorbed by IECs.
]]></description>
<dc:creator>Zhang, Y.</dc:creator>
<dc:creator>Kuhn, M.</dc:creator>
<dc:creator>Yung, C.</dc:creator>
<dc:creator>Armstrong, R. J.</dc:creator>
<dc:creator>Olyaei, A.</dc:creator>
<dc:creator>Aloia, M.</dc:creator>
<dc:creator>Scottoline, B.</dc:creator>
<dc:creator>Andres, S. F.</dc:creator>
<dc:date>2023-09-03</dc:date>
<dc:identifier>doi:10.1101/2023.09.03.556067</dc:identifier>
<dc:title><![CDATA[Human milk extracellular vesicles survive neonatal digestion in vivo to be absorbed by neonatal enteroids]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-09-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.09.05.556390v1?rss=1">
<title>
<![CDATA[
Normative Birth Weight Variation is Associated with White Matter Connectivity in Full-Term Neonates 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.09.05.556390v1?rss=1</link>
<description><![CDATA[
ImportanceVariation in birth weight, an indicator of fetal growth, has been prospectively associated with both alterations in brain development and developmental delays in cognitive and language domains. However, studies examining birth weight variation and white matter development in the brain typically test these associations in infants that are preterm or very low birth weight, leaving potential normative associations in full term infants unclear.

ObjectiveHere, we test prospective associations between birth weight variation in the  normative range (> 2.5 kg and <4.5 kg) and white matter connectivity in full-term neonates. Further, the main objective includes testing associations between normative birth weight variation and cognitive and language developmental scores at 18 months, and if white matter connectivity that is related to birth weight variation is further associated with cognitive language development. We hypothesized, greater normative birth weight would be associated with higher white matter connectivity controlling for gestational age, particularly in thalamic, inferior frontal, and middle temporal connectivity. Further, we predicted greater connectivity for these tracts would be associated with higher scores for cognitive (thalamic) and language development (inferior and middle temporal) at 18 months of age.

DesignThe study was an observational longitudinal design of data from the Developing Human Connectome Project (dHCP). Birth weight was measured at the birth of the child, white matter connectivity was measured as neonates (mean=40.07 weeks, SD=1.14), and cognitive/language outcomes were measured at 18 months of age.

SettingThe dHCP data was collected at the Evelina Newborn Imaging Centre, Evalina London Childrens Hospital between 2015 and 2019.

ParticipantsA sub-sample of the full dHCP was tested. These participants were full-term neonates with birth weight variation in the  normative range (> 2.5 kg and < 4.5 kg). Participants also had to have usable diffusion-weighted imaging data as neonates and cognitive/language developmental assessments collected at 18 months (n=323).

Exposure(s)The study participants were all born at full-term and in the normative birth weight range.

Main Outcome(s) and Measure(s)The study had two main outcomes: white matter connectivity at the neonatal timepoint and cognitive/language developmental scores at 18 months. White matter connectivity was calculated from diffusion-weighted data for the whole-brain. Cognitive/language developmental scores were measured using the Bayley Scales of Infant and Toddler Development, Third Edition (Bayley-III) at 18 months.

ResultsUsing a Network-Based Statistic (NBS) approach, we found widespread associations between normative birth weight variation and white matter connectivity in full-term neonates, primarily in the positive direction for the right middle occipital gyrus and left supplementary motor area.

Conclusions and RelevanceWhile investigations have been focused on the extreme ends of the birth weight spectrum, we find evidence that there is a robust association between birth weight and white matter connectivity even within the normative birth weight range. As normative birth weight variation and regions of white matter associated with birth weight were further associated with language development scores at 18 months, our results suggest the birth weight to white matter pathway may be an underlying pathway between birth weights association with language development.

Key PointsO_ST_ABSQuestionC_ST_ABSAre variations in normative birth weight associated with white matter connectivity and cognitive/language outcomes in infancy?

FindingsGreater normative birth weight is associated with greater white matter organization across a widespread network of connections in the neonatal brain. Greater white matter organization in this network for neonates has a positive prospective with expressive language development at 18 months of age.

MeaningEven variations within the normative birth weight range have robust associations with early white matter development and can be prospectively linked to language development.
]]></description>
<dc:creator>Dufford, A. J.</dc:creator>
<dc:creator>Dai, W.</dc:creator>
<dc:creator>Scheinost, D.</dc:creator>
<dc:date>2023-09-05</dc:date>
<dc:identifier>doi:10.1101/2023.09.05.556390</dc:identifier>
<dc:title><![CDATA[Normative Birth Weight Variation is Associated with White Matter Connectivity in Full-Term Neonates]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-09-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.09.05.555810v1?rss=1">
<title>
<![CDATA[
Enriched Single-Nucleus RNA-Sequencing reveals unique attributes of distal convoluted tubule cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.09.05.555810v1?rss=1</link>
<description><![CDATA[
BackgroundThe distal convoluted tubule (DCT) comprises two subsegments, DCT1 and DCT2, with different functional and molecular characteristics. The functional and molecular distinction between these segments, however, has been controversial.

MethodsTo understand the heterogeneity within the DCT population with better clarity, we enriched for DCT nuclei by using a mouse line combining "Isolation of Nuclei TAgged in specific Cell Types" and NCC (sodium chloride cotransporter)-driven inducible Cre recombinase. We sorted the fluorescently labeled DCT nuclei using Fluorescence-Activated Nucleus Sorting, and performed single nucleus transcriptomics.

ResultsAmong 25,183 DCT cells, 70% were from DCT1 and 30% from DCT2. Additionally, there was a small population (<1%) enriched in proliferation-related genes, such asTop2a, Cenpp, and Mki67. Both DCT1 and DCT2 express NCC, magnesium transport genes are more abundant along DCT1; whereas calcium, electrogenic sodium and potassium transport genes are more abundant along DCT2. The transition between these two segments are gradual with a transitional zone where DCT1 and DCT2 cells are interspersed. The expression of the homeobox genes is not consistent between all DCT cells, suggesting that they develop along different trajectories.

ConclusionTranscriptomics analysis of an enriched rare cell population using genetically targeted approach offers better clarification of the function and classification. The DCT segment is short, yet, can be separated into two sub-types that serve distinct functions, and are speculated to derive from different origins during development.

Significance StatementHigh-resolution snRNAseq data indicate a clear separation between primary sites of calcium and magnesium handling within DCT. Both DCT1 and DCT2 express Slc12a3, but these subsegments serve distinctive functions, with more abundant magnesium handling genes along DCT1 and more calcium handling genes along DCT2. The data also provides insight into the plasticity of the distal nephron-collecting duct junction, formed from cells of separate embryonic origins. By focusing/changing gradients of gene expression, the DCT can morph into different physiological cell states on demand.
]]></description>
<dc:creator>Su, X.-T.</dc:creator>
<dc:creator>Reyes, J. V.</dc:creator>
<dc:creator>Lackey, A. E.</dc:creator>
<dc:creator>Maeoka, Y.</dc:creator>
<dc:creator>Cornelius, R. J.</dc:creator>
<dc:creator>McCormick, J. A.</dc:creator>
<dc:creator>Yang, C.-L.</dc:creator>
<dc:creator>Jung, H. J.</dc:creator>
<dc:creator>Welling, P. A.</dc:creator>
<dc:creator>Nelson, J. W.</dc:creator>
<dc:creator>Ellison, D. H.</dc:creator>
<dc:date>2023-09-06</dc:date>
<dc:identifier>doi:10.1101/2023.09.05.555810</dc:identifier>
<dc:title><![CDATA[Enriched Single-Nucleus RNA-Sequencing reveals unique attributes of distal convoluted tubule cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-09-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.09.05.556437v1?rss=1">
<title>
<![CDATA[
PKA catalytic subunits dissociate from regulatory subunits to regulate neuronal functions 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.09.05.556437v1?rss=1</link>
<description><![CDATA[
Protein kinase A (PKA) plays essential roles in diverse cellular functions. However, the spatiotemporal dynamics of endogenous PKA upon activation remain debated. The classical model predicts that PKA catalytic subunits dissociate from regulatory subunits in the presence of cAMP, whereas a second model proposes that catalytic subunits remain associated with regulatory subunits following physiological activation. Here we report that different PKA subtypes, as defined by the regulatory subunit, exhibit distinct subcellular localization at rest in CA1 neurons of cultured hippocampal slices. Nevertheless, when all tested PKA subtypes are activated by norepinephrine, presumably via the {beta}-adrenergic receptor, catalytic subunits translocate to dendritic spines but regulatory subunits remain unmoved. These differential spatial dynamics between the subunits indicate that at least a significant fraction of PKA dissociates. Furthermore, PKA-dependent regulation of synaptic plasticity and transmission can be supported only by wildtype, dissociable PKA, but not by inseparable PKA. These results indicate that endogenous PKA regulatory and catalytic subunits dissociate to achieve PKA function in neurons.
]]></description>
<dc:creator>Xiong, W.</dc:creator>
<dc:creator>Qin, M.</dc:creator>
<dc:creator>Zhong, H.</dc:creator>
<dc:date>2023-09-07</dc:date>
<dc:identifier>doi:10.1101/2023.09.05.556437</dc:identifier>
<dc:title><![CDATA[PKA catalytic subunits dissociate from regulatory subunits to regulate neuronal functions]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-09-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.09.08.556943v1?rss=1">
<title>
<![CDATA[
Mass spectrometry and split luciferase complementation assays reveal the MecA protein interactome of Streptococcus mutans 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.09.08.556943v1?rss=1</link>
<description><![CDATA[
MecA is a highly conserved adaptor protein encoded by prokaryotes from the Bacillota phylum. MecA mutants exhibit similar pleiotropic defects in a variety of organisms, although most of these phenotypes currently lack a mechanistic basis. MecA mediates ClpCP-dependent proteolysis of its substrates, but only several such substrates have been reported in the literature and there are suggestions that proteolysis-independent regulatory mechanisms may also exist. Here, we provide the first comprehensive characterization of the MecA interactome and further assess its regulatory role in Clp-dependent proteolysis. Untargeted coimmunoprecipitation assays coupled with mass spectrometry revealed that the MecA ortholog from the oral pathobiont Streptococcus mutans likely serves as a major protein interaction network hub by potentially complexing with >100 distinct protein substrates, most of which function in highly conserved metabolic pathways. The interactome results were independently verified using a newly developed prokaryotic split luciferase complementation assay (SLCA) to detect MecA protein-protein interactions in vivo. In addition, we further develop a new application of SLCA to support in vivo measurements of MecA relative protein binding affinities. SLCA results were independently verified using targeted coimmunoprecipitation assays, suggesting the general utility of this approach for prokaryotic protein-protein interaction studies. Our results indicate that MecA indeed regulates its interactome through both Clp-dependent proteolysis as well as through an as yet undefined proteolysis-independent mechanism that may affect more than half of its protein interactome. This suggests a significant aspect of MecA regulatory function still has yet to be discovered.
]]></description>
<dc:creator>Qin, H.</dc:creator>
<dc:creator>Anderson, D.</dc:creator>
<dc:creator>Zou, Z.</dc:creator>
<dc:creator>Higashi, D.</dc:creator>
<dc:creator>Borland, C.</dc:creator>
<dc:creator>Kreth, J. L.</dc:creator>
<dc:creator>Merritt, J.</dc:creator>
<dc:date>2023-09-09</dc:date>
<dc:identifier>doi:10.1101/2023.09.08.556943</dc:identifier>
<dc:title><![CDATA[Mass spectrometry and split luciferase complementation assays reveal the MecA protein interactome of Streptococcus mutans]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-09-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.09.13.556416v1?rss=1">
<title>
<![CDATA[
Cardiac glycosides restore autophagy flux in an iPSC-derived neuronal model of WDR45 deficiency 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.09.13.556416v1?rss=1</link>
<description><![CDATA[
Beta-Propeller Protein-Associated Neurodegeneration (BPAN) is one of the commonest forms of Neurodegeneration with Brain Iron Accumulation, caused by mutations in the gene encoding the autophagy-related protein, WDR45. The mechanisms linking autophagy, iron overload and neurodegeneration in BPAN are poorly understood and, as a result, there are currently no disease-modifying treatments for this progressive disorder. We have developed a patient-derived, induced pluripotent stem cell (iPSC)-based midbrain dopaminergic neuronal cell model of BPAN (3 patient, 2 age-matched controls and 2 isogenic control lines) which shows defective autophagy and aberrant gene expression in key neurodegenerative, neurodevelopmental and collagen pathways. A high content imaging-based medium-throughput blinded drug screen using the FDA-approved Prestwick library identified 5 cardiac glycosides that both corrected disease-related defective autophagosome formation and restored BPAN-specific gene expression profiles. Our findings have clear translational potential and emphasise the utility of iPSC-based modelling in elucidating disease pathophysiology and identifying targeted therapeutics for early-onset monogenic disorders.
]]></description>
<dc:creator>Papandreou, A.</dc:creator>
<dc:creator>Singh, N.</dc:creator>
<dc:creator>Gianfrancesco, L.</dc:creator>
<dc:creator>Budinger, D.</dc:creator>
<dc:creator>Barwick, K.</dc:creator>
<dc:creator>Agrotis, A.</dc:creator>
<dc:creator>Luft, C.</dc:creator>
<dc:creator>Shao, Y.</dc:creator>
<dc:creator>Lenaerts, A.-S.</dc:creator>
<dc:creator>Gregory, A.</dc:creator>
<dc:creator>Jeong, S. Y.</dc:creator>
<dc:creator>Hogarth, P.</dc:creator>
<dc:creator>Hayflick, S.</dc:creator>
<dc:creator>Barral, S.</dc:creator>
<dc:creator>Kriston-Vizi, J.</dc:creator>
<dc:creator>Gissen, P.</dc:creator>
<dc:creator>Kurian, M. A. A.</dc:creator>
<dc:creator>Ketteler, R.</dc:creator>
<dc:date>2023-09-13</dc:date>
<dc:identifier>doi:10.1101/2023.09.13.556416</dc:identifier>
<dc:title><![CDATA[Cardiac glycosides restore autophagy flux in an iPSC-derived neuronal model of WDR45 deficiency]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-09-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.09.12.557449v1?rss=1">
<title>
<![CDATA[
Extracellular matrix scaffold-assisted tumor vaccines induce tumor regression and long-term immune memory 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.09.12.557449v1?rss=1</link>
<description><![CDATA[
Injectable scaffold delivery is an immune engineering strategy to enhance the efficacy and reliability of cancer vaccine immunotherapy. The composition and structure of the biomaterial scaffold determines both vaccine release kinetics and inherent immune stimulation via the scaffold host response. Extracellular matrix (ECM) scaffolds prepared from decellularized tissues initiate an acute alternative inflammatory response following implantation, which facilitates wound healing following tumor resection and promotes local cancer immune surveillance. However, it remains unknown whether this environment is compatible with generating protective anti-tumor cytotoxic immunity with local immunotherapy delivery. Here, we engineered an ECM scaffold-assisted therapeutic cancer vaccine that maintained an immune microenvironment consistent with tissue reconstruction. Immune adjuvants MPLA, GM-CSF, and CDA were screened in a cancer vaccine formulated for decellularized small intestinal submucosa (SIS) ECM scaffold co-delivery. Though MPLA and GM-CSF showed the greatest increase in local myeloid cell infiltration, we found that the STING pathway adjuvant CDA was the most potent inducer of cytotoxic immunity with SIS-ECM scaffold delivery. Further, CDA did not diminish hallmark ECM immune responses needed in wound healing such as high Il4 cytokine expression. SIS scaffold delivery enhanced therapeutic vaccine efficacy using CDA and the antigen ovalbumin, curing greater than 50% of established EG.7 tumors in young mice and 75% in 24-week-old mature mice, compared to soluble components alone (0% cured). SIS-ECM scaffold assisted vaccination extended antigen exposure, was dependent on CD8+ cytotoxic T cells, and generated long term anti-tumor memory at least 7 months post-vaccination in both young and mature-aged mice. This study shows that an ECM scaffold is a promising delivery vehicle to enhance cancer vaccine efficacy while being orthogonal to characteristics of pro-healing immune hallmarks.
]]></description>
<dc:creator>Pal, S.</dc:creator>
<dc:creator>Chaudhari, R.</dc:creator>
<dc:creator>Baurceanu, I.</dc:creator>
<dc:creator>Hill, B. J.</dc:creator>
<dc:creator>Nagy, B. A.</dc:creator>
<dc:creator>Wolf, M.</dc:creator>
<dc:date>2023-09-15</dc:date>
<dc:identifier>doi:10.1101/2023.09.12.557449</dc:identifier>
<dc:title><![CDATA[Extracellular matrix scaffold-assisted tumor vaccines induce tumor regression and long-term immune memory]]></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/10.1101/2023.09.13.556997v1?rss=1">
<title>
<![CDATA[
Intracellular K+ limits T cell exhaustion and preserves antitumor function 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.09.13.556997v1?rss=1</link>
<description><![CDATA[
The cancer-killing activity of T cells is often compromised within tumors, allowing disease progression. We previously found that intratumoral elevations in extracellular K+ related to ongoing cell death constrained CD8+ T cell Akt-mTOR signaling and effector function (1,2). To alleviate K+ mediated T cell suppression, we pursued genetic means to lower intracellular K+.

Transcriptomic analysis of CD8+ T cells demonstrated the Na+/K+ ATPase to be robustly and dynamically expressed. CRISPR-Cas9 mediated deletion of the catalytic alpha subunit of the Na+/K+ ATPase lowered intracellular K+ but produced tonic hyperactivity in multiple signal transduction cascades along with the acquisition of co-inhibitory receptors and terminal differentiation in mouse and human CD8+ T cells. Mechanistically, Na+/K+ ATPase disruption led to ROS accumulation due to depletion of intracellular K+ in T cells. Antioxidant treatment or high K+ media prevented Atp1a1 deficient T cells from exhausted T (TEx) cell formation. Consistent with transcriptional and proteomic data suggesting a TEx cell phenotype, T cells lacking Atp1a1 had compromised persistence and antitumor activity in a syngeneic model of orthotopic murine melanoma. Translational application of these findings will include efforts to lower intracellular K+ while limiting ROS accumulation within tumor specific T cells.

SynopsisHigh extracellular K+ ({uparrow}[K+]e) is found within tumors and suppresses T cell effector function. Collier et al. find that deletion of the Na+/K+ ATPase in T cells lowers intracellular K+ and promotes ROS accumulation, tonic signal transduction and T cell exhaustion owing to ROS accumulation. Engineering T cell ion transport is an important consideration for cancer immunotherapy.
]]></description>
<dc:creator>Collier, C.</dc:creator>
<dc:creator>Wucherer, K.</dc:creator>
<dc:creator>Mcwhorter, M.</dc:creator>
<dc:creator>Jenkins, C.</dc:creator>
<dc:creator>Bartlett, A.</dc:creator>
<dc:creator>Roychoudhuri, R.</dc:creator>
<dc:creator>Eil, R.</dc:creator>
<dc:date>2023-09-15</dc:date>
<dc:identifier>doi:10.1101/2023.09.13.556997</dc:identifier>
<dc:title><![CDATA[Intracellular K+ limits T cell exhaustion and preserves antitumor function]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-09-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.09.15.557996v1?rss=1">
<title>
<![CDATA[
Androgen blockade primes NLRP3 in macrophages to induce tumor phagocytosis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.09.15.557996v1?rss=1</link>
<description><![CDATA[
Immune-based therapies induce durable remissions in subsets of patients across multiple malignancies. However, there is limited efficacy of immunotherapy in metastatic castrate-resistant prostate cancer (mCRPC), manifested by an enrichment of immunosuppressive (M2) tumor- associated macrophages (TAM) in the tumor immune microenvironment (TME). Therefore, therapeutic strategies to overcome TAM-mediated immunosuppression are critically needed in mCRPC. Here we discovered that NLR family pyrin domain containing 3 (NLRP3), an innate immune sensing protein, is highly expressed in TAM from metastatic PC patients treated with standard-of-care androgen deprivation therapy (ADT). Importantly, ex vivo studies revealed that androgen receptor (AR) blockade in TAM upregulates NLRP3 expression, but not inflammasome activity, and concurrent AR blockade/NLRP3 agonist (NLRP3a) treatment promotes cancer cell phagocytosis by immunosuppressive M2 TAM. In contrast, NLRP3a monotherapy was sufficient to enhance phagocytosis of cancer cells in anti-tumor (M1) TAM, which exhibit high de novo NLRP3 expression. Critically, combinatorial treatment with ADT/NLRP3a in a murine model of advanced PC resulted in significant tumor control, with tumor clearance in 55% of mice via TAM phagocytosis. Collectively, our results demonstrate NLRP3 as an AR-regulated "macrophage phagocytic checkpoint", inducibly expressed in TAM by ADT and activated by NLRP3a treatment, the combination resulting in TAM-mediated phagocytosis and tumor control.
]]></description>
<dc:creator>Chaudagar, K. K.</dc:creator>
<dc:creator>Srikrishnan, R.</dc:creator>
<dc:creator>Mei, S.</dc:creator>
<dc:creator>Hirz, T.</dc:creator>
<dc:creator>Hu, Y.-M.</dc:creator>
<dc:creator>Argulian, A.</dc:creator>
<dc:creator>Labadie, B.</dc:creator>
<dc:creator>Desai, K.</dc:creator>
<dc:creator>Grimaldo, S.</dc:creator>
<dc:creator>Kahramangil, D.</dc:creator>
<dc:creator>Nair, R.</dc:creator>
<dc:creator>D Souza, S.</dc:creator>
<dc:creator>Zhou, D.</dc:creator>
<dc:creator>Li, M.</dc:creator>
<dc:creator>Doughan, F.</dc:creator>
<dc:creator>Chen, R.</dc:creator>
<dc:creator>Shafran, J.</dc:creator>
<dc:creator>Loyd, M.</dc:creator>
<dc:creator>Xia, Z.</dc:creator>
<dc:creator>Sykes, D. B.</dc:creator>
<dc:creator>Moran, A. E.</dc:creator>
<dc:creator>Patnaik, A.</dc:creator>
<dc:date>2023-09-17</dc:date>
<dc:identifier>doi:10.1101/2023.09.15.557996</dc:identifier>
<dc:title><![CDATA[Androgen blockade primes NLRP3 in macrophages to induce tumor phagocytosis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-09-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.09.18.557989v1?rss=1">
<title>
<![CDATA[
Fetal Membrane Inflammasome Activation by Choriodecidual Ureaplasma parvum Infection without Intra-Amniotic Infection in an NHP Model 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.09.18.557989v1?rss=1</link>
<description><![CDATA[
Intrauterine infection is a significant cause of preterm labor and neonatal morbidity and mortality. Ureaplasma parvum is the micro-organism most commonly isolated from cases of preterm birth and preterm premature rupture of membranes (pPROM). However, the mechanisms during the early stages of ascending reproductive tract infection that initiate maternal-fetal inflammatory pathways, preterm birth and pPROM remain poorly understood. To examine inflammation in fetal (chorioamnionic) membranes in response to Ureaplasma parvum infection, we utilized a novel in vivo non-human primate model of early choriodecidual infection. Eight chronically catheterized pregnant rhesus macaques underwent maternal-fetal catheterization surgery at 105-112 days gestation and choriodecidual inoculation with Ureaplasma parvum (105cfu/mL of a low passaged clinical isolate, serovar 1; n=4) or saline/sterile media (Controls; n=4) starting at 115-119 days gestation, repeated every 5 days until scheduled cesarean-section at 136-140d gestation (term=167d). The average inoculation to delivery interval was 21 days and Ureaplasma infection of the amniotic fluid was undetectable by culture and PCR in all animals. Inflammatory mediators in amniotic fluid (AF) were assessed by Luminex, ELISA and multiplex assays. RNA was extracted from the chorion and amnionic membranes for single gene analysis (qRT-PCR) and protein expression was determined by Western blot and immunohistochemistry. Our NHP model of choriodecidual Ureaplasma infection, representing an early-stage ascending reproductive tract infection without microbial invasion of the amniotic cavity, resulted in increased fetal membrane protein and gene expression of MMP-9 and PTGS2, but did not result in preterm labor (no increase in uterine contractility) or increased concentrations of amniotic fluid pro-inflammatory cytokines (IL-1{beta}, IL-6, IL-8, IL-18, TNF-). However, membrane expression of inflammasome sensor molecules, NLRP3, NLRC4, AIM2 and NOD2, and the adaptor protein ASC (PYCARD) gene expression were significantly increased in the Ureaplasma group when compared to non-infected controls. Gene expression of IL-1{beta}, IL-18, the IL-18R1 receptor, CASPASE-1 and pro-CASPASE-1 protein were also increased in the fetal membranes with Ureaplasma infection. Downstream inflammatory signaling genes MYD88 was also significantly upregulated in both the amnion and chorion, along with a significant increase in NFKB in the chorion. These results demonstrate that even at the early stages of ascending reproductive tract Ureaplasma infection, activation of inflammasome complexes and pathways associated with degradation of chorioamnionic membrane integrity are present. This study therefore provides experimental evidence for the importance of the early stages of ascending Ureaplasma infection in initiating processes of pPROM and preterm labor. These findings have implications for the identification of intrauterine inflammation before microbes are detectable in the amniotic fluid (sterile inflammation) and the timing of potential treatments for preterm labor and fetal injury caused by intrauterine infection.
]]></description>
<dc:creator>Tripathy, S.</dc:creator>
<dc:creator>Burd, I.</dc:creator>
<dc:creator>Kelleher, M. A.</dc:creator>
<dc:date>2023-09-22</dc:date>
<dc:identifier>doi:10.1101/2023.09.18.557989</dc:identifier>
<dc:title><![CDATA[Fetal Membrane Inflammasome Activation by Choriodecidual Ureaplasma parvum Infection without Intra-Amniotic Infection in an NHP Model]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-09-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.09.19.558512v1?rss=1">
<title>
<![CDATA[
Leukemic mutation FLT3-ITD is retained in dendritic cells and disrupts their homeostasis leading to expanded Th17 frequency 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.09.19.558512v1?rss=1</link>
<description><![CDATA[
Dendritic cells (DC) are mediators of adaptive immune responses to pathogens and tumors. DC development is determined by signaling through the receptor tyrosine kinase Fms-like tyrosine kinase 3 (FLT3) in bone marrow myeloid progenitors. Recently the naming conventions for DC phenotypes have been updated to distinguish between "Conventional" DCs (cDCs) and plasmacytoid DCs (pDCs). Activating mutations of FLT3, including Internal Tandem Duplication (FLT3-ITD), are associated with poor prognosis for leukemia patients. To date, there is little information on the effects of FLT3-ITD in DC biology. We examined the cDC phenotype and frequency in bone marrow aspirates from patients with acute myeloid leukemia (AML) to understand the changes to cDCs associated with FLT3-ITD. When compared to healthy donor (HD) we found that a subset of FLT3-ITD+ AML patient samples have overrepresented populations of cDCs and disrupted phenotypes. Using a mouse model of FLT3-ITD+ AML, we found that cDCs were increased in percentage and number compared to control wild-type (WT) mice. Single cell RNA-seq identified FLT3-ITD+ cDCs as skewed towards a cDC2 T-bet-phenotype, previously shown to promote Th17 T cells. We assessed the phenotypes of CD4+ T cells in the AML mice and found significant enrichment of both Treg and Th17 CD4+ T cells. Furthermore, co-culture of AML mouse- derived DCs and naive OT-II cells preferentially skewed T cells into a Th17 phenotype. Together, our data suggests that FLT3-ITD+ leukemia-associated cDCs polarize CD4+ T cells into Th17 subsets, a population that has been shown to be negatively associated with survival in solid tumor contexts. This illustrates the complex tumor microenvironment of AML and highlights the need for further investigation into the effects of FLT3-ITD mutations on DC phenotypes.
]]></description>
<dc:creator>Flynn, P. A.</dc:creator>
<dc:creator>Long, M. D.</dc:creator>
<dc:creator>Kosaka, Y.</dc:creator>
<dc:creator>Mulkey, J. S.</dc:creator>
<dc:creator>Coy, J. L.</dc:creator>
<dc:creator>Agarwal, A.</dc:creator>
<dc:creator>Lind, E. F.</dc:creator>
<dc:date>2023-09-22</dc:date>
<dc:identifier>doi:10.1101/2023.09.19.558512</dc:identifier>
<dc:title><![CDATA[Leukemic mutation FLT3-ITD is retained in dendritic cells and disrupts their homeostasis leading to expanded Th17 frequency]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-09-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.09.23.558896v1?rss=1">
<title>
<![CDATA[
The Human Infertility Single-cell Testis Atlas (HISTA): An interactive molecular scRNA-Seq reference of the human testis. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.09.23.558896v1?rss=1</link>
<description><![CDATA[
BackgroundThe Human Infertility Single-cell Testis Atlas (HISTA) is an interactive web tool and a reference for navigating the transcriptome of the human testis. It was developed using joint analyses of scRNA-Seq datasets derived from a dozen donors, including healthy adult controls, juveniles, and several infertility cases. HISTA is very different than other websites of testis scRNA-seq data, providing visualization and hypothesis testing tools on a batch-removed and integrated dataset of 23429 genes measured across 26093 cells using.

ObjectiveThe main goal of this manuscript is to describe HISTA in detail and highlight its unique and novel features.

MethodsTherefore, we used HISTA as a guide for its application and demonstrated HISTAs translational capacity to follow up on two observations of biological relevance.

ResultsOur first analytical vignette identifies novel groupings of tightly regulated long non-coding RNA (lncRNA) molecules throughout spermatogenesis, suggesting specific functional genomics of these groupings. This analysis also found highly controlled expression of pairs of sense and antisense transcripts, suggesting conjoined regulatory mechanisms. In the next investigative vignette, we examined gene patterns in undifferentiated spermatogonia (USgs). We found the NANOS family of genes function as key drivers of transcriptomic signatures involved in human spermatogonial self-renewal programming; for the first time, demonstrating the relationship of NANOS1/2/3 transcripts in humans with scRNA-seq.

Discussion and ConclusionsUsing HISTA, we found new observations that contribute to unraveling the mechanisms behind transcriptional regulation and maintenance germ cells across spermatogenesis. Furthermore, our findings provide guidance on future validation studies and experimental direction. Overall, HISTA continues to be utilized in testis-related research, and thus is updated regularly with new analytical methods, visualizations, and data. We aim to have it serve as a research environment for a broad range of investigators looking to explore the testis tissue and male infertility.

Availability and ImplementationHISTA is available as an interactive web tool: https://conradlab.shinyapps.io/HISTA

Source code and documentation for HISTA are provided on GitHub: https://github.com/eisascience/HISTA
]]></description>
<dc:creator>Mahyari, E.</dc:creator>
<dc:creator>Vigh-Conrad, K. A.</dc:creator>
<dc:creator>Daube, C.</dc:creator>
<dc:creator>Lima, A. C.</dc:creator>
<dc:creator>Guo, J.</dc:creator>
<dc:creator>Carell, D.</dc:creator>
<dc:creator>Hotaling, J. M.</dc:creator>
<dc:creator>Aston, K. I.</dc:creator>
<dc:creator>Conrad, D.</dc:creator>
<dc:date>2023-09-24</dc:date>
<dc:identifier>doi:10.1101/2023.09.23.558896</dc:identifier>
<dc:title><![CDATA[The Human Infertility Single-cell Testis Atlas (HISTA): An interactive molecular scRNA-Seq reference of the human testis.]]></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/10.1101/2023.09.23.559125v1?rss=1">
<title>
<![CDATA[
Recurrent activity within microcircuits of macaque dorsolateral prefrontal cortex tracks cognitive flexibility 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.09.23.559125v1?rss=1</link>
<description><![CDATA[
Human and non-human primate data clearly implicate the dorsolateral prefrontal cortex (dlPFC) as critical for advanced cognitive functions1,2. It is thought that intracortical synaptic architectures within dlPFC are the integral neurobiological substrate that gives rise to these processes, including working memory, inferential reasoning, and decision-making3-7. In the prevailing model, each cortical column makes up one fundamental processing unit composed of dense intrinsic connectivity, conceptualized as the  canonical cortical microcircuit3,8. Each cortical microcircuit receives sensory and cognitive information from a variety of sources which are represented by sustained activity within the microcircuit, referred to as persistent or recurrent activity4,9. Via recurrent connections within the microcircuit, activity can propagate for a variable length of time, thereby allowing temporary storage and computations to occur locally before ultimately passing a transformed representation to a downstream output4,5,10. Competing theories regarding how microcircuit activity is coordinated have proven difficult to reconcile in vivo where intercortical and intracortical computations cannot be fully dissociated5,9,11,12. Here, we interrogated the intrinsic features of isolated microcircuit networks using high-density calcium imaging of macaque dlPFC ex vivo. We found that spontaneous activity is intrinsically maintained by microcircuit architecture, persisting at a high rate in the absence of extrinsic connections. Further, using perisulcal stimulation to evoke persistent activity in deep layers, we found that activity propagates through stochastically assembled intracortical networks, creating predictable population-level events from largely non-overlapping ensembles. Microcircuit excitability covaried with individual cognitive performance, thus anchoring heuristic models of abstract cortical functions within quantifiable constraints imposed by the underlying synaptic architecture.
]]></description>
<dc:creator>Nolan, S. O.</dc:creator>
<dc:creator>Melugin, P. R.</dc:creator>
<dc:creator>Erickson, K. R.</dc:creator>
<dc:creator>Adams, W. R.</dc:creator>
<dc:creator>Farahbakhsh, Z. Z.</dc:creator>
<dc:creator>Mcgonigle, C. E.</dc:creator>
<dc:creator>Kwon, M. H.</dc:creator>
<dc:creator>Costa, V. D.</dc:creator>
<dc:creator>Lapish, C. C.</dc:creator>
<dc:creator>Hackett, T. A.</dc:creator>
<dc:creator>Cuzon Carlson, V. C.</dc:creator>
<dc:creator>Constantinidis, C.</dc:creator>
<dc:creator>Grant, K. A.</dc:creator>
<dc:creator>Siciliano, C. A.</dc:creator>
<dc:date>2023-09-24</dc:date>
<dc:identifier>doi:10.1101/2023.09.23.559125</dc:identifier>
<dc:title><![CDATA[Recurrent activity within microcircuits of macaque dorsolateral prefrontal cortex tracks cognitive flexibility]]></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/10.1101/2023.09.22.559019v1?rss=1">
<title>
<![CDATA[
Immune Correlates of Hyperglycemia and Vaccination in a Non-human Primate Model of Long-COVID 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.09.22.559019v1?rss=1</link>
<description><![CDATA[
Hyperglycemia, and exacerbation of pre-existing deficits in glucose metabolism, are major manifestations of the post-acute sequelae of SARS-CoV-2 (PASC). Our understanding of lasting glucometabolic disruptions after acute COVID-19 remains unclear due to the lack of animal models for metabolic PASC. Here, we report a non-human primate model of metabolic PASC using SARS-CoV-2 infected African green monkeys (AGMs). Using this model, we have identified a dysregulated chemokine signature and hypersensitive T cell population during acute COVID-19 that correlates with elevated and persistent hyperglycemia four months post-infection. This persistent hyperglycemia correlates with elevated hepatic glycogen, but there was no evidence of long-term SARS-CoV-2 replication in the liver and pancreas. Finally, we report a favorable glycemic effect of the SARS-CoV-2 mRNA vaccine, administered on day 4 post-infection. Together, these data suggest that the AGM metabolic PASC model exhibits important similarities to human metabolic PASC and can be utilized to assess therapeutic candidates to combat this syndrome.
]]></description>
<dc:creator>Palmer, C. S.</dc:creator>
<dc:creator>Perdios, C.</dc:creator>
<dc:creator>Abdel-Mohsen, M.</dc:creator>
<dc:creator>Mudd, J.</dc:creator>
<dc:creator>Datta, P.</dc:creator>
<dc:creator>Maness, N.</dc:creator>
<dc:creator>Lehmicke, G.</dc:creator>
<dc:creator>Golden, N.</dc:creator>
<dc:creator>Hellmer, L.</dc:creator>
<dc:creator>Coyne, C.</dc:creator>
<dc:creator>Moore Green, K.</dc:creator>
<dc:creator>Midkiff, C.</dc:creator>
<dc:creator>Williams, K.</dc:creator>
<dc:creator>Tiburcio, R.</dc:creator>
<dc:creator>Fahlberg, M.</dc:creator>
<dc:creator>Boykin, K.</dc:creator>
<dc:creator>Kenway, C.</dc:creator>
<dc:creator>Russell-Lodrigue, K.</dc:creator>
<dc:creator>Bohm, R.</dc:creator>
<dc:creator>Blair, R. V.</dc:creator>
<dc:creator>Dufour, J.</dc:creator>
<dc:creator>Fischer, T.</dc:creator>
<dc:creator>Saied, A.</dc:creator>
<dc:creator>Rappaport, J.</dc:creator>
<dc:date>2023-09-25</dc:date>
<dc:identifier>doi:10.1101/2023.09.22.559019</dc:identifier>
<dc:title><![CDATA[Immune Correlates of Hyperglycemia and Vaccination in a Non-human Primate Model of Long-COVID]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-09-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.09.26.559528v1?rss=1">
<title>
<![CDATA[
An enhancer RNA recruits MLL1 to regulate transcription of Myb 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.09.26.559528v1?rss=1</link>
<description><![CDATA[
The Myb proto-oncogene encodes the transcription factor c-MYB, which is critical for hematopoiesis. Distant enhancers of Myb form a hub of interactions with the Myb promoter. We identified a long non-coding RNA (Myrlin) originating from the -81 kb murine Myb enhancer. Myrlin and Myb are coordinately regulated during erythroid differentiation. Myrlin TSS deletion using CRISPR/Cas9 reduced Myrlin and Myb expression and LDB1 complex occupancy at the Myb enhancers, compromising enhancer contacts and reducing RNA Pol II occupancy in the locus. In contrast, CRISPRi silencing of Myrlin left LDB1 and the Myb enhancer hub unperturbed, although Myrlin and Myb expression were downregulated, decoupling transcription and chromatin looping. Myrlin interacts with the MLL1 complex. Myrlin CRISPRi compromised MLL1 occupancy in the Myb locus, decreasing CDK9 and RNA Pol II binding and resulting in Pol II pausing in the Myb first exon/intron. Thus, Myrlin directly participates in activating Myb transcription by recruiting MLL1.
]]></description>
<dc:creator>Kim, J.</dc:creator>
<dc:creator>Diaz, L. F.</dc:creator>
<dc:creator>Miller, M. J.</dc:creator>
<dc:creator>Leadem, B.</dc:creator>
<dc:creator>Krivega, I.</dc:creator>
<dc:creator>Dean, A.</dc:creator>
<dc:date>2023-09-26</dc:date>
<dc:identifier>doi:10.1101/2023.09.26.559528</dc:identifier>
<dc:title><![CDATA[An enhancer RNA recruits MLL1 to regulate transcription of Myb]]></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/10.1101/2023.09.26.559633v1?rss=1">
<title>
<![CDATA[
Dysregulation of astrocyte-secreted pleiotrophin contributes to neuronal structural and functional phenotypes in Down Syndrome 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.09.26.559633v1?rss=1</link>
<description><![CDATA[
Neuronal dendrite patterning and synapse formation are tightly regulated during development to promote proper connectivity. Astrocyte-secreted proteins act as guidance and pro-synaptogenic factors during development, but little is known about how astrocytes may contribute to neurodevelopmental disorders. Here we identify down-regulation of the astrocyte-secreted molecule pleiotrophin as a major contributor to neuronal morphological alterations in the Ts65Dn mouse model of Down Syndrome. We find overlapping deficits in neuronal dendrites, spines and intracortical synapses in Ts65Dn mutant and pleiotrophin knockout mice. By targeting pleiotrophin overexpression to astrocytes in adult Ts65Dn mutant mice in vivo, we show that pleiotrophin can rescue dendrite morphology and spine density and increase excitatory synapse number. We further demonstrate functional improvements in behavior. Our findings identify pleiotrophin as a molecule that can be used in Down Syndrome to promote proper circuit connectivity, importantly at later stages of development after typical periods of circuit refinement have completed.
]]></description>
<dc:creator>Brandebura, A. N.</dc:creator>
<dc:creator>Asbell, Q. N.</dc:creator>
<dc:creator>Micael, M. K. B.</dc:creator>
<dc:creator>Allen, N. J.</dc:creator>
<dc:date>2023-09-27</dc:date>
<dc:identifier>doi:10.1101/2023.09.26.559633</dc:identifier>
<dc:title><![CDATA[Dysregulation of astrocyte-secreted pleiotrophin contributes to neuronal structural and functional phenotypes in Down Syndrome]]></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/10.1101/2023.09.27.559779v1?rss=1">
<title>
<![CDATA[
Engineered Lactiplantibacillus plantarum as a Biosensor Probe for the Lungs 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.09.27.559779v1?rss=1</link>
<description><![CDATA[
Lungs are a frequent site for disease but are difficult to image and probe, potentially exacerbating lung disease, delaying diagnoses and impacting survival. Here, we demonstrate a low cost, minimally invasive method to probe the lungs for disease, using genetically engineered Lactiplantibacillus plantarum strain WCFS1. Genetically modified WCFS1 was delivered specifically to the lungs of mice, where it was shown to remain transcriptionally active for several hours and then be cleared without further colonization. WCFS1 was further modified to secrete nanoluciferase as a synthetic biomarker, which traveled from the bacteria in the lung to the urine where it was easily detected. The administered bacteria also secreted nano-luciferase upon detecting a specific peptide secreted by a mouse lung cancer cell line in vitro or in vivo from syngeneic tumors.
]]></description>
<dc:creator>Brasino, M.</dc:creator>
<dc:creator>Wagnell, E.</dc:creator>
<dc:creator>Manalo, E. C.</dc:creator>
<dc:creator>Drennan, S.</dc:creator>
<dc:creator>Fischer, J. M.</dc:creator>
<dc:creator>Merritt, J.</dc:creator>
<dc:date>2023-09-27</dc:date>
<dc:identifier>doi:10.1101/2023.09.27.559779</dc:identifier>
<dc:title><![CDATA[Engineered Lactiplantibacillus plantarum as a Biosensor Probe for the Lungs]]></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/10.1101/2023.09.27.557464v1?rss=1">
<title>
<![CDATA[
Longitudinal and multimodal auditing of tumor adaptation to CDK4/6 inhibitors in HR+ metastatic breast cancers 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.09.27.557464v1?rss=1</link>
<description><![CDATA[
CDK4/6 inhibitors (CDK4/6i) have transformed the treatment of hormone receptor-positive (HR+), HER2-negative (HR+) breast cancers as they are effective across all clinicopathological, age, and ethnicity subgroups for metastatic HR+ breast cancer. In metastatic ER+ breast cancer, CDK4/6i lead to strong and consistent improvement in survival across different lines of therapy. To improve understanding of how metastatic HR+ breast cancers become refractory to CDK4/6i, we have created a multimodal and longitudinal tumor atlas to investigate therapeutic adaptations in malignant cells and in the tumor immune microenvironment. This atlas is part of the NCI Cancer Moonshot Human Tumor Atlas Network and includes seven pairs of pre- and on-progression biopsies from five metastatic HR+ breast cancer patients treated with CDK4/6i. Biopsies were profiled with bulk genomics, transcriptomics, and proteomics as well as single-cell ATAC-seq and multiplex tissue imaging for spatial, single-cell resolution. These molecular datasets were then linked with detailed clinical metadata to create an atlas for understanding tumor adaptations during therapy. Analysis of our atlas datasets suggests a diverse set of tumor adaptations to CDK4/6i therapy. Malignant cells may adapt to therapy via mTORC1 activation, cell cycle bypass, and increased replication stress. The tumor immune microenvironment displayed evidence of both immune activation and immune suppression during therapy. Together, our metastatic ER+ breast cancer atlas represents a rich multimodal resource to better understand HR+ breast cancer tumor therapeutic adaptations to CDK4/6i therapy.
]]></description>
<dc:creator>Creason, A. L.</dc:creator>
<dc:creator>Egger, J.</dc:creator>
<dc:creator>Watson, C.</dc:creator>
<dc:creator>Sivagnanam, S.</dc:creator>
<dc:creator>Chin, K.</dc:creator>
<dc:creator>MacPherson, K.</dc:creator>
<dc:creator>Lin, J.-R.</dc:creator>
<dc:creator>Chen, Y.-A.</dc:creator>
<dc:creator>Johnson, B. E.</dc:creator>
<dc:creator>Feiler, H. S.</dc:creator>
<dc:creator>Galipeau, D.</dc:creator>
<dc:creator>Navin, N. E.</dc:creator>
<dc:creator>Demir, E.</dc:creator>
<dc:creator>Chang, Y. H.</dc:creator>
<dc:creator>Corless, C. L.</dc:creator>
<dc:creator>Mitri, Z. I.</dc:creator>
<dc:creator>Thomas, G.</dc:creator>
<dc:creator>Sorger, P. K.</dc:creator>
<dc:creator>Adey, A. C.</dc:creator>
<dc:creator>Coussens, L. M.</dc:creator>
<dc:creator>Gray, J. W.</dc:creator>
<dc:creator>Mills, G. B.</dc:creator>
<dc:creator>Goecks, J.</dc:creator>
<dc:date>2023-09-29</dc:date>
<dc:identifier>doi:10.1101/2023.09.27.557464</dc:identifier>
<dc:title><![CDATA[Longitudinal and multimodal auditing of tumor adaptation to CDK4/6 inhibitors in HR+ metastatic breast cancers]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-09-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.09.27.559834v1?rss=1">
<title>
<![CDATA[
MultipleXed Population Selection and Enrichment single nucleus RNA sequencing (XPoSE-seq) enables sample identity retention during transcriptional profiling of rare populations 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.09.27.559834v1?rss=1</link>
<description><![CDATA[
Single nucleus RNA-sequencing is critical in deciphering tissue heterogeneity and identifying rare populations. However, current high throughput techniques are not optimized for rare target populations and require tradeoffs in design due to feasibility. We provide a novel snRNA pipeline, MulipleXed Population Selection and Enrichment snRNA-sequencing (XPoSE-seq), to enable targeted snRNA-seq experiments and in-depth transcriptomic characterization of rare target populations while retaining individual sample identity.
]]></description>
<dc:creator>Savell, K. E.</dc:creator>
<dc:creator>Madangopal, R.</dc:creator>
<dc:creator>Saravanan, P.</dc:creator>
<dc:creator>Palaganas, R. G.</dc:creator>
<dc:creator>Woods, K. D.</dc:creator>
<dc:creator>Thompson, D. J.</dc:creator>
<dc:creator>Drake, O. R.</dc:creator>
<dc:creator>Brenner, M. B.</dc:creator>
<dc:creator>Weber, S. J.</dc:creator>
<dc:creator>Van Leer, E.</dc:creator>
<dc:creator>Choi, J. H.</dc:creator>
<dc:creator>Martin, T. L.</dc:creator>
<dc:creator>Martin, J. C.</dc:creator>
<dc:creator>Steinberg, M. K.</dc:creator>
<dc:creator>Austin, J. W.</dc:creator>
<dc:creator>Charendoff, C. I.</dc:creator>
<dc:creator>Hope, B. T.</dc:creator>
<dc:date>2023-09-29</dc:date>
<dc:identifier>doi:10.1101/2023.09.27.559834</dc:identifier>
<dc:title><![CDATA[MultipleXed Population Selection and Enrichment single nucleus RNA sequencing (XPoSE-seq) enables sample identity retention during transcriptional profiling of rare populations]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-09-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.09.29.560039v1?rss=1">
<title>
<![CDATA[
Commensal myeloid crosstalk in neonatal skin regulates long-term cutaneous type 17 inflammation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.09.29.560039v1?rss=1</link>
<description><![CDATA[
Early life microbe-immune interactions at barrier surfaces have lasting impacts on the trajectory towards health versus disease. Monocytes, macrophages and dendritic cells are primary sentinels in barrier tissues, yet the salient contributions of commensal-myeloid crosstalk during tissue development remain poorly understood. Here, we identify that commensal microbes facilitate accumulation of a population of monocytes in neonatal skin. Transient postnatal depletion of these monocytes resulted in heightened IL-17A production by skin T cells, which was particularly sustained among CD4+ T cells and sufficient to exacerbate inflammatory skin pathologies. Neonatal skin monocytes were enriched in expression of negative regulators of the IL-1 pathway. Functional in vivo experiments confirmed a key role for excessive IL-1R1 signaling in T cells as contributing to the dysregulated type 17 response in neonatal monocyte-depleted mice. Thus, a commensal-driven wave of monocytes into neonatal skin critically facilitates immune homeostasis in this prominent barrier tissue.
]]></description>
<dc:creator>Dhariwala, M. O.</dc:creator>
<dc:creator>DeRogatis, A. M.</dc:creator>
<dc:creator>Okoro, J. N.</dc:creator>
<dc:creator>Weckel, A. A.</dc:creator>
<dc:creator>Tran, V. M.</dc:creator>
<dc:creator>Habrylo, I.</dc:creator>
<dc:creator>Ojewumi, O. T.</dc:creator>
<dc:creator>Tammen, A. E.</dc:creator>
<dc:creator>Leech, J. M.</dc:creator>
<dc:creator>Merana, G. R.</dc:creator>
<dc:creator>Carale, R. O.</dc:creator>
<dc:creator>Barrere-Cain, R.</dc:creator>
<dc:creator>Hiam-Galvez, K. J.</dc:creator>
<dc:creator>Spitzer, M. H.</dc:creator>
<dc:creator>Scharschmidt, T. C.</dc:creator>
<dc:date>2023-10-02</dc:date>
<dc:identifier>doi:10.1101/2023.09.29.560039</dc:identifier>
<dc:title><![CDATA[Commensal myeloid crosstalk in neonatal skin regulates long-term cutaneous type 17 inflammation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.09.30.560267v1?rss=1">
<title>
<![CDATA[
Remyelination protects neurons from DLK-mediated neurodegeneration 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.09.30.560267v1?rss=1</link>
<description><![CDATA[
Chronic demyelination and oligodendrocyte loss deprive neurons of crucial support. It is the degeneration of neurons and their connections that drives progressive disability in demyelinating disease. However, whether chronic demyelination triggers neurodegeneration and how it may do so remain unclear. We characterize two genetic mouse models of inducible demyelination, one distinguished by effective remyelination and the other by remyelination failure and chronic demyelination. While both demyelinating lines feature axonal damage, mice with blocked remyelination have elevated neuronal apoptosis and altered microglial inflammation, whereas mice with efficient remyelination do not feature neuronal apoptosis and have improved functional recovery. Remyelination incapable mice show increased activation of kinases downstream of dual leucine zipper kinase (DLK) and phosphorylation of c-Jun in neuronal nuclei. Pharmacological inhibition or genetic disruption of DLK block c-Jun phosphorylation and the apoptosis of demyelinated neurons. Together, we demonstrate that remyelination is associated with neuroprotection and identify DLK inhibition as protective strategy for chronically demyelinated neurons.

HighlightsO_LICharacterization of a transgenic mouse model of demyelination without subsequent remyelination
C_LIO_LIRemyelination protects neurons from axon loss and neuronal apoptosis
C_LIO_LIMAPK and c-Jun phosphorylation are increased in mice featuring remyelination failure
C_LIO_LIDLK is necessary for the apoptosis of chronically demyelinated neurons
C_LI
]]></description>
<dc:creator>Duncan, G. J.</dc:creator>
<dc:creator>Ingram, S. D.</dc:creator>
<dc:creator>Emberley, K.</dc:creator>
<dc:creator>Hill, J.</dc:creator>
<dc:creator>Cordano, C.</dc:creator>
<dc:creator>Abdelhak, A.</dc:creator>
<dc:creator>McCane, M.</dc:creator>
<dc:creator>Jabassini, N.</dc:creator>
<dc:creator>Ananth, K.</dc:creator>
<dc:creator>Ferrara, S. J.</dc:creator>
<dc:creator>Stedelin, B.</dc:creator>
<dc:creator>Aicher, S. A.</dc:creator>
<dc:creator>Watkins, T. A.</dc:creator>
<dc:creator>Mishra, A.</dc:creator>
<dc:creator>Nelson, J.</dc:creator>
<dc:creator>Green, A. J.</dc:creator>
<dc:creator>Emery, B.</dc:creator>
<dc:date>2023-10-02</dc:date>
<dc:identifier>doi:10.1101/2023.09.30.560267</dc:identifier>
<dc:title><![CDATA[Remyelination protects neurons from DLK-mediated neurodegeneration]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.10.02.560575v1?rss=1">
<title>
<![CDATA[
Microbiome imaging goes &agrave la carte: Incorporating click chemistry into the fluorescence-activating and absorption-shifting tag (FAST) imaging platform 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.02.560575v1?rss=1</link>
<description><![CDATA[
The human microbiome is predominantly composed of facultative and obligate anaerobic bacteria that live in hypoxic/anoxic polymicrobial biofilm communities. Given the oxidative sensitivity of large fractions of the human microbiota, green fluorescent protein (GFP) and related genetically-encoded fluorophores only offer limited utility for live cell imaging due the oxygen requirement for chromophore maturation. Consequently, new fluorescent imaging modalities are needed to study polymicrobial interactions and microbiome-host interactions within anaerobic environments. The fluorescence-activating and absorption shifting tag (FAST) is a rapidly developing genetically-encoded fluorescent imaging technology that exhibits tremendous potential to address this need. In the FAST system, fluorescence only occurs when the FAST protein is complexed with one of a suite of cognate small molecule fluorogens. To expand the utility of FAST imaging, we sought to develop a modular platform (Click-FAST) to democratize fluorogen engineering for personalized use cases. Using Click-FAST, investigators can quickly and affordably sample a vast chemical space of compounds, potentially imparting a broad range of desired functionalities to the parental fluorogen. In this work, we demonstrate the utility of the Click-FAST platform using a novel fluorogen, PLBlaze-alkyne, which incorporates the widely available small molecule ethylvanillin as the hydroxybenzylidine head group. Different azido reagents were clicked onto PLBlaze-alkyne and shown to impart useful characteristics to the fluorogen, such as selective bacterial labeling in mixed populations as well as fluorescent signal enhancement. Conjugation of an 80 [A] PEG molecule to PLBlaze-alkyne illustrates the broad size range of functional fluorogen chimeras that can be employed. This PEGylated fluorogen also functions as an exquisitely selective membrane permeability marker capable of outperforming propidium iodide as a fluorescent marker of cell viability.
]]></description>
<dc:creator>Anderson, D. M.</dc:creator>
<dc:creator>Logan, M. G.</dc:creator>
<dc:creator>Patty, S. S.</dc:creator>
<dc:creator>Kendall, A. J.</dc:creator>
<dc:creator>Borland, C. Z.</dc:creator>
<dc:creator>Pfeifer, C. S.</dc:creator>
<dc:creator>Kreth, J.</dc:creator>
<dc:creator>Merritt, J. L.</dc:creator>
<dc:date>2023-10-02</dc:date>
<dc:identifier>doi:10.1101/2023.10.02.560575</dc:identifier>
<dc:title><![CDATA[Microbiome imaging goes &agrave la carte: Incorporating click chemistry into the fluorescence-activating and absorption-shifting tag (FAST) imaging platform]]></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/10.1101/2023.10.03.560675v1?rss=1">
<title>
<![CDATA[
Targeting the Hippo pathway in cancers via ubiquitination dependent TEAD degradation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.03.560675v1?rss=1</link>
<description><![CDATA[
The Hippo pathway is among the most frequently altered key signaling pathways in cancer. TEAD1-4 are essential transcription factors and key downstream effectors in the Hippo pathway in human cells. Here, we identified RNF146 as a ubiquitin ligase (E3) of TEADs, which negatively regulates their stability in cells through proteasome-mediated degradation. We show that RNF146-mediated TEAD ubiquitination is dependent on the TEAD PARylation state. We further validated the genetic interaction between RNF146 and the Hippo pathway in cancer cell lines and the model organism Drosophila melanogaster. Despite the RNF146 and proteasome-mediated degradation mechanisms, TEADs are stable proteins with a long half-life in cells. We demonstrate that degradation of TEADs can be greatly enhanced pharmacologically with heterobifunctional chemical inducers of protein degradation (CIDEs). These TEAD-CIDEs can effectively suppress activation of YAP/TAZ target genes in a dose-dependent manner and exhibit significant anti-proliferative effects in YAP/TAZ-dependent tumor cells, thus phenocopying the effect of genetic ablation of TEAD protein. Collectively, this study demonstrates that the ubiquitin-proteasome system plays an important role in regulating TEAD functions and provides a proof-of-concept demonstration that pharmacologically induced TEAD ubiquitination could be leveraged to target YAP/TAZ-driven cancers.
]]></description>
<dc:creator>Pham, T. H.</dc:creator>
<dc:creator>Pahuja, K. B.</dc:creator>
<dc:creator>Hagenbeek, T. J.</dc:creator>
<dc:creator>Zbieg, J.</dc:creator>
<dc:creator>Noland, C. L.</dc:creator>
<dc:creator>Pham, V. C.</dc:creator>
<dc:creator>Yao, X.</dc:creator>
<dc:creator>Rose, C. M.</dc:creator>
<dc:creator>Browder, K. C.</dc:creator>
<dc:creator>Lee, H.-J.</dc:creator>
<dc:creator>Yu, M.</dc:creator>
<dc:creator>Liang-Chu, M.</dc:creator>
<dc:creator>Martin, S. E.</dc:creator>
<dc:creator>Verschueren, E.</dc:creator>
<dc:creator>Li, J.</dc:creator>
<dc:creator>Kubala, M. H.</dc:creator>
<dc:creator>Fong, R.</dc:creator>
<dc:creator>Lorenzo, M.</dc:creator>
<dc:creator>Beroza, P.</dc:creator>
<dc:creator>Hsu, P.</dc:creator>
<dc:creator>Paul, S.</dc:creator>
<dc:creator>Villemure, E.</dc:creator>
<dc:creator>Lee, W.</dc:creator>
<dc:creator>Cheung, T.</dc:creator>
<dc:creator>Clausen, S.</dc:creator>
<dc:creator>Lacap, J.</dc:creator>
<dc:creator>Liang, Y.</dc:creator>
<dc:creator>Cheng, J.</dc:creator>
<dc:creator>Schmidt, S.</dc:creator>
<dc:creator>Modrusan, Z.</dc:creator>
<dc:creator>Cohen, M. S.</dc:creator>
<dc:creator>Crawford, J.</dc:creator>
<dc:creator>Jasper, H.</dc:creator>
<dc:creator>Ashworth, A.</dc:creator>
<dc:creator>Lill, J. R.</dc:creator>
<dc:creator>Malek, S.</dc:creator>
<dc:creator>Rudolph, J.</dc:creator>
<dc:creator>Wertz, I. E.</dc:creator>
<dc:creator>Chang, M.</dc:creator>
<dc:creator>Ye, X.</dc:creator>
<dc:creator>Dey, A.</dc:creator>
<dc:date>2023-10-04</dc:date>
<dc:identifier>doi:10.1101/2023.10.03.560675</dc:identifier>
<dc:title><![CDATA[Targeting the Hippo pathway in cancers via ubiquitination dependent TEAD degradation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.10.04.560576v1?rss=1">
<title>
<![CDATA[
Analysis of the Diverse Antigenic Landscape of the Malaria Invasion Protein RH5 Identifies a Potent Vaccine-Induced Human Public Antibody Clonotype 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.04.560576v1?rss=1</link>
<description><![CDATA[
The highly conserved and essential Plasmodium falciparum reticulocyte-binding protein homolog 5 (PfRH5) has emerged as the leading target for vaccines that seek to protect against the disease-causing blood-stage of malaria. However, the features of the human vaccine-induced antibody response that confer highly potent inhibition of malaria parasite invasion into red blood cells are not well defined. Here we characterize over 200 human IgG monoclonal antibodies induced by the most advanced PfRH5 vaccine. We define the antigenic landscape of this molecule, and establish epitope specificity, antibody association rate and intra-PfRH5 antibody interactions are key determinants of functional anti-parasitic potency. In addition, we identify a germline gene combination that results in an exceptionally potent class of antibody and demonstrate its prophylactic potential to protect against P. falciparum parasite challenge in vivo. This comprehensive dataset provides a framework to guide rational design of next-generation vaccines and prophylactic antibodies to protect against blood-stage malaria.
]]></description>
<dc:creator>Barrett, J. R.</dc:creator>
<dc:creator>Pipini, D.</dc:creator>
<dc:creator>Wright, N. D.</dc:creator>
<dc:creator>Cooper, A. J. R.</dc:creator>
<dc:creator>Gorini, G.</dc:creator>
<dc:creator>Quinkert, D.</dc:creator>
<dc:creator>Lias, A. M.</dc:creator>
<dc:creator>Davies, H.</dc:creator>
<dc:creator>Rigby, C.</dc:creator>
<dc:creator>Aleshnick, M.</dc:creator>
<dc:creator>Williams, B. G.</dc:creator>
<dc:creator>Bradshaw, W. J.</dc:creator>
<dc:creator>Paterson, N. G.</dc:creator>
<dc:creator>Martinson, T.</dc:creator>
<dc:creator>Kirtley, P.</dc:creator>
<dc:creator>Picard, L.</dc:creator>
<dc:creator>Wiggins, C. D.</dc:creator>
<dc:creator>Donnellan, F. R.</dc:creator>
<dc:creator>King, L. D. W.</dc:creator>
<dc:creator>Wang, L. T.</dc:creator>
<dc:creator>Popplewell, J. F.</dc:creator>
<dc:creator>Silk, S. E.</dc:creator>
<dc:creator>Swain, J. d. R.</dc:creator>
<dc:creator>Skinner, K.</dc:creator>
<dc:creator>Kotraiah, V.</dc:creator>
<dc:creator>Noe, A. R.</dc:creator>
<dc:creator>MacGill, R. S.</dc:creator>
<dc:creator>King, C. R.</dc:creator>
<dc:creator>Birkett, A. J.</dc:creator>
<dc:creator>Soisson, L. A.</dc:creator>
<dc:creator>Minassian, A. M.</dc:creator>
<dc:creator>Lauffenburger, D. A.</dc:creator>
<dc:creator>Miura, K.</dc:creator>
<dc:creator>Long, C. A.</dc:creator>
<dc:creator>Wilder, B. K.</dc:creator>
<dc:creator>Koekemoer, L.</dc:creator>
<dc:creator>Tan, J.</dc:creator>
<dc:creator>Nielsen, C. M.</dc:creator>
<dc:creator>McHugh, K.</dc:creator>
<dc:creator>Draper, S. J.</dc:creator>
<dc:date>2023-10-04</dc:date>
<dc:identifier>doi:10.1101/2023.10.04.560576</dc:identifier>
<dc:title><![CDATA[Analysis of the Diverse Antigenic Landscape of the Malaria Invasion Protein RH5 Identifies a Potent Vaccine-Induced Human Public Antibody Clonotype]]></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/10.1101/2023.10.04.560975v1?rss=1">
<title>
<![CDATA[
Robust CNV detection using single-cell ATAC-seq 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.04.560975v1?rss=1</link>
<description><![CDATA[
Copy number variation (CNV) is a widely studied type of structural variation seen in the genomes of cancerous and other dysfunctional cells. CNVs can have direct and indirect effects on gene dosage, and are thought to drive cancer progression and other disorders. Advancements in single-cell assays such as sc-ATAC-seq and sc-RNA-seq, along with their ubiquitous use, allows for the identification of CNVs at single cell resolution. While there are a variety of available tools for CNV detection in sc-RNA-seq, development of sc-ATAC-seq based accurate and reliable CNV callers is in the early stages, with only two available algorithms so far. We present RIDDLER, a single-cell ATAC-seq CNV detection algorithm based on outlier aware generalized linear modeling. By utilizing tools from robust statistics, we developed an extensible model that is able to identify single-cell CNVs from sc-ATAC-seq data in an unsupervised fashion, while providing probabilistic justification for results. Our statistical approach also allows us to estimate when loss of signal is likely caused by drop-out or a true genome deletion event, as well as predict reliable CNVs without the need for normative reference cells. We demonstrate the effectiveness of our algorithm on cancer cell line models where it achieves better agreement with bulk WGS derived CNVs than competing methods. We also compare our approach on 10x multimone data, where it shows better agreement and integration with RNA derived CNV estimates.
]]></description>
<dc:creator>Moore, T. W.</dc:creator>
<dc:creator>Yardimci, G. G.</dc:creator>
<dc:date>2023-10-06</dc:date>
<dc:identifier>doi:10.1101/2023.10.04.560975</dc:identifier>
<dc:title><![CDATA[Robust CNV detection using single-cell ATAC-seq]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.10.05.560352v1?rss=1">
<title>
<![CDATA[
Deciphering Cell-types and Gene Signatures Associated with Disease Activity in Rheumatoid Arthritis using Single Cell RNA-sequencing 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.05.560352v1?rss=1</link>
<description><![CDATA[
ObjectiveSingle cell profiling of synovial tissue has previously identified gene signatures associated with rheumatoid arthritis (RA) pathophysiology, but synovial tissue is difficult to obtain. This study leverages single cell sequencing of peripheral blood mononuclear cells (PBMCs) from patients with RA and matched healthy controls to identify disease relevant cell subsets and cell type specific signatures of disease.

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

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

ConclusionsWe provide a novel single-cell transcriptomics dataset of PBMCs from patients with RA, and identify insights into the systemic cellular and molecular mechanisms underlying RA disease activity.
]]></description>
<dc:creator>Binvignat, M.</dc:creator>
<dc:creator>Miao, B.</dc:creator>
<dc:creator>Wibrand, C.</dc:creator>
<dc:creator>Yang, M.</dc:creator>
<dc:creator>Rychkov, D.</dc:creator>
<dc:creator>Flynn, E.</dc:creator>
<dc:creator>Nititham, J.</dc:creator>
<dc:creator>Tamaki, W.</dc:creator>
<dc:creator>Khan, U.</dc:creator>
<dc:creator>Carvidi, A.</dc:creator>
<dc:creator>Krueger, M.</dc:creator>
<dc:creator>Niemi, E.</dc:creator>
<dc:creator>Sun, Y.</dc:creator>
<dc:creator>Fragiadakis, G. K.</dc:creator>
<dc:creator>Sellam, J.</dc:creator>
<dc:creator>Mariotti-Ferrandiz, E.</dc:creator>
<dc:creator>Klatzmann, D.</dc:creator>
<dc:creator>Gross, A.</dc:creator>
<dc:creator>Ye, C. J.</dc:creator>
<dc:creator>Butte, A. J.</dc:creator>
<dc:creator>Criswell, L.</dc:creator>
<dc:creator>Nakamura, M.</dc:creator>
<dc:creator>Sirota, M.</dc:creator>
<dc:date>2023-10-06</dc:date>
<dc:identifier>doi:10.1101/2023.10.05.560352</dc:identifier>
<dc:title><![CDATA[Deciphering Cell-types and Gene Signatures Associated with Disease Activity in Rheumatoid Arthritis using Single Cell RNA-sequencing]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.10.08.561378v1?rss=1">
<title>
<![CDATA[
Thymically imprinted differential responsivity of naive CD4 T cells to IL2 results in a heterogeneity of both Treg induction and subsequent effector functioning 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.08.561378v1?rss=1</link>
<description><![CDATA[
Thymic selection predisposes naive T cells to particular outcomes when challenged later with cognate antigen, whether the antigen is self or foreign. This suggests that there is an inherent heterogeneity of functioning among T cells within the naive population (both CD4 and CD8s), and that each T cell, as part of its thymic development, is given a certain  programming which will affect its eventual fate decisions. In this project, we looked at the primary effects of this thymic imprinting on the conversion of naive CD4 T cells into Tregs. Further, using an induced-Treg-reporter system, we exam the impact of thymic imprinted heterogeneity on effector functionality and identity stability. We report that naive T cell differential responsivity to cytokines leads to the observed difference in Treg induction, and that the Tregs induced from T cells of different self-affinities maintain a heterogeneity of effector function and identity.
]]></description>
<dc:creator>Pennock, N. D.</dc:creator>
<dc:creator>Qian, Y.</dc:creator>
<dc:creator>Ishihara, K.</dc:creator>
<dc:creator>Nakamura, Y.</dc:creator>
<dc:creator>Cross, E. W.</dc:creator>
<dc:creator>Sakaguchi, S.</dc:creator>
<dc:creator>White, J. T.</dc:creator>
<dc:date>2023-10-10</dc:date>
<dc:identifier>doi:10.1101/2023.10.08.561378</dc:identifier>
<dc:title><![CDATA[Thymically imprinted differential responsivity of naive CD4 T cells to IL2 results in a heterogeneity of both Treg induction and subsequent effector functioning]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.10.10.560919v1?rss=1">
<title>
<![CDATA[
In vitro development and optimization of cell-laden injectable bioprinted gelatin methacryloyl (GelMA) microgels mineralized on the nanoscale 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.10.560919v1?rss=1</link>
<description><![CDATA[
Bone defects may occur in different sizes and shapes due to trauma, infections, and cancer resection. Autografts are still considered the primary treatment choice for bone regeneration. However, they are hard to source and often create donor-site morbidity. Injectable microgels have attracted much attention in tissue engineering and regenerative medicine due to their ability to replace inert implants with a minimally invasive delivery. Here, we developed novel cell-laden bioprinted gelatin methacrylate (GelMA) injectable microgels, with controllable shapes and sizes that can be controllably mineralized on the nanoscale, while stimulating the response of cells embedded within the matrix. The injectable microgels were mineralized using a calcium and phosphate-rich medium that resulted in nanoscale crystalline hydroxyapatite deposition and increased stiffness within the crosslinked matrix of bioprinted GelMA microparticles. Next, we studied the effect of mineralization in osteocytes, a key bone homeostasis regulator. Viability stains showed that osteocytes were maintained at 98% viability after mineralization with elevated expression of sclerostin in mineralized compared to non-mineralized microgels, indicating that mineralization effectively enhances osteocyte maturation. Based on our findings, bioprinted mineralized GelMA microgels appear to be an efficient material to approximate the bone microarchitecture and composition with desirable control of sample injectability and polymerization. These bone-like bioprinted mineralized biomaterials are exciting platforms for potential minimally invasive translational methods in bone regenerative therapies.
]]></description>
<dc:creator>Sousa, M. G. C.</dc:creator>
<dc:creator>Balbinot, G. d. S.</dc:creator>
<dc:creator>Subbiah, R.</dc:creator>
<dc:creator>Visalakshan, R.</dc:creator>
<dc:creator>Tahayeri, A.</dc:creator>
<dc:creator>Verde, M. E. Q. L.</dc:creator>
<dc:creator>Athirasala, A.</dc:creator>
<dc:creator>Romanowicz, G.</dc:creator>
<dc:creator>Guldberg, R. E.</dc:creator>
<dc:creator>Bertassoni, L. E.</dc:creator>
<dc:date>2023-10-12</dc:date>
<dc:identifier>doi:10.1101/2023.10.10.560919</dc:identifier>
<dc:title><![CDATA[In vitro development and optimization of cell-laden injectable bioprinted gelatin methacryloyl (GelMA) microgels mineralized on the nanoscale]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.10.16.562528v1?rss=1">
<title>
<![CDATA[
Trifunctional sphinganine: a new tool to dissect sphingolipid function 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.16.562528v1?rss=1</link>
<description><![CDATA[
Functions of the sphingolipids sphingosine and sphinganine in cells are not well established. While some signaling roles for sphingosine have been elucidated, the closely related sphinganine has been described only insofar as it does not elicit many of the same signaling responses. The underlying mechanisms behind the cell biological differences between these lipids are not well understood. Here, we prepared multifunctionalized derivatives of the two lipid species that only differ in a single double bond of the carbon backbone. Using these novel probes, we were able to define their spatiotemporal distribution within cells. Furthermore, we used these tools to systematically map the protein interactomes of both lipids. The lipid-protein conjugates, prepared through photo-crosslinking in live cells and extraction via click chemistry to azide beads, revealed significant differences in the captured proteins, highlighting their distinct roles in various cellular processes. This work elucidates mechanistic differences between these critical lipids and sets the foundation for further studies on the functions of sphingosine and sphinganine.
]]></description>
<dc:creator>Farley, S. E.</dc:creator>
<dc:creator>Stein, F.</dc:creator>
<dc:creator>Haberkant, P.</dc:creator>
<dc:creator>Tafesse, F. G.</dc:creator>
<dc:creator>Schultz, C.</dc:creator>
<dc:date>2023-10-16</dc:date>
<dc:identifier>doi:10.1101/2023.10.16.562528</dc:identifier>
<dc:title><![CDATA[Trifunctional sphinganine: a new tool to dissect sphingolipid function]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.10.17.562839v1?rss=1">
<title>
<![CDATA[
Membrane Lipids Augment Cell Envelope Stress Signaling and Resistance to Antibiotics and Antimicrobial Peptides in Enterococcus faecalis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.17.562839v1?rss=1</link>
<description><![CDATA[
Enterococci have evolved resistance mechanisms to protect their cell envelopes against bacteriocins and host cationic antimicrobial peptides (CAMPs) produced in the gastrointestinal environment. Activation of the membrane stress response has also been tied to resistance to the lipopeptide antibiotic daptomycin. However, the actual effectors mediating resistance have not been elucidated. Here, we show that the MadRS (formerly YxdJK) membrane antimicrobial peptide defense system controls a network of genes, including a previously uncharacterized three gene operon (madEFG) that protects the E. faecalis cell envelope from antimicrobial peptides. Constitutive activation of the system confers protection against CAMPs and daptomycin in the absence of a functional LiaFSR system and leads to persistence of cardiac microlesions in vivo. Moreover, changes in the lipid cell membrane environment alter CAMP susceptibility and expression of the MadRS system. Thus, we provide a framework supporting a multilayered envelope defense mechanism for resistance and survival coupled to virulence.
]]></description>
<dc:creator>Miller, W. R.</dc:creator>
<dc:creator>Nguyen, A.</dc:creator>
<dc:creator>Singh, K. V.</dc:creator>
<dc:creator>Rizvi, S.</dc:creator>
<dc:creator>Khan, A.</dc:creator>
<dc:creator>Erickson, S. G.</dc:creator>
<dc:creator>Egge, S. L.</dc:creator>
<dc:creator>Cruz, M.</dc:creator>
<dc:creator>Dinh, A. Q.</dc:creator>
<dc:creator>Diaz, L.</dc:creator>
<dc:creator>Zhang, R.</dc:creator>
<dc:creator>Xu, L.</dc:creator>
<dc:creator>Garsin, D. A.</dc:creator>
<dc:creator>Shamoo, Y.</dc:creator>
<dc:creator>Arias, C. A.</dc:creator>
<dc:date>2023-10-19</dc:date>
<dc:identifier>doi:10.1101/2023.10.17.562839</dc:identifier>
<dc:title><![CDATA[Membrane Lipids Augment Cell Envelope Stress Signaling and Resistance to Antibiotics and Antimicrobial Peptides in Enterococcus faecalis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.10.17.562653v1?rss=1">
<title>
<![CDATA[
A syngeneic spontaneous zebrafish model of tp53-deficient, EGFRviii, and PI3KCA H1047R-driven glioblastoma reveals inhibitory roles for inflammation during tumor initiation and relapse in vivo 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.17.562653v1?rss=1</link>
<description><![CDATA[
High-throughput vertebrate animal model systems for the study of patient-specific biology and new therapeutic approaches for aggressive brain tumors are currently lacking, and new approaches are urgently needed. Therefore, to build a patient-relevant in vivo model of human glioblastoma, we expressed common oncogenic variants including activated human EGFRvIII and PI3KCAH1047R under the control of the radial glial-specific promoter her4.1 in syngeneic tp53 loss-of-function mutant zebrafish. Robust tumor formation was observed prior to 45 days of life, and tumors had a gene expression signature similar to human glioblastoma of the mesenchymal subtype, with a strong inflammatory component. Within early stage tumor lesions, and in an in vivo and endogenous tumor microenvironment, we visualized infiltration of phagocytic cells, as well as internalization of tumor cells by mpeg1.1:EGFP+ microglia/macrophages, suggesting negative regulatory pressure by pro-inflammatory cell types on tumor growth at early stages of glioblastoma initiation. Furthermore, CRISPR/Cas9-mediated gene targeting of master inflammatory transcription factors irf7 or irf8 led to increased tumor formation in the primary context, while suppression of phagocyte activity led to enhanced tumor cell engraftment following transplantation into otherwise immune-competent zebrafish hosts. Altogether, we developed a genetically relevant model of aggressive human glioblastoma and harnessed the unique advantages of zebrafish including live imaging, high-throughput genetic and chemical manipulations to highlight important tumor suppressive roles for the innate immune system on glioblastoma initiation, with important future opportunities for therapeutic discovery and optimizations.
]]></description>
<dc:creator>Weiss, A.</dc:creator>
<dc:creator>D'Amata, C.</dc:creator>
<dc:creator>Pearson, B. J.</dc:creator>
<dc:creator>Hayes, M. N.</dc:creator>
<dc:date>2023-10-20</dc:date>
<dc:identifier>doi:10.1101/2023.10.17.562653</dc:identifier>
<dc:title><![CDATA[A syngeneic spontaneous zebrafish model of tp53-deficient, EGFRviii, and PI3KCA H1047R-driven glioblastoma reveals inhibitory roles for inflammation during tumor initiation and relapse in vivo]]></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/10.1101/2023.10.18.562964v1?rss=1">
<title>
<![CDATA[
Pathway-based, reaction-specific annotation of disease variants for elucidation of molecular phenotypes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.18.562964v1?rss=1</link>
<description><![CDATA[
Disease variant annotation in the context of biological reactions and pathways can provide a standardized overview of molecular phenotypes of pathogenic mutations that is amenable to computational mining and mathematical modeling. Reactome, an open source, manually curated, peer-reviewed database of human biological pathways, provides annotations for over 4000 disease variants of close to 400 genes in the context of [~]800 disease reactions constituting [~]400 disease pathways. Functional annotation of disease variants proceeds from normal gene functions, through disease variants whose divergence from normal molecular behaviors has been experimentally verified, to extrapolation from molecular phenotypes of characterized variants to variants of unknown significance using criteria of the American College of Medical Genetics and Genomics (ACMG). Reactomes pathway-based, reaction-specific disease variant dataset and data model provide a platform to infer pathway output impacts of numerous human disease variants and model organism orthologs, complementing computational predictions of variant pathogenicity.
]]></description>
<dc:creator>Orlic-Milacic, M.</dc:creator>
<dc:creator>Rothfels, K.</dc:creator>
<dc:creator>Matthews, L.</dc:creator>
<dc:creator>Wright, A.</dc:creator>
<dc:creator>Jassal, B.</dc:creator>
<dc:creator>Shamovsky, V.</dc:creator>
<dc:creator>Trinh, Q.</dc:creator>
<dc:creator>Gillespie, M.</dc:creator>
<dc:creator>Sevilla, C.</dc:creator>
<dc:creator>Tiwari, K.</dc:creator>
<dc:creator>Ragueneau, E.</dc:creator>
<dc:creator>Gong, C.</dc:creator>
<dc:creator>Stephan, R.</dc:creator>
<dc:creator>May, B.</dc:creator>
<dc:creator>Haw, R.</dc:creator>
<dc:creator>Weiser, J.</dc:creator>
<dc:creator>Beavers, D.</dc:creator>
<dc:creator>Conley, P.</dc:creator>
<dc:creator>Hermjakob, H.</dc:creator>
<dc:creator>Stein, L. D.</dc:creator>
<dc:creator>D'Eustachio, P.</dc:creator>
<dc:creator>Wu, G.</dc:creator>
<dc:date>2023-10-22</dc:date>
<dc:identifier>doi:10.1101/2023.10.18.562964</dc:identifier>
<dc:title><![CDATA[Pathway-based, reaction-specific annotation of disease variants for elucidation of molecular phenotypes]]></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/10.1101/2023.10.20.563335v1?rss=1">
<title>
<![CDATA[
Machine learning links T cell function and spatial localization to neoadjuvant immunotherapy and clinical outcome in pancreatic cancer 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.20.563335v1?rss=1</link>
<description><![CDATA[
Tumor molecular datasets are becoming increasingly complex, making it nearly impossible for humans alone to effectively analyze them. Here, we demonstrate the power of using machine learning to analyze a single-cell, spatial, and highly multiplexed proteomic dataset from human pancreatic cancer and reveal underlying biological mechanisms that may contribute to clinical outcome. A novel multiplex immunohistochemistry antibody panel was used to audit T cell functionality and spatial localization in resected tumors from treatment-naive patients with localized pancreatic ductal adenocarcinoma (PDAC) compared to a second cohort of patients treated with neoadjuvant agonistic CD40 (CD40) monoclonal antibody therapy. In total, nearly 2.5 million cells from 306 tissue regions collected from 29 patients across both treatment cohorts were assayed, and more than 1,000 tumor microenvironment (TME) features were quantified. We then trained machine learning models to accurately predict CD40 treatment status and disease-free survival (DFS) following CD40 therapy based upon TME features. Through downstream interpretation of the machine learning models predictions, we found CD40 therapy to reduce canonical aspects of T cell exhaustion within the TME, as compared to treatment-naive TMEs. Using automated clustering approaches, we found improved DFS following CD40 therapy to correlate with the increased presence of CD44+ CD4+ Th1 cells located specifically within cellular spatial neighborhoods characterized by increased T cell proliferation, antigen-experience, and cytotoxicity in immune aggregates. Overall, our results demonstrate the utility of machine learning in molecular cancer immunology applications, highlight the impact of CD40 therapy on T cells within the TME, and identify potential candidate biomarkers of DFS for CD40-treated patients with PDAC.
]]></description>
<dc:creator>Blise, K. E.</dc:creator>
<dc:creator>Sivagnanam, S.</dc:creator>
<dc:creator>Betts, C. B.</dc:creator>
<dc:creator>Betre, K.</dc:creator>
<dc:creator>Kirchberger, N.</dc:creator>
<dc:creator>Tate, B.</dc:creator>
<dc:creator>Furth, E. E.</dc:creator>
<dc:creator>Dias Costa, A.</dc:creator>
<dc:creator>Nowak, J. A.</dc:creator>
<dc:creator>Wolpin, B. M.</dc:creator>
<dc:creator>Vonderheide, R. H.</dc:creator>
<dc:creator>Goecks, J.</dc:creator>
<dc:creator>Coussens, L. M.</dc:creator>
<dc:creator>Byrne, K.</dc:creator>
<dc:date>2023-10-23</dc:date>
<dc:identifier>doi:10.1101/2023.10.20.563335</dc:identifier>
<dc:title><![CDATA[Machine learning links T cell function and spatial localization to neoadjuvant immunotherapy and clinical outcome in pancreatic cancer]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.10.23.563546v1?rss=1">
<title>
<![CDATA[
Fine-tuning TrailMap: The utility of transfer learning to improve the performance of deep learning in axon segmentation of light-sheet microscopy images 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.23.563546v1?rss=1</link>
<description><![CDATA[
Light-sheet microscopy has made possible the 3D imaging of both fixed and live biological tissue, with samples as large as the entire mouse brain. However, segmentation and quantification of that data remains a time-consuming manual undertaking. Machine learning methods promise the possibility of automating this process. This study seeks to advance the performance of prior models through optimizing transfer learning. We fine-tuned the existing TrailMap model using expert-labeled data from noradrenergic axonal structures in the mouse brain. By fine-tuning the final two layers of the neural network at a lower learning rate of the TrailMap model, we demonstrate an improved recall and an occasionally improved adjusted F1- score within our test dataset over using the originally trained TrailMap model.

Availability and implementation: The software and data are freely available at https://github.com/pnnl/brain_ohsu and https://data.pnl.gov/group/204/nodes/dataset/35673, respectively.
]]></description>
<dc:creator>Oostrom, M.</dc:creator>
<dc:creator>Muniak, M.</dc:creator>
<dc:creator>Eichler West, R. M.</dc:creator>
<dc:creator>Akers, S.</dc:creator>
<dc:creator>Pande, P.</dc:creator>
<dc:creator>Obiri, M.</dc:creator>
<dc:creator>Wang, W.</dc:creator>
<dc:creator>Bowyer, K.</dc:creator>
<dc:creator>Wu, Z.</dc:creator>
<dc:creator>Bramer, L. M.</dc:creator>
<dc:creator>Mao, T.</dc:creator>
<dc:creator>Webb-Robertson, B.-J. M.</dc:creator>
<dc:date>2023-10-23</dc:date>
<dc:identifier>doi:10.1101/2023.10.23.563546</dc:identifier>
<dc:title><![CDATA[Fine-tuning TrailMap: The utility of transfer learning to improve the performance of deep learning in axon segmentation of light-sheet microscopy images]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.10.23.563614v1?rss=1">
<title>
<![CDATA[
Sex and species associated differences in Complement-mediated immunity in Humans and Rhesus macaques 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.23.563614v1?rss=1</link>
<description><![CDATA[
The complement system can be viewed as a  moderator of innate immunity,  instructor of humoral immunity, and  regulator of adaptive immunity. While sex and aging are known to affect humoral and cellular immune systems, their impact on the complement pathway in humans and rhesus macaques, a commonly used non-human primate model system, have not been well-studied. To address this knowledge gap, we analyzed serum samples from 90 humans and 75 rhesus macaques for the abundance and activity of the complement system components. While sequences of cascade proteins were highly conserved, dramatically different levels were observed between species. Whereas the low levels detected in rhesus samples raised questions about the suitability of the test, differences in levels of complement proteins were observed in male and female humans. Levels of total and antibody-dependent deposition of C1q and C3b on a glycosylated antigen differed between human and rhesus, suggesting differential recognition of glycans. Functional differences in complement-mediated lysis of antibody-sensitized cells were observed in multiple assays and showed that human females frequently exhibited higher lytic activity than human males or rhesus macaques, which typically did not exhibit such sexual dimorphism. Other differences between species and sexes were observed in more narrow contexts--for only certain antibodies, antigens, or assays. Collectively, these results expand our knowledge of sexual dimorphism in the complement system in humans, identifying differences that appear to be absent from rhesus macaques.
]]></description>
<dc:creator>Kelkar, N. S.</dc:creator>
<dc:creator>Goldberg, B. S.</dc:creator>
<dc:creator>Dufloo, J.</dc:creator>
<dc:creator>Bruel, T.</dc:creator>
<dc:creator>Schwartz, O.</dc:creator>
<dc:creator>Hessell, A. J.</dc:creator>
<dc:creator>Ackerman, M. E.</dc:creator>
<dc:date>2023-10-25</dc:date>
<dc:identifier>doi:10.1101/2023.10.23.563614</dc:identifier>
<dc:title><![CDATA[Sex and species associated differences in Complement-mediated immunity in Humans and Rhesus macaques]]></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/10.1101/2023.10.25.563997v1?rss=1">
<title>
<![CDATA[
The prevalence of Fusobacterium nucleatum subspecies in the oral cavity stratifies by local health status 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.25.563997v1?rss=1</link>
<description><![CDATA[
The ubiquitous inflammophilic pathobiont Fusobacterium nucleatum is widely recognized for its strong association with a variety of human dysbiotic diseases such as periodontitis and oral/extraoral abscesses, as well as multiple types of cancer. F. nucleatum is currently subdivided into four subspecies: F. nucleatum subspecies nucleatum (Fn. nucleatum), animalis (Fn. animalis), polymorphum (Fn. polymorphum), and vincentii/fusiforme (Fn. vincentii). Although these subspecies have been historically considered as functionally interchangeable in the oral cavity, direct clinical evidence is largely lacking for this assertion. Consequently, we assembled a collection of oral clinical specimens to determine whether F. nucleatum subspecies prevalence in the oral cavity stratifies by local oral health status. Patient-matched clinical specimens of both disease-free dental plaque and odontogenic abscess were analyzed with newly developed culture-dependent and culture-independent approaches using 44 and 60 oral biofilm/tooth abscess paired specimens, respectively. Most oral cavities were found to simultaneously harbor multiple F. nucleatum subspecies, with a greater diversity present within dental plaque compared to abscesses. In dental plaque, Fn. polymorphum is clearly the dominant organism, but this changes dramatically within odontogenic abscesses where Fn. animalis is heavily favored over all other fusobacteria. Surprisingly, the most commonly studied F. nucleatum subspecies, Fn. nucleatum, is only a minor constituent in the oral cavity. To gain further insights into the genetic basis for these phenotypes, we subsequently performed pangenome, phylogenetic, and functional enrichment analyses of oral fusobacterial genomes using the Anvio platform, which revealed significant genotypic distinctions among F. nucleatum subspecies. Accordingly, our results strongly support a taxonomic reassignment of each F. nucleatum subspecies into distinct Fusobacterium species. Of these, Fn. animalis should be considered as the most clinically relevant at sites of active inflammation, despite being among the least characterized oral fusobacteria.
]]></description>
<dc:creator>Krieger, M.</dc:creator>
<dc:creator>AbdelRahman, Y. M.</dc:creator>
<dc:creator>Choi, D.</dc:creator>
<dc:creator>Palmer, E. A.</dc:creator>
<dc:creator>Yoo, A.</dc:creator>
<dc:creator>McGuire, S.</dc:creator>
<dc:creator>Kreth, J.</dc:creator>
<dc:creator>Merritt, J.</dc:creator>
<dc:date>2023-10-26</dc:date>
<dc:identifier>doi:10.1101/2023.10.25.563997</dc:identifier>
<dc:title><![CDATA[The prevalence of Fusobacterium nucleatum subspecies in the oral cavity stratifies by local health status]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.10.24.563815v1?rss=1">
<title>
<![CDATA[
Analysis of uveal melanoma scRNA sequencing data identifies neoplastic-immune hybrid cells that exhibit metastatic potential 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.24.563815v1?rss=1</link>
<description><![CDATA[
Uveal melanoma (UM) is the most common non-cutaneous melanoma and is an intraocular malignancy that affects nearly 7,000 individuals per year worldwide. Of these, nearly 50% will progress to metastatic disease for which there are currently no effective therapies. Despite advances in the molecular profiling and metastatic stratification of class 1 and 2 UM tumors, little is known regarding the underlying biology of UM metastasis. Our group has identified a disseminated tumor cell population characterized by co-expression of immune and melanoma proteins, (circulating hybrid cells (CHCs), in patients with UM. Compared to circulating tumor cells, CHCs are detected at an increased prevalence in peripheral blood and can be used as a non-invasive biomarker to predict metastatic progression. To identify mechanisms underlying enhanced hybrid cell dissemination we sought to identify hybrid cells within a primary UM single cell RNA-seq dataset. Using rigorous doublet discrimination approaches, we identified UM hybrids and evaluated their gene expression, predicted ligand-receptor status, and cell-cell communication state in relation to other melanoma and immune cells within the primary tumor. We identified several genes and pathways upregulated in hybrid cells, including those involved in enhancing cell motility and cytoskeleton rearrangement, evading immune detection, and altering cellular metabolism. In addition, we identified that hybrid cells express ligand-receptor signaling pathways implicated in promoting cancer metastasis including IGF1-IGFR1, GAS6-AXL, LGALS9-P4HB, APP-CD74 and CXCL12-CXCR4. These results contribute to our understanding of tumor progression and interactions between tumor cells and immune cells in the UM microenvironment that may promote metastasis.
]]></description>
<dc:creator>Anderson, A. N.</dc:creator>
<dc:creator>Conley, P.</dc:creator>
<dc:creator>Klocke, C. D.</dc:creator>
<dc:creator>Sengupta, S. K.</dc:creator>
<dc:creator>Giske, N. R.</dc:creator>
<dc:creator>Robinson, T. L.</dc:creator>
<dc:creator>Fan, Y.</dc:creator>
<dc:creator>Jones, J. A.</dc:creator>
<dc:creator>Gibbs, S. L.</dc:creator>
<dc:creator>Skalet, A. H.</dc:creator>
<dc:creator>Wu, G.</dc:creator>
<dc:creator>Wong, M. H.</dc:creator>
<dc:date>2023-10-27</dc:date>
<dc:identifier>doi:10.1101/2023.10.24.563815</dc:identifier>
<dc:title><![CDATA[Analysis of uveal melanoma scRNA sequencing data identifies neoplastic-immune hybrid cells that exhibit metastatic potential]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.10.26.564271v1?rss=1">
<title>
<![CDATA[
Dock1 acts cell-autonomously in Schwann cells to regulate the development, maintenance, and repair of peripheral myelin 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.26.564271v1?rss=1</link>
<description><![CDATA[
Schwann cells, the myelinating glia of the peripheral nervous system (PNS), are critical for myelin development, maintenance, and repair. Rac1 is a known regulator of radial sorting, a key step in developmental myelination, and we previously showed in zebrafish that loss of Dock1, a Rac1-specific guanine nucleotide exchange factor, results in delayed peripheral myelination in development. We demonstrate here that Dock1 is necessary for myelin maintenance and remyelination after injury in adult zebrafish. Furthermore, it performs an evolutionary conserved role in mice, acting cell-autonomously in Schwann cells to regulate peripheral myelin development, maintenance, and repair. Additionally, manipulating Rac1 levels in larval zebrafish reveals that dock1 mutants are sensitized to inhibition of Rac1, suggesting an interaction between the two proteins during PNS development. We propose that the interplay between Dock1 and Rac1 signaling in Schwann cells is required to establish, maintain, and facilitate repair and remyelination within the peripheral nervous system.
]]></description>
<dc:creator>Doan, R. A.</dc:creator>
<dc:creator>Monk, K. R.</dc:creator>
<dc:date>2023-10-27</dc:date>
<dc:identifier>doi:10.1101/2023.10.26.564271</dc:identifier>
<dc:title><![CDATA[Dock1 acts cell-autonomously in Schwann cells to regulate the development, maintenance, and repair of peripheral myelin]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.10.27.564401v1?rss=1">
<title>
<![CDATA[
Gardenin A improves cognitive and motor function in A53T-α-syn mice 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.27.564401v1?rss=1</link>
<description><![CDATA[
Oxidative stress and neuroinflammation are widespread in the Parkinsons disease (PD) brain and contribute to the synaptic degradation and dopaminergic cell loss that result in cognitive impairment and motor dysfunction. The polymethoxyflavone Gardenin A (GA) has been shown to activate the NRF2-regulated antioxidant pathway and inhibit the NFkB-dependent pro-inflammatory pathway in a Drosophila model of PD. Here, we evaluate the effects of GA on A53T alpha-synuclein overexpressing (A53TSyn) mice.

A53TSyn mice were treated orally for 4 weeks with 0, 25, or 100 mg/kg GA. In the fourth week, mice underwent behavioral testing and tissue was harvested for immunohistochemical analysis of tyrosine hydroxylase (TH) and phosphorylated alpha synuclein (pSyn) expression, and quantification of synaptic, antioxidant and inflammatory gene expression. Results were compared to vehicle-treated C57BL6 mice.

Treatment with 100 mg/kg GA improved associative memory and decreased abnormalities in mobility and gait in A53TSyn mice. GA treatment also reduced cortical and hippocampal levels of pSyn and attenuated the reduction in TH expression in the striatum. Additionally, GA increased cortical expression of NRF2-regulated antioxidant genes and decreased expression of NFkB-dependent pro-inflammatory genes. GA was readily detectable in the brains of treated mice and modulated the lipid profile in the deep gray brain tissue of those animals.

While the beneficial effects of GA on cognitive deficits, motor dysfunction and PD pathology are promising, future studies are needed to further fully elucidate the mechanism of action of GA, optimizing dosing and confirm these effects in other PD models.

Significance StatementThe polymethoxyflavone Gardenin A can improve cognitive and motor function and attenuate both increases in phosphorylated alpha synuclein and reductions in tyrosine hydroxylase expression in A53T alpha synuclein overexpressing mice. These effects may be related to activation of the NRF2-regulated antioxidant response and downregulation of NFkB-dependent inflammatory response by Gardenin A in treated animals. The study also showed excellent brain bioavailability of Gardenin A and modifications of the lipid profile, possibly through interactions between Gardenin A with the lipid bilayer, following oral administration. The study confirms neuroprotective activity of Gardenin A previously reported in toxin induced Drosophila model of Parkinsons disease.
]]></description>
<dc:creator>Hack, W.</dc:creator>
<dc:creator>Gladen-Kolarsky, N.</dc:creator>
<dc:creator>Chatterjee, S.</dc:creator>
<dc:creator>Lang, Q.</dc:creator>
<dc:creator>Maitra, U.</dc:creator>
<dc:creator>Ciesla, L.</dc:creator>
<dc:creator>Gray, N. E.</dc:creator>
<dc:date>2023-11-01</dc:date>
<dc:identifier>doi:10.1101/2023.10.27.564401</dc:identifier>
<dc:title><![CDATA[Gardenin A improves cognitive and motor function in A53T-α-syn mice]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-11-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.10.30.563998v1?rss=1">
<title>
<![CDATA[
Efficient semi-supervised semantic segmentation of electron microscopy cancer images with sparse annotations 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.30.563998v1?rss=1</link>
<description><![CDATA[
Electron microscopy (EM) enables imaging at nanometer resolution and can shed light on how cancer evolves to develop resistance to therapy. Acquiring these images has become a routine task; however, analyzing them is now the bottleneck, as manual structure identification is very time-consuming and can take up to several months for a single sample. Deep learning approaches offer a suitable solution to speed up the analysis. In this work, we present a study of several state-of-the-art deep learning models for the task of segmenting nuclei and nucleoli in volumes from tumor biopsies. We compared previous results obtained with the ResUNet architecture to the more recent UNet++, FracTALResNet, SenFormer, and CEECNet models. In addition, we explored the utilization of unlabeled images through semi-supervised learning with Cross Pseudo Supervision. We have trained and evaluated all of the models on sparse manual labels from three fully annotated in-house datasets that we have made available on demand, demonstrating improvements in terms of 3D Dice score. From the analysis of these results, we drew conclusions on the relative gains of using more complex models, semi-supervised learning as well as next steps for the mitigation of the manual segmentation bottleneck.
]]></description>
<dc:creator>Pagano, L.</dc:creator>
<dc:creator>Thibault, G.</dc:creator>
<dc:creator>Bousselham, W.</dc:creator>
<dc:creator>Riesterer, J. L.</dc:creator>
<dc:creator>Song, X.</dc:creator>
<dc:creator>Gray, J. W.</dc:creator>
<dc:date>2023-11-01</dc:date>
<dc:identifier>doi:10.1101/2023.10.30.563998</dc:identifier>
<dc:title><![CDATA[Efficient semi-supervised semantic segmentation of electron microscopy cancer images with sparse annotations]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-11-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.10.31.565026v1?rss=1">
<title>
<![CDATA[
From average transient transporter currents to microscopic mechanism -- A Bayesian analysis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.10.31.565026v1?rss=1</link>
<description><![CDATA[
Electrophysiology studies of secondary active transporters have revealed quantitative, mechanistic insights over many decades of research. However, the emergence of new experimental and analysis approaches calls for investigation of the capabilities and limitations of the newer methods. We examine the ability of solid-supported membrane electrophysiology (SSME) to characterize discrete-state kinetic models with > 10 rate constants. We use a Bayesian framework applied to synthetic data for three tasks: to quantify and check (i) the precision of parameter estimates under different assumptions, (ii) the ability of computation to guide selection of experimental conditions, and (iii) the ability of SSME data to distinguish among mechanisms. When the general mechanism - event order - is known in advance, we show that a subset of kinetic parameters can be "practically identified" within[~] 1 order of magnitude, based on SSME current traces that visually appear to exhibit simple exponential behavior. This remains true even when accounting for systematic measurement bias and realistic uncertainties in experimental inputs (concentrations) are incorporated into the analysis. When experimental conditions are optimized or different experiments are combined, the number of practically identifiable parameters can be increased substantially. Some parameters remain intrinsically difficult to estimate through SSME data alone, suggesting additional experiments are required to fully characterize parameters. We additionally demonstrate the ability to perform model selection and determine the order of events when that is not known in advance, comparing Bayesian and maximum-likelihood approaches. Finally, our studies elucidate good practices for the increasingly popular, but subtly challenging, Bayesian calculations for structural and systems biology.
]]></description>
<dc:creator>George, A.</dc:creator>
<dc:creator>Zuckerman, D. M.</dc:creator>
<dc:date>2023-11-02</dc:date>
<dc:identifier>doi:10.1101/2023.10.31.565026</dc:identifier>
<dc:title><![CDATA[From average transient transporter currents to microscopic mechanism -- A Bayesian analysis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-11-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.11.02.565346v1?rss=1">
<title>
<![CDATA[
CG-SLENP: A chemical genetics strategy to selectively label existing proteins and newly synthesized proteins 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.11.02.565346v1?rss=1</link>
<description><![CDATA[
Protein synthesis and subsequent delivery to the target locations in cells are essential for their proper functions. Methods to label and distinguish newly synthesized proteins from existing ones are critical to assess their differential properties, but such methods are lacking. We describe the first chemical genetics-based approach for selective labeling of existing and newly synthesized proteins that we termed as CG-SLENP. Using HaloTag in-frame fusion with lamin A (LA), we demonstrate that the two pools of proteins can be selectively labeled using CG-SLENP in living cells. We further employ our recently developed selective small molecule ligand LBL1 for LA to probe the potential differences between newly synthesized and existing LA. Our results show that LBL1 can differentially modulate these two pools of LA. These results indicate that the assembly states of newly synthesized LA are distinct from existing LA in living cells. The CG-SLENP method is potentially generalizable to study any cellular proteins.



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]]></description>
<dc:creator>Wang, J.</dc:creator>
<dc:creator>Chao, B.</dc:creator>
<dc:creator>Piesner, J.</dc:creator>
<dc:creator>Kelly, F.</dc:creator>
<dc:creator>Petrie, S. K.</dc:creator>
<dc:creator>Xiao, X.</dc:creator>
<dc:creator>Li, B. X.</dc:creator>
<dc:date>2023-11-02</dc:date>
<dc:identifier>doi:10.1101/2023.11.02.565346</dc:identifier>
<dc:title><![CDATA[CG-SLENP: A chemical genetics strategy to selectively label existing proteins and newly synthesized proteins]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-11-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.11.02.565050v1?rss=1">
<title>
<![CDATA[
AXDND1 is required to balance spermatogonial commitment and for sperm tail formation in mice and humans 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.11.02.565050v1?rss=1</link>
<description><![CDATA[
Dynein complexes are large, multi-unit assemblies involved in many biological processes including male fertility via their critical roles in protein transport and axoneme motility. Previously we identified a pathogenic variant in the dynein gene AXDND1 in an infertile man. Subsequently we identified an additional four potentially compound heterozygous variants of unknown significance in AXDND1 in two additional infertile men. We thus tested the role of AXDND1 in mammalian male fertility by generating a knockout mouse model. Axdnd1-/- males were sterile at all ages but could undergo one round of histologically complete spermatogenesis. Subsequently, a progressive imbalance of spermatogonial commitment to spermatogenesis over self-renewal occurred, ultimately leading to catastrophic germ cell loss, loss of blood-testis barrier patency and immune cell infiltration. Sperm produced during the first wave of spermatogenesis were immotile due to abnormal axoneme structure, including the presence of ectopic vesicles and abnormalities in outer dense fibres and microtubule doublet structures. Sperm output was additionally compromised by a severe spermiation defect and abnormal sperm individualisation. Collectively, our data highlight the essential roles of AXDND1 as a regulator of spermatogonial commitment to spermatogenesis and during the processes of spermiogenesis where it is essential for sperm tail development, release and motility.
]]></description>
<dc:creator>Houston, B. J.</dc:creator>
<dc:creator>Nguyen, J.</dc:creator>
<dc:creator>Merriner, D. J.</dc:creator>
<dc:creator>O'Connor, A. E.</dc:creator>
<dc:creator>Lopes, A. M.</dc:creator>
<dc:creator>Nagirnaja, L.</dc:creator>
<dc:creator>Friedrich, C.</dc:creator>
<dc:creator>Kliesch, S.</dc:creator>
<dc:creator>Tuettelmann, F.</dc:creator>
<dc:creator>Aston, K. I.</dc:creator>
<dc:creator>Conrad, D.</dc:creator>
<dc:creator>Hobbs, R. M.</dc:creator>
<dc:creator>Dunleavy, J. E.</dc:creator>
<dc:creator>O'Bryan, M. K.</dc:creator>
<dc:date>2023-11-04</dc:date>
<dc:identifier>doi:10.1101/2023.11.02.565050</dc:identifier>
<dc:title><![CDATA[AXDND1 is required to balance spermatogonial commitment and for sperm tail formation in mice and humans]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-11-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.11.02.565358v1?rss=1">
<title>
<![CDATA[
Rapid nongenomic estrogen signaling controls alcohol drinking behavior 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.11.02.565358v1?rss=1</link>
<description><![CDATA[
Ovarian-derived estrogen is a key modulator of numerous physiological processes via genomic and nongenomic mechanisms, including signaling non-canonically at membrane-associated estrogen receptors in the brain to rapidly regulate neuronal function. However, the mechanisms mediating estrogen regulation of behaviors such as alcohol consumption remain unclear. Early alcohol drinking confers greater risk for alcohol use disorder in women than men, and binge alcohol drinking is correlated with high circulating estrogen levels, but a causal role for estrogen signaling in driving alcohol drinking in gonadally-intact animals has not been established. We found that female mice displayed greater binge alcohol drinking and reduced avoidance behavior when circulating estrogen was high during the proestrus phase of the estrous cycle than when it was low, contributing to sex differences in these behaviors. The pro-drinking, but not anxiolytic, effect of high endogenous estrogen state occurred via rapid estrogen signaling at membrane-associated estrogen receptor alpha in the bed nucleus of the stria terminalis, which promoted synaptic excitation of corticotropin-releasing factor neurons and facilitated their activity during alcohol drinking behavior. This study is the first to demonstrate a rapid, nongenomic signaling mechanism for ovarian-derived estrogen signaling in the brain controlling behavior in gonadally intact females, and it establishes a causal role for estrogen in an intact hormonal context for driving alcohol consumption that contributes to known sex differences in this behavior.
]]></description>
<dc:creator>Zallar, L. J.</dc:creator>
<dc:creator>Rivera-Irizarry, J. K.</dc:creator>
<dc:creator>Hamor, P. U.</dc:creator>
<dc:creator>Pigulevskiy, I.</dc:creator>
<dc:creator>Liu, D.</dc:creator>
<dc:creator>Welday, J. P.</dc:creator>
<dc:creator>Rico Rozo, A.-S.</dc:creator>
<dc:creator>Bender, R.</dc:creator>
<dc:creator>Asfouri, J.</dc:creator>
<dc:creator>Skelly, M. J.</dc:creator>
<dc:creator>Fecteau, K. M.</dc:creator>
<dc:creator>Hadley, C. K.</dc:creator>
<dc:creator>Levine, O. B.</dc:creator>
<dc:creator>Mehanna, H.</dc:creator>
<dc:creator>Nelson, S.</dc:creator>
<dc:creator>Miller, J.</dc:creator>
<dc:creator>Ghazal, P.</dc:creator>
<dc:creator>Bellotti, P.</dc:creator>
<dc:creator>Erikson, D. W.</dc:creator>
<dc:creator>Geri, J.</dc:creator>
<dc:creator>Pleil, K. E.</dc:creator>
<dc:date>2023-11-05</dc:date>
<dc:identifier>doi:10.1101/2023.11.02.565358</dc:identifier>
<dc:title><![CDATA[Rapid nongenomic estrogen signaling controls alcohol drinking behavior]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-11-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.11.06.565932v1?rss=1">
<title>
<![CDATA[
Tweek-dependent formation of ER-PM contact sites enables astrocyte phagocytic function and remodeling of neurons 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.11.06.565932v1?rss=1</link>
<description><![CDATA[
Neuronal remodeling generates an enormous amount of cellular debris, which is cleared from the nervous system by glia. At the larva-to-adult transition, Drosophila astrocytes transform into phagocytes and engulf degenerating larval synapses, axonal and dendritic debris. Here we show Tweek, a member of the bridge-like lipid transfer protein family, is upregulated in astrocytes as they ramp up their phagocytic function early in metamorphosis, and is essential for internalization and degradation of neuronal debris. Tweek forms a bridge between the endoplasmic reticulum (ER) and plasma membrane (PM), and loss of Tweek disrupts ER-PM contact formation and membrane lipid distribution. Patient-identified mutations in the human homolog associated with Alkuraya-Kucinskas syndrome resulted in similar defects in neuronal remodeling, indicating these are loss of function mutations. We propose Tweek helps establish and maintain ER-PM contacts during astrocyte phagocytic function and drives bulk lipid transfer to the plasma membrane for continued efficient internalization and degradation of neuronal debris.
]]></description>
<dc:creator>Kang, Y.</dc:creator>
<dc:creator>Jefferson, A.</dc:creator>
<dc:creator>Sheehan, A.</dc:creator>
<dc:creator>De La Torre, R.</dc:creator>
<dc:creator>Jay, T.</dc:creator>
<dc:creator>Chiao, L.</dc:creator>
<dc:creator>Hulegaard, A.</dc:creator>
<dc:creator>Corty, M. M.</dc:creator>
<dc:creator>Baconguis, I.</dc:creator>
<dc:creator>Zhou, Z.</dc:creator>
<dc:creator>Freeman, M.</dc:creator>
<dc:date>2023-11-07</dc:date>
<dc:identifier>doi:10.1101/2023.11.06.565932</dc:identifier>
<dc:title><![CDATA[Tweek-dependent formation of ER-PM contact sites enables astrocyte phagocytic function and remodeling of neurons]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-11-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.11.08.566195v1?rss=1">
<title>
<![CDATA[
ChatGPT usage in the Reactome curation process 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.11.08.566195v1?rss=1</link>
<description><![CDATA[
Appreciating the rapid advancement and ubiquity of generative AI, particularly ChatGPT, a chatbot using large language models like GPT, we endeavour to explore the potential application of ChatGPT in the data collection and annotation stages within the Reactome curation process. This exploration aimed to create an automated or semi-automated framework to mitigate the extensive manual effort traditionally required for gathering and annotating information pertaining to biological pathways, adopting a Reactome "reaction-centric" approach. In this pilot study, we used ChatGPT/GPT4 to address gaps in the pathway annotation and enrichment in parallel with the conventional manual curation process. This approach facilitated a comparative analysis, where we assessed the outputs generated by ChatGPT against manually extracted information. The primary objective of this comparison was to ascertain the efficiency of integrating ChatGPT or other large language models into the Reactome curation workflow and helping plan our annotation pipeline, ultimately improving our protein-to-pathway association in a reliable and automated or semi-automated way. In the process, we identified some promising capabilities and inherent challenges associated with the utilisation of ChatGPT/GPT4 in general and also specifically in the context of Reactome curation processes. We describe approaches and tools for refining the output given by ChatGPT/GPT4 that aid in generating more accurate and detailed output.
]]></description>
<dc:creator>Tiwari, K. K.</dc:creator>
<dc:creator>Matthews, L.</dc:creator>
<dc:creator>May, B.</dc:creator>
<dc:creator>Shamovsky, V.</dc:creator>
<dc:creator>Orlic-Milacic, M.</dc:creator>
<dc:creator>Rothfels, K.</dc:creator>
<dc:creator>Ragueneau, E.</dc:creator>
<dc:creator>Gong, C.</dc:creator>
<dc:creator>Stephan, R.</dc:creator>
<dc:creator>Li, N.</dc:creator>
<dc:creator>Wu, G.</dc:creator>
<dc:creator>Stein, L. D.</dc:creator>
<dc:creator>D'eustachio, P.</dc:creator>
<dc:creator>Hermjakob, H.</dc:creator>
<dc:date>2023-11-08</dc:date>
<dc:identifier>doi:10.1101/2023.11.08.566195</dc:identifier>
<dc:title><![CDATA[ChatGPT usage in the Reactome curation process]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-11-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.11.08.565511v1?rss=1">
<title>
<![CDATA[
Altered hippocampal activation in seizure-prone CACNA2D2 knockout mice 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.11.08.565511v1?rss=1</link>
<description><![CDATA[
The voltage-gated calcium channel subunit 2{delta}-2 controls calcium-dependent signaling in neurons, and loss of this subunit causes epilepsy in both mice and humans. To determine whether mice without 2{delta}-2 demonstrate hippocampal activation or histopathological changes associated with seizure activity, we measured expression of the activity-dependent gene c-fos and various histopathological correlates of temporal lobe epilepsy in hippocampal tissue from wildtype (WT) and 2{delta}-2 knockout (CACNA2D2 KO) mice using immunohistochemical staining and confocal microscopy. Both genotypes demonstrated similarly sparse c-fos expression within the hippocampal dentate granule cell layer (GCL) at baseline, consistent with no difference in basal activity of granule cells between genotypes. Surprisingly, when mice were assayed 1 hour after handling-associated convulsions, KO mice had fewer c-fos-positive cells in the dentate gyrus, indicating that activity in the dentate gyrus actually decreased. However, the dentate was significantly more active in KO mice compared to WT after administration of a subthreshold pentylenetetrazole dose, consistent with increased susceptibility to proconvulsant stimuli. Other histopathological markers of temporal lobe epilepsy in these mice, including markers of neurogenesis, glial activation, and mossy fiber sprouting, were similar in WT and KO mice, apart from a small but significant increase in hilar mossy cell density, opposite to what is typically found in mice with temporal lobe epilepsy. This suggests that the differences in seizure-associated hippocampal function in the absence of 2{delta}-2 protein are likely due to altered functional properties of the network without associated structural changes in the hippocampus at the typical age of seizure onset.

Significance StatementCalcium channel 2{delta} subunits play important roles in controlling neuronal circuit structure and function, and mutation of the 2{delta}-2 isoform of this protein is associated with spontaneous seizures. In this study, we find that seizures in 2{delta}-2 mutant mice involve altered hippocampal activation without substantial histopathological changes in hippocampal structure. This suggests that the differences in seizure-associated hippocampal function are likely due to altered functional properties of the network as well as to the contribution of additional brain regions to seizures in mice lacking this protein.
]]></description>
<dc:creator>Danis, A.</dc:creator>
<dc:creator>Gallagher, A. A.</dc:creator>
<dc:creator>Anderson, A. N.</dc:creator>
<dc:creator>Isakharov, A.</dc:creator>
<dc:creator>Beeson, K. A.</dc:creator>
<dc:creator>Schnell, E.</dc:creator>
<dc:date>2023-11-10</dc:date>
<dc:identifier>doi:10.1101/2023.11.08.565511</dc:identifier>
<dc:title><![CDATA[Altered hippocampal activation in seizure-prone CACNA2D2 knockout mice]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-11-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.11.09.566384v1?rss=1">
<title>
<![CDATA[
Identification of Cellular Interactions in the Tumor Immune Microenvironment Underlying CD8 T Cell Exhaustion 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.11.09.566384v1?rss=1</link>
<description><![CDATA[
While immune checkpoint inhibitors show success in treating a subset of patients with certain late-stage cancers, these treatments fail in many other patients as a result of mechanisms that have yet to be fully characterized. The process of CD8 T cell exhaustion, by which T cells become dysfunctional in response to prolonged antigen exposure, has been implicated in immunotherapy resistance. Single-cell RNA sequencing (scRNA-seq) produces an abundance of data to analyze this process; however, due to the complexity of the process, contributions of other cell types to a process within a single cell type cannot be simply inferred. We constructed an analysis framework to first rank human skin tumor samples by degree of exhaustion in tumor-infiltrating CD8 T cells and then identify immune cell type-specific gene-regulatory network patterns significantly associated with T cell exhaustion. Using this framework, we further analyzed scRNA-seq data from human tumor and chronic viral infection samples to compare the T cell exhaustion process between these two contexts. In doing so, we identified transcription factor activity in the macrophages of both tissue types associated with this process. Our framework can be applied beyond the tumor immune microenvironment to any system involving cell-cell communication, facilitating insights into key biological processes that underpin the effective treatment of cancer and other complicated diseases.
]]></description>
<dc:creator>Klocke, C. D.</dc:creator>
<dc:creator>Moran, A.</dc:creator>
<dc:creator>Adey, A.</dc:creator>
<dc:creator>McWeeney, S.</dc:creator>
<dc:creator>Wu, G.</dc:creator>
<dc:date>2023-11-13</dc:date>
<dc:identifier>doi:10.1101/2023.11.09.566384</dc:identifier>
<dc:title><![CDATA[Identification of Cellular Interactions in the Tumor Immune Microenvironment Underlying CD8 T Cell Exhaustion]]></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/10.1101/2023.11.14.567059v1?rss=1">
<title>
<![CDATA[
Prior Fc Receptor activation primes macrophage for increased sensitivity to IgG via long term and short term mechanisms 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.11.14.567059v1?rss=1</link>
<description><![CDATA[
Macrophages measure the  eat-me signal IgG to identify targets for phagocytosis. We wondered if prior encounters with IgG influence macrophage appetite. IgG is recognized by the Fc Receptor. To temporally control Fc Receptor activation, we engineered an Fc Receptor that is activated by light-induced oligomerization of Cry2, triggering phagocytosis. Using this tool, we demonstrate that Fc Receptor activation primes macrophages to be more sensitive to IgG in future encounters. Macrophages that have previously experienced Fc Receptor activation eat more IgG-bound cancer cells. Increased phagocytosis occurs by two discrete mechanisms - a short- and long-term priming. Long term priming requires new protein synthesis and Erk activity. Short term priming does not require new protein synthesis and correlates with an increase in Fc Receptor mobility. Our work demonstrates that IgG primes macrophages for increased phagocytosis, suggesting that therapeutic antibodies may become more effective after initial priming doses.
]]></description>
<dc:creator>Bond, A.</dc:creator>
<dc:creator>Fiaz, S.</dc:creator>
<dc:creator>Rollins, K. R.</dc:creator>
<dc:creator>Nario, J. E. Q.</dc:creator>
<dc:creator>Rosen, S. J.</dc:creator>
<dc:creator>Granados, A.</dc:creator>
<dc:creator>Wilson, M. Z.</dc:creator>
<dc:creator>Morrissey, M. A.</dc:creator>
<dc:date>2023-11-14</dc:date>
<dc:identifier>doi:10.1101/2023.11.14.567059</dc:identifier>
<dc:title><![CDATA[Prior Fc Receptor activation primes macrophage for increased sensitivity to IgG via long term and short term mechanisms]]></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/10.1101/2023.11.15.567132v1?rss=1">
<title>
<![CDATA[
The Impact of SIV-Induced Immunodeficiency on Clinical Manifestation, Immune Response, and Viral Dynamics in SARS-CoV-2 Coinfection 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.11.15.567132v1?rss=1</link>
<description><![CDATA[
Persistent and uncontrolled SARS-CoV-2 replication in immunocompromised individuals has been observed and may be a contributing source of novel viral variants that continue to drive the pandemic. Importantly, the effects of immunodeficiency associated with chronic HIV infection on COVID-19 disease and viral persistence have not been directly addressed in a controlled setting. Here we conducted a pilot study wherein two pigtail macaques (PTM) chronically infected with SIVmac239 were exposed to SARS-CoV-2 and monitored for six weeks for clinical disease, viral replication, and viral evolution, and compared to our previously published cohort of SIV-naive PTM infected with SARS-CoV-2. At the time of SARS-CoV-2 infection, one PTM had minimal to no detectable CD4+ T cells in gut, blood, or bronchoalveolar lavage (BAL), while the other PTM harbored a small population of CD4+ T cells in all compartments. Clinical signs were not observed in either PTM; however, the more immunocompromised PTM exhibited a progressive increase in pulmonary infiltrating monocytes throughout SARS-CoV-2 infection. Single-cell RNA sequencing (scRNAseq) of the infiltrating monocytes revealed a less activated/inert phenotype. Neither SIV-infected PTM mounted detectable anti-SARS-CoV-2 T cell responses in blood or BAL, nor anti-SARS-CoV-2 neutralizing antibodies. Interestingly, despite the diminished cellular and humoral immune responses, SARS-CoV-2 viral kinetics and evolution were indistinguishable from SIV-naive PTM in all sampled mucosal sites (nasal, oral, and rectal), with clearance of virus by 3-4 weeks post infection. SIV-induced immunodeficiency significantly impacted immune responses to SARS-CoV-2 but did not alter disease progression, viral kinetics or evolution in the PTM model. SIV-induced immunodeficiency alone may not be sufficient to drive the emergence of novel viral variants.
]]></description>
<dc:creator>Melton, A.</dc:creator>
<dc:creator>Rowe, L. A.</dc:creator>
<dc:creator>Penney, T.</dc:creator>
<dc:creator>Krzykwa, C.</dc:creator>
<dc:creator>Goff, K.</dc:creator>
<dc:creator>Scheuermann, S. E.</dc:creator>
<dc:creator>Melton, H. J.</dc:creator>
<dc:creator>Williams, K.</dc:creator>
<dc:creator>Golden, N.</dc:creator>
<dc:creator>Moore Green, K.</dc:creator>
<dc:creator>Smith, B.</dc:creator>
<dc:creator>Russell-Lodrigue, K.</dc:creator>
<dc:creator>Dufour, J. P.</dc:creator>
<dc:creator>Doyle-Meyers, L. A.</dc:creator>
<dc:creator>Schiro, F.</dc:creator>
<dc:creator>Aye, P. P.</dc:creator>
<dc:creator>Lifson, J. D.</dc:creator>
<dc:creator>Beddingfield, B. J.</dc:creator>
<dc:creator>Blair, R. V.</dc:creator>
<dc:creator>Bohm, R. P.</dc:creator>
<dc:creator>Kolls, J. K.</dc:creator>
<dc:creator>Rappaport, J.</dc:creator>
<dc:creator>Hoxie, J. A.</dc:creator>
<dc:creator>Maness, N. J.</dc:creator>
<dc:date>2023-11-16</dc:date>
<dc:identifier>doi:10.1101/2023.11.15.567132</dc:identifier>
<dc:title><![CDATA[The Impact of SIV-Induced Immunodeficiency on Clinical Manifestation, Immune Response, and Viral Dynamics in SARS-CoV-2 Coinfection]]></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/10.1101/2023.11.16.567415v1?rss=1">
<title>
<![CDATA[
Water inside the selectivity filter of a K+ ion channel: structural heterogeneity, picosecond dynamics, and hydrogen-bonding 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.11.16.567415v1?rss=1</link>
<description><![CDATA[
Water inside biological ion channels regulates the key properties of these proteins such as selectivity, ion conductance, and gating. In this Article we measure the picosecond spectral diffusion of amide I vibrations of an isotope labeled KcsA potassium channel using two-dimensional infrared (2D IR) spectroscopy. By combining waiting time (100 - 2000 fs) 2D IR measurements of the KcsA channel including 13C18O isotope labeled Val76 and Gly77 residues with molecular dynamics simulations, we elucidated the site-specific dynamics of water and K+ ions inside the selectivity filter of KcsA. We observe inhomogeneous 2D lineshapes with extremely slow spectral diffusion. Our simulations quantitatively reproduce the experiments and show that water is the only component with any appreciable dynamics, whereas K+ ions and the protein are essentially static on a picosecond timescale. By analyzing simulated and experimental vibrational frequencies, we find that water in the selectivity filter can be oriented to form hydrogen bonds with adjacent, or non-adjacent carbonyl groups with the reorientation timescales being three times slower and comparable to that of water molecules in liquid, respectively. Water molecules can reside in the cavity sufficiently far from carbonyls and behave essentially like "free" gas-phase-like water with fast reorientation times. Remarkably, no interconversion between these configurations were observed on a picosecond timescale. These dynamics are in stark contrast with liquid water that remains highly dynamic even in the presence of ions at high concentrations.
]]></description>
<dc:creator>Ryan, M. J.</dc:creator>
<dc:creator>Gao, L.</dc:creator>
<dc:creator>Valiyaveetil, F. I.</dc:creator>
<dc:creator>Kananenka, A. A.</dc:creator>
<dc:creator>Zanni, M. T.</dc:creator>
<dc:date>2023-11-17</dc:date>
<dc:identifier>doi:10.1101/2023.11.16.567415</dc:identifier>
<dc:title><![CDATA[Water inside the selectivity filter of a K+ ion channel: structural heterogeneity, picosecond dynamics, and hydrogen-bonding]]></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/10.1101/2023.11.27.568889v1?rss=1">
<title>
<![CDATA[
Towards Generalizability and Robustness in Biological Object Detection in Electron Microscopy Images 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.11.27.568889v1?rss=1</link>
<description><![CDATA[
Machine learning approaches have the potential for meaningful impact in the biomedical field. However, there are often challenges unique to biomedical data that prohibits the adoption of these innovations. For example, limited data, data volatility, and data shifts all compromise model robustness and generalizability. Without proper tuning and data management, deploying machine learning models in the presence of unaccounted for corruptions leads to reduced or misleading performance. This study explores techniques to enhance model generalizability through iterative adjustments. Specifically, we investigate a detection tasks using electron microscopy images and compare models trained with different normalization and augmentation techniques. We found that models trained with Group Normalization or texture data augmentation outperform other normalization techniques and classical data augmentation, enabling them to learn more generalized features. These improvements persist even when models are trained and tested on disjoint datasets acquired through diverse data acquisition protocols. Results hold true for transformerand convolution-based detection architectures. The experiments show an impressive 29% boost in average precision, indicating significant enhancements in the models generalizibality. This underscores the models capacity to effectively adapt to diverse datasets and demonstrates their increased resilience in real-world applications.
]]></description>
<dc:creator>GIANNIOS, K.</dc:creator>
<dc:creator>Chaurasia, A.</dc:creator>
<dc:creator>Bueno, C.</dc:creator>
<dc:creator>Riesterer, J. L.</dc:creator>
<dc:creator>Pagano, L.</dc:creator>
<dc:creator>Lo, T. P.</dc:creator>
<dc:creator>Thibault, G.</dc:creator>
<dc:creator>Gray, J. W.</dc:creator>
<dc:creator>Song, X.</dc:creator>
<dc:creator>DeLaRosa, B.</dc:creator>
<dc:date>2023-11-27</dc:date>
<dc:identifier>doi:10.1101/2023.11.27.568889</dc:identifier>
<dc:title><![CDATA[Towards Generalizability and Robustness in Biological Object Detection in Electron Microscopy Images]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-11-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.11.30.569167v1?rss=1">
<title>
<![CDATA[
Natural and age-related variation in circulating human hematopoietic stem cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.11.30.569167v1?rss=1</link>
<description><![CDATA[
Hematopoietic stem and progenitor cells (HSPCs) deliver life-long multi-lineage output. However, with aging, we exhibit certain characteristic blood count changes and accumulation of clonal disorders. Better understanding of inter-individual variation in HSPC behavior is needed to understand these age-related phenomena and the transition from health to chronic and acute hematological malignancies. Here we study 627K single circulating CD34+ HSPCs (cHSPCs) from 148 healthy individuals, along with their clinical information and clonal hematopoiesis (CH) profiles, to characterize population-wide and age-related hematopoietic variability. Individuals with CH were linked with reduced frequencies of lymphocyte progenitors and higher RDW. An age-related decrease in lymphoid progenitors was observed, predominantly in males. Inter-individual transcriptional variation in expression of a Lamin-A signature and stemness gene programs were linked with aging and presence of macrocytic anemia. Based on our model for healthy cHSPC variation we construct the normal reference for cHSPC subtype frequencies. We show how compositional and expression deviations from this normal reference can robustly identify myeloid malignancies and pre-malignant states. Together, our data and methodologies present a novel resource, shedding light on various age-related hematopoietic processes, and a comprehensive normal cHSPC reference, which can serve as a tool for diagnosing and characterizing hematological disorders.
]]></description>
<dc:creator>Furer, N.</dc:creator>
<dc:creator>Rappoport, N.</dc:creator>
<dc:creator>Lifshitz, A.</dc:creator>
<dc:creator>bercovich, A.</dc:creator>
<dc:creator>Ben-Kiki, O.</dc:creator>
<dc:creator>Danin, A.</dc:creator>
<dc:creator>Kedmi, M.</dc:creator>
<dc:creator>Shipony, Z.</dc:creator>
<dc:creator>Lipson, D.</dc:creator>
<dc:creator>Meiri, E.</dc:creator>
<dc:creator>Yanai, G.</dc:creator>
<dc:creator>Shapira, S.</dc:creator>
<dc:creator>Arber, N.</dc:creator>
<dc:creator>Berdichevsky, S.</dc:creator>
<dc:creator>Tavor, S.</dc:creator>
<dc:creator>Tyner, J.</dc:creator>
<dc:creator>Joshi, S. K.</dc:creator>
<dc:creator>Landau, D.</dc:creator>
<dc:creator>Ganesan, S.</dc:creator>
<dc:creator>Dusaj, N.</dc:creator>
<dc:creator>Chamely, P.</dc:creator>
<dc:creator>Kaushansky, N.</dc:creator>
<dc:creator>Chapal Ilani, N.</dc:creator>
<dc:creator>Shamir, R.</dc:creator>
<dc:creator>Tanay, A.</dc:creator>
<dc:creator>Shlush, L.</dc:creator>
<dc:date>2023-11-30</dc:date>
<dc:identifier>doi:10.1101/2023.11.30.569167</dc:identifier>
<dc:title><![CDATA[Natural and age-related variation in circulating human hematopoietic stem cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-11-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.11.29.569274v1?rss=1">
<title>
<![CDATA[
Metabolic abnormalities in the bone marrow cells of young offspring born to obese mothers 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.11.29.569274v1?rss=1</link>
<description><![CDATA[
Intrauterine metabolic reprogramming occurs in obese mothers during gestation, putting the offspring at high risk of developing obesity and associated metabolic disorders even before birth. We have generated a mouse model of maternal high-fat diet-induced obesity that recapitulates the metabolic changes seen in humans. Here, we profiled and compared the metabolic characteristics of bone marrow cells of newly weaned 3-week-old offspring of dams fed either a high-fat (Off-HFD) or a regular diet (Off-RD). We utilized a state-of-the-art targeted metabolomics approach coupled with a Seahorse metabolic analyzer. We revealed significant metabolic perturbation in the offspring of HFD-fed vs. RD-fed dams, including utilization of glucose primarily via oxidative phosphorylation, and reduction in levels of amino acids, a phenomenon previously linked to aging. Furthermore, in the bone marrow of three-week-old offspring of high-fat diet-fed mothers, we identified a unique B cell population expressing CD19 and CD11b, and found increased expression of Cyclooxygenase-2 (COX-2) on myeloid CD11b, and on CD11bhi B cells, with all the populations being significantly more abundant in offspring of dams fed HFD but not a regular diet. Altogether, we demonstrate that the offspring of obese mothers show metabolic and immune changes in the bone marrow at a very young age and prior to any symptomatic metabolic disease.
]]></description>
<dc:creator>Phillips, E. A.</dc:creator>
<dc:creator>Alharithi, Y. J.</dc:creator>
<dc:creator>Kadam, L.</dc:creator>
<dc:creator>Coussens, L. M.</dc:creator>
<dc:creator>Kumar, S.</dc:creator>
<dc:creator>Maloyan, A.</dc:creator>
<dc:date>2023-12-01</dc:date>
<dc:identifier>doi:10.1101/2023.11.29.569274</dc:identifier>
<dc:title><![CDATA[Metabolic abnormalities in the bone marrow cells of young offspring born to obese mothers]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-12-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.11.30.569198v1?rss=1">
<title>
<![CDATA[
The Complete Sequence and Comparative Analysis of Ape Sex Chromosomes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.11.30.569198v1?rss=1</link>
<description><![CDATA[
Apes possess two sex chromosomes--the male-specific Y and the X shared by males and females. The Y chromosome is crucial for male reproduction, with deletions linked to infertility1. The X chromosome carries genes vital for reproduction and cognition2. Variation in mating patterns and brain function among great apes suggests corresponding differences in their sex chromosomes. However, due to their highly repetitive nature and incomplete reference assemblies, ape sex chromosomes have been challenging to study. Here, using the methodology developed for the telomere-to-telomere (T2T) human genome, we produced gapless assemblies of the X and Y chromosomes for five great apes (chimpanzee, bonobo, gorilla, Bornean and Sumatran orangutans) and a lesser ape, the siamang gibbon. These assemblies allowed us to untangle the intricacies of ape sex chromosome evolution. We found that, compared to the Xs, the ape Ys vary greatly in size and have low alignability and high levels of structural rearrangements. This divergence on the Y arises from the accumulation of lineage-specific ampliconic regions, palindromes, transposable elements, and satellites. Our analysis of Y chromosome genes revealed expansions of multi-copy gene families and signatures of purifying selection. Thus, the Y exhibits dynamic evolution, while the X is more stable. Mapping short-read sequencing data to these assemblies revealed diversity and selection patterns on sex chromosomes of >100 great ape individuals. These reference assemblies are expected to inform human evolution and conservation genetics of nonhuman apes, all of which are endangered species.
]]></description>
<dc:creator>Makova, K. D.</dc:creator>
<dc:creator>Pickett, B. D.</dc:creator>
<dc:creator>Harris, R. S.</dc:creator>
<dc:creator>Hartley, G. A.</dc:creator>
<dc:creator>Cechova, M.</dc:creator>
<dc:creator>Pal, K.</dc:creator>
<dc:creator>Nurk, S.</dc:creator>
<dc:creator>Yoo, D.</dc:creator>
<dc:creator>Li, Q.</dc:creator>
<dc:creator>Hebbar, P.</dc:creator>
<dc:creator>McGrath, B. C.</dc:creator>
<dc:creator>Antonacci, F.</dc:creator>
<dc:creator>Aubel, M.</dc:creator>
<dc:creator>Biddanda, A.</dc:creator>
<dc:creator>Borchers, M.</dc:creator>
<dc:creator>Bornberg, E.</dc:creator>
<dc:creator>Bouffard, G. G.</dc:creator>
<dc:creator>Brooks, S. Y.</dc:creator>
<dc:creator>Carbone, L.</dc:creator>
<dc:creator>Carrel, L.</dc:creator>
<dc:creator>Carroll, A.</dc:creator>
<dc:creator>Chang, P.-C.</dc:creator>
<dc:creator>Chin, C.-S.</dc:creator>
<dc:creator>Cook, D. E.</dc:creator>
<dc:creator>Craig, S. J. C.</dc:creator>
<dc:creator>de Gennaro, L.</dc:creator>
<dc:creator>Diekhans, M.</dc:creator>
<dc:creator>Dutra, A.</dc:creator>
<dc:creator>Garcia, G. H.</dc:creator>
<dc:creator>Grady, P. G. S.</dc:creator>
<dc:creator>Green, R. E.</dc:creator>
<dc:creator>Haddad, D.</dc:creator>
<dc:creator>Hallast, P.</dc:creator>
<dc:creator>Harvey, W. T.</dc:creator>
<dc:creator>Hickey, G.</dc:creator>
<dc:creator>Hillis, D. A.</dc:creator>
<dc:creator>Hoyt, S. J.</dc:creator>
<dc:creator>Jeong, H.</dc:creator>
<dc:creator>Kamali, K.</dc:creator>
<dc:creator>Pond, S. L. K.</dc:creator>
<dc:creator>LaPolice, T. M.</dc:creator>
<dc:creator>Lee, C.</dc:creator>
<dc:creator>Lewis, A. P.</dc:creator>
<dc:creator>Loh, Y.-H. E.</dc:creator>
<dc:creator>Maste</dc:creator>
<dc:date>2023-12-01</dc:date>
<dc:identifier>doi:10.1101/2023.11.30.569198</dc:identifier>
<dc:title><![CDATA[The Complete Sequence and Comparative Analysis of Ape Sex Chromosomes]]></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/10.1101/2023.12.02.569621v1?rss=1">
<title>
<![CDATA[
The Parkinson's Disease related mutant VPS35 (D620N) amplifies the LRRK2 response to endolysosomal stress 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.12.02.569621v1?rss=1</link>
<description><![CDATA[
The identification of multiple genes linked to Parkinsons Disease invites the question as to how they may cooperate. We have generated isogenic cell lines that inducibly express either wild-type or a mutant form of the retromer component VPS35 (D620N), which has been linked to Parkinsons Disease. This has enabled us to test proposed effects of this mutation in a setting where the relative expression reflects the physiological occurrence. We confirm that this mutation compromises VPS35 association with the WASH complex, but find no defect in WASH recruitment to endosomes, nor in the distribution of lysosomal receptors, cation-independent mannose-6-phosphate receptor and Sortilin. We show VPS35 (D620N) enhances the activity of the Parkinsons associated kinase LRRK2 towards RAB12 under basal conditions. Furthermore, VPS35 (D620N) amplifies the LRRK2 response to endolysosomal stress resulting in enhanced phosphorylation of RABs 10 and 12. By comparing different types of endolysosomal stresses such as the ionophore nigericin and the membranolytic agent LLOMe, we are able to dissociate phospho-RAB accumulation from membrane rupture.
]]></description>
<dc:creator>McCarron, K. R.</dc:creator>
<dc:creator>Elcocks, H.</dc:creator>
<dc:creator>Mortiboys, H. J.</dc:creator>
<dc:creator>Urbe, S.</dc:creator>
<dc:creator>Clague, M. J.</dc:creator>
<dc:date>2023-12-03</dc:date>
<dc:identifier>doi:10.1101/2023.12.02.569621</dc:identifier>
<dc:title><![CDATA[The Parkinson's Disease related mutant VPS35 (D620N) amplifies the LRRK2 response to endolysosomal stress]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-12-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.12.05.570129v1?rss=1">
<title>
<![CDATA[
A non-conducting role of the Cav1.4 Ca2+ channel drives homeostatic plasticity at the cone photoreceptor synapse 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.12.05.570129v1?rss=1</link>
<description><![CDATA[
In congenital stationary night blindness type 2 (CSNB2)--a disorder involving the Cav1.4 (L-type) Ca2+ channel--visual impairment is mild considering that Cav1.4 mediates synaptic release from rod and cone photoreceptors. Here, we addressed this conundrum using a Cav1.4 knockout (KO) mouse and a knock-in (G369i KI) mouse expressing a non-conducting Cav1.4. Surprisingly, Cav3 (T-type) Ca2+ currents were detected in cones of G369i KI mice and Cav1.4 KO mice but not in cones of wild-type mouse, ground squirrel, and macaque retina. Whereas Cav1.4 KO mice are blind, G369i KI mice exhibit normal photopic (i.e., cone-mediated) visual behavior. Cone synapses, which fail to form in Cav1.4 KO mice, are present, albeit enlarged, and with some errors in postsynaptic wiring in G369i KI mice. While Cav1.4 KO mice lack evidence of cone synaptic responses, electrophysiological recordings in G369i KI mice revealed nominal transmission from cones to horizontal cells and bipolar cells. In CSNB2, we propose that Cav3 channels maintain cone synaptic output provided that the nonconducting role of Cav1.4 in cone synaptogenesis remains intact. Our findings reveal an unexpected form of homeostatic plasticity that relies on a non-canonical role of an ion channel.
]]></description>
<dc:creator>Maddox, W.</dc:creator>
<dc:creator>Ordemann, G. J.</dc:creator>
<dc:creator>de la Rosa Vazquez, J.</dc:creator>
<dc:creator>Huang, A.</dc:creator>
<dc:creator>Gault, C.</dc:creator>
<dc:creator>Wisner, S. R.</dc:creator>
<dc:creator>Randall, K.</dc:creator>
<dc:creator>Futagi, D.</dc:creator>
<dc:creator>DeVries, S.</dc:creator>
<dc:creator>Hoon, M.</dc:creator>
<dc:creator>Lee, A.</dc:creator>
<dc:date>2023-12-06</dc:date>
<dc:identifier>doi:10.1101/2023.12.05.570129</dc:identifier>
<dc:title><![CDATA[A non-conducting role of the Cav1.4 Ca2+ channel drives homeostatic plasticity at the cone photoreceptor synapse]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-12-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.12.05.570324v1?rss=1">
<title>
<![CDATA[
Astrocyte-dependent local neurite pruning and Hox gene-mediated cell death in Beat-Va neurons 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.12.05.570324v1?rss=1</link>
<description><![CDATA[
Neuronal remodeling is extensive and mechanistically diverse across the nervous systems of complex metazoans. To explore circuit refinement mechanisms, we screened for new neuronal subtypes in the Drosophila nervous system that undergo remodeling early in metamorphosis. We find Beat-VaM neurons elaborate a highly branched neurite network during larval stages that undergoes local neurite pruning during early metamorphosis. Surprisingly, Beat-VaM neurons remodel their branches despite blockade of steroid hormone signaling and instead depend on signaling from astrocytes to activate pruning. We show Beat-VaL neurons undergo steroid hormone-dependent cell death in posterior but not anterior abdominal segments. Correct activation of apoptotic cell death relies on segment-specific expression of the hox gene Abd-B, which is capable of activating cell death in any Beat-VaL neuron. Our work provides new model cells in which to study neuronal remodeling, highlights an important role for astrocytes in activating local pruning in Drosophila independent of steroid signaling, and defines a Hox gene-mediated mechanism for segment-specific cell elimination.

SummaryLehmann et al. characterize two new populations of neurons that undergo remodeling during Drosophila metamorphosis. Beat-VaM neurons undergo drastic neurite pruning that is largely independent of ecdysone signaling and instead is driven by astrocytes. Beat-VaL neurons undergo Abd-B mediated, caspase driven cell death in a segmentally restricted manner.
]]></description>
<dc:creator>Lehmann, K. S.</dc:creator>
<dc:creator>Hupp, M. T.</dc:creator>
<dc:creator>Jefferson, A.</dc:creator>
<dc:creator>Cheng, Y.-C.</dc:creator>
<dc:creator>Sheehan, A. E.</dc:creator>
<dc:creator>Kang, Y.</dc:creator>
<dc:creator>Freeman, M.</dc:creator>
<dc:date>2023-12-06</dc:date>
<dc:identifier>doi:10.1101/2023.12.05.570324</dc:identifier>
<dc:title><![CDATA[Astrocyte-dependent local neurite pruning and Hox gene-mediated cell death in Beat-Va neurons]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-12-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2023.12.07.570622v1?rss=1">
<title>
<![CDATA[
Three-dimensional reconstruction of fetal rhesus macaque kidneys at single-cell resolution reveals complex inter-relation of structures 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.12.07.570622v1?rss=1</link>
<description><![CDATA[
Kidneys are among the most structurally complex organs in the body. Their architecture is critical to ensure proper function and is often impacted by diseases such as diabetes and hypertension. Understanding the spatial interplay between the different structures of the nephron and renal vasculature is crucial. Recent efforts have demonstrated the value of three-dimensional (3D) imaging in revealing new insights into the various components of the kidney; however, these studies used antibodies or autofluorescence to detect structures and so were limited in their ability to compare the many subtle structures of the kidney at once. Here, through 3D reconstruction of fetal rhesus macaque kidneys at cellular resolution, we demonstrate the power of deep learning in exhaustively labelling seventeen microstructures of the kidney. Using these tissue maps, we interrogate the spatial distribution and spatial correlation of the glomeruli, renal arteries, and the nephron. This work demonstrates the power of deep learning applied to 3D tissue images to improve our ability to compare many microanatomical structures at once, paving the way for further works investigating renal pathologies.
]]></description>
<dc:creator>Dequiedt, L.</dc:creator>
<dc:creator>Forjaz, A.</dc:creator>
<dc:creator>Lo, J. O.</dc:creator>
<dc:creator>McCarty, O. J.</dc:creator>
<dc:creator>Wu, P.-H.</dc:creator>
<dc:creator>Rosenberg, A.</dc:creator>
<dc:creator>Wirtz, D.</dc:creator>
<dc:creator>Kiemen, A. L.</dc:creator>
<dc:date>2023-12-08</dc:date>
<dc:identifier>doi:10.1101/2023.12.07.570622</dc:identifier>
<dc:title><![CDATA[Three-dimensional reconstruction of fetal rhesus macaque kidneys at single-cell resolution reveals complex inter-relation of structures]]></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/10.1101/2023.12.14.571758v1?rss=1">
<title>
<![CDATA[
BMI and BPH correlate with urinary microbiome diversity and lower urinary tract symptoms in men 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.12.14.571758v1?rss=1</link>
<description><![CDATA[
Several studies have identified bacteria and other microbes in the bladder and lower urinary tract in the absence of infection. In women, the urinary microbiome has been associated with lower urinary tract symptoms (LUTS), however, similar studies have not been undertaken in large cohorts of men. Here we examine the urinary microbiome and its association with LUTS in a subset of 500 men aged 65 to 90 years from the Osteoporotic Fractures in Men (MrOS) study. We identified significant associations between benign prostatic hyperplasia (BPH), age, and body mass index (BMI) with several diversity metrics. Our analysis revealed complex relationships between BMI, BPH, LUTS, and alpha diversity which give insight into the intricate dynamics of the urinary microbiome. By beginning to uncover the interrelationships of BPH, BMI, LUTS, and the urinary microbiome, these results can inform future study design to better understand the heterogeneity of the male urinary microbiome.
]]></description>
<dc:creator>Bowie, K.</dc:creator>
<dc:creator>Garzotto, M.</dc:creator>
<dc:creator>Orwoll, E.</dc:creator>
<dc:creator>Karstens, L.</dc:creator>
<dc:date>2023-12-15</dc:date>
<dc:identifier>doi:10.1101/2023.12.14.571758</dc:identifier>
<dc:title><![CDATA[BMI and BPH correlate with urinary microbiome diversity and lower urinary tract symptoms in men]]></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/10.1101/2023.12.31.573735v1?rss=1">
<title>
<![CDATA[
Rapid turnover of CTLA4 is associated with a complex architecture of reversible ubiquitylation. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2023.12.31.573735v1?rss=1</link>
<description><![CDATA[
The immune checkpoint regulator CTLA4 is an unusually short-lived membrane protein. Here we show that its lysosomal degradation is dependent on ubiquitylation at Lysine residues 203 and 213. Inhibition of the v-ATPase partially restores CTLA4 levels following cycloheximide treatment, but also reveals a fraction that is secreted in exosomes. The endosomal deubiquitylase, USP8, interacts with CTLA4 and its loss enhances CTLA4 ubiquitylation in cancer cells, mouse CD4+ T cells and in cancer cell-derived exosomes. Depletion of the USP8 adapter protein, HD-PTP, but not ESCRT-0 recapitulates this cellular phenotype, but shows distinct properties vis-a-vis exosome incorporation. Re-expression of wild-type USP8, but neither a catalytically inactive, nor a localisation-compromised {Delta}MIT domain mutant can rescue delayed degradation of CTLA4, or counteract its accumulation in clustered endosomes. UbiCRest analysis of CTLA4-associated ubiquitin chain linkages identifies a complex mixture of conventional Lys63- and more unusual Lys27- and Lys29-linked polyubiquitin chains that may underly the rapidity of protein turnover.
]]></description>
<dc:creator>Tey, P. Y.</dc:creator>
<dc:creator>Dufner, A.</dc:creator>
<dc:creator>Knobeloch, K.-P.</dc:creator>
<dc:creator>Pruneda, J. N.</dc:creator>
<dc:creator>Clague, M. J.</dc:creator>
<dc:creator>Urbe, S.</dc:creator>
<dc:date>2024-01-01</dc:date>
<dc:identifier>doi:10.1101/2023.12.31.573735</dc:identifier>
<dc:title><![CDATA[Rapid turnover of CTLA4 is associated with a complex architecture of reversible ubiquitylation.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-01-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.01.13.575526v1?rss=1">
<title>
<![CDATA[
Alpha-synuclein regulates nucleolar DNA double-strand break repair in melanoma 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.01.13.575526v1?rss=1</link>
<description><![CDATA[
Although an increased risk of the skin cancer melanoma in people with Parkinsons Disease (PD) has been shown in multiple studies, the mechanisms involved are poorly understood, but increased expression of the PD-associated protein alpha-synuclein (Syn) in melanoma cells may be important. Our previous work suggests that Syn can facilitate DNA double-strand break (DSB) repair, promoting genomic stability. We now show that Syn is preferentially enriched within the nucleolus in the SK-MEL28 melanoma cell line, where it colocalizes with DNA damage markers and DSBs. Inducing DSBs specifically within nucleolar ribosomal DNA (rDNA) increases Syn levels near sites of damage. Syn knockout increases DNA damage within the nucleolus at baseline, after specific rDNA DSB induction, and prolongs the rate of recovery from this induced damage. Syn is important downstream of ATM signaling to facilitate 53BP1 recruitment to DSBs, reducing micronuclei formation and promoting cellular proliferation, migration, and invasion.
]]></description>
<dc:creator>Arnold, M. R.</dc:creator>
<dc:creator>Cohn, G. M.</dc:creator>
<dc:creator>Oxe, K. C.</dc:creator>
<dc:creator>Elliott, S. N.</dc:creator>
<dc:creator>Moore, C.</dc:creator>
<dc:creator>Laraia, P. V.</dc:creator>
<dc:creator>Shekoohi, S.</dc:creator>
<dc:creator>Brownell, D.</dc:creator>
<dc:creator>Meshul, C. K.</dc:creator>
<dc:creator>Witt, S. N.</dc:creator>
<dc:creator>Larsen, D. H.</dc:creator>
<dc:creator>Unni, V. K.</dc:creator>
<dc:date>2024-01-14</dc:date>
<dc:identifier>doi:10.1101/2024.01.13.575526</dc:identifier>
<dc:title><![CDATA[Alpha-synuclein regulates nucleolar DNA double-strand break repair in melanoma]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-01-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.01.25.577207v1?rss=1">
<title>
<![CDATA[
Mutation of the peptide-regulated transcription factor ComR for amidated peptide specificity and heterologous function in Lactiplantibacillus plantarum WCFS1 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.01.25.577207v1?rss=1</link>
<description><![CDATA[
There is a growing interest in the use of probiotic bacteria as biosensors for the detection of disease. However, there is a lack of bacterial receptors developed for specific disease biomarkers. Here, we have investigated the use of the peptide-regulated transcription factor ComR from Streptococcus spp. for specific peptide biomarker detection. ComR exhibits a number of attractive features that are potentially exploitable to create an exquisitely sensitive biomolecular switch for engineered biosensor circuitry within the probiotic organism Lactiplantibacillus plantarum WCFS1. By screening a library of ComR mutant protein variants, we identified mutations that increased the specificity of ComR toward an amidated version of its cognate extracellular signaling peptide, demonstrating the potential for ComR to detect this important class of biomarker.
]]></description>
<dc:creator>Brasino, M.</dc:creator>
<dc:creator>Wagnell, E.</dc:creator>
<dc:creator>Ozdemir, S.</dc:creator>
<dc:creator>Ranganathan, S.</dc:creator>
<dc:creator>Merritt, J.</dc:creator>
<dc:date>2024-01-26</dc:date>
<dc:identifier>doi:10.1101/2024.01.25.577207</dc:identifier>
<dc:title><![CDATA[Mutation of the peptide-regulated transcription factor ComR for amidated peptide specificity and heterologous function in Lactiplantibacillus plantarum WCFS1]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-01-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.02.20.581121v1?rss=1">
<title>
<![CDATA[
Estradiol elicits distinct firing patterns in arcuate nucleus kisspeptin neurons of females through altering ion channel conductances 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.02.20.581121v1?rss=1</link>
<description><![CDATA[
Hypothalamic kisspeptin (Kiss1) neurons are vital for pubertal development and reproduction. Arcuate nucleus Kiss1 (Kiss1ARH) neurons are responsible for the pulsatile release of Gonadotropin-releasing Hormone (GnRH). In females, the behavior of Kiss1ARH neurons, expressing Kiss1, Neurokinin B (NKB), and Dynorphin (Dyn), varies throughout the ovarian cycle. Studies indicate that 17{beta}-estradiol (E2) reduces peptide expression but increases Vglut2 mRNA and glutamate neurotransmission in these neurons, suggesting a shift from peptidergic to glutamatergic signaling. To investigate this shift, we combined transcriptomics, electrophysiology, and mathematical modeling. Our results demonstrate that E2 treatment upregulates the mRNA expression of voltage-activated calcium channels, elevating the whole-cell calcium current and that contribute to high-frequency burst firing. Additionally, E2 treatment decreased the mRNA levels of Canonical Transient Receptor Potential (TPRC) 5 and G protein-coupled K+ (GIRK) channels. When TRPC5 channels in Kiss1ARH neurons were deleted using CRISPR, the slow excitatory postsynaptic potential (sEPSP) was eliminated. Our data enabled us to formulate a biophysically realistic mathematical model of the Kiss1ARH neuron, suggesting that E2 modifies ionic conductances in Kiss1ARH neurons, enabling the transition from high frequency synchronous firing through NKB-driven activation of TRPC5 channels to a short bursting mode facilitating glutamate release. In a low E2 milieu, synchronous firing of Kiss1ARH neurons drives pulsatile release of GnRH, while the transition to burst firing with high, preovulatory levels of E2 would facilitate the GnRH surge through its glutamatergic synaptic connection to preoptic Kiss1 neurons.
]]></description>
<dc:creator>Qiu, J.</dc:creator>
<dc:creator>Voliotis, M.</dc:creator>
<dc:creator>Bosch, M. A.</dc:creator>
<dc:creator>Li, X. F.</dc:creator>
<dc:creator>Zweifel, L. S.</dc:creator>
<dc:creator>Tsaneva-Atanasova, K.</dc:creator>
<dc:creator>O'Byrne, K.</dc:creator>
<dc:creator>Ronnekleiv, O. K.</dc:creator>
<dc:creator>Kelly, M. J.</dc:creator>
<dc:date>2024-02-23</dc:date>
<dc:identifier>doi:10.1101/2024.02.20.581121</dc:identifier>
<dc:title><![CDATA[Estradiol elicits distinct firing patterns in arcuate nucleus kisspeptin neurons of females through altering ion channel conductances]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-02-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.02.21.580762v1?rss=1">
<title>
<![CDATA[
Peptide Amphiphiles Hitchhike on Endogenous Biomolecules for Enhanced Cancer Imaging and Therapy 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.02.21.580762v1?rss=1</link>
<description><![CDATA[
The interactions of nanomaterials with biomolecules in vivo determine their biological fate. Here, we show that self-assembled peptide amphiphile (PA) nanostructures can dynamically interact with endogenous biomolecules and take advantage of naturally occurring processes to target a broad range of solid tumors. In circulation, self-assembled PA nanostructures disassemble and reassemble mainly with lipoproteins, which prolongs blood circulation and dramatically improves tumor accumulation and retention. Mechanistic studies suggested that PAs internalize into cancer cells by assembling with their cell membranes and independently of specific receptors. By exploiting these interactions, a PA developed in this study (namely SA-E) demonstrated specific accumulation in various xenograft, syngeneic, patient-derived xenograft, or transgenic rodent models. In addition, SA-E enabled the effective delivery of highly potent chemotherapy to different syngeneic and xenografted tumors with reduced side effects. With its simple and modular design and universal tumor accumulation mechanism, SA-E represents a promising platform for broad applications in cancer imaging and therapy.
]]></description>
<dc:creator>Fischer, J. M.</dc:creator>
<dc:creator>Stewart, M.</dc:creator>
<dc:creator>Dai, M.</dc:creator>
<dc:creator>Drennan, S.</dc:creator>
<dc:creator>Holland, S.</dc:creator>
<dc:creator>Quentel, A.</dc:creator>
<dc:creator>Sabuncu, S.</dc:creator>
<dc:creator>Kingston, B. R.</dc:creator>
<dc:creator>Dengos, I.</dc:creator>
<dc:creator>Xiang, L.</dc:creator>
<dc:creator>Bonic, K.</dc:creator>
<dc:creator>Goncalves, F.</dc:creator>
<dc:creator>Yi, X.</dc:creator>
<dc:creator>Ranganathan, S.</dc:creator>
<dc:creator>Branchaud, B. P.</dc:creator>
<dc:creator>Muldoon, L. L.</dc:creator>
<dc:creator>Barajas, R. F.</dc:creator>
<dc:creator>Yildirim, A.</dc:creator>
<dc:date>2024-02-23</dc:date>
<dc:identifier>doi:10.1101/2024.02.21.580762</dc:identifier>
<dc:title><![CDATA[Peptide Amphiphiles Hitchhike on Endogenous Biomolecules for Enhanced Cancer Imaging and Therapy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-02-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.02.21.581451v1?rss=1">
<title>
<![CDATA[
Development of novel tools for dissection of central versus peripheral dopamine D2-like receptor signaling in dysglycemia 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.02.21.581451v1?rss=1</link>
<description><![CDATA[
Dopamine (DA) D2-like receptors in both the central nervous system (CNS) and the periphery are key modulators of metabolism. Moreover, disruption of D2-like receptor signaling is implicated in dysglycemia. Yet, the respective metabolic contributions of CNS versus peripheral D2-like receptors including D2 (D2R) and D3 (D3R) receptors remain poorly understood. To address this, we developed new pharmacological tools, D2-like receptor agonists with diminished and delayed blood-brain barrier capability, to selectively manipulate D2R/D3R signaling in the periphery. We designated bromocriptine methiodide (BrMeI), a quaternary methiodide analogue of D2/3R agonist and diabetes drug bromocriptine, as our lead compound based on preservation of D2R/D3R binding and functional efficacy. We then used BrMeI and unmodified bromocriptine to dissect relative contributions of CNS versus peripheral D2R/D3R signaling in treating dysglycemia. Systemic administration of bromocriptine, with unrestricted access to CNS and peripheral targets, significantly improved both insulin sensitivity and glucose tolerance in obese, dysglycemic mice in vivo. In contrast, metabolic improvements were attenuated when access to bromocriptine was restricted either to the CNS through intracerebroventricular administration or delayed access to the CNS via BrMeI. Our findings demonstrate that the coordinated actions of both CNS and peripheral D2-like receptors are required for correcting dysglycemia. Ultimately, the development of a first-generation of drugs designed to selectively target the periphery provides a blueprint for dissecting mechanisms of central versus peripheral DA signaling and paves the way for novel strategies to treat dysglycemia.
]]></description>
<dc:creator>Bonifazi, A.</dc:creator>
<dc:creator>Ellenberger, M.</dc:creator>
<dc:creator>Farino, Z. J.</dc:creator>
<dc:creator>Aslanoglou, D.</dc:creator>
<dc:creator>Rais, R.</dc:creator>
<dc:creator>Pereira, S.</dc:creator>
<dc:creator>Mantilla-Rivas, J. O.</dc:creator>
<dc:creator>Boateng, C. A.</dc:creator>
<dc:creator>Eshleman, A. J.</dc:creator>
<dc:creator>Janowsky, A.</dc:creator>
<dc:creator>Hahn, M. K.</dc:creator>
<dc:creator>Schwartz, G. J.</dc:creator>
<dc:creator>Slusher, B. S.</dc:creator>
<dc:creator>Freyberg, Z.</dc:creator>
<dc:date>2024-02-23</dc:date>
<dc:identifier>doi:10.1101/2024.02.21.581451</dc:identifier>
<dc:title><![CDATA[Development of novel tools for dissection of central versus peripheral dopamine D2-like receptor signaling in dysglycemia]]></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/10.1101/2024.03.03.583210v1?rss=1">
<title>
<![CDATA[
Maternal cannabis use alters excitatory inputs to corticostriatal efferent neurons in rat offspring 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.03.03.583210v1?rss=1</link>
<description><![CDATA[
With the recent surge in cannabis legalization across North America, there is legitimate concern that rates of cannabis use during pregnancy will dramatically increase in the coming years. However, the long-term impacts of prenatal cannabis exposure (PCE) on the brain and behavior remain poorly understood. Using a model of passive cannabis vapor exposure, we have previously shown that PCE impairs behavioral flexibility in an attentional set-shifting task in adult offspring, which is orchestrated in part by excitatory inputs from the medial prefrontal cortex (mPFC) to the nucleus accumbens (NAc). Given the fundamental role of these corticostriatal inputs in coordinating flexible reward-seeking strategies, we used a combination of retrograde tracing and ex vivo electrophysiology to test the hypothesis that maternal cannabis use alters the synaptic and intrinsic membrane properties of corticostriatal efferent neurons in exposed male and female rat offspring. Specifically, pregnant rat dams were trained to self-administer vaporized cannabis (69.7% THC; 150 mg/ml) twice daily throughout mating and gestation and offspring were subsequently injected with fluorescent retrobeads into the NAc core prior to conducting whole-cell ex vivo recordings of spontaneous excitatory and inhibitory post-synaptic currents (EPSC and IPSC, respectively) in retrolabeled mPFC neurons in adulthood. Our results indicate that PCE increases the frequency of spontaneous glutamatergic events (EPSCs) in NAc-projecting mPFC neurons in a sex-specific manner, which drives changes in excitatory to inhibitory (EPSC/IPSC) ratio, particularly in females. Furthermore, the amplitude of phasic glutamatergic events was reduced in cannabis-exposed offspring of both sexes, suggesting changes in postsynaptic receptor function. Altogether, these data demonstrate that PCE shifts the balance of excitatory/inhibitory inputs onto NAc-projecting mPFC neurons with limited effects on membrane conductance in females, resulting in reduced sex differences following maternal cannabis self-administration. These results provide putative neurophysiological mechanisms mediating previously observed behavioral changes, and future studies will need to test if these cannabis-induced changes are causal to long-term deficits in behavioral flexibility that have been previously documented in exposed offspring.
]]></description>
<dc:creator>Ginder, D. E.</dc:creator>
<dc:creator>Weimar, H. V.</dc:creator>
<dc:creator>Lindberg, J. E. M.</dc:creator>
<dc:creator>Fisher, Z. D. G.</dc:creator>
<dc:creator>Lim, M. M.</dc:creator>
<dc:creator>Peters, J. H.</dc:creator>
<dc:creator>McLaughlin, R. J.</dc:creator>
<dc:date>2024-03-06</dc:date>
<dc:identifier>doi:10.1101/2024.03.03.583210</dc:identifier>
<dc:title><![CDATA[Maternal cannabis use alters excitatory inputs to corticostriatal efferent neurons in rat offspring]]></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/10.1101/2024.03.07.583582v1?rss=1">
<title>
<![CDATA[
Deletion of Robo4 worsens neuroinflammation and motor coordination in a mouse model of Alzheimer's disease 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.03.07.583582v1?rss=1</link>
<description><![CDATA[
Declines in vascular integrity are potential contributors to Alzheimers disease (AD) as these result in increased blood-brain barrier permeability and, as a consequence, accelerate neuroinflammation and cognitive impairment. Roundabout guidance receptor 4 (Robo4) is primarily expressed in endothelial cells and stabilizes the vasculature, and thus, has the potential to protect the brain in AD. To study the effect of Robo4 on neuroinflammation and cognitive function in the context of AD, we compared Robo4 knockout and wildtype mice crossed with mice with and without AD mutations (APP/tau). We found that the knockout of Robo4 led to greater astrocyte activation, as demonstrated by GFAP content, but this was dependent on the brain region studied. The knockout of Robo4 also led to greater activated microglia, as assessed by Iba1 content, but only in the presence of AD-related mutations. We found that AD mutations, but not Robo4, were associated with cognitive dysfunction measured by a nest-building test. In contrast, Robo4 deletion, but not AD mutations, was associated with impaired motor coordination. Lastly, Robo4 deletion was associated with greater arterial stiffness, but this trend did not reach statistical significance. In summary, these results demonstrate that Robo4 impacts neuroinflammation, motor coordination, and arterial stiffness, however, the impact on neuroinflammation is dependent on the presence/absence of AD-related mutations and the brain region examined.
]]></description>
<dc:creator>Cullen, A. E.</dc:creator>
<dc:creator>Winder, N.</dc:creator>
<dc:creator>Lee, B.</dc:creator>
<dc:creator>Krishna Kumaran, S.</dc:creator>
<dc:creator>Arora, N.</dc:creator>
<dc:creator>Wolf, J.</dc:creator>
<dc:creator>Woltjer, R.</dc:creator>
<dc:creator>Walker, A.</dc:creator>
<dc:date>2024-03-12</dc:date>
<dc:identifier>doi:10.1101/2024.03.07.583582</dc:identifier>
<dc:title><![CDATA[Deletion of Robo4 worsens neuroinflammation and motor coordination in a mouse model of Alzheimer's disease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-03-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.03.07.583989v1?rss=1">
<title>
<![CDATA[
Fatiguing Exercise Reduces Cellular Passive Young's Modulus in Human Vastus Lateralis Muscle 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.03.07.583989v1?rss=1</link>
<description><![CDATA[
Previous studies demonstrated that acute, exercise-induced fatigue transiently reduces whole-muscle stiffness. Because reduced muscle stiffness at fatigue may contribute to increased injury risk and impaired contractile performance, the present study seeks to elucidate potential intracellular mechanisms underlying these reductions. To that end, cellular passive Youngs Modulus was measured in single, permeabilized muscle fibers from healthy, recreationally-active males and females. Eight volunteers (4 male, 4 female) completed unilateral, repeated maximal voluntary knee extensions until fatigue, after which percutaneous needle biopsies were performed on the fatigued (F) and non-fatigued (NF) Vastus Lateralis muscles. Muscle samples were processed for mechanical assessment and separately for imaging and phosphoproteomics. Single fibers were passively (pCa 8.0), incrementally stretched to 156% of the initial sarcomere length to assess Youngs Modulus, calculated as the slope of the resulting stress-strain curve at short (strain = 1.00-1.24 %Lo) and long (strain = 1.32-1.56 %Lo) fiber lengths. Titin phosphorylation was assessed by liquid chromatography followed by high-resolution mass spectrometry (LC-MS). Passive modulus was significantly reduced by fatigue at short and long lengths in male, but not female, participants. Fatigue increased phosphorylation of four serine residues located within the elastic region of titin and reduced phosphorylation at one serine residue but did not impact active tension nor sarcomere ultrastructure. Collectively, these results suggest muscle fatigue reduces cellular passive modulus in young males, but not females, concurrent with altered titin phosphorylation. These results provide mechanistic insight contributing to the understanding of sex-based differences in soft tissue injury and falls risk.

Key Points SummaryO_LIPrevious studies have shown that skeletal muscle stiffness is reduced following a single bout of fatiguing exercise.
C_LIO_LILower muscle stiffness at fatigue may increase risk for soft-tissue injury, however, the underlying mechanisms of this change are unclear.
C_LIO_LIOur findings show that fatiguing exercise reduces passive Youngs modulus in skeletal muscle cells from males but not females, suggesting that intracellular proteins contribute to reduced muscle stiffness with fatigue in a sex-dependent manner.
C_LIO_LIThe phosphorylation status of the intracellular protein titin is modified by fatiguing exercise in a way that may contribute to altered muscle stiffness after fatiguing exercise.
C_LIO_LIThese results provide important mechanistic insight that may help explain why biological sex impacts risk for soft tissue injury in with repeated or high intensity mechanical loading in athletes and falls risk in older adults.
C_LI

New and NoteworthyMuscle fatigue has previously been shown to reduce musculotendinous stiffness, but the underlying mechanisms remain unclear. Our study presents novel evidence of fatigue-induced reductions in passive cellular Youngs Modulus in skeletal muscle from males, but not females, in conjunction with fatigue-induced alterations in titin phosphorylation. Collectively, these results suggest that intracellular mechanisms including titin phosphorylation may contribute to altered skeletal muscle stiffness following fatiguing exercise, and that this response is mediated by biological sex.
]]></description>
<dc:creator>Privett, G. E.</dc:creator>
<dc:creator>Ricci, A. W.</dc:creator>
<dc:creator>David, L.</dc:creator>
<dc:creator>Needham, K. W.</dc:creator>
<dc:creator>Tan, Y. H.</dc:creator>
<dc:creator>Nakayama, K. H.</dc:creator>
<dc:creator>Callahan, D. M.</dc:creator>
<dc:date>2024-03-12</dc:date>
<dc:identifier>doi:10.1101/2024.03.07.583989</dc:identifier>
<dc:title><![CDATA[Fatiguing Exercise Reduces Cellular Passive Young's Modulus in Human Vastus Lateralis Muscle]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-03-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.03.09.584228v1?rss=1">
<title>
<![CDATA[
Structure-based probe reveals the presence of large transthyretin aggregates in plasma of ATTR amyloidosis patients 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.03.09.584228v1?rss=1</link>
<description><![CDATA[
ATTR amyloidosis is a relentlessly progressive disease caused by the misfolding and systemic accumulation of amyloidogenic transthyretin into amyloid fibrils. These fibrils cause diverse clinical phenotypes, mainly cardiomyopathy and/or polyneuropathy. Little is known about the aggregation of transthyretin during disease development and whether this has implications for diagnosis and treatment. Using the cryogenic electron microscopy structures of mature ATTR fibrils, we developed a peptide probe for fibril detection. With this probe, we have identified previously unknown aggregated transthyretin species in plasma of patients with ATTR amyloidosis. These species are large, non-native, and distinct from monomeric and tetrameric transthyretin. Observations from our study open many questions about the biology of ATTR amyloidosis and reveals a potential diagnostic and therapeutic target.
]]></description>
<dc:creator>Pedretti, R.</dc:creator>
<dc:creator>Wang, L.</dc:creator>
<dc:creator>Yakubovska, A.</dc:creator>
<dc:creator>Zhang, Q. S.</dc:creator>
<dc:creator>Nguyen, B.</dc:creator>
<dc:creator>Grodin, J. L.</dc:creator>
<dc:creator>Masri, A.</dc:creator>
<dc:creator>Saelices, L.</dc:creator>
<dc:date>2024-03-10</dc:date>
<dc:identifier>doi:10.1101/2024.03.09.584228</dc:identifier>
<dc:title><![CDATA[Structure-based probe reveals the presence of large transthyretin aggregates in plasma of ATTR amyloidosis patients]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-03-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.03.12.584558v1?rss=1">
<title>
<![CDATA[
Multi-modal comparison of molecular programs driving nurse cell death and clearance in Drosophila melanogaster oogenesis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.03.12.584558v1?rss=1</link>
<description><![CDATA[
The death and clearance of nurse cells is a consequential milestone in Drosophila melanogaster oogenesis. In preparation for oviposition, the germline-derived nurse cells bequeath to the developing oocyte all their cytoplasmic contents and undergo programmed cell death. The death of the nurse cells is controlled non-autonomously and is precipitated by epithelial follicle cells of somatic origin acquiring a squamous morphology and acidifying the nurse cells externally. Alternatively, stressors such as starvation can induce the death of nurse cells earlier in mid-oogenesis, manifesting apoptosis signatures, followed by their engulfment by epithelial follicle cells. To identify and contrast the molecular pathways underlying these morphologically and genetically distinct cell death paradigms, both mediated by follicle cells, we compared their genome-wide transcriptional, translational, and secretion profiles before and after differentiating to acquire a phagocytic capability, as well as during well-fed and nutrient-deprived conditions. By coupling the GAL4-UAS system to Translating Ribosome Affinity Purification (TRAP-seq) and proximity labeling (HRP-KDEL) followed by Liquid Chromatography tandem mass-spectrometry, we performed high-throughput screens to identify pathways selectively activated or repressed by follicle cells to employ nurse cell-clearance routines contextually and preferentially. We also integrated two publicly available single-cell RNAseq atlases of the Drosophila ovary to define the transcriptomic profiles of follicle cells. In this report, we describe the genes and major pathways identified in the screens and the striking consequences to Drosophila melanogaster oogenesis caused by RNAi perturbation of prioritized candidates. To the best of our knowledge, our study is the first of its kind to comprehensively characterize two distinct apoptotic and non-apoptotic cell death paradigms in the same multi-cellular system. Beyond molecular differences in cell death, our investigation may also provide insights into how key systemic trade-offs are made between survival and reproduction when faced with physiological stress.
]]></description>
<dc:creator>Bandyadka, S.</dc:creator>
<dc:creator>Lebo, D. P.</dc:creator>
<dc:creator>Mondragon, A.</dc:creator>
<dc:creator>Serizier, S. B.</dc:creator>
<dc:creator>Kwan, J.</dc:creator>
<dc:creator>Peterson, J. S.</dc:creator>
<dc:creator>Chasse, A. Y.</dc:creator>
<dc:creator>Jenkins, V.</dc:creator>
<dc:creator>Calikyan, A.</dc:creator>
<dc:creator>Ortega, A.</dc:creator>
<dc:creator>Campbell, J. D.</dc:creator>
<dc:creator>Emili, A.</dc:creator>
<dc:creator>McCall, K.</dc:creator>
<dc:date>2024-03-14</dc:date>
<dc:identifier>doi:10.1101/2024.03.12.584558</dc:identifier>
<dc:title><![CDATA[Multi-modal comparison of molecular programs driving nurse cell death and clearance in Drosophila melanogaster oogenesis]]></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/10.1101/2024.03.15.584711v1?rss=1">
<title>
<![CDATA[
Synchrony between midbrain gene transcription and dopamine terminal regulation is modulated by chronic alcohol drinking 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.03.15.584711v1?rss=1</link>
<description><![CDATA[
Alcohol use disorder is marked by disrupted behavioral and emotional states which persist into abstinence. The enduring synaptic alterations that remain despite the absence of alcohol are of interest for interventions to prevent relapse. Here, 28 male rhesus macaques underwent over 20 months of alcohol drinking interspersed with three 30-day forced abstinence periods. After the last abstinence period, we paired direct sub-second dopamine monitoring via ex vivo voltammetry in nucleus accumbens slices with RNA-sequencing of the ventral tegmental area. We found persistent augmentation of dopamine transporter function, kappa opioid receptor sensitivity, and dynorphin release - all inhibitory regulators which act to decrease extracellular dopamine. Surprisingly, though transcript expression was not altered, the relationship between gene expression and functional readouts of these encoded proteins was highly dynamic and altered by drinking history. These results outline the long-lasting synaptic impact of alcohol use and suggest that assessment of transcript-function relationships is critical for the rational design of precision therapeutics.
]]></description>
<dc:creator>Farahbakhsh, Z. Z.</dc:creator>
<dc:creator>Holleran, K. M.</dc:creator>
<dc:creator>Sens, J. P.</dc:creator>
<dc:creator>Fordahl, S. C.</dc:creator>
<dc:creator>Mauterer, M. I.</dc:creator>
<dc:creator>Lopez, A. J.</dc:creator>
<dc:creator>Cuzon Carlson, V. C.</dc:creator>
<dc:creator>Kiraly, D. D.</dc:creator>
<dc:creator>Grant, K. A.</dc:creator>
<dc:creator>Jones, S. R.</dc:creator>
<dc:creator>Siciliano, C. A.</dc:creator>
<dc:date>2024-03-17</dc:date>
<dc:identifier>doi:10.1101/2024.03.15.584711</dc:identifier>
<dc:title><![CDATA[Synchrony between midbrain gene transcription and dopamine terminal regulation is modulated by chronic alcohol drinking]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-03-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.03.18.585621v1?rss=1">
<title>
<![CDATA[
Structural diversity and clustering of bacterial flagellar outer domains 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.03.18.585621v1?rss=1</link>
<description><![CDATA[
Supercoiled flagellar filaments function as mechanical propellers within the bacterial flagellum complex, playing a crucial role in motility. Flagellin, the building block of the filament, features a conserved inner D0/D1 core domain across different bacterial species. In contrast, approximately half of the flagellins possess additional, highly divergent outer domain(s), suggesting varied functional potential. In this study, we elucidate atomic structures of flagellar filaments from three distinct bacterial species: Cupriavidus gilardii, Stenotrophomonas maltophilia, and Geovibrio thiophilus. Our findings reveal that the flagella from the facultative anaerobic G. thiophilus possesses a significantly more negatively charged surface, potentially enabling adhesion to positively charged minerals. Furthermore, we analyzed all AlphaFold predicted structures for annotated bacterial flagellins, categorizing the flagellin outer domains into 682 structural clusters. This classification provides insights into the prevalence and experimental verification of these outer domains. Remarkably, two of the flagellar structures reported herein belong to a previously unexplored cluster, indicating new opportunities on the study of the functional diversity of flagellar outer domains. Our findings underscore the complexity of bacterial flagellins and open up possibilities for future studies into their varied roles beyond motility.
]]></description>
<dc:creator>Fields, J. L.</dc:creator>
<dc:creator>Zhang, H.</dc:creator>
<dc:creator>Bellis, N. F.</dc:creator>
<dc:creator>Petersen, H. A.</dc:creator>
<dc:creator>Halder, S. K.</dc:creator>
<dc:creator>Rich-New, S. T.</dc:creator>
<dc:creator>Wu, H.</dc:creator>
<dc:creator>Wang, F.</dc:creator>
<dc:date>2024-03-18</dc:date>
<dc:identifier>doi:10.1101/2024.03.18.585621</dc:identifier>
<dc:title><![CDATA[Structural diversity and clustering of bacterial flagellar outer domains]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-03-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.03.21.585773v1?rss=1">
<title>
<![CDATA[
Trace Amines are Essential Metabolites for the Autocrine Regulation of β-Cell Signaling and Insulin Secretion 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.03.21.585773v1?rss=1</link>
<description><![CDATA[
Secretion of insulin in response to extracellular stimuli, such as elevated glucose levels and small molecules that act on G-protein coupled receptors (GPCRs), is the hallmark of {beta}-cell physiology. Trace amines (TAs) are small aromatic metabolites that were identified as low-abundant ligands of the trace amine-associated receptor 1 (TAAR1) in the central nervous system (CNS), a GPCR that is also expressed by pancreatic {beta}-cells. In the present work, we identify TAs as essential autocrine signaling factors for {beta}-cell activity and insulin secretion. We find that {beta}-cells are producing TAs in significant amounts and that the modulation of endogenous TA levels by the selective inhibition of TA biosynthetic pathways directly translated into changes of oscillations of the intracellular Ca2+ concentration ([Ca2+]i oscillations) and insulin secretion. Selective TAAR1 agonists or inhibitors of monoamine oxidases increased [Ca2+]i oscillations and insulin secretion. Opposite effects were mediated by selective TAAR1 antagonists, by recombinant monoamine oxidase action and by the inhibition of amino acid decarboxylase. As the modulation of TA biochemical pathways immediately translated into changes of [Ca2+]i oscillations, we inferred high metabolic turnover rates of TAs and autocrine feedback. We found that psychotropic drugs modulate [Ca2+]i oscillations and insulin secretion, either directly acting on TAAR1 or by altering endogenous TA levels. Our combined data support the hypothesis of TAs as essential autocrine signaling factors for {beta}-cell activity and insulin secretion as well as TAAR1 as an important mediator of amine-modulated insulin secretion.
]]></description>
<dc:creator>Keutler, K.</dc:creator>
<dc:creator>Hauke, S.</dc:creator>
<dc:creator>Laguerre, A.</dc:creator>
<dc:creator>Andreu-Carbo, M.</dc:creator>
<dc:creator>Rada, J.</dc:creator>
<dc:creator>Grandy, D.</dc:creator>
<dc:creator>Yushchenko, D. A.</dc:creator>
<dc:creator>Schultz, C.</dc:creator>
<dc:date>2024-03-23</dc:date>
<dc:identifier>doi:10.1101/2024.03.21.585773</dc:identifier>
<dc:title><![CDATA[Trace Amines are Essential Metabolites for the Autocrine Regulation of β-Cell Signaling and Insulin Secretion]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-03-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.03.21.586146v1?rss=1">
<title>
<![CDATA[
Cannabinoid Receptor Signaling is Dependent on Sub-Cellular Location 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.03.21.586146v1?rss=1</link>
<description><![CDATA[
G protein-coupled receptors (GPCRs) are membrane bound signaling molecules that regulate many aspects of human physiology. Recent advances have demonstrated that GPCR signaling can occur both at the cell surface and internal cellular membranes. Our findings suggest that cannabinoid receptor 1 (CB1) signaling is highly dependent on its subcellular location. We find that intracellular CB1 receptors predominantly couple to Gi while plasma membrane receptors couple to Gs. Here we show subcellular location of CB1, and its signaling, is contingent on the choice of promoters and receptor tags. Heterologous expression with a strong promoter or N-terminal tag resulted in CB1 predominantly localizing to the plasma membrane and signaling through Gs. Conversely, CB1 driven by low expressing promoters and lacking N-terminal genetic tags largely localized to internal membranes and signals via Gi. Lastly, we demonstrate that genetically encodable non-canonical amino acids (ncAA) offer a solution to the problem of non-native N-terminal tags disrupting CB1 signaling. We identified sites in CB1R and CB2R which can be tagged with fluorophores without disrupting CB signaling or trafficking using (trans-cyclooctene attached to lysine (TCO*A)) and copper-free click chemistry to attach fluorophores in live cells. Together, our data demonstrate the origin of location bias in cannabinoid signaling which can be experimentally controlled and tracked in living cells through promoters and novel CBR tagging strategies.
]]></description>
<dc:creator>Thomas, A.</dc:creator>
<dc:creator>Lobingier, B.</dc:creator>
<dc:creator>Schultz, C.</dc:creator>
<dc:creator>Laguerre, A.</dc:creator>
<dc:date>2024-03-22</dc:date>
<dc:identifier>doi:10.1101/2024.03.21.586146</dc:identifier>
<dc:title><![CDATA[Cannabinoid Receptor Signaling is Dependent on Sub-Cellular Location]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-03-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.03.22.586341v1?rss=1">
<title>
<![CDATA[
Discovery of a cell-active chikungunya virus nsP2 protease inhibitor using a covalent fragment-based screening approach 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.03.22.586341v1?rss=1</link>
<description><![CDATA[
Chikungunya virus (CHIKV) is a mosquito-borne alphavirus that has been responsible for numerous large-scale outbreaks in the last twenty years. Currently, there are no FDA-approved therapeutics for any alphavirus infection. CHIKV non-structural protein 2 (nsP2), which contains a cysteine protease domain, is essential for viral replication, making it an attractive target for a drug discovery campaign. Here, we optimized a CHIKV nsP2 protease (nsP2pro) biochemical assay for the screening of a 6,120-compound cysteine-directed covalent fragment library. Using a 50% inhibition threshold, we identified 153 hits (2.5% hit rate). In dose-response follow up, RA-0002034, a covalent fragment that contains a vinyl sulfone warhead, inhibited CHIKV nsP2pro with an IC50 of 58 {+/-} 17 nM, and further analysis with time-dependent inhibition studies yielded a kinact/KI of 6.4 x 103 M-1s-1. LC-MS/MS analysis determined that RA-0002034 covalently modified the catalytic cysteine in a site-specific manner. Additionally, RA-0002034 showed no significant off-target reactivity against a panel of cysteine proteases. In addition to the potent biochemical inhibition of CHIKV nsP2pro activity and exceptional selectivity, RA-0002034 was tested in cellular models of alphavirus infection and effectively inhibited viral replication of both CHIKV and related alphaviruses. This study highlights the discovery and characterization of the chemical probe RA-0002034 as a promising hit compound from covalent fragment-based screening for development toward a CHIKV or pan-alphavirus therapeutic.

Significance StatementChikungunya virus is one of the most prominent and widespread alphaviruses and has caused explosive outbreaks of arthritic disease. Currently, there are no FDA-approved drugs to treat disease caused by chikungunya virus or any other alphavirus-caused infection. Here, we report the discovery of a covalent small molecule inhibitor of chikungunya virus nsP2 protease activity and viral replication of four diverse alphaviruses. This finding highlights the utility of covalent fragment screening for inhibitor discovery and represents a starting point towards the development of alphavirus therapeutics targeting nsP2 protease.
]]></description>
<dc:creator>Merten, E. M.</dc:creator>
<dc:creator>Sears, J. D.</dc:creator>
<dc:creator>Leisner, T. M.</dc:creator>
<dc:creator>Hardy, P. B.</dc:creator>
<dc:creator>Ghoshal, A.</dc:creator>
<dc:creator>Hossain, M. A.</dc:creator>
<dc:creator>Asressu, K. H.</dc:creator>
<dc:creator>Brown, P. J.</dc:creator>
<dc:creator>Stashko, M. A.</dc:creator>
<dc:creator>Herring, L. E.</dc:creator>
<dc:creator>Mordant, A. L.</dc:creator>
<dc:creator>Webb, T. S.</dc:creator>
<dc:creator>Mills, C. A.</dc:creator>
<dc:creator>Barker, N. K.</dc:creator>
<dc:creator>Arnold, J. J.</dc:creator>
<dc:creator>Cameron, C. E.</dc:creator>
<dc:creator>Streblow, D. N.</dc:creator>
<dc:creator>Moorman, N. J.</dc:creator>
<dc:creator>Heise, M.</dc:creator>
<dc:creator>Willson, T. M.</dc:creator>
<dc:creator>Popov, K. I.</dc:creator>
<dc:creator>Pearce, K. H.</dc:creator>
<dc:date>2024-03-22</dc:date>
<dc:identifier>doi:10.1101/2024.03.22.586341</dc:identifier>
<dc:title><![CDATA[Discovery of a cell-active chikungunya virus nsP2 protease inhibitor using a covalent fragment-based screening approach]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-03-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.03.22.586355v1?rss=1">
<title>
<![CDATA[
Dynamic fibrillar assembly of αB-crystallin induced by perturbation of the conserved NT-IXI motif resolved by cryo-EM. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.03.22.586355v1?rss=1</link>
<description><![CDATA[
B-crystallin is an archetypical member of the small heat-shock proteins (sHSPs) vital for cellular proteostasis and mitigating protein misfolding diseases. Gaining insights into the principles defining their molecular organization and chaperone function have been hindered by intrinsic dynamic properties and limited high-resolution structural analysis. To disentangle the mechanistic underpinnings of these dynamical properties, we mutated a conserved IXI-motif located within the N-terminal (NT) domain of human B-crystallin. This resulted in a profound structural transformation, from highly polydispersed caged-like native assemblies into a comparatively well-ordered helical fibril state amenable to high-resolution cryo-EM analysis. The reversible nature of the induced fibrils facilitated interrogation of functional effects due to perturbation of the NT-IXI motif in both the native-like oligomer and fibril states. Together, our investigations unveiled several features thought to be key mechanistic attributes to sHSPs and point to a critical significance of the NT-IXI motif in B-crystallin assembly, dynamics and chaperone activity.
]]></description>
<dc:creator>Reichow, S. L.</dc:creator>
<dc:creator>McFarland, R.</dc:creator>
<dc:date>2024-03-27</dc:date>
<dc:identifier>doi:10.1101/2024.03.22.586355</dc:identifier>
<dc:title><![CDATA[Dynamic fibrillar assembly of αB-crystallin induced by perturbation of the conserved NT-IXI motif resolved by cryo-EM.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-03-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.03.27.587069v1?rss=1">
<title>
<![CDATA[
Prevalent co-release of glutamate and GABA throughout the mouse brain 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.03.27.587069v1?rss=1</link>
<description><![CDATA[
Several neuronal populations in the brain transmit both the excitatory and inhibitory neurotransmitters, glutamate, and GABA, to downstream neurons. However, it remains largely unknown whether these opposing neurotransmitters are co-released onto the same postsynaptic neuron simultaneously or are independently transmitted at different time and locations (called co-transmission). Here, using whole-cell patch-clamp recording on acute mouse brain slices, we observed biphasic miniature postsynaptic currents, i.e., minis with time-locked excitatory and inhibitory currents, in striatal spiny projection neurons (SPNs). This observation cannot be explained by accidental coincidence of monophasic miniature excitatory and inhibitory postsynaptic currents (mEPSCs and mIPSCs, respectively), arguing for the co-release of glutamate and GABA. Interestingly, these biphasic minis could either be an mEPSC leading an mIPSC or vice versa. Although dopaminergic axons release both glutamate and GABA in the striatum, deletion of dopamine neurons did not eliminate biphasic minis, indicating that the co-release originates from another neuronal type. Importantly, we found that both types of biphasic minis were detected in other neuronal subtypes in the striatum as well as in nine out of ten additionally tested brain regions. Our results suggest that co-release of glutamate and GABA is a prevalent mode of neurotransmission in the brain.
]]></description>
<dc:creator>Ceballos, C. C.</dc:creator>
<dc:creator>Ma, L.</dc:creator>
<dc:creator>Qin, M.</dc:creator>
<dc:creator>Zhong, H.</dc:creator>
<dc:date>2024-03-29</dc:date>
<dc:identifier>doi:10.1101/2024.03.27.587069</dc:identifier>
<dc:title><![CDATA[Prevalent co-release of glutamate and GABA throughout the mouse brain]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-03-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.03.28.587263v1?rss=1">
<title>
<![CDATA[
Tools for Cre-mediated conditional deletion of floxed alleles from developing cerebellar Purkinje cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.03.28.587263v1?rss=1</link>
<description><![CDATA[
The Cre-lox system is an indispensable tool in neuroscience research for targeting gene deletions to specific cellular populations. Here we assess the utility of several transgenic Cre lines, along with a viral approach, for targeting cerebellar Purkinje cells. Using a combination of a fluorescent reporter line (Ai14) to indicate Cre-mediated recombination and a floxed Dystroglycan line (Dag1flox) we show that reporter expression does not always align precisely with loss of protein. The commonly used Pcp2Cre line exhibits a gradual mosaic pattern of Cre recombination in Purkinje cells from P7-P14, while loss of Dag1 protein is not complete until P30. Ptf1aCre drives recombination in precursor cells that give rise to GABAergic neurons in the embryonic cerebellum, including Purkinje cells and molecular layer interneurons. However, due to its transient expression in precursors, Ptf1aCre results in stochastic loss of Dag1 protein in these neurons. NestinCre, which is often described as a "pan-neuronal" Cre line for the central nervous system, does not drive Cre-mediated recombination in Purkinje cells. We identify a Calb1Cre line that drives efficient and complete recombination in embryonic Purkinje cells, resulting in loss of Dag1 protein before the period of synaptogenesis. AAV8-mediated delivery of Cre at P0 results in gradual transduction of Purkinje cells during the second postnatal week, with loss of Dag1 protein not reaching appreciable levels until P35. These results characterize several tools for targeting conditional deletions in cerebellar Purkinje cells at different developmental stages and illustrate the importance of validating the loss of protein following recombination.

Significance StatementThe development of Cre lines for targeting gene deletions to defined cellular populations has led to important discoveries in neuroscience. As with any tool, there are inherent limitations that must be carefully considered. Here we describe several Cre lines available for targeting of cerebellar Purkinje cells at various developmental stages. We use the combination of a Cre-dependent fluorescent reporter line and conditional deletion of the synaptic scaffolding molecule Dystroglycan as an example to highlight the potential disconnect between the presence of a fluorescent reporter and the loss of protein.
]]></description>
<dc:creator>Jahncke, J. N.</dc:creator>
<dc:creator>Wright, K. M.</dc:creator>
<dc:date>2024-03-30</dc:date>
<dc:identifier>doi:10.1101/2024.03.28.587263</dc:identifier>
<dc:title><![CDATA[Tools for Cre-mediated conditional deletion of floxed alleles from developing cerebellar Purkinje cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-03-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.04.01.587595v1?rss=1">
<title>
<![CDATA[
Cryo-electron tomographic investigation of native hippocampal glutamatergic synapses 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.04.01.587595v1?rss=1</link>
<description><![CDATA[
Chemical synapses are the major sites of communication between neurons in the nervous system and mediate either excitatory or inhibitory signaling [1]. At excitatory synapses, glutamate is the primary neurotransmitter and upon release from presynaptic vesicles, is detected by postsynaptic glutamate receptors, which include ionotropic AMPA and NMDA receptors [2, 3]. Here we have developed methods to identify glutamatergic synapses in brain tissue slices, label AMPA receptors with small gold nanoparticles (AuNPs), and prepare lamella for cryo-electron tomography studies. The targeted imaging of glutamatergic synapses in the lamella is facilitated by fluorescent pre- and postsynaptic signatures, and the subsequent tomograms allow for identification of key features of chemical synapses, including synaptic vesicles, the synaptic cleft and AuNP-labeled AMPA receptors. These methods pave the way for imaging brain regions at high resolution, using unstained, unfixed samples preserved under near-native conditions.
]]></description>
<dc:creator>Matsui, A.</dc:creator>
<dc:creator>Spangler, C.</dc:creator>
<dc:creator>Elferich, J.</dc:creator>
<dc:creator>Shiozaki, M.</dc:creator>
<dc:creator>Jean, N.</dc:creator>
<dc:creator>Zhao, X.</dc:creator>
<dc:creator>Qin, M.</dc:creator>
<dc:creator>Zhong, H.</dc:creator>
<dc:creator>Yu, Z.</dc:creator>
<dc:creator>Gouaux, E.</dc:creator>
<dc:date>2024-04-01</dc:date>
<dc:identifier>doi:10.1101/2024.04.01.587595</dc:identifier>
<dc:title><![CDATA[Cryo-electron tomographic investigation of native hippocampal glutamatergic synapses]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-04-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.04.01.587649v1?rss=1">
<title>
<![CDATA[
Biologically informed machine learning modeling of immune cells to reveal physiological and pathological aging process 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.04.01.587649v1?rss=1</link>
<description><![CDATA[
The immune system undergoes progressive functional remodeling from neonatal stages to old age. Therefore, understanding how aging shapes immune cell function is vital for precise treatment of patients at different life stages. Here, we constructed the first transcriptomic atlas of immune cells encompassing human lifespan, ranging from newborns to supercentenarians, and comprehensively examined gene expression signatures involving cell signaling, metabolism, differentiation, and functions in all cell types to investigate immune aging changes. By comparing immune cell composition among different age groups, HLA highly expressing NK cells and CD83 positive B cells were identified with high percentages exclusively in the teenager (Tg) group, whereas CD4_CTL precursors were exclusively enriched in the supercentenarian (Sc) group. Notably, we found that the biological age (BA) of pediatric COVID-19 patients with multisystem inflammatory syndrome accelerated aging according to their chronological age (CA). Besides, we proved that inflammatory shift-myeloid abundance and signature correlate with the progression of complications in Kawasaki disease (KD). Finally, based on those age-related immune cell compositions, we developed a novel BA prediction model, PHARE (https://xiazlab.org/phare/), which applies to both scRNA-seq and bulk RNA-seq data. Overall, our study revealed changes in immune cell proportions and function associated with aging, both in health and disease, and provided a novel tool for successfully capturing features that accelerate or delay aging.
]]></description>
<dc:creator>Xia, Z.</dc:creator>
<dc:creator>Zhang, B.</dc:creator>
<dc:creator>Sun, L.</dc:creator>
<dc:creator>Zhang, C.</dc:creator>
<dc:creator>Ren, T.</dc:creator>
<dc:creator>Zhao, X.</dc:creator>
<dc:creator>Su, Y.</dc:creator>
<dc:creator>Wang, Q.</dc:creator>
<dc:creator>Zhang, T.</dc:creator>
<dc:creator>He, B.</dc:creator>
<dc:creator>Wu, L.-Y.</dc:creator>
<dc:date>2024-04-02</dc:date>
<dc:identifier>doi:10.1101/2024.04.01.587649</dc:identifier>
<dc:title><![CDATA[Biologically informed machine learning modeling of immune cells to reveal physiological and pathological aging process]]></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/10.1101/2024.04.02.587799v1?rss=1">
<title>
<![CDATA[
Development of BODIPY-based fluorescent probes for highly selective amino acid identification 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.04.02.587799v1?rss=1</link>
<description><![CDATA[
Amino acid sensing is a powerful tool for intra- and extra-cellular assays. We report herein the generation and optimization of bioconjugatable fluorescent reporter for pan-amino acid sensing, the OMNI probe, that through inductive electronics produces a unique photophysical response upon coupling to the -amino group of an amino acid depending on the identity of the adjacent side chain. Using an auto-photocatalyzed Meerwein arylation reaction, we created a library of bis-arylated 8-methylthio-BODIPY dye derivatives exhibiting improved spectral variation, brightness, dynamic range, robustness, water solubility, and reactivity. Aryl groups with optimal photophysical behaviors were combined to create asymmetric versions conferring high spectral range, brightness, and water solubility. Combining aryl group modifications yielded a set of engineered asymmetric probes that achieved complete spectral resolution in a multi-variable analysis (fluorescent emission, lifetime, brightness) for all twenty native amino acid-dye conjugates while increasing reactivity toward amino acids in aqueous buffer. With our robust OMNI probe design, pan-amino acid resolution and identification through high sensitivity fluorescence down to single molecules is now routinely possible for diverse potential applications including protein sequencing.
]]></description>
<dc:creator>Hendrick, N.</dc:creator>
<dc:creator>Martin, A.</dc:creator>
<dc:creator>Chan, A.</dc:creator>
<dc:creator>Lenihan, J.</dc:creator>
<dc:creator>Reiter, H.</dc:creator>
<dc:creator>Clough, M.</dc:creator>
<dc:creator>Jiang, L.</dc:creator>
<dc:creator>Whaley, E.</dc:creator>
<dc:creator>Jeong Huh, Y.</dc:creator>
<dc:creator>McNeely, J.</dc:creator>
<dc:creator>Chen, A.</dc:creator>
<dc:creator>Emili, A.</dc:creator>
<dc:creator>Beeler, A. B.</dc:creator>
<dc:date>2024-04-02</dc:date>
<dc:identifier>doi:10.1101/2024.04.02.587799</dc:identifier>
<dc:title><![CDATA[Development of BODIPY-based fluorescent probes for highly selective amino acid identification]]></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/10.1101/2024.04.04.588147v1?rss=1">
<title>
<![CDATA[
Detailed phenotyping of Tbr1-2A-CreER knock-in mice demonstrates significant impacts on TBR1 protein levels and axon development 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.04.04.588147v1?rss=1</link>
<description><![CDATA[
Spatiotemporal control of Cre-mediated recombination has been an invaluable tool for understanding key developmental processes. For example, knock-in of Cre into cell type marker gene loci drives Cre expression under endogenous promoter and enhancer sequences, greatly facilitating the study of diverse neuronal subtypes in the cerebral cortex. However, insertion of exogenous DNA into the genome can have unintended effects on local gene regulation or protein function that must be carefully considered. Here, we analyze a recently generated Tbr1-2A-CreER knock-in mouse line, where a 2A-CreER cassette was inserted in-frame just before the stop codon of the transcription factor gene Tbr1. Heterozygous TBR1 mutations in humans and mice are known to cause autism or autism-like behavioral phenotypes accompanied by structural brain malformations, most frequently a reduction of the anterior commissure. Thus, it is critical for modified versions of Tbr1 to exhibit true wild-type-like activity. We evaluated the Tbr1-2A-CreER allele for its potential impact on Tbr1 function and complementation to Tbr1 loss-of-function alleles. In mice with one copy of the Tbr1-2A-CreER allele, we identified reduction of TBR1 protein in early postnatal cortex along with thinning of the anterior commissure, suggesting hypersensitivity of this structure to TBR1 dosage. Comparing Tbr1-2A-CreER and Tbr1-null heterozygous and homozygous mice to Tbr1-null complementation crosses showed reductions of TBR1 dosage ranging from 28.4% to 95.9%. Using these combinatorial genotypes, we found that low levels of TBR1 protein ([~]16%) are sufficient to establish cortical layer positioning, while greater levels (>50%) are required for normal suppression of layer 5 identity. In total, these results strongly support the conclusion that Tbr1-2A-CreER is a hypomorphic allele. We advise caution when interpreting experiments using this allele, such as transcriptomic studies, considering the sensitivity of various corticogenic processes to TBR1 dosage and the association of heterozygous TBR1 mutations with complex neurodevelopmental disorders.
]]></description>
<dc:creator>Co, M.</dc:creator>
<dc:creator>O'Brien, G. K.</dc:creator>
<dc:creator>Wright, K. M.</dc:creator>
<dc:creator>O'Roak, B. J.</dc:creator>
<dc:date>2024-04-04</dc:date>
<dc:identifier>doi:10.1101/2024.04.04.588147</dc:identifier>
<dc:title><![CDATA[Detailed phenotyping of Tbr1-2A-CreER knock-in mice demonstrates significant impacts on TBR1 protein levels and axon development]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-04-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.04.11.589120v1?rss=1">
<title>
<![CDATA[
Chronic alcohol consumption alters sex-dependent BNST neuron function in rhesus macaques 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.04.11.589120v1?rss=1</link>
<description><![CDATA[
Repeated alcohol drinking contributes to a number of neuropsychiatric diseases, including alcohol use disorder and co-expressed anxiety and mood disorders. Women are more susceptible to the development and expression of these diseases with the same history of alcohol exposure as men, suggesting they may be more sensitive to alcohol-induced plasticity in limbic brain regions controlling alcohol drinking, stress responsivity, and reward processing, among other behaviors. Using a translational model of alcohol drinking in rhesus monkeys, we examined sex differences in the basal function and plasticity of neurons in the bed nucleus of the stria terminalis (BNST), a brain region in the extended amygdala shown to be a hub circuit node dysregulated in individuals with anxiety and alcohol use disorder. We performed slice electrophysiology recordings from BNST neurons in male and female monkeys following daily "open access" (22 hr/day) to 4% ethanol and water for more than one year or control conditions. We found that BNST neurons from control females had reduced overall current density, hyperpolarization-activated depolarizing current (Ih), and inward rectification, as well as higher membrane resistance and greater synaptic glutamatergic release and excitatory drive, than those from control males, suggesting that female BNST neurons are more basally excited than those from males. Chronic alcohol drinking produced a shift in these measures in both sexes, decreasing current density, Ih, and inward rectification and increasing synaptic excitation. In addition, network activity-dependent synaptic inhibition was basally higher in BNST neurons of males than females, and alcohol exposure increased this in both sexes, a putative homeostatic mechanism to counter hyperexcitability. Altogether, these results suggest that the rhesus BNST is more basally excited in females than males and chronic alcohol drinking produces an overall increase in excitability and synaptic excitation. These results shed light on the mechanisms contributing to the female-biased susceptibility to neuropsychiatric diseases including co-expressed anxiety and alcohol use disorder.
]]></description>
<dc:creator>Pleil, K. E.</dc:creator>
<dc:creator>Grant, K. A.</dc:creator>
<dc:creator>Carlson, V. C. C.</dc:creator>
<dc:creator>Kash, T. L.</dc:creator>
<dc:date>2024-04-15</dc:date>
<dc:identifier>doi:10.1101/2024.04.11.589120</dc:identifier>
<dc:title><![CDATA[Chronic alcohol consumption alters sex-dependent BNST neuron function in rhesus macaques]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-04-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.04.15.589654v1?rss=1">
<title>
<![CDATA[
Relaxin Modulates the Genomic Actions and Biological Effects of Estrogen in the Myometrium by Reducing Estrogen Receptor Alpha Phosphorylation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.04.15.589654v1?rss=1</link>
<description><![CDATA[
Estradiol (E2) and relaxin (Rln) are steroid and polypeptide hormones, respectively, with important roles in the female reproductive tract, including myometrium. Some actions of Rln, which are mediated by its membrane receptor RXFP1, require or are augmented by E2 signaling through its cognate nuclear steroid receptor, estrogen receptor alpha (ER). In contrast, other actions of Rln act in opposition to the effects of E2. Here we explored the molecular and genomic mechanisms that underlie the functional interplay between E2 and Rln in the myometrium. We used both ovariectomized female mice and immortalized human myometrial cells expressing wild-type or mutant ER (hTERT-HM-ER cells). Our results indicate that Rln modulates the genomic actions and biological effects of estrogen in the myometrium and myometrial cells by reducing phosphorylation of ER on serine 118 (S118), as well as by reducing the E2-dependent binding of ER across the genome. These effects were associated with changes in the hormone-regulated transcriptome, including a decrease in the E2-dependent expression of some genes and enhanced expression of others. The inhibitory effects of Rln cotreatment on the E2-dependent phosphorylation of ER required the nuclear dual-specificity phosphatases DUSP1 and DUSP5. Moreover, the inhibitory effects of Rln were reflected in a concomitant inhibition of the E2-dependent contraction of myometrial cells. Collectively, our results identify a pathway that integrates Rln/RXFP1 and E2/ER signaling, resulting in a convergence of membrane and nuclear signaling pathways to control genomic and biological outcomes.
]]></description>
<dc:creator>Sudeshna Tripathy, S.</dc:creator>
<dc:creator>Nagari, A.</dc:creator>
<dc:creator>Chiu, S.-P.</dc:creator>
<dc:creator>Nandu, T.</dc:creator>
<dc:creator>Camacho, C. V.</dc:creator>
<dc:creator>Mahendroo, M.</dc:creator>
<dc:creator>Kraus, W. L.</dc:creator>
<dc:date>2024-04-16</dc:date>
<dc:identifier>doi:10.1101/2024.04.15.589654</dc:identifier>
<dc:title><![CDATA[Relaxin Modulates the Genomic Actions and Biological Effects of Estrogen in the Myometrium by Reducing Estrogen Receptor Alpha Phosphorylation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-04-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.04.16.589814v1?rss=1">
<title>
<![CDATA[
Massively parallel jumping assay decodes Alu retrotransposition activity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.04.16.589814v1?rss=1</link>
<description><![CDATA[
The human genome contains millions of retrotransposons, several of which could become active due to somatic mutations having phenotypic consequences, including disease. However, it is not thoroughly understood how nucleotide changes in retrotransposons affect their jumping activity. Here, we developed a novel massively parallel jumping assay (MPJA) that can test the jumping potential of thousands of transposons en masse. We generated nucleotide variant library of selected four Alu retrotransposons containing 165,087 different haplotypes and tested them for their jumping ability using MPJA. We found 66,821 unique jumping haplotypes, allowing us to pinpoint domains and variants vital for transposition. Mapping these variants to the Alu-RNA secondary structure revealed stem-loop features that contribute to jumping potential. Combined, our work provides a novel high-throughput assay that assesses the ability of retrotransposons to jump and identifies nucleotide changes that have the potential to reactivate them in the human genome.
]]></description>
<dc:creator>Ahituv, N.</dc:creator>
<dc:creator>Matharu, N.</dc:creator>
<dc:creator>Zhao, J.</dc:creator>
<dc:creator>Sohota, A.</dc:creator>
<dc:creator>Deng, L.</dc:creator>
<dc:creator>Hung, Y.</dc:creator>
<dc:creator>Li, Z.</dc:creator>
<dc:creator>Sims, J.</dc:creator>
<dc:creator>Rattanasopha, S.</dc:creator>
<dc:creator>Meyer, J.</dc:creator>
<dc:creator>Carbone, L.</dc:creator>
<dc:creator>Kircher, M.</dc:creator>
<dc:date>2024-04-19</dc:date>
<dc:identifier>doi:10.1101/2024.04.16.589814</dc:identifier>
<dc:title><![CDATA[Massively parallel jumping assay decodes Alu retrotransposition activity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-04-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.04.17.589812v1?rss=1">
<title>
<![CDATA[
A Futile Cycle? Tissue Homeostatic Trans-Membrane Water Co-Transport: Kinetics, Thermodynamics, Metabolic Consequences 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.04.17.589812v1?rss=1</link>
<description><![CDATA[
The phenomenon of active trans-membrane water cycling (AWC) has emerged in little over a decade. Here, we consider H2O transport across cell membranes from the origins of its study. Historically, trans-membrane water transport processes were classified into: A) compensating bidirectional fluxes ("exchange"), and B) unidirectional flux ("net flow") categories. Recent literature molecular structure determinations and molecular dynamic (MD) simulations indicate probably all the many different hydrophilic substrate membrane co-transporters have membrane-spanning hydrophilic pathways and co-transport water along with their substrates, and that they individually catalyze category A and/or B water flux processes, although usually not simultaneously. The AWC name signifies that, integrated over the all the cells co-transporters, the rate of homeostatic, bidirectional trans-cytolemmal water exchange (category A) is synchronized with the metabolic rate of the crucial Na+,K+-ATPase (NKA) enzyme. A literature survey indicates the stoichiometric (category B) water/substrate ratios of individual co-transporters are often very large. The MD simulations also suggest how different co-transporter reactions can be kinetically coupled molecularly.

Is this (Na+,K+-ATPase rate-synchronized) cycling futile, or is it consequential? Conservatively representative literature metabolomic and proteinomic results enable comprehensive free energy analyses of the many transport reactions with known water stoichiometries. Free energy calculations, using literature intracellular pressure (Pi) values reveals there is an outward trans-membrane H2O barochemical gradient of magnitude comparable to that of the well-known inward Na+ electrochemical gradient. For most co-influxers, these gradients are finely balanced to maintain intracellular metabolite concentration values near their consuming enzyme Michaelis constants. The thermodynamic analyses include glucose, glutamate-, gamma-aminobutyric acid (GABA), and lactate- transporters. 2%-4% Pi alterations can lead to disastrous concentration levels. For the neurotransmitters glutamate- and GABA, very small astrocytic Pi changes can allow/disallow synaptic transmission. Unlike the Na+ and K+ electrochemical steady-states, the H2O barochemical steady-state is in (or near) chemical equilibrium. The analyses show why the presence of aquaporins (AQPs) does not dissipate the trans-membrane pressure gradient. A feedback loop inherent in the opposing Na+ electrochemical and H2O barochemical gradients regulates AQP-catalyzed water flux as an integral AWC aspect. These results also require a re-consideration of the underlying nature of Pi. Active trans-membrane water cycling is not futile, but is inherent to the cells "NKA system" - a new, fundamental aspect of biology.

SYNOPSISVia intracellular pressure, membrane co-transported water influences thermodynamic control of cell metabolite maintenance.
]]></description>
<dc:creator>Springer, C. S.</dc:creator>
<dc:creator>Pike, M. M.</dc:creator>
<dc:creator>Barbara, T. M.</dc:creator>
<dc:date>2024-04-19</dc:date>
<dc:identifier>doi:10.1101/2024.04.17.589812</dc:identifier>
<dc:title><![CDATA[A Futile Cycle? Tissue Homeostatic Trans-Membrane Water Co-Transport: Kinetics, Thermodynamics, Metabolic Consequences]]></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/10.1101/2024.04.19.590252v1?rss=1">
<title>
<![CDATA[
Proteomic Discovery of RNA-Protein Molecular Clamps Using a Thermal Shift Assay with ATP and RNA (TSAR) 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.04.19.590252v1?rss=1</link>
<description><![CDATA[
Uncompetitive inhibition is an effective strategy for suppressing dysregulated enzymes and their substrates, but discovery of suitable ligands depends on often-unavailable structural knowledge and serendipity. Hence, despite surging interest in mass spectrometry-based target identification, proteomic studies of substrate-dependent target engagement remain sparse. Herein, we describe a strategy for the discovery of substrate-dependent ligand binding. Using proteome integral solubility alteration (PISA) assays, we show that simple biochemical additives can enable detection of RNA-protein-small molecule complexes in native cell lysates. We apply our approach to rocaglates, molecules that specifically clamp RNA to eukaryotic translation initiation factor 4A (eIF4A), DEAD-box helicase 3X (DDX3X), and potentially other members of the DEAD-box (DDX) helicase family. To identify unexpected interactions, we used a target class-specific thermal window and compared ATP analog and RNA base dependencies for key rocaglate-DDX interactions. We report and validate novel DDX targets of high-profile rocaglates - including the clinical candidate Zotatifin - using limited proteolysis-mass spectrometry and fluorescence polarization (FP) experiments. We also provide structural insight into divergent DDX3X affinities between synthetic rocaglates. Taken together, our study provides a model for screening uncompetitive inhibitors using a chemical proteomics approach and uncovers actionable DDX clamping targets, clearing a path towards characterization of novel molecular clamps and associated RNA helicases.
]]></description>
<dc:creator>Goldstein, S. I.</dc:creator>
<dc:creator>Fan, A. C.</dc:creator>
<dc:creator>Wang, Z.</dc:creator>
<dc:creator>Naineni, S. K.</dc:creator>
<dc:creator>Lengqvist, J.</dc:creator>
<dc:creator>Chernobrovkin, A.</dc:creator>
<dc:creator>Garcia-Gutierrez, S. B.</dc:creator>
<dc:creator>Cencic, R.</dc:creator>
<dc:creator>Patel, K.</dc:creator>
<dc:creator>Huang, S.</dc:creator>
<dc:creator>Brown, L. E.</dc:creator>
<dc:creator>Emili, A.</dc:creator>
<dc:creator>Porco, J. A.</dc:creator>
<dc:date>2024-04-19</dc:date>
<dc:identifier>doi:10.1101/2024.04.19.590252</dc:identifier>
<dc:title><![CDATA[Proteomic Discovery of RNA-Protein Molecular Clamps Using a Thermal Shift Assay with ATP and RNA (TSAR)]]></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/10.1101/2024.04.25.591167v1?rss=1">
<title>
<![CDATA[
Divergent Subregional Information Processing in Mouse Prefrontal Cortex During Working Memory 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.04.25.591167v1?rss=1</link>
<description><![CDATA[
Working memory (WM) is a critical cognitive function allowing recent information to be temporarily held in mind to inform future action. This process depends on coordination between key subregions in prefrontal cortex (PFC) and other connected brain areas. However, few studies have examined the degree of functional specialization between these subregions throughout the phases of WM using electrophysiological recordings in freely-moving animals, particularly mice. To this end, we recorded single-units in three neighboring medial PFC (mPFC) subregions in mouse - supplementary motor area (MOs), dorsomedial PFC (dmPFC), and ventromedial (vmPFC) - during a freely-behaving non-match-to-position WM task. We found divergent patterns of task-related activity across the phases of WM. The MOs is most active around task phase transitions and encodes the starting sample location most selectively. Dorsomedial PFC contains a more stable population code, including persistent sample-location-specific firing during a five second delay period. Finally, the vmPFC responds most strongly to reward-related information during the choice phase. Our results reveal anatomically and temporally segregated computation of WM task information in mPFC and motivate more precise consideration of the dynamic neural activity required for WM.
]]></description>
<dc:creator>Sonneborn, A.</dc:creator>
<dc:creator>Bartlett, L.</dc:creator>
<dc:creator>Olson, R. J.</dc:creator>
<dc:creator>Milton, R.</dc:creator>
<dc:creator>Abbas, A. I.</dc:creator>
<dc:date>2024-04-28</dc:date>
<dc:identifier>doi:10.1101/2024.04.25.591167</dc:identifier>
<dc:title><![CDATA[Divergent Subregional Information Processing in Mouse Prefrontal Cortex During Working Memory]]></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/10.1101/2024.04.26.591186v1?rss=1">
<title>
<![CDATA[
PARP14 is pro- and anti-viral host factor that promotes IFN production and affects the replication of multiple viruses 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.04.26.591186v1?rss=1</link>
<description><![CDATA[
PARP14 is a 203 kDa multi-domain protein that is primarily known as an ADP-ribosyltransferase, and is involved in a variety of cellular functions including DNA damage, microglial activation, inflammation, and cancer progression. In addition, PARP14 is upregulated by interferon (IFN), indicating a role in the antiviral response. Furthermore, PARP14 has evolved under positive selection, again indicating that it is involved in host-pathogen conflict. We found that PARP14 is required for increased IFN-I production in response to coronavirus infection lacking ADP-ribosylhydrolase (ARH) activity and poly(I:C), however, whether it has direct antiviral function remains unclear. Here we demonstrate that the catalytic activity of PARP14 enhances IFN-{beta} and IFN-{lambda} responses and restricts ARH-deficient murine hepatitis virus (MHV) and severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) replication. To determine if PARP14s antiviral functions extended beyond CoVs, we tested the ability of herpes simplex virus 1 (HSV-1), a DNA virus, vesicular stomatitis virus (VSV), a negative-sense RNA virus, and lymphocytic choriomeningitis virus (LCMV), an ambisense RNA virus, to infect A549 PARP14 knockout (KO) cells. While LCMV infection was unaffected, HSV-1 replication was increased in PARP14 KO cells and VSV replication was decreased. These results indicate that PARP14 restricts HSV-1 replication but enhances the replication of VSV. A PARP14 active site inhibitor had no impact on HSV-1 or VSV replication, indicating that its effect on these viruses was independent of its catalytic activity. These data demonstrate that PARP14 promotes IFN production and has both proviral and antiviral functions targeting multiple viruses.

IMPORTANCEThe antiviral response is largely regulated by post-translation modifications (PTM), including ADP-ribosylation. PARP14 is an ADP-ribosyltransferase that is upregulated by interferon and is under positive selection, indicating that it is involved in host-pathogen conflict. However, no anti-viral function has been described for PARP14. Here, we found that PARP14 represses both coronavirus and HSV-1 replication, demonstrating that PARP14 has antiviral functions. Surprisingly, we also found that PARP14 also has pro-viral functions, as it was critical for the efficient replication of VSV. These data indicate that PARP14 has both proviral and antiviral functions. Defining the mechanisms used by PARP14 to both repress and promote virus replication will provide new insights into how PARPs regulate virus infection.
]]></description>
<dc:creator>Parthasarathy, S.</dc:creator>
<dc:creator>Saenjamsai, P.</dc:creator>
<dc:creator>Hao, H.</dc:creator>
<dc:creator>Ferkul, A.</dc:creator>
<dc:creator>Pfannenstiel, J. J.</dc:creator>
<dc:creator>Suder, E. L.</dc:creator>
<dc:creator>Bejan, D. S.</dc:creator>
<dc:creator>Chen, Y.</dc:creator>
<dc:creator>Schwarting, N.</dc:creator>
<dc:creator>Aikawa, M.</dc:creator>
<dc:creator>Muhlberger, E.</dc:creator>
<dc:creator>Orozco, R. C.</dc:creator>
<dc:creator>Sullivan, C. S.</dc:creator>
<dc:creator>Cohen, M. S.</dc:creator>
<dc:creator>Davido, D.</dc:creator>
<dc:creator>Hume, A. J.</dc:creator>
<dc:creator>Fehr, A.</dc:creator>
<dc:date>2024-04-26</dc:date>
<dc:identifier>doi:10.1101/2024.04.26.591186</dc:identifier>
<dc:title><![CDATA[PARP14 is pro- and anti-viral host factor that promotes IFN production and affects the replication of multiple viruses]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-04-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.04.26.591387v1?rss=1">
<title>
<![CDATA[
IL-12/IL23 blockade reveals patterns of asynchronous inflammation in pyoderma gangrenosum 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.04.26.591387v1?rss=1</link>
<description><![CDATA[
Pyoderma gangrenosum (PG) is a rare neutrophilic dermatosis causing chronic and recalcitrant painful ulcerations. Pathogenic mechanisms are yet poorly understood limiting therapeutic options, however, IL-12/IL-23 inhibition via ustekinumab has previously been associated with positive outcomes. We aimed to elucidate the dysregulated immune landscape of PG and lesional skin changes associated with IL-12/IL-23 blockade. We applied spatial transcriptomics and comparative computation analysis on lesional biopsies from two patients obtained before and after IL-12/IL-23 blockade with ustekinumab. Our data indicate lesional PG skin exhibits complex patterns of inflammation, including a not previously described major infiltration of B cells and establishment of tertiary lymphoid structures. In both patients, IL-12/IL-23 blockade led to marked clinical improvement but was associated with amelioration of contrasting inflammatory pathways. Notably, plasma cell markers and tertiary structures were recalcitrant to the treatment regime suggesting that B cells might play a role in the refractory nature of PG.
]]></description>
<dc:creator>Yadav, R.</dc:creator>
<dc:creator>Vague, M.</dc:creator>
<dc:creator>Rettig, M.</dc:creator>
<dc:creator>Loo, C. P.</dc:creator>
<dc:creator>Brown, K.</dc:creator>
<dc:creator>Samiea, A.</dc:creator>
<dc:creator>Moreau, J. M.</dc:creator>
<dc:creator>Ortega-Loayza, A. G.</dc:creator>
<dc:date>2024-04-29</dc:date>
<dc:identifier>doi:10.1101/2024.04.26.591387</dc:identifier>
<dc:title><![CDATA[IL-12/IL23 blockade reveals patterns of asynchronous inflammation in pyoderma gangrenosum]]></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/10.1101/2024.04.27.591485v1?rss=1">
<title>
<![CDATA[
Investigating the Triple Code Model in Numerical Cognition Using Stereotactic Electroencephalography 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.04.27.591485v1?rss=1</link>
<description><![CDATA[
The ability to conceptualize numerical quantities is an essential human trait. According to the "Triple Code Model" (TCM) in numerical cognition, distinct neural substrates encode the processing of visual, auditory, and nonsymbolic numerical representations. While our contemporary understanding of human number cognition has benefited greatly from advances in clinical imaging, limited studies have investigated the intracranial electrophysiological correlates of number processing. In this study, 13 subjects undergoing stereotactic electroencephalography for epilepsy participated in a number recognition task. Drawing upon postulates of the TCM, we presented subjects with numerical stimuli varying in representation type (symbolic vs. non-symbolic) and mode of stimuli delivery (visual vs. auditory). Time-frequency spectrograms were dimensionally reduced with principal component analysis and passed into a linear support vector machine classification algorithm to identify regions associated with number perception compared to inter-trial periods. Across representation formats, the highest classification accuracy was observed in the bilateral parietal lobes. Auditory (spoken and beeps) and visual (Arabic) number formats preferentially engaged the superior temporal cortices and the frontoparietal regions, respectively. The left parietal cortex was found to have the highest classification for number dots. Notably, the putamen exhibited robust classification accuracies in response to numerical stimuli. Analyses of spectral feature maps revealed that non-gamma frequency below 30 Hz held greater than chance classification value and could be potentially used to characterize format specific number representations. Taken together, our findings obtained from intracranial recordings provide further support and expand on the TCM model for numerical cognition.
]]></description>
<dc:creator>Tan, H.</dc:creator>
<dc:creator>Rockhill, A. P.</dc:creator>
<dc:creator>Lopez Ramos, C. G.</dc:creator>
<dc:creator>Nerison, C.</dc:creator>
<dc:creator>Shafie, B.</dc:creator>
<dc:creator>Shahin, M. N.</dc:creator>
<dc:creator>Fecker, A. L.</dc:creator>
<dc:creator>Ismail, M.</dc:creator>
<dc:creator>Cleary, D. R.</dc:creator>
<dc:creator>Collins, K. L.</dc:creator>
<dc:creator>Raslan, A. M.</dc:creator>
<dc:date>2024-04-29</dc:date>
<dc:identifier>doi:10.1101/2024.04.27.591485</dc:identifier>
<dc:title><![CDATA[Investigating the Triple Code Model in Numerical Cognition Using Stereotactic Electroencephalography]]></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/10.1101/2024.05.02.592167v1?rss=1">
<title>
<![CDATA[
Elucidating the Impact of Hypericum alpestre Extract and L-NAME on the PI3K/Akt Signaling Pathway in A549 Lung Adenocarcinoma Cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.05.02.592167v1?rss=1</link>
<description><![CDATA[
Plants within the Hypericaceae family have been traditionally used for their medicinal properties, showcasing a wide range of effects such as antibacterial, antiviral, and antioxidant qualities. Hypericum alpestre (HA) extracts have exhibited significant cytotoxicity against various cancer cell lines. The phenolic compounds found in HA extracts have attracted attention for their potential in cancer prevention. L-NAME, known for its ability to inhibit nitric oxide synthase (NOS) activity, has emerged as a promising approach in cancer therapy. However, the precise molecular mechanisms underlying the anticancer effects of HA and L-NAME remain unclear. This study aims to clarify the impact of HA and L-NAME on the PI3K/Akt signaling pathway in A549 lung adenocarcinoma cells, with a specific focus on TNFa/COX-2 and VEGFa/MMP-2 pathways. In silico analysis, they identified the compounds with the highest affinity for PI3K/Akt, a finding validated by subsequent in vitro experiments. Furthermore, the combination of herbs and L-NAME exhibited superior efficacy compared to the herb and 5-FU combination, as evidenced by the promotion of apoptosis. Both the herb alone and the combination of the herb with L-NAME demonstrated inhibitory effects on the TNFa/COX-2 and VEGFa/MMP-2 pathways. This therapeutic approach is hypothesized to operate through the PI3k/Akt cell signaling pathway. A better understanding of the interaction between HA polyphenols and PI3K/Akt signaling could pave the way for novel therapeutic strategies against cancer, including drug-resistant tumors.
]]></description>
<dc:creator>Javrushyan, H.</dc:creator>
<dc:creator>Ginovyan, M.</dc:creator>
<dc:creator>Petrosyan, G.</dc:creator>
<dc:creator>Qocharyan, M.</dc:creator>
<dc:creator>Harutyunyan, T.</dc:creator>
<dc:creator>Gevorgyan, S.</dc:creator>
<dc:creator>Karabekian, Z.</dc:creator>
<dc:creator>Maloyan, A.</dc:creator>
<dc:creator>Avtandilyan, N.</dc:creator>
<dc:date>2024-05-04</dc:date>
<dc:identifier>doi:10.1101/2024.05.02.592167</dc:identifier>
<dc:title><![CDATA[Elucidating the Impact of Hypericum alpestre Extract and L-NAME on the PI3K/Akt Signaling Pathway in A549 Lung Adenocarcinoma Cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-05-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.05.02.592287v1?rss=1">
<title>
<![CDATA[
A novel genetic mouse model of fatal aortic dissection reveals massive inflammatory cell infiltration in the thoracic aorta 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.05.02.592287v1?rss=1</link>
<description><![CDATA[
BackgroundAortic dissection (AD) is the separation of medial layers of the aorta and is a major cause of death in patients with connective tissue disorders such as Marfan syndrome. However, molecular triggers instigating AD, its temporospatial progression, and how vascular cells in each vessel layer interact and participate in the pathological process remain incompletely understood. To unravel the underlying molecular mechanism of AD, we generated a spontaneous AD mouse model.

MethodsWe incorporated a novel missense variant (p.G234D) in FBN1, the gene for fibrillin-1, identified in a non-syndromic familial AD patient into mice using CRISPR/Cas9 system. We performed histopathological analyses of the aortic lesions by histology, immunofluorescence staining, electron microscopy, synchrotron-based imaging and single-cell (sc)RNA-sequencing. Biochemical analysis was performed to examine the binding capacity of mutant human FBN1G234D protein to latent Tgf{beta} binding proteins (LTBPs), and signaling pathways in the mutant aortic wall were examined by western blot analysis.

Results50% of the Fbn1G234D/G234D mutant mice died within 5 weeks of age from multiple intimomedial tears that expanded longitudinally and progressed to aortic rupture accompanied by massive immune cell infiltration. scRNA-sequencing, validated by immunostaining, revealed a significant increase in MHC class II-positive pro-inflammatory macrophages and monocytes at the site of intima tears with upregulation of MMP2/9 and marked disruption of elastic lamina. Subendothelial matrices, such as type IV collagen and laminin, expanded into the medial layer, where fibronectin expression was highly upregulated. Fbn1G234D/G234D endothelial cells exhibited altered mechanosensing with loss of parallel alignment to blood flow and upregulation of VCAM-1 and ICAM-1, all of which likely contributed to the infiltration of immune cells. Biochemically, FBN1G234D lost the ability to bind to latent TGF{beta} binding protein (LTBP)-1, -2, and -4, resulting in the downregulation of TGF{beta} signaling in the aortic wall.

ConclusionsWe show that dynamic interactions involving endothelial cells (ECs) and macrophages/monocytes in the intima, where the ECM microenvironment is altered with the reduced TGF{beta} signaling, contributes to the initiation of AD. Our novel AD mouse model provides a unique opportunity to identify target molecules involved in the intimomedial tears that can be utilized for development of therapeutic strategies.
]]></description>
<dc:creator>Kimura, K.</dc:creator>
<dc:creator>Motoyama, E.</dc:creator>
<dc:creator>Kanki, S.</dc:creator>
<dc:creator>Asano, K.</dc:creator>
<dc:creator>Sheikh, M. A. A.</dc:creator>
<dc:creator>Clarin, M. T. R. D. C.</dc:creator>
<dc:creator>Raja, E.</dc:creator>
<dc:creator>Takeda, M.</dc:creator>
<dc:creator>Ishii, R.</dc:creator>
<dc:creator>Murata, K.</dc:creator>
<dc:creator>Deleeuw, V.</dc:creator>
<dc:creator>Sips, P.</dc:creator>
<dc:creator>Muino-Mosquera, L.</dc:creator>
<dc:creator>De Backer, J.</dc:creator>
<dc:creator>Mizuno, S.</dc:creator>
<dc:creator>Sakai, L. Y.</dc:creator>
<dc:creator>Nakamura, T.</dc:creator>
<dc:creator>Yanagisawa, H.</dc:creator>
<dc:date>2024-05-07</dc:date>
<dc:identifier>doi:10.1101/2024.05.02.592287</dc:identifier>
<dc:title><![CDATA[A novel genetic mouse model of fatal aortic dissection reveals massive inflammatory cell infiltration in the thoracic aorta]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-05-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.05.05.592573v1?rss=1">
<title>
<![CDATA[
COEXIST: Coordinated single-cell integration of serial multiplexed tissue images 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.05.05.592573v1?rss=1</link>
<description><![CDATA[
Multiplexed tissue imaging (MTI) and other spatial profiling technologies commonly utilize serial tissue sectioning to comprehensively profile samples by imaging each section with unique biomarker panels or assays. The dependence on serial sections is attributed to technological limitations of MTI panel size or incompatible multi-assay protocols. Although image registration can align serially sectioned MTIs, integration at the single-cell level poses a challenge due to inherent biological heterogeneity. Existing computational methods overlook both cell population heterogeneity across modalities and spatial information, which are critical for effectively completing this task. To address this problem, we first use Monte-Carlo simulations to estimate the overlap between serial 5m-thick sections. We then introduce COEXIST, a novel algorithm that synergistically combines shared molecular profiles with spatial information to seamlessly integrate serial sections at the single-cell level. We demonstrate COEXIST necessity and performance across several applications. These include combining MTI panels for improved spatial single-cell profiling, rectification of miscalled cell phenotypes using a single MTI panel, and the comparison of MTI platforms at single-cell resolution. COEXIST not only elevates MTI platform validation but also overcomes the constraints of MTIs panel size and the limitation of full nuclei on a single slide, capturing more intact nuclei in consecutive sections and thus enabling deeper profiling of cell lineages and functional states.
]]></description>
<dc:creator>Heussner, R. T.</dc:creator>
<dc:creator>Watson, C. F.</dc:creator>
<dc:creator>Eddy, C. Z.</dc:creator>
<dc:creator>Wang, K.</dc:creator>
<dc:creator>Cramer, E. M.</dc:creator>
<dc:creator>Creason, A. L.</dc:creator>
<dc:creator>Mills, G. B.</dc:creator>
<dc:creator>Chang, Y. H.</dc:creator>
<dc:date>2024-05-07</dc:date>
<dc:identifier>doi:10.1101/2024.05.05.592573</dc:identifier>
<dc:title><![CDATA[COEXIST: Coordinated single-cell integration of serial multiplexed tissue images]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-05-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.05.08.593181v1?rss=1">
<title>
<![CDATA[
Androgen receptor activity inversely correlates with immune cell infiltration and immunotherapy response across multiple cancer lineages 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.05.08.593181v1?rss=1</link>
<description><![CDATA[
There is now increasing recognition of the important role of androgen receptor (AR) in modulating immune function. To gain a comprehensive understanding of the effects of AR activity on cancer immunity, we employed a computational approach to profile AR activity in 33 human tumor types using RNA-Seq datasets from The Cancer Genome Atlas. Our pan-cancer analysis revealed that the genes most negatively correlated with AR activity across cancers are involved in active immune system processes. Importantly, we observed a significant negative correlation between AR activity and IFN{gamma} pathway activity at the pan-cancer level. Indeed, using a matched biopsy dataset from subjects with prostate cancer before and after AR-targeted treatment, we verified that inhibiting AR enriches immune cell abundances and is associated with higher IFN{gamma} pathway activity. Furthermore, by analyzing immunotherapy datasets in multiple cancers, our results demonstrate that low AR activity was significantly associated with a favorable response to immunotherapy. Together, our data provide a comprehensive assessment of the relationship between AR signaling and tumor immunity.
]]></description>
<dc:creator>Hu, Y.-M.</dc:creator>
<dc:creator>Zhao, F.</dc:creator>
<dc:creator>Graff, J.</dc:creator>
<dc:creator>Chen, C.</dc:creator>
<dc:creator>Zhao, X.</dc:creator>
<dc:creator>Thomas, G. V.</dc:creator>
<dc:creator>Wu, H.</dc:creator>
<dc:creator>Kardosh, A.</dc:creator>
<dc:creator>Mills, G.</dc:creator>
<dc:creator>Alumkal, J.</dc:creator>
<dc:creator>Moran, A. E.</dc:creator>
<dc:creator>Xia, Z.</dc:creator>
<dc:date>2024-05-14</dc:date>
<dc:identifier>doi:10.1101/2024.05.08.593181</dc:identifier>
<dc:title><![CDATA[Androgen receptor activity inversely correlates with immune cell infiltration and immunotherapy response across multiple cancer lineages]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-05-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.05.09.593410v1?rss=1">
<title>
<![CDATA[
O-GalNAc glycans enrich in white matter tracts and regulate nodes of Ranvier 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.05.09.593410v1?rss=1</link>
<description><![CDATA[
Protein O-glycosylation is a critical modification in the brain, as genetic variants in the pathway are associated with both common and severe neuropsychiatric phenotypes. However, little is known about the most abundant type of O-glycan in the mammalian brain, which are O-GalNAc linked. Here, we determined the spatial localization, protein carriers, and cellular function of O-GalNAc glycans in mouse brain. We observed striking spatial enrichment of O-GalNAc glycans in white matter tracts and at nodes of Ranvier. Glycoproteomic analysis revealed that more than half of the identified O-GalNAc glycans were present on chondroitin sulfate proteoglycans termed lecticans, and display both domain enrichment and site-specific heterogeneity. Though genetic ablation in a single cell type failed to replicate the severe phenotypes seen in congenital disorders, inhibition of O-GalNAc synthesis in neurons reduced binding of Siglec-4, a known regulator neurite growth, and shortened the length of nodes of Ranvier. This work highlights a new function of O-GalNAc glycans in the brain and will inform future studies on their role in development and disease.
]]></description>
<dc:creator>Noel, M.</dc:creator>
<dc:creator>Suttapitugsakul, S.</dc:creator>
<dc:creator>Cummings, R. D.</dc:creator>
<dc:creator>Mealer, R. G.</dc:creator>
<dc:date>2024-05-09</dc:date>
<dc:identifier>doi:10.1101/2024.05.09.593410</dc:identifier>
<dc:title><![CDATA[O-GalNAc glycans enrich in white matter tracts and regulate nodes of Ranvier]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-05-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.05.10.593572v1?rss=1">
<title>
<![CDATA[
A phosphorylation code coordinating transcription condensate dynamics with DNA replication 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.05.10.593572v1?rss=1</link>
<description><![CDATA[
Chromatin is organized into compartments enriched with functionally-related proteins driving non-linear biochemical activities. Some compartments, e.g. transcription foci, behave as liquid condensates. While the principles governing the enrichment of proteins within condensates are being elucidated, mechanisms that coordinate condensate dynamics with other nuclear processes like DNA replication have not been identified. We show that at the G1/S cell cycle transition, large transcription condensates form at histone locus bodies (HLBs) in a cyclin-dependent kinase 1 and 2 (CDK1/2)-dependent manner. As cells progress through S phase, ataxia-telangiectasia and Rad3-related (ATR) accumulates within HLBs and dissolves the associated transcription condensates. Integration of CDK1/2 and ATR signaling creates a phosphorylation code within the intrinsically-disordered region of mediator subunit 1 (MED1) coordinating condensate dynamics with DNA replication. Disruption of this code results in imbalanced histone biosynthesis, and consequently, global DNA damage. We propose the spatiotemporal dynamics of transcription condensates are actively controlled via phosphorylation and essential for viability of proliferating cells.
]]></description>
<dc:creator>Marmolejo, C. O.</dc:creator>
<dc:creator>Sanchez, C.</dc:creator>
<dc:creator>Lee, J.</dc:creator>
<dc:creator>Werner, M.</dc:creator>
<dc:creator>Roberts, P.</dc:creator>
<dc:creator>Hamperl, S.</dc:creator>
<dc:creator>Saldivar, J. C.</dc:creator>
<dc:date>2024-05-11</dc:date>
<dc:identifier>doi:10.1101/2024.05.10.593572</dc:identifier>
<dc:title><![CDATA[A phosphorylation code coordinating transcription condensate dynamics with DNA replication]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-05-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.05.12.592631v1?rss=1">
<title>
<![CDATA[
Structural basis of autoantibody binding in Neuromyelitis Optica 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.05.12.592631v1?rss=1</link>
<description><![CDATA[
Neuromyelitis Optica (NMO) is an autoimmune disease of the central nervous system where pathogenic autoantibodies target the human astrocyte water channel aquaporin-4 causing neurological impairment. Autoantibody binding leads to complement dependent and complement independent cytotoxicity, ultimately resulting in astrocyte death, demyelination, and neuronal loss. Aquaporin-4 assembles in astrocyte plasma membranes as symmetric tetramers or as arrays of tetramers. We report molecular structures of aquaporin-4 alone and bound to Fab fragments from patient-derived NMO autoantibodies using cryogenic electron microscopy. Each antibody binds to epitopes comprised of three extracellular loops of aquaporin-4 with contributions from multiple molecules in the assembly. The structures distinguish between antibodies that bind to the tetrameric form of aquaporin-4, and those targeting higher order orthogonal arrays of tetramers that provide more diverse bridging epitopes.

One-Sentence SummaryPathogenic autoantibodies in Neuromyelitis Optica distinguish multiply presented epitopes on arrays of human aquaporin-4.
]]></description>
<dc:creator>Gupta, M.</dc:creator>
<dc:creator>Khandelwal, N. K.</dc:creator>
<dc:creator>Nelson, A.</dc:creator>
<dc:creator>Hwang, P.</dc:creator>
<dc:creator>Pourmal, S.</dc:creator>
<dc:creator>Bennett, J. L.</dc:creator>
<dc:creator>Stroud, R. M.</dc:creator>
<dc:date>2024-05-14</dc:date>
<dc:identifier>doi:10.1101/2024.05.12.592631</dc:identifier>
<dc:title><![CDATA[Structural basis of autoantibody binding in Neuromyelitis Optica]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-05-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.05.12.593749v1?rss=1">
<title>
<![CDATA[
Coumarin as a general switching auxiliary to prepare photochromic and spontaneously blinking fluorophores 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.05.12.593749v1?rss=1</link>
<description><![CDATA[
Single-molecule localization microscopy (SMLM) uses activatable or switchable fluorophores to create non-diffraction limited maps of molecular location in biological samples. Despite the utility of this imaging technique, the portfolio of appropriate labels for SMLM remains limited. Here, we describe a general strategy for the construction of "glitter bomb" labels by simply combining rhodamine and coumarin dyes though an amide bond. Condensation of the ortho-carboxyl group on the pendant phenyl ring of rhodamine dyes with a 7-aminocoumarin yields photochromic or spontaneously blinking fluorophores depending on the parent rhodamine structure. We apply this strategy to prepare labels useful super-resolution experiments in fixed cells using different attachment techniques. This general glitter bomb strategy should lead to improved labels for SMLM, ultimately enabling the creation of detailed molecular maps in biological samples.
]]></description>
<dc:creator>Jradi, F.</dc:creator>
<dc:creator>English, B. P.</dc:creator>
<dc:creator>Brown, T. A.</dc:creator>
<dc:creator>Arron, J.</dc:creator>
<dc:creator>Khuon, S.</dc:creator>
<dc:creator>Galbraith, J. A.</dc:creator>
<dc:creator>Galbraith, C. G.</dc:creator>
<dc:creator>Lavis, L. D.</dc:creator>
<dc:date>2024-05-12</dc:date>
<dc:identifier>doi:10.1101/2024.05.12.593749</dc:identifier>
<dc:title><![CDATA[Coumarin as a general switching auxiliary to prepare photochromic and spontaneously blinking fluorophores]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-05-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.05.13.593968v1?rss=1">
<title>
<![CDATA[
Detection of ATTR aggregates in plasma of polyneuropathic ATTR-V30M amyloidosis patients 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.05.13.593968v1?rss=1</link>
<description><![CDATA[
ATTR amyloidosis is caused by the deposition of transthyretin amyloid fibrils in tissues often leading to organ failure and death. The clinical spectrum of this disease is highly diverse and dependent on many factors including the presence or absence of mutations within the transthyretin protein and/or an individuals ancestry. The phenotypic variability of ATTR amyloidosis makes it difficult to diagnose, delaying treatment and worsening patient prognosis. Our lab has recently developed a peptide probe that detects transthyretin aggregates in plasma of ATTR amyloidosis patients with cardiomyopathy but has not been tested in plasma from polyneuropathic patients. Here we evaluate our probe in a cohort of Portuguese patients carrying the ATTR-V30M mutation and having no cardiac phenotype. We found that we could indeed detect aggregates in their plasma, and there appeared to be no relationship between the presence of aggregates and patient age or gender. Our work has broad implications on the pathobiology of ATTR amyloidosis and contribute to the validation of our probe as a novel detection tool for this disease.
]]></description>
<dc:creator>Pedretti, R.</dc:creator>
<dc:creator>Wang, L.</dc:creator>
<dc:creator>Grodin, J. L.</dc:creator>
<dc:creator>Masri, A.</dc:creator>
<dc:creator>Kelly, J.</dc:creator>
<dc:creator>Saelices, L.</dc:creator>
<dc:date>2024-05-13</dc:date>
<dc:identifier>doi:10.1101/2024.05.13.593968</dc:identifier>
<dc:title><![CDATA[Detection of ATTR aggregates in plasma of polyneuropathic ATTR-V30M amyloidosis patients]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-05-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.05.15.594383v1?rss=1">
<title>
<![CDATA[
Trifunctional fatty acid derivatives demonstrate the impact of diazirine placement 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.05.15.594383v1?rss=1</link>
<description><![CDATA[
Functionalized lipid probes are a critical new tool to interrogate the function of individual lipid species, but the structural parameters that constrain their utility have not been thoroughly described. Here, we synthesize three palmitic acid derivatives with a diazirine at different positions on the acyl chain and examine their metabolism, subcellular localization, and protein interactions. We demonstrate that while they produce very similar metabolites and subcellular distributions, probes with the diazirine at either end pulldown distinct subsets of proteins after photo-crosslinking. This highlights the importance of thoughtful diazirine placement when developing probes based on biological molecules.
]]></description>
<dc:creator>Farley, S. E.</dc:creator>
<dc:creator>Hashimoto, R.</dc:creator>
<dc:creator>Evangelista, J.</dc:creator>
<dc:creator>Stein, F.</dc:creator>
<dc:creator>Haberkant, P.</dc:creator>
<dc:creator>Kikuchi, K.</dc:creator>
<dc:creator>Tafesse, F.</dc:creator>
<dc:creator>Schultz, C.</dc:creator>
<dc:date>2024-05-15</dc:date>
<dc:identifier>doi:10.1101/2024.05.15.594383</dc:identifier>
<dc:title><![CDATA[Trifunctional fatty acid derivatives demonstrate the impact of diazirine placement]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-05-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.05.16.589954v1?rss=1">
<title>
<![CDATA[
Genomic loss of GPR108 disrupts AAV transduction in birds 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.05.16.589954v1?rss=1</link>
<description><![CDATA[
The G protein-coupled receptor 108 (GPR108) gene encodes a protein factor identified as critical for adeno-associated virus (AAV) entry into mammalian cells, but whether it is universally involved in AAV transduction is unknown. Remarkably, we have discovered that GPR108 is absent in the genomes of birds and in most other sauropsids, providing a likely explanation for the overall lower AAV transduction efficacy of common AAV serotypes in birds compared to mammals. Importantly, transgenic expression of human GPR108 and manipulation of related glycan binding sites in the viral capsid significantly boost AAV transduction in zebra finch cells. These findings contribute to a more in depth understanding of the mechanisms and evolution of AAV transduction, with potential implications for the design of efficient tools for gene manipulation in experimental animal models, and a range of gene therapy applications in humans.
]]></description>
<dc:creator>Nevue, A. A.</dc:creator>
<dc:creator>Sairavi, A.</dc:creator>
<dc:creator>Huang, S. J.</dc:creator>
<dc:creator>Nakai, H.</dc:creator>
<dc:creator>Mello, C. V.</dc:creator>
<dc:date>2024-05-17</dc:date>
<dc:identifier>doi:10.1101/2024.05.16.589954</dc:identifier>
<dc:title><![CDATA[Genomic loss of GPR108 disrupts AAV transduction in birds]]></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/10.1101/2024.05.16.594391v1?rss=1">
<title>
<![CDATA[
Protein CoAlation on TXNRD2 regulates mitochondrial thioredoxin system to protect against ferroptosis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.05.16.594391v1?rss=1</link>
<description><![CDATA[
The Cystine-xCT transporter-Glutathione (GSH)-GPX4 axis is the canonical pathway to protect against ferroptosis. While not required for ferroptosis-inducing compounds (FINs) targeting GPX4, FINs targeting the xCT transporter require mitochondria and its lipid peroxidation to trigger ferroptosis. However, the mechanism underlying the difference between these FINs is still unknown. Given that cysteine is also required for coenzyme A (CoA) biosynthesis, here we show that CoA supplementation specifically prevents ferroptosis induced by xCT inhibitors but not GPX4 inhibitors. We find that, auranofin, a thioredoxin reductase inhibitor, abolishes the protective effect of CoA. We also find that CoA availability determines the enzymatic activity of thioredoxin reductase, but not thioredoxin. Importantly, the mitochondrial thioredoxin system, but not the cytosolic thioredoxin system, determines CoA-mediated ferroptosis inhibition. Our data show that the CoA regulates the in vitro enzymatic activity of mitochondrial thioredoxin reductase (TXNRD2) by covalently modifying the thiol group of cysteine (CoAlation) on Cys-483. Replacing Cys-483 with alanine on TXNRD2 abolishes its in vitro enzymatic activity and ability to protect cells from ferroptosis. Targeting xCT to limit cysteine import and, therefore, CoA biosynthesis reduced CoAlation on TXNRD2, an effect that was rescued by CoA supplementation. Furthermore, the fibroblasts from patients with disrupted CoA metabolism demonstrate increased mitochondrial lipid peroxidation. In organotypic brain slice cultures, inhibition of CoA biosynthesis leads to an oxidized thioredoxin system, mitochondrial lipid peroxidation, and loss in cell viability, which were all rescued by ferrostatin-1. These findings identify CoA-mediated post-translation modification to regulate the thioredoxin system as an alternative ferroptosis protection pathway with potential clinical relevance for patients with disrupted CoA metabolism.
]]></description>
<dc:creator>Lin, C.-C.</dc:creator>
<dc:creator>Lin, Y.-T.</dc:creator>
<dc:creator>Chen, S.-Y.</dc:creator>
<dc:creator>Setayeshpour, Y.</dc:creator>
<dc:creator>Chen, Y.</dc:creator>
<dc:creator>Dunn, D. E.</dc:creator>
<dc:creator>Soderblom, E. J.</dc:creator>
<dc:creator>Zhang, G.</dc:creator>
<dc:creator>Filonenko, V.</dc:creator>
<dc:creator>Jeong, S. Y.</dc:creator>
<dc:creator>Floyd, S. R.</dc:creator>
<dc:creator>Hayflick, S.</dc:creator>
<dc:creator>Gout, I.</dc:creator>
<dc:creator>Chi, J.-T. A.</dc:creator>
<dc:date>2024-05-18</dc:date>
<dc:identifier>doi:10.1101/2024.05.16.594391</dc:identifier>
<dc:title><![CDATA[Protein CoAlation on TXNRD2 regulates mitochondrial thioredoxin system to protect against ferroptosis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-05-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.05.16.594555v1?rss=1">
<title>
<![CDATA[
In vivo Targeting MEK and TNK2/SRC pathways in PTPN11 driven leukemia 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.05.16.594555v1?rss=1</link>
<description><![CDATA[
PTPN11 encodes for a tyrosine phosphatase implicated in the pathogenesis of hematologic malignancies such as Juvenile Myelomonocytic Leukemia (JMML), Acute Myeloid Leukemia (AML), and Acute Lymphoblastic Leukemia (ALL). Since activating mutations of PTPN11 increase proliferative signaling and cell survival through the RAS/MAPK proliferative pathway there is significant interest in using MEK inhibitors for clinical benefit. Yet, single agent clinical activity has been minimal. Previously, we showed that PTPN11 is further activated by upstream tyrosine kinases TNK2/SRC, and that PTPN11-mutant JMML and AML cells are sensitive to TNK2 inhibition using dasatinib. In order to validate these findings, we adopted a genetically engineered mouse model of PTPN11 driven leukemia using the mouse strain 129S/Sv-Ptpn11tm6Bgn/Mmucd crossed with B6.129P2-Lyz2tm1(cre)Ifo/J. The F1 progeny expressing Ptpn11D61Y within hematopoietic cells destined along the granulocyte-monocyte progenitor lineage developed a fatal myeloproliferative disorder characterized by neutrophilia and monocytosis, and infiltration of myeloid cells into the liver and spleen. Cohorts of Ptpn11D61Y expressing animals treated with combination of dasatinib and trametinib for an extended period of time was well tolerated and had a significant effect in mitigating disease parameters compared to single agents. Finally, a primary patient-derived xenograft model using a myeloid leukemia with PTPN11 F71L also displayed improved disease response to combination. Collectively, these studies point to combined therapies targeting MEK and TNK2/SRC as a promising therapeutic potential for PTPN11-mutant leukemias.

Key PointsO_LICombining MEK and TNK2/SRC inhibitors has therapeutic potential in PTPN11 mutant JMML and AML
C_LI
]]></description>
<dc:creator>Chang, B. H.</dc:creator>
<dc:creator>Thiel-Klare, K.</dc:creator>
<dc:creator>Tyner, J.</dc:creator>
<dc:date>2024-05-18</dc:date>
<dc:identifier>doi:10.1101/2024.05.16.594555</dc:identifier>
<dc:title><![CDATA[In vivo Targeting MEK and TNK2/SRC pathways in PTPN11 driven leukemia]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-05-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.05.17.594191v1?rss=1">
<title>
<![CDATA[
Maternal Western-style diet programs skeletal muscle gene expression in lean adolescent Japanese macaque offspring 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.05.17.594191v1?rss=1</link>
<description><![CDATA[
Early-life exposure to maternal obesity or a maternal calorically dense Western-style diet (WSD) is strongly associated with a greater risk of metabolic diseases in offspring, most notably insulin resistance and metabolic dysfunction-associated steatotic liver disease (MASLD). Prior studies in our well-characterized Japanese macaque model demonstrated that offspring of dams fed a WSD, even when weaned onto a control (CTR) diet, had reductions in skeletal muscle mitochondrial metabolism and increased skeletal muscle insulin resistance compared to offspring of dams on CTR diet. In the current study, we employed a nested design to test for differences in gene expression in skeletal muscle from lean 3-year-old adolescent offspring from dams fed a maternal WSD in both the presence and absence of maternal obesity or lean dams fed a CTR diet. We included offspring weaned to both a WSD or CTR diet to further account for differences in response to post-weaning diet and interaction effects between diets. Overall, we found that a maternal WSD fed to dams during pregnancy and lactation was the principal driver of differential gene expression (DEG) in offspring muscle at this time point. We identified key gene pathways important in insulin signaling including PI3K-Akt and MAP-kinase, regulation of muscle regeneration, and transcription-translation feedback loops, in both male and female offspring. Muscle DEG showed no measurable difference between offspring of obese dams on WSD compared to those of lean dams fed WSD. A post-weaning WSD effected offspring transcription only in individuals from the maternal CTR diet group but not in maternal WSD group. Collectively, we identify that maternal diet composition has a significant and lasting impact on offspring muscle transcriptome and influences later transcriptional response to WSD in muscle, which may underlie the increased metabolic disease risk in offspring.
]]></description>
<dc:creator>Beck, E. A.</dc:creator>
<dc:creator>Hetrick, B.</dc:creator>
<dc:creator>Nassar, L.</dc:creator>
<dc:creator>Turnball, D. W.</dc:creator>
<dc:creator>Dean, T. A.</dc:creator>
<dc:creator>Gannon, M.</dc:creator>
<dc:creator>Aagaard, K. M.</dc:creator>
<dc:creator>Wesolowski, S. R.</dc:creator>
<dc:creator>Friedman, J. E.</dc:creator>
<dc:creator>Kievit, P.</dc:creator>
<dc:creator>McCurdy, C. E.</dc:creator>
<dc:date>2024-05-21</dc:date>
<dc:identifier>doi:10.1101/2024.05.17.594191</dc:identifier>
<dc:title><![CDATA[Maternal Western-style diet programs skeletal muscle gene expression in lean adolescent Japanese macaque offspring]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-05-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.05.21.594597v1?rss=1">
<title>
<![CDATA[
Viral and host network analysis of the human cytomegalovirus transcriptome in latency. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.05.21.594597v1?rss=1</link>
<description><![CDATA[
HCMV genes UL135 and UL138 play opposing roles regulating latency and reactivation in CD34+ human progenitor cells (HPCs). Using the THP-1 cell line model for latency and reactivation, we designed an RNA sequencing study to compare the transcriptional profile of HCMV infection in the presence and absence of these genes. The loss of UL138 results in elevated levels of viral gene expression and increased differentiation of cell populations that support HCMV gene expression and genome synthesis. The loss of UL135 results in diminished viral gene expression during an initial burst that occurs as latency is established and no expression of eleven viral genes from the ULb region even following stimulation for differentiation and reactivation. Transcriptional network analysis revealed host transcription factors with potential to regulate the ULb genes in coordination with pUL135. These results reveal roles for UL135 and UL138 in regulation of viral gene expression and potentially hematopoietic differentiation.
]]></description>
<dc:creator>Collins-McMillen, D. K.</dc:creator>
<dc:creator>De Oliveira Pessoa, D.</dc:creator>
<dc:creator>Zarrella, K.</dc:creator>
<dc:creator>Parkins, C. J.</dc:creator>
<dc:creator>Daily, M.</dc:creator>
<dc:creator>Moorman, N. K.</dc:creator>
<dc:creator>Kamil, J. P.</dc:creator>
<dc:creator>Caposio, P.</dc:creator>
<dc:creator>Padi, M.</dc:creator>
<dc:creator>Goodrum, F.</dc:creator>
<dc:date>2024-05-21</dc:date>
<dc:identifier>doi:10.1101/2024.05.21.594597</dc:identifier>
<dc:title><![CDATA[Viral and host network analysis of the human cytomegalovirus transcriptome in latency.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-05-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.05.24.595793v1?rss=1">
<title>
<![CDATA[
Transcriptomic landscape of mammalian ventral pallidum at single-cell resolution 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.05.24.595793v1?rss=1</link>
<description><![CDATA[
The ventral pallidum (VP) is critical for motivated behaviors. While contemporary work has begun to elucidate the functional diversity of VP neurons, the molecular heterogeneity underlying this functional diversity remains incompletely understood. We used snRNA-seq and in situ hybridization to define the transcriptional taxonomy of VP cell types in mice, macaques, and baboons. We found transcriptional conservation between all three species, within the broader neurochemical cell types. Unique dopaminoceptive and cholinergic subclusters were identified and conserved across both primate species but had no homolog in mice. This harmonized consensus VP cellular atlas will pave the way for understanding the structure and function of the VP and identified key neuropeptides, neurotransmitters, and neuro receptors that could be targeted within specific VP cell types for functional investigations.

TeaserGenetic identity of ventral pallidum cell types is conserved across rodents and primates at the transcriptional level
]]></description>
<dc:creator>Fang, L. Z.</dc:creator>
<dc:creator>Yang, L.</dc:creator>
<dc:creator>Lynch, M. R.</dc:creator>
<dc:creator>Xu, C. S.</dc:creator>
<dc:creator>Hahm, H.</dc:creator>
<dc:creator>Zhang, Y.</dc:creator>
<dc:creator>Heitmeier, M. R.</dc:creator>
<dc:creator>Costa, V.</dc:creator>
<dc:creator>Samineni, V. K.</dc:creator>
<dc:creator>Creed, M. C.</dc:creator>
<dc:date>2024-05-26</dc:date>
<dc:identifier>doi:10.1101/2024.05.24.595793</dc:identifier>
<dc:title><![CDATA[Transcriptomic landscape of mammalian ventral pallidum at single-cell resolution]]></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/10.1101/2024.05.24.595655v1?rss=1">
<title>
<![CDATA[
Clonal spheroids capture functional and genetic heterogeneity of head and neck cancer 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.05.24.595655v1?rss=1</link>
<description><![CDATA[
Head and neck cancer squamous cell carcinoma (HNSCC) cells exhibit both structural and functional diversity, making them valuable models for understanding tumor heterogeneity at clinical levels. In this study, we generated single-cell-derived spheroids (SCDS) from HNSCC cell lines and patient tumor cells using scaffold- and non-scaffold-based methods to assess this variability. A distinct structural variability among these SCDS, categorized as hypo- and hyperproliferative spheroids based on size, was observed. Hyperproliferative spheroids demonstrated heightened proliferative and tumorigenic potential and increased sensitivity to cisplatin and radiation, while hypoproliferative spheroids exhibited enhanced migratory capabilities. Single-cell RNA sequencing (scRNA-seq) of hypo- and hyperproliferative spheroids provided insights into the transcriptional landscape of HNSCC cells, validating the observed structural and functional heterogeneities within primary tumors. These functionally and genetically characterized spheroids offer valuable tools for the development of next-generation therapeutics.

Statement of SignificanceEstablishment and characterization of single-cell-derived spheroids from head and neck cancer cells, employing scaffold and non-scaffold materials, demonstrate functional and genetic heterogeneity. Single-cell analysis reveals correlations between genetic diversity and spheroid functionality. These characterized spheroids offer potential for advancing therapeutics development.
]]></description>
<dc:creator>Pandey, J.</dc:creator>
<dc:creator>Malik, M. Z.</dc:creator>
<dc:creator>Shyanti, R. k.</dc:creator>
<dc:creator>Parashar, P.</dc:creator>
<dc:creator>kujur, P. K.</dc:creator>
<dc:creator>Mishra, D.</dc:creator>
<dc:creator>Tailor, D.</dc:creator>
<dc:creator>Lee, J. M.</dc:creator>
<dc:creator>Kataria, T.</dc:creator>
<dc:creator>Gupta, D.</dc:creator>
<dc:creator>Verma, H.</dc:creator>
<dc:creator>Malhotra, S. V.</dc:creator>
<dc:creator>Kateriya, S.</dc:creator>
<dc:creator>Tandon, V.</dc:creator>
<dc:creator>Chaturvedi, R.</dc:creator>
<dc:creator>Singh, R. P.</dc:creator>
<dc:date>2024-05-28</dc:date>
<dc:identifier>doi:10.1101/2024.05.24.595655</dc:identifier>
<dc:title><![CDATA[Clonal spheroids capture functional and genetic heterogeneity of head and neck cancer]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-05-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.05.24.595690v1?rss=1">
<title>
<![CDATA[
Comprehensive multi-site profiling of the malignant pleural mesothelioma micro-environment identifies candidate molecular determinants of histopathologic type 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.05.24.595690v1?rss=1</link>
<description><![CDATA[
Pleural mesothelioma (PM) comprises sarcomatoid, epithelioid and biphasic histologic subtypes. Bulk PM RNA-sequencing identifies a histology-associated molecular gradient with features of epithelial-mesenchymal (EM) transition but cannot parse malignant, stromal, and immune tumor components. The mechanisms driving PM malignant cell phenotype and associated histology is not well-characterized. Here, we use single-cell RNA-sequencing (scRNA-seq) paired with exome, bulk RNA-sequencing, and histologic analysis of adjacent samples to characterize malignant cell EM state, parse the tumor microenvironment (TME), and identify candidate drivers of PM cell fate. We observe EM variation in malignant cells analogous to bulk samples. We characterize epithelioid and sarcomatoid malignant cell programs and identify a new uncommitted malignant cell EM phenotype enriched in biphasic histology samples. Using inferred CNVs we observe that single individual PM clones consist of cells exhibiting all three EM cell states. We find that distinct non-malignant microenvironments associated with tumors consisting of mostly cells in each state, and identify WNT inhibition, GAS6-AXL, and HBEGF-EGFR signaling as pathways associated with distinct EM cell states. These findings provide deeper insight into the molecular drivers of PM malignant cells and identify non-malignant cell signals as potential EMT and growth drivers in PM.
]]></description>
<dc:creator>Severson, D. T.</dc:creator>
<dc:creator>Freyaldenhoven, S.</dc:creator>
<dc:creator>Wadowski, B.</dc:creator>
<dc:creator>Hughes, T.</dc:creator>
<dc:creator>Hung, Y. P.</dc:creator>
<dc:creator>Chirieac, L.</dc:creator>
<dc:creator>Jensen, R. V.</dc:creator>
<dc:creator>Richards, W. G.</dc:creator>
<dc:creator>Sedak, A. A.</dc:creator>
<dc:creator>Gustafson, C. E.</dc:creator>
<dc:creator>Vermilya, K.</dc:creator>
<dc:creator>Innocenti, S.</dc:creator>
<dc:creator>Korle, S.</dc:creator>
<dc:creator>Barlow, J. S.</dc:creator>
<dc:creator>Couger, M. B.</dc:creator>
<dc:creator>Anderson, J.</dc:creator>
<dc:creator>Meyervitz, J.</dc:creator>
<dc:creator>Wang, V.</dc:creator>
<dc:creator>Dao, M. N.</dc:creator>
<dc:creator>Shalek, A. K.</dc:creator>
<dc:creator>De Rienzo, A.</dc:creator>
<dc:creator>Bueno, R.</dc:creator>
<dc:date>2024-05-28</dc:date>
<dc:identifier>doi:10.1101/2024.05.24.595690</dc:identifier>
<dc:title><![CDATA[Comprehensive multi-site profiling of the malignant pleural mesothelioma micro-environment identifies candidate molecular determinants of histopathologic type]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-05-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.05.28.596304v1?rss=1">
<title>
<![CDATA[
Altered liver metabolism post-wean abolishes efficacy of vitamin D for breast cancer prevention in a mouse model 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.05.28.596304v1?rss=1</link>
<description><![CDATA[
Young women have increased risk of vitamin D deficiency, which may increase breast cancer incidence. Here, we assessed the anti-cancer efficacy of vitamin D in mouse models of young-onset breast cancer. In never-pregnant mice, vitamin D supplementation increased serum 25(OH)D and hepatic 1,25(OH)2D3, reduced tumor size, and associated with anti-tumor immunity. These anti-tumor effects were not replicated in a mouse model of postpartum breast cancer, where hepatic metabolism of vitamin D was suppressed post-wean, which resulted in deficient serum 25(OH)D and reduced hepatic 1,25(OH)2D3. Treatment with active 1,25(OH)2D3 induced hypercalcemia exclusively in post-wean mice, highlighting metabolic imbalance post-wean. RNAseq revealed suppressed CYP450 expression postpartum. In sum, we provide evidence that vitamin D anti-tumor activity is mediated through immunomodulatory mechanisms and is ineffective in the post-wean window due to altered hepatic metabolism. These findings have implications for suppressed xenobiotic metabolism in postpartum women beyond vitamin D.

Statement of SignificanceIn a rodent model of postpartum breast cancer, weaning suppresses hepatic CYP450 activity and renders vitamin D supplementation ineffective, with implications for xenobiotic drug efficacy and safety. A tailored approach to therapy based on reproductive history is crucial for young breast cancer patients, and for healthcare strategies for postpartum women.
]]></description>
<dc:creator>Bernhardt, S. M.</dc:creator>
<dc:creator>Ozaki, M. K.</dc:creator>
<dc:creator>Betts, C.</dc:creator>
<dc:creator>Bleyle, L. A.</dc:creator>
<dc:creator>DeBarber, A. E.</dc:creator>
<dc:creator>Fornetti, J.</dc:creator>
<dc:creator>Liberty, A. L.</dc:creator>
<dc:creator>De Wilde, E.</dc:creator>
<dc:creator>Zhang, Y.</dc:creator>
<dc:creator>Xia, Z.</dc:creator>
<dc:creator>Schedin, P.</dc:creator>
<dc:date>2024-06-02</dc:date>
<dc:identifier>doi:10.1101/2024.05.28.596304</dc:identifier>
<dc:title><![CDATA[Altered liver metabolism post-wean abolishes efficacy of vitamin D for breast cancer prevention in a mouse model]]></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/10.1101/2024.06.04.596911v1?rss=1">
<title>
<![CDATA[
TNBC response to paclitaxel phenocopies interferon response which reveals cell cycle-associated resistance mechanisms 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.06.04.596911v1?rss=1</link>
<description><![CDATA[
Paclitaxel is a standard of care neoadjuvant therapy for patients with triple negative breast cancer (TNBC); however, it shows limited benefit for locally advanced or metastatic disease. Here we used a coordinated experimental-computational approach to explore the influence of paclitaxel on the cellular and molecular responses of TNBC cells. We found that escalating doses of paclitaxel resulted in multinucleation, promotion of senescence, and initiation of DNA damage induced apoptosis. Single-cell RNA sequencing (scRNA-seq) of TNBC cells after paclitaxel treatment revealed upregulation of innate immune programs canonically associated with interferon response and downregulation of cell cycle progression programs. Systematic exploration of transcriptional responses to paclitaxel and cancer-associated microenvironmental factors revealed common gene programs induced by paclitaxel, IFNB, and IFNG. Transcription factor (TF) enrichment analysis identified 13 TFs that were both enriched based on activity of downstream targets and also significantly upregulated after paclitaxel treatment. Functional assessment with siRNA knockdown confirmed that the TFs FOSL1, NFE2L2 and ELF3 mediate cellular proliferation and also regulate nuclear structure. We further explored the influence of these TFs on paclitaxel-induced cell cycle behavior via live cell imaging, which revealed altered progression rates through G1, S/G2 and M phases. We found that ELF3 knockdown synergized with paclitaxel treatment to lock cells in a G1 state and prevent cell cycle progression. Analysis of publicly available breast cancer patient data showed that high ELF3 expression was associated with poor prognosis and enrichment programs associated with cell cycle progression. Together these analyses disentangle the diverse aspects of paclitaxel response and identify ELF3 upregulation as a putative biomarker of paclitaxel resistance in TNBC.
]]></description>
<dc:creator>Calistri, N. L.</dc:creator>
<dc:creator>Liby, T. A.</dc:creator>
<dc:creator>Hu, Z.</dc:creator>
<dc:creator>Zhang, H.</dc:creator>
<dc:creator>Dane, M. A.</dc:creator>
<dc:creator>Gross, S. M.</dc:creator>
<dc:creator>Heiser, L. M.</dc:creator>
<dc:date>2024-06-06</dc:date>
<dc:identifier>doi:10.1101/2024.06.04.596911</dc:identifier>
<dc:title><![CDATA[TNBC response to paclitaxel phenocopies interferon response which reveals cell cycle-associated resistance mechanisms]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-06-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.06.06.597774v1?rss=1">
<title>
<![CDATA[
A Nanobody Interaction with SARS-CoV-2 Spike Allows the Versatile Targeting of Lentivirus Vectors 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.06.06.597774v1?rss=1</link>
<description><![CDATA[
While investigating methods to target gene delivery vectors to specific cell types, we examined the potential of using a nanobody against the SARS-CoV-2 Spike protein receptor binding domain to direct lentivirus infection of Spike-expressing cells. Using three different approaches, we found that lentiviruses with surface-exposed nanobody domains selectively infect Spike-expressing cells. The targeting is dependent on the fusion function of Spike, and conforms to a model in which nanobody binding to the Spike protein triggers the Spike fusion machinery. The nanobody-Spike interaction also is capable of directing cell-cell fusion, and the selective infection of nanobody-expressing cells by Spike-pseudotyped lentivirus vectors. Significantly, cells infected with SARS-CoV-2 are efficiently and selectively infected by lentivirus vectors pseudotyped with a chimeric nanobody protein. Our results suggest that cells infected by any virus that forms syncytia may be targeted for gene delivery using an appropriate nanobody or virus receptor mimic. Vectors modified in this fashion may prove useful in the delivery of immunomodulators to infected foci to mitigate the effects of viral infections.

IMPORTANCEWe have discovered that lentiviruses decorated on their surfaces with a nanobody against the SARS-CoV-2 Spike protein selectively infect Spike-expressing cells. Infection is dependent on the specificity of the nanobody and the fusion function of the Spike protein, and conforms to a reverse fusion model, in which nanobody binding to Spike triggers the Spike fusion machinery. The nanobody-Spike interaction also can drive cell-cell fusion, and infection of nanobody-expressing cells with viruses carrying the Spike protein. Importantly, cells infected with SARS-CoV-2 are selectively infected with nanobody-decorated lentiviruses. These results suggest that cells infected by any virus that expresses an active receptor-binding fusion protein may be targeted by vectors for delivery of cargoes to mitigate infections.
]]></description>
<dc:creator>Alfadhli, A.</dc:creator>
<dc:creator>Bates, T. A.</dc:creator>
<dc:creator>Barklis, R. L.</dc:creator>
<dc:creator>Romanaggi, C.</dc:creator>
<dc:creator>Tafesse, F.</dc:creator>
<dc:creator>Barklis, E.</dc:creator>
<dc:date>2024-06-07</dc:date>
<dc:identifier>doi:10.1101/2024.06.06.597774</dc:identifier>
<dc:title><![CDATA[A Nanobody Interaction with SARS-CoV-2 Spike Allows the Versatile Targeting of Lentivirus Vectors]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-06-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.06.11.598497v1?rss=1">
<title>
<![CDATA[
Synthesis of ionizable lipopolymers using split-Ugi reaction for pulmonary delivery of various size RNAs and gene editing 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.06.11.598497v1?rss=1</link>
<description><![CDATA[
We present an efficient approach for synthesizing cationic poly(ethylene imine) derivatives using the multicomponent split-Ugi reaction to rapidly create a library of complex functional ionizable lipopolymers. We synthesized a diverse library of 155 polymers, formulated them into polyplexes to establish structure-activity relationships crucial for endosomal escape and efficient transfection. After discovering a lead structure, lipopolymer-lipid hybrid nanoparticles are introduced to preferentially deliver to and elicit effective mRNA transfection in lung endothelium and immune cells, including T cells with low in vivo toxicity. The lipopolymer-lipid hybrid nanoparticles showed 300-fold improvement in systemic mRNA delivery to the lung compared to in vivo-JetPEI(R). Lipopolymer-lipid hybrid nanoparticles demonstrated efficient delivery of mRNA-based therapeutics for treatment of two different disease models. Lewis Lung cancer progression was significantly delayed after treatment with loaded IL-12 mRNA in U155@lipids after repeated i.v. administration. Systemic delivery of human CFTR (hCFTR) mRNA resulted in production of functional form of CFTR protein in the lungs. The functionality of hCFTR protein was confirmed by restoration of CFTR- mediated chloride secretion in conductive airway epithelia in CFTR knockout mice after nasal instillation of hCFTR mRNA loaded U155@lipids. We further showed that, U155@lipids nanoparticles can deliver complex CRISPR-Cas9 based RNA cargo to the lung, achieving 5.6 {+/-} 2.4 % gene editing in lung tissue. Moreover, we demonstrated successful PD-1 gene knockout of T cells in vivo. Our results highlight a versatile delivery platform for systemic delivering of mRNA of various sizes for gene therapy for a variety of therapeutics.
]]></description>
<dc:creator>Vlasova, K. Y.</dc:creator>
<dc:creator>Kerr, A.</dc:creator>
<dc:creator>Pennock, N. D.</dc:creator>
<dc:creator>Jozic, A.</dc:creator>
<dc:creator>Sahel, D.</dc:creator>
<dc:creator>Gautam, M.</dc:creator>
<dc:creator>Murthy, N.</dc:creator>
<dc:creator>Roberts, A.</dc:creator>
<dc:creator>Ali, M. W.</dc:creator>
<dc:creator>MacDonald, K. D.</dc:creator>
<dc:creator>Walker, J.</dc:creator>
<dc:creator>Luxenhofer, R.</dc:creator>
<dc:creator>Sahay, G.</dc:creator>
<dc:date>2024-06-13</dc:date>
<dc:identifier>doi:10.1101/2024.06.11.598497</dc:identifier>
<dc:title><![CDATA[Synthesis of ionizable lipopolymers using split-Ugi reaction for pulmonary delivery of various size RNAs and gene editing]]></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/10.1101/2024.06.11.598597v1?rss=1">
<title>
<![CDATA[
A genetically encoded sensor for real-time monitoring of poly-ADP-ribosylation dynamics in-vitro and in cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.06.11.598597v1?rss=1</link>
<description><![CDATA[
ADP-ribosylation, the transfer of ADP-ribose (ADPr) from nicotinamide adenine dinucleotide (NAD+) groups to proteins, is a conserved post-translational modification (PTM) that occurs most prominently in response to DNA damage. ADP-ribosylation is a dynamic PTM regulated by writers (PARPs), erasers (ADPr hydrolases), and readers (ADPR binders). PARP1 is the primary DNA damage-response writer responsible for adding a polymer of ADPR to proteins (PARylation). Real-time monitoring of PARP1-mediated PARylation, especially in live cells, is critical for understanding the spatial and temporal regulation of this unique PTM. Here, we describe a genetically encoded FRET probe (pARS) for semi-quantitative monitoring of PARylation dynamics. pARS feature a PAR-binding WWE domain flanked with turquoise and Venus. With a ratiometric readout and excellent signal-to-noise characteristics, we show that pARS can monitor PARP1-dependent PARylation temporally and spatially in real-time. pARS provided unique insights into PARP1-mediated PARylation kinetics in vitro and high-sensitivity detection of PARylation in live cells, even under mild DNA damage. We also show that pARS can be used to determine the potency of PARP inhibitors in vitro and, for the first time, in live cells in response to DNA damage. The robustness and ease of use of pARS make it an important tool for the PARP field.
]]></description>
<dc:creator>Thomas, A.</dc:creator>
<dc:creator>Upadhyaya, K.</dc:creator>
<dc:creator>Bejan, D.</dc:creator>
<dc:creator>Adoff, H.</dc:creator>
<dc:creator>Cohen, M. S.</dc:creator>
<dc:creator>Schultz, C.</dc:creator>
<dc:date>2024-06-13</dc:date>
<dc:identifier>doi:10.1101/2024.06.11.598597</dc:identifier>
<dc:title><![CDATA[A genetically encoded sensor for real-time monitoring of poly-ADP-ribosylation dynamics in-vitro and in cells]]></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/10.1101/2024.06.14.598695v1?rss=1">
<title>
<![CDATA[
Transcriptomic comparison of in vitro models of the human placenta 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.06.14.598695v1?rss=1</link>
<description><![CDATA[
Studying the human placenta through in vitro cell culture methods is necessary due to limited access and amenability of human placental tissue to certain experimental methods as well as distinct anatomical and physiological differences between animal and human placentas. Selecting an in vitro culture model of the human placenta is challenging due to representation of different trophoblast cell types with distinct biological roles and limited comparative studies that define key characteristics of these models. Therefore, the aim of this research was to create a comprehensive transcriptomic comparison of common in vitro models of the human placenta compared to bulk placental tissue from the CANDLE and GAPPS cohorts (N=1083). We performed differential gene expression analysis on publicly available RNA sequencing data from 6 common in vitro models of the human placenta (HTR-8/SVneo, BeWo, JEG-3, JAR, Primary Trophoblasts, and Villous Explants) and compared to CANDLE and GAPPS bulk placental tissue or cytotrophoblast, syncytiotrophoblast, and extravillous trophoblast cell types derived from bulk placental tissue. All in vitro placental models had a substantial number of differentially expressed genes (DEGs, FDR<0.01) compared to the CANDLE and GAPPS placentas (Average DEGs=10,873), and the individual trophoblast cell types (Average DEGs=5,346), indicating that there are vast differences in gene expression compared to bulk and cell-type specific human placental tissue. Hierarchical clustering identified 53 gene clusters with distinct expression profiles across placental models, with 22 clusters enriched for specific KEGG pathways, 7 clusters enriched for high-expression placental genes, and 7 clusters enriched for absorption, distribution, metabolism, and excretion genes. In vitro placental models were classified by fetal sex based on expression of Y-chromosome genes that identified HTR-8/SVneo cells as being of female origin, while JEG-3, JAR, and BeWo cells are of male origin. Overall, none of the models were a close approximation of the transcriptome of bulk human placental tissue, highlighting the challenges with model selection. To enable researchers to select appropriate models, we have compiled data on differential gene expression, clustering, and fetal sex into an accessible web application: "Comparative Transcriptomic Placental Model Atlas (CTPMA)" which can be utilized by researchers to make informed decisions about their selection of in vitro placental models.
]]></description>
<dc:creator>Lapehn, S.</dc:creator>
<dc:creator>Nair, S.</dc:creator>
<dc:creator>Firsick, E. J.</dc:creator>
<dc:creator>MacDonald, J.</dc:creator>
<dc:creator>Thoreson, C.</dc:creator>
<dc:creator>Litch, J. A.</dc:creator>
<dc:creator>Bush, N. R.</dc:creator>
<dc:creator>Kadam, L.</dc:creator>
<dc:creator>Girard, S.</dc:creator>
<dc:creator>Myatt, L.</dc:creator>
<dc:creator>Prasad, B.</dc:creator>
<dc:creator>Sathyanarayana, S.</dc:creator>
<dc:creator>Paquette, A. G.</dc:creator>
<dc:date>2024-06-16</dc:date>
<dc:identifier>doi:10.1101/2024.06.14.598695</dc:identifier>
<dc:title><![CDATA[Transcriptomic comparison of in vitro models of the human placenta]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-06-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.06.27.600929v1?rss=1">
<title>
<![CDATA[
Discovery of ester-linked ubiquitylation of PARP10 mono-ADP-ribosylation in cells: a dual post-translational modification on Glu/Asp side chains 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.06.27.600929v1?rss=1</link>
<description><![CDATA[
The prevailing view on post-translational modifications (PTMs) is that amino acid side chains in proteins are modified with a single PTM at any given time. However, a growing body of work has demonstrated crosstalk between different PTMs, some occurring on the same residue. Such interplay is seen with ADP-ribosylation and ubiquitylation, where specialized E3 ligases ubiquitylate targets for proteasomal degradation in an ADP-ribosylation-dependent manner. More recently, the DELTEX family of E3 ligases was reported to catalyze ubiquitylation of the 3- hydroxy group of the adenine-proximal ribose of free NAD+ and ADP-ribose in vitro, generating a non-canonical ubiquitin ester-linked species. In this report, we show, for the first time, that this dual PTM occurs in cells on mono-ADP-ribosylated (MARylated) PARP10 on Glu/Asp sites to form a MAR ubiquitin ester (MARUbe). We term this process mono-ADP-ribosyl ubiquitylation or MARUbylation. Using chemical and enzymatic treatments, including a newly characterized bacterial deubiquitinase with esterase-specific activity, we discovered that PARP10 MARUbylation is extended with K11-linked polyubiquitin chains. Finally, mechanistic studies using proteasomal and ubiquitin-activating enzyme inhibitors demonstrated that PARP10 MARUbylation leads to its proteasomal degradation, providing a functional role for this new PTM in regulating protein turnover.
]]></description>
<dc:creator>Bejan, D. S.</dc:creator>
<dc:creator>Lacoursiere, R. E.</dc:creator>
<dc:creator>Pruneda, J. N.</dc:creator>
<dc:creator>Cohen, M. S.</dc:creator>
<dc:date>2024-06-27</dc:date>
<dc:identifier>doi:10.1101/2024.06.27.600929</dc:identifier>
<dc:title><![CDATA[Discovery of ester-linked ubiquitylation of PARP10 mono-ADP-ribosylation in cells: a dual post-translational modification on Glu/Asp side chains]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-06-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.06.26.600812v1?rss=1">
<title>
<![CDATA[
Dopamine transmission at D1 and D2 receptors in the nucleus accumbens contributes to the expression of incubation of cocaine craving 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.06.26.600812v1?rss=1</link>
<description><![CDATA[
Relapse represents a consistent clinical problem for individuals with substance use disorder. In the incubation of craving model of persistent craving and relapse, cue-induced drug seeking progressively intensifies or  incubates during the first weeks of abstinence from drug self-administration and then remains high for months. Previously, we and others have demonstrated that expression of incubated cocaine craving requires strengthening of excitatory synaptic transmission in the nucleus accumbens core (NAcc). However, despite the importance of dopaminergic signaling in the NAcc for motivated behavior, little is known about the role that dopamine (DA) plays in the incubation of cocaine craving. Here we used fiber photometry to measure DA transients in the NAcc of male and female rats during cue-induced seeking tests conducted in early abstinence from cocaine self-administration, prior to incubation, and late abstinence, after incubation of craving has plateaued. We observed DA transients time-locked to cue-induced responding but their magnitude did not differ significantly when measured during early versus late abstinence seeking tests. Next, we tested for a functional role of these DA transients by injecting DA receptor antagonists into the NAcc just before the cue-induced seeking test. Blockade of either D1 or D2 DA receptors reduced cue-induced cocaine seeking after but not before incubation. We found no main effect of sex in our experiments. These results suggest that DA contributes to incubated cocaine seeking but the emergence of this role reflects changes in postsynaptic responsiveness to DA rather than presynaptic alterations.
]]></description>
<dc:creator>Weber, S. J.</dc:creator>
<dc:creator>Kawa, A. B.</dc:creator>
<dc:creator>Moutier, A. L.</dc:creator>
<dc:creator>Beutler, M. M.</dc:creator>
<dc:creator>Koyshman, L. M.</dc:creator>
<dc:creator>Moreno, C. D.</dc:creator>
<dc:creator>Westlake, J. G.</dc:creator>
<dc:creator>Wunsch, A. M.</dc:creator>
<dc:creator>Wolf, M. E.</dc:creator>
<dc:date>2024-06-30</dc:date>
<dc:identifier>doi:10.1101/2024.06.26.600812</dc:identifier>
<dc:title><![CDATA[Dopamine transmission at D1 and D2 receptors in the nucleus accumbens contributes to the expression of incubation of cocaine craving]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-06-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.07.07.602350v1?rss=1">
<title>
<![CDATA[
Quantifying social roles in multi-animal videos using subject-aware deep-learning 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.07.07.602350v1?rss=1</link>
<description><![CDATA[
Analyzing social behaviors is critical for many fields, including neuroscience, psychology, and ecology. While computational tools have been developed to analyze videos containing animals engaging in limited social interactions under specific experimental conditions, automated identification of the social roles of freely moving individuals in a multi-animal group remains unresolved. Here we describe a deep-learning-based system - named LabGym2 - for identifying and quantifying social roles in multi-animal groups. This system uses a subject-aware approach: it evaluates the behavioral state of every individual in a group of two or more animals while factoring in its social and environmental surroundings. We demonstrate the performance of subject-aware deep-learning in different species and assays, from partner preference in freely-moving insects to primate social interactions in the field. Our subject-aware deep learning approach provides a controllable, interpretable, and efficient framework to enable new experimental paradigms and systematic evaluation of interactive behavior in individuals identified within a group.
]]></description>
<dc:creator>Goss, K.</dc:creator>
<dc:creator>Bueno-Junior, L. S.</dc:creator>
<dc:creator>Stangis, K.</dc:creator>
<dc:creator>Ardoin, T.</dc:creator>
<dc:creator>Carmon, H.</dc:creator>
<dc:creator>Zhou, J.</dc:creator>
<dc:creator>Satapathy, R.</dc:creator>
<dc:creator>Baker, I.</dc:creator>
<dc:creator>Jones-Tinsley, C. E.</dc:creator>
<dc:creator>Lim, M. M.</dc:creator>
<dc:creator>Watson, B. O.</dc:creator>
<dc:creator>Sueur, C.</dc:creator>
<dc:creator>Ferrario, C. R.</dc:creator>
<dc:creator>Murphy, G. G.</dc:creator>
<dc:creator>Ye, B.</dc:creator>
<dc:creator>Hu, Y.</dc:creator>
<dc:date>2024-07-10</dc:date>
<dc:identifier>doi:10.1101/2024.07.07.602350</dc:identifier>
<dc:title><![CDATA[Quantifying social roles in multi-animal videos using subject-aware deep-learning]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-07-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.07.08.602530v1?rss=1">
<title>
<![CDATA[
Goal-directed action preparation in humans entails a mixture of corticospinal neural computations 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.07.08.602530v1?rss=1</link>
<description><![CDATA[
The seemingly effortless ability of humans to transition from thinking about actions to initiating them relies on sculpting corticospinal output from primary motor cortex. This study tested whether canonical additive and multiplicative neural computations, well-described in sensory systems, generalize to the corticospinal pathway during human action preparation. We used non-invasive brain stimulation to measure corticospinal input-output across varying action preparation contexts during instructed-delay finger response tasks. Goal-directed action preparation was marked by increased multiplicative gain of corticospinal projections to task-relevant muscles and additive suppression of corticospinal projections to non-selected and task-irrelevant muscles. Individuals who modulated corticospinal gain to a greater extent were faster to initiate prepared responses. Our findings provide physiological evidence of combined additive suppression and gain modulation in the human motor system. We propose these computations support action preparation by enhancing the contrast between selected motor representations and surrounding background activity to facilitate response selection and execution.



O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=108 SRC="FIGDIR/small/602530v4_figa1.gif" ALT="Figure 1">
View larger version (25K):
org.highwire.dtl.DTLVardef@9f2444org.highwire.dtl.DTLVardef@5f6eforg.highwire.dtl.DTLVardef@196f41corg.highwire.dtl.DTLVardef@1746035_HPS_FORMAT_FIGEXP  M_FIG O_FLOATNOC_FLOATNO Goal-directed action preparation shapes corticospinal output across selected, nonselected, and task-irrelevant motor representations. This study examined whether additive and multiplicative neural computations, common in sensory systems, occur within the corticospinal pathway during action preparation. We probed corticospinal input-output during the performance of various instructed-delay response tasks by applying a range of transcranial magnetic stimulation (TMS) intensities (input) over the primary motor cortex and measuring the resultant motor-evoked potentials (output) from the hand. We found that goal-directed action preparation increases corticospinal gain multiplicatively in task-relevant motor representations while additively suppressing nonselected and irrelevant representations. Greater gain modulation predicted faster responses, highlighting how these computations can enhance signal-to-noise (SNR) to enable efficient action selection and execution in the human motor system.

C_FIG Key pointsO_LINeural computations determine what information is transmitted through brain circuits.
C_LIO_LIWe investigated whether the motor system uses computations similar to those observed in sensory systems by noninvasively stimulating the corticospinal pathway in humans during goal-directed action preparation.
C_LIO_LIWe discovered physiological evidence that corticospinal projections to behaviorally relevant muscles exhibit nonlinear gain computations, while projections to behaviorally irrelevant muscles exhibit linear suppression.
C_LIO_LIOur findings suggest that certain computational principles generalize to the human motor system and serve to enhance the contrast between relevant and background neural activity.
C_LIO_LIThese results indicate that neural computations during goal-directed action preparation may support motor control by increasing signal-to-noise within the corticospinal pathway.
C_LI
]]></description>
<dc:creator>Wadsley, C. G.</dc:creator>
<dc:creator>Nguyen, T.</dc:creator>
<dc:creator>Horton, C.</dc:creator>
<dc:creator>Greenhouse, I.</dc:creator>
<dc:date>2024-07-11</dc:date>
<dc:identifier>doi:10.1101/2024.07.08.602530</dc:identifier>
<dc:title><![CDATA[Goal-directed action preparation in humans entails a mixture of corticospinal neural computations]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-07-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.07.10.602558v1?rss=1">
<title>
<![CDATA[
Mild neonatal hypoxia disrupts adult hippocampal learning and memory and is associated with CK2-mediated dysregulation of synaptic calcium-activated potassium channel KCNN2 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.07.10.602558v1?rss=1</link>
<description><![CDATA[
ObjectiveAlthough nearly half of preterm survivors display persistent neurobehavioral dysfunction including memory impairment without overt gray matter injury, the underlying mechanisms of neuronal or glial dysfunction, and their relationship to commonly observed cerebral white matter injury are unclear. We developed a mouse model to test the hypothesis that mild hypoxia during preterm equivalence is sufficient to persistently disrupt hippocampal neuronal maturation related to adult cellular mechanisms of learning and memory.

Methods: Neonatal (P2) mice were exposed to mild hypoxia (8%O2) for 30 min and evaluated for acute injury responses or survived until adulthood for assessment of learning and memory and hippocampal neurodevelopment.

ResultsNeonatal mild hypoxia resulted in clinically relevant oxygen desaturation and tachycardia without bradycardia and was not accompanied by cerebral gray or white matter injury. Neonatal hypoxia exposure was sufficient to cause hippocampal learning and memory deficits and abnormal maturation of CA1 neurons that persisted into adulthood. This was accompanied by reduced hippocampal CA3-CA1 synaptic strength and LTP and reduced synaptic activity of calcium-sensitive SK2 channels, key regulators of spike timing dependent neuroplasticity, including LTP. Structural illumination microscopy revealed reduced synaptic density, but intact SK2 localization at the synapse. Persistent loss of SK2 activity was mediated by altered casein kinase 2 (CK2) signaling.

InterpretationClinically relevant mild hypoxic exposure in the neonatal mouse is sufficient to produce morphometric and functional disturbances in hippocampal neuronal maturation independently of white matter injury. Additionally, we describe a novel persistent mechanism of potassium channel dysregulation after neonatal hypoxia. Collectively our findings suggest an unexplored explanation for the broad spectrum of neurobehavioral, cognitive and learning disabilities that paradoxically persist into adulthood without overt gray matter injury after preterm birth.
]]></description>
<dc:creator>Riddle, A.</dc:creator>
<dc:creator>Srivastava, T.</dc:creator>
<dc:creator>Wang, K.</dc:creator>
<dc:creator>Tellez, E.</dc:creator>
<dc:creator>O'neill, H.</dc:creator>
<dc:creator>Gong, X.</dc:creator>
<dc:creator>O'Neil, A.</dc:creator>
<dc:creator>Bell, J. A.</dc:creator>
<dc:creator>Raber, J.</dc:creator>
<dc:creator>Lattal, M.</dc:creator>
<dc:creator>Maylie, J.</dc:creator>
<dc:creator>Back, S. A.</dc:creator>
<dc:date>2024-07-16</dc:date>
<dc:identifier>doi:10.1101/2024.07.10.602558</dc:identifier>
<dc:title><![CDATA[Mild neonatal hypoxia disrupts adult hippocampal learning and memory and is associated with CK2-mediated dysregulation of synaptic calcium-activated potassium channel KCNN2]]></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/10.1101/2024.07.17.604013v1?rss=1">
<title>
<![CDATA[
Cytoplasmic ribosomes on mitochondria alter the local membrane environment for protein import 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.07.17.604013v1?rss=1</link>
<description><![CDATA[
Most of the mitochondria proteome is nuclear-encoded, synthesized by cytoplasmic ribosomes, and targeted to mitochondria post-translationally. However, a subset of mitochondrial-targeted proteins is imported co-translationally, although the molecular mechanisms governing this process remain unclear. We employ cellular cryo-electron tomography to visualize interactions between cytoplasmic ribosomes and mitochondria in Saccharomyces cerevisiae. We use surface morphometrics tools to identify a subset of ribosomes optimally oriented on mitochondrial membranes for protein import. This allows us to establish the first subtomogram average structure of a cytoplasmic ribosome on the surface of the mitochondria in the native cellular context, which showed three distinct connections with the outer mitochondrial membrane surrounding the peptide exit tunnel. Further, this analysis demonstrated that cytoplasmic ribosomes primed for mitochondrial protein import cluster on the outer mitochondrial membrane at sites of local constrictions of the outer and inner mitochondrial membrane. Overall, our study reveals the architecture and the spatial organization of cytoplasmic ribosomes at the mitochondrial surface, providing a native cellular context to define the mechanisms that mediate efficient mitochondrial co-translational protein import.

SUMMARYChang et al. present a membrane-guided approach for identifying a subset of cytoplasmic ribosomes oriented for protein import on the mitochondrial surface in Saccharomyces cerevisiae using cryo-electron tomography. They show that ribosomes cluster, make multiple contacts with, and induce local changes to the mitochondrial membrane ultrastructure at import sites.
]]></description>
<dc:creator>Chang, Y.-T.</dc:creator>
<dc:creator>Barad, B. A.</dc:creator>
<dc:creator>Rahmani, H.</dc:creator>
<dc:creator>Zid, B. M.</dc:creator>
<dc:creator>Grotjahn, D. A.</dc:creator>
<dc:date>2024-07-19</dc:date>
<dc:identifier>doi:10.1101/2024.07.17.604013</dc:identifier>
<dc:title><![CDATA[Cytoplasmic ribosomes on mitochondria alter the local membrane environment for protein import]]></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/10.1101/2024.07.19.604275v1?rss=1">
<title>
<![CDATA[
Quantifiable Blood TCR Repertoire Components Associated with Immune Aging 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.07.19.604275v1?rss=1</link>
<description><![CDATA[
T cell senescence results in decayed adaptive immune protection in older individuals, with decreased or increased abundance of certain T cell phenotypic subpopulations. However, no study has linked aging to the dynamic changes of T cell clones. Through a newly develop computational framework, Repertoire Functional Units (RFU), we investigated over 6,500 TCR repertoire sequencing samples from multiple human cohorts. Our analysis identified age-associated RFUs repeatedly and consistently across different cohorts. Quantification of RFU decreases with aging revealed accelerated loss under immunosuppressive conditions. Systematic analysis of age-associated RFUs in clinical samples manifested a potential link between these RFUs and improved clinical outcomes during acute viral infections, such as lower ICU admission and reduced risk of developing complications. Finally, our investigation of bone-marrow transplantation patients indicated a secondary expansion of the age-associated clones upon receiving stem cells from younger donors. Together, our results suggest the existence of certain clones or a  TCR clock that could reflect the immune functions in aging populations.
]]></description>
<dc:creator>Li, B.</dc:creator>
<dc:creator>Hu, J.</dc:creator>
<dc:creator>Pan, M.</dc:creator>
<dc:creator>Reid, B.</dc:creator>
<dc:creator>Tworoger, S.</dc:creator>
<dc:date>2024-07-19</dc:date>
<dc:identifier>doi:10.1101/2024.07.19.604275</dc:identifier>
<dc:title><![CDATA[Quantifiable Blood TCR Repertoire Components Associated with Immune Aging]]></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/10.1101/2024.07.17.603957v1?rss=1">
<title>
<![CDATA[
Spike inference from mouse spinal cord calcium imaging data 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.07.17.603957v1?rss=1</link>
<description><![CDATA[
Calcium imaging is a key method to record the spiking activity of identified and genetically targeted neurons. However, the observed calcium signals are only an indirect readout of the underlying electrophysiological events (single spikes or bursts of spikes) and require dedicated algorithms to recover the spike rate. These algorithms for spike inference can be optimized using ground truth data from combined electrical and optical recordings, but it is not clear how such optimized algorithms perform on cell types and brain regions for which ground truth does not exist. Here, we use a state-of-the-art algorithm based on supervised deep learning (CASCADE) and a non-supervised algorithm based on non-negative deconvolution (OASIS) to test spike rate inference in spinal cord neurons. To enable these tests, we recorded specific ground truth from glutamatergic and GABAergic somatosensory neurons in the superficial dorsal horn of spinal cord in mice of both sexes. We find that CASCADE and OASIS algorithms that were designed for cortical excitatory neurons generalize well to both spinal cord cell types. However, CASCADE models re-trained on our ground truth further improved the performance, resulting in a more accurate inference of spiking activity from spinal cord neurons. We openly provide re-trained models that can be applied to spinal cord data of variable noise levels and frame rates. Together, our ground-truth recordings and analyses provide a solid foundation for the interpretation of calcium imaging data from spinal cord dorsal horn and showcase how spike rate inference can generalize between different regions of the nervous system.

Significance StatementCalcium imaging is a powerful method for measuring the activity of genetically identified neurons. However, accurate interpretation of calcium transients depends on having a detailed understanding of how neuronal activity correlates with fluorescence. Such calibration recordings have been performed for cerebral cortex but not yet for most other CNS regions and neuron types. Here, we obtained ground truth data in spinal cord by conducting simultaneous calcium and electrophysiology recordings in excitatory and inhibitory neurons. We systematically investigated the transferability of cortical algorithms to spinal neuron subpopulations and generated inference algorithms optimized to excitatory and inhibitory neurons. Our study provides a foundation for the rigorous interpretation of calcium imaging data from spinal cord.

Conflict of interest statementThe authors declare no competing financial conflicts of interest.
]]></description>
<dc:creator>Rupprecht, P.</dc:creator>
<dc:creator>Fan, W.</dc:creator>
<dc:creator>Sullivan, S. J.</dc:creator>
<dc:creator>Helmchen, F.</dc:creator>
<dc:creator>Sdrulla, A.</dc:creator>
<dc:date>2024-07-22</dc:date>
<dc:identifier>doi:10.1101/2024.07.17.603957</dc:identifier>
<dc:title><![CDATA[Spike inference from mouse spinal cord calcium imaging data]]></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/10.1101/2024.07.22.604443v1?rss=1">
<title>
<![CDATA[
Smooth Muscle Cells-Expressed Bmal1 Regulates Vascular Calcification Independent of the Canonical Circadian Pathway 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.07.22.604443v1?rss=1</link>
<description><![CDATA[
Vascular calcification is a major cardiovascular issue that increases morbidity and mortality in diabetes patients. While dysregulation of the circadian master regulator Basic Helix-Loop-Helix ARNT-Like Protein 1 (Bmal1) in vascular smooth muscle cells (VSMC) under diabetic conditions has been suggested, its role in vascular calcification is unclear. In VSMC, Bmal1 was upregulated under high glucose treatment and in aortic tissues from a diabetic mouse model. RNA sequencing from isolated VSMC between Bmal1 deletion and wildtype mice indicated Bmal1s pro-calcification role. Indeed, reduced levels of the osteogenic master regulator, Runt-Related Transcription Factor 2 (Runx2), were found in Bmal1 deletion VSMC under diabetic conditions. Alizarin red staining showed reduced calcification in Bmal1 deletion VSMC in vitro and vascular rings ex vivo. Furthermore, in a diabetic mouse model, SMC-Bmal1 deletion showed reduced calcium deposition in aortas. Collectively, diabetes-upregulated circadian regulator Bmal1 in VSMC contributes to vascular calcification. Maintaining normal circadian regulation may improve vascular health in diabetes.
]]></description>
<dc:creator>He, M.</dc:creator>
<dc:creator>Sun, Y.</dc:creator>
<dc:creator>Huang, F.</dc:creator>
<dc:creator>Zhu, Z.</dc:creator>
<dc:creator>Velazquez-Miranda, E.</dc:creator>
<dc:creator>Chong, Z.</dc:creator>
<dc:creator>Chen, Y.</dc:creator>
<dc:date>2024-07-22</dc:date>
<dc:identifier>doi:10.1101/2024.07.22.604443</dc:identifier>
<dc:title><![CDATA[Smooth Muscle Cells-Expressed Bmal1 Regulates Vascular Calcification Independent of the Canonical Circadian Pathway]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-07-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.07.19.604387v1?rss=1">
<title>
<![CDATA[
A high-performance genetically encoded sensor for cellular imaging of PKC activity in vivo 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.07.19.604387v1?rss=1</link>
<description><![CDATA[
We report a genetically encoded fluorescence lifetime sensor for protein kinase C (PKC) activity, named CKAR3, based on Forster resonance energy transfer. CKAR3 exhibits a 10-fold increased dynamic range compared to its parental sensors and enables in vivo imaging of PKC activity during animal behavior. Our results reveal robust PKC activity in a sparse neuronal subset in the motor cortex during locomotion, in part mediated by muscarinic acetylcholine receptors.
]]></description>
<dc:creator>Yahiro, T.</dc:creator>
<dc:creator>Bayless-Edwards, L.</dc:creator>
<dc:creator>Jones, J. A.</dc:creator>
<dc:creator>Ma, L.</dc:creator>
<dc:creator>Qin, M.</dc:creator>
<dc:creator>Mao, T.</dc:creator>
<dc:creator>Zhong, H.</dc:creator>
<dc:date>2024-07-23</dc:date>
<dc:identifier>doi:10.1101/2024.07.19.604387</dc:identifier>
<dc:title><![CDATA[A high-performance genetically encoded sensor for cellular imaging of PKC activity in vivo]]></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/10.1101/2024.07.24.605016v1?rss=1">
<title>
<![CDATA[
Changes in maternal blood and placental lipidomic profile in obesity and gestational diabetes: Evidence for sexual dimorphism 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.07.24.605016v1?rss=1</link>
<description><![CDATA[
IntroductionObesity and gestational diabetes (GDM) are associated with adverse pregnancy outcomes and program the offspring for cardiometabolic disease in a sexually dimorphic manner. The placenta transfers lipids to the fetus and uses these substrates to support its own metabolism impacting the amount of substrate available to the growing fetus.

MethodsWe collected maternal plasma and placental villous tissue following elective cesarean section at term from women who were lean (pre-pregnancy BMI 18.5-24.9), obese (BMI>30) and type A2 GDM (matched to obese BMI) with male or female fetus (n=4 each group). Lipids were extracted and fatty acid composition of different lipid classes were analyzed by LC-MS/MS analysis. Significant changes in GDM vs obese, GDM vs lean, and obese vs lean were determined using t-test with a Tukey correction set at p<0.05.

ResultsIn placental samples 436 lipids were identified, among which 85 showed significant changes. Of note only in male placentas significant decreases in C22:6 - docosahexaenoic acid (DHA) in phosphatidylcholine (PC) and triglyceride lipid species were seen when comparing tissue from GDM women to lean. In maternal plasma we observed no effect of obesity. GDM or fetal sex.

ConclusionThis is the first study assessing fatty acid composition of lipids in matched maternal plasma and placental tissue from lean, obese, and GDM women stratified by fetal sex. It highlights how GDM affects distribution of fatty acids in lipid classes changes in a sexually dimorphic manner in the placenta.
]]></description>
<dc:creator>Kadam, L.</dc:creator>
<dc:creator>Velickovic, M.</dc:creator>
<dc:creator>Straton, K.</dc:creator>
<dc:creator>Nicora, C. D.</dc:creator>
<dc:creator>Kyle, J.</dc:creator>
<dc:creator>Wang, E.</dc:creator>
<dc:creator>Monroe, M.</dc:creator>
<dc:creator>Bramer, L.</dc:creator>
<dc:creator>Myatt, L.</dc:creator>
<dc:creator>Burnum-Johnson, K.</dc:creator>
<dc:date>2024-07-25</dc:date>
<dc:identifier>doi:10.1101/2024.07.24.605016</dc:identifier>
<dc:title><![CDATA[Changes in maternal blood and placental lipidomic profile in obesity and gestational diabetes: Evidence for sexual dimorphism]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-07-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.07.25.605151v1?rss=1">
<title>
<![CDATA[
Human P2X4 receptor gating is modulated by a stable cytoplasmic cap and a unique allosteric pocket 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.07.25.605151v1?rss=1</link>
<description><![CDATA[
P2X receptors (P2XRs) are a family of ATP-gated ion channels comprising homomeric and heteromeric trimers of seven subunits (P2X1 - P2X7) that confer different rates of desensitization. The helical recoil model of P2XR desensitization proposes the stability of the cytoplasmic cap sets the rate of desensitization, but timing of its formation is unclear for slow-desensitizing P2XRs. We report cryo-EM structures of full-length, wild-type human P2X4 receptor in apo, antagonist-bound, and desensitized states. Because the apo and antagonist-bound structures of this slow-desensitizing P2XR include an intact cytoplasmic cap while the desensitized state structure does not, the cytoplasmic cap forms before agonist binding. Furthermore, structural and functional data suggests the cytoplasmic cap is stabilized by lipids to slow desensitization and that P2X4 is further modified by glycosylation and palmitoylation. Finally, our antagonist-bound inhibited state structure reveals features specific to the allosteric ligand-binding pocket in human receptors that empower the development of small-molecule modulators.
]]></description>
<dc:creator>Shi, H.</dc:creator>
<dc:creator>Ditter, I. A.</dc:creator>
<dc:creator>Oken, A. C.</dc:creator>
<dc:creator>Mansoor, S. E.</dc:creator>
<dc:date>2024-07-25</dc:date>
<dc:identifier>doi:10.1101/2024.07.25.605151</dc:identifier>
<dc:title><![CDATA[Human P2X4 receptor gating is modulated by a stable cytoplasmic cap and a unique allosteric pocket]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-07-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.07.29.605485v1?rss=1">
<title>
<![CDATA[
Stromal KITL/SCF promotes pancreas tissue homeostasis and restrains tumor progression 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.07.29.605485v1?rss=1</link>
<description><![CDATA[
Components of normal tissue architecture serve as barriers to tumor progression. Inflammatory and wound-healing programs are requisite features of solid tumorigenesis, wherein alterations to immune and non-immune stromal elements enable loss of homeostasis during tumor evolution. The precise mechanisms by which normal stromal cell states limit tissue plasticity and tumorigenesis, and which are lost during tumor progression, remain largely unknown. Here we show that healthy pancreatic mesenchyme expresses the paracrine signaling molecule KITL, also known as stem cell factor, and identify loss of stromal KITL during tumorigenesis as tumor-promoting. Genetic inhibition of mesenchymal KITL in the contexts of homeostasis, injury, and cancer together indicate a role for KITL signaling in maintenance of pancreas tissue architecture, such that loss of the stromal KITL pool increased tumor growth and reduced survival of tumor-bearing mice. Together, these findings implicate loss of mesenchymal KITL as a mechanism for establishing a tumor-permissive microenvironment.

Statement of significanceBy analyzing transcriptional programs in healthy and tumor-associated pancreatic mesenchyme, we find that a sub-population of mesenchymal cells in healthy pancreas tissue express the paracrine signaling factor KITL. Loss of mesenchymal KITL is an accompanying and permissive feature of pancreas tumor evolution, with potential implications for cancer interception.
]]></description>
<dc:creator>Onate, M. K.</dc:creator>
<dc:creator>Oon, C.</dc:creator>
<dc:creator>Bhattacharyya, S.</dc:creator>
<dc:creator>Low, V.</dc:creator>
<dc:creator>Chen, C.</dc:creator>
<dc:creator>Zhao, X.</dc:creator>
<dc:creator>Yan, Z.</dc:creator>
<dc:creator>Hang, Y.</dc:creator>
<dc:creator>Kim, S. K.</dc:creator>
<dc:creator>Xia, Z.</dc:creator>
<dc:creator>Sherman, M. H.</dc:creator>
<dc:date>2024-07-30</dc:date>
<dc:identifier>doi:10.1101/2024.07.29.605485</dc:identifier>
<dc:title><![CDATA[Stromal KITL/SCF promotes pancreas tissue homeostasis and restrains tumor progression]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-07-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.07.31.605654v1?rss=1">
<title>
<![CDATA[
Complete sequencing of ape genomes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.07.31.605654v1?rss=1</link>
<description><![CDATA[
We present haplotype-resolved reference genomes and comparative analyses of six ape species, namely: chimpanzee, bonobo, gorilla, Bornean orangutan, Sumatran orangutan, and siamang. We achieve chromosome-level contiguity with unparalleled sequence accuracy (<1 error in 500,000 base pairs), completely sequencing 215 gapless chromosomes telomere-to-telomere. We resolve challenging regions, such as the major histocompatibility complex and immunoglobulin loci, providing more in-depth evolutionary insights. Comparative analyses, including human, allow us to investigate the evolution and diversity of regions previously uncharacterized or incompletely studied without bias from mapping to the human reference. This includes newly minted gene families within lineage-specific segmental duplications, centromeric DNA, acrocentric chromosomes, and subterminal heterochromatin. This resource should serve as a definitive baseline for all future evolutionary studies of humans and our closest living ape relatives.
]]></description>
<dc:creator>Yoo, D.</dc:creator>
<dc:creator>Rhie, A.</dc:creator>
<dc:creator>Hebbar, P.</dc:creator>
<dc:creator>Antonacci, F.</dc:creator>
<dc:creator>Logsdon, G. A.</dc:creator>
<dc:creator>Solar, S. J.</dc:creator>
<dc:creator>Antipov, D.</dc:creator>
<dc:creator>Pickett, B. D.</dc:creator>
<dc:creator>Safonova, Y.</dc:creator>
<dc:creator>Montinaro, F.</dc:creator>
<dc:creator>Luo, Y.</dc:creator>
<dc:creator>Malukiewicz, J.</dc:creator>
<dc:creator>Storer, J. M.</dc:creator>
<dc:creator>Lin, J.</dc:creator>
<dc:creator>Sequeira, A. N.</dc:creator>
<dc:creator>Mangan, R. J.</dc:creator>
<dc:creator>Hickey, G.</dc:creator>
<dc:creator>Anez, G. M.</dc:creator>
<dc:creator>Balachandran, P.</dc:creator>
<dc:creator>Bankevich, A.</dc:creator>
<dc:creator>Beck, C. R.</dc:creator>
<dc:creator>Biddanda, A.</dc:creator>
<dc:creator>Borchers, M.</dc:creator>
<dc:creator>Bouffard, G. G.</dc:creator>
<dc:creator>Brannan, E.</dc:creator>
<dc:creator>Brooks, S. Y.</dc:creator>
<dc:creator>Carbone, L.</dc:creator>
<dc:creator>Carrel, L.</dc:creator>
<dc:creator>Chan, A. P.</dc:creator>
<dc:creator>Crawford, J.</dc:creator>
<dc:creator>Diekhans, M.</dc:creator>
<dc:creator>Engelbrecht, E.</dc:creator>
<dc:creator>Feschotte, C.</dc:creator>
<dc:creator>Formenti, G.</dc:creator>
<dc:creator>Garcia, G. H.</dc:creator>
<dc:creator>Gennaro, L. d.</dc:creator>
<dc:creator>Gilbert, D.</dc:creator>
<dc:creator>Green, R. E.</dc:creator>
<dc:creator>Guarracino, A.</dc:creator>
<dc:creator>Gupta, I.</dc:creator>
<dc:creator>Haddad, D.</dc:creator>
<dc:creator>Han, J.</dc:creator>
<dc:creator>Harris, R. S.</dc:creator>
<dc:creator>Hartley</dc:creator>
<dc:date>2024-07-31</dc:date>
<dc:identifier>doi:10.1101/2024.07.31.605654</dc:identifier>
<dc:title><![CDATA[Complete sequencing of ape genomes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-07-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.07.31.606010v1?rss=1">
<title>
<![CDATA[
Comparative lifespan and healthspan of nonhuman primate species common to biomedical research 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.07.31.606010v1?rss=1</link>
<description><![CDATA[
There is a critical need to generate age- and sex-specific survival curves to characterize chronological aging consistently across nonhuman primates (NHP) used in biomedical research. Sex-specific Kaplan-Meier survival curves were computed in 12 translational aging models: baboon, bonnet macaque, chimpanzee, common marmoset, coppery titi monkey, cotton-top tamarin, cynomolgus macaque, Japanese macaque, pigtail macaque, rhesus macaque, squirrel monkey, and vervet/African green. After employing strict inclusion criteria, primary results are based on 12,269 NHP that survived to adulthood and died of natural/health-related causes. A secondary analysis was completed for 32,616 NHP that died of any cause. Results show a pattern of reduced male survival among catarrhines (African and Asian primates), especially macaques, but not platyrrhines (Central and South American primates). For many species, median lifespans were lower than previously reported. An important consideration is that these analyses may offer a better reflection of healthspan than lifespan since research NHP are typically euthanized for humane welfare reasons before their natural end of life. This resource represents the most comprehensive characterization of sex-specific lifespan and age-at-death distributions for 12 biomedically relevant species, to date. These results clarify relationships among NHP ages and provide a valuable resource for the aging research community, improving human-NHP age equivalencies, informing investigators of expected survival rates, providing a metric for comparisons in future studies, and contributing to understanding of factors driving lifespan differences within and among species.
]]></description>
<dc:creator>Huber, H. F.</dc:creator>
<dc:creator>Ainsworth, H. C.</dc:creator>
<dc:creator>Quillen, E. E.</dc:creator>
<dc:creator>Salmon, A.</dc:creator>
<dc:creator>Ross, C.</dc:creator>
<dc:creator>Azhar, A. D.</dc:creator>
<dc:creator>Bales, K.</dc:creator>
<dc:creator>Basso, M. A.</dc:creator>
<dc:creator>Coleman, K.</dc:creator>
<dc:creator>Colman, R.</dc:creator>
<dc:creator>Darusman, H. S.</dc:creator>
<dc:creator>Hopkins, B.</dc:creator>
<dc:creator>Hotchkiss, C. E.</dc:creator>
<dc:creator>Jorgensen, M. J.</dc:creator>
<dc:creator>Kavanagh, K.</dc:creator>
<dc:creator>Li, C.</dc:creator>
<dc:creator>Mattison, J. A.</dc:creator>
<dc:creator>Nathanielsz, P. W.</dc:creator>
<dc:creator>Saputro, S.</dc:creator>
<dc:creator>Scorpio, D. G.</dc:creator>
<dc:creator>Sosa, P.-M.</dc:creator>
<dc:creator>Vallender, E. J.</dc:creator>
<dc:creator>Wang, Y.</dc:creator>
<dc:creator>Zeiss, C. J.</dc:creator>
<dc:creator>Shively, C. A.</dc:creator>
<dc:creator>Cox, L. A.</dc:creator>
<dc:date>2024-08-01</dc:date>
<dc:identifier>doi:10.1101/2024.07.31.606010</dc:identifier>
<dc:title><![CDATA[Comparative lifespan and healthspan of nonhuman primate species common to biomedical research]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-08-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.08.13.607670v1?rss=1">
<title>
<![CDATA[
Single-cell DNA methylation analysis tool Amethyst reveals distinct noncanonical methylation patterns in human glial cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.08.13.607670v1?rss=1</link>
<description><![CDATA[
Single-cell sequencing technologies have revolutionized biomedical research by enabling deconvolution of cell type-specific properties in highly heterogeneous tissue. While robust tools have been developed to handle bioinformatic challenges posed by single-cell RNA and ATAC data, options for emergent modalities such as methylation are much more limited, impeding the utility of results. Here we present Amethyst, a comprehensive R package for atlas-scale single-cell methylation sequencing data analysis. Amethyst begins with base-level methylation calls and expedites batch integration, doublet detection, dimensionality reduction, clustering, cell type annotation, differentially methylated region calling, and interpretation of results, facilitating rapid data interaction in a local environment. We introduce the workflow using published single-cell methylation human peripheral blood mononuclear cell (PBMC) and human cortex data. We further leverage Amethyst on an atlas-scale brain dataset to describe a noncanonical methylation pattern in human astrocytes and oligodendrocytes, challenging the notion that this form of methylation is principally relevant to neurons in the brain. Tools such as Amethyst will increase accessibility to single-cell methylation data analysis, catalyzing research progress across diverse contexts.
]]></description>
<dc:creator>Rylaarsdam, L. E.</dc:creator>
<dc:creator>Nichols, R. V.</dc:creator>
<dc:creator>O'Connell, B. L.</dc:creator>
<dc:creator>Coleman, S.</dc:creator>
<dc:creator>Yardimci, G.</dc:creator>
<dc:creator>Adey, A. C.</dc:creator>
<dc:date>2024-08-16</dc:date>
<dc:identifier>doi:10.1101/2024.08.13.607670</dc:identifier>
<dc:title><![CDATA[Single-cell DNA methylation analysis tool Amethyst reveals distinct noncanonical methylation patterns in human glial cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-08-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.08.14.607951v1?rss=1">
<title>
<![CDATA[
An epigenetic memory at the CYP1A gene in cancer-resistant, pollution-adapted killifish 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.08.14.607951v1?rss=1</link>
<description><![CDATA[
Human exposure to polycyclic aromatic hydrocarbons (PAH) is a significant and growing public health problem. Frequent, high dose exposures are likely to increase due to a warming climate and increased frequency of large-scale wildfires. Here, we characterize an epigenetic memory at the cytochrome P450 1A (CYP1A) gene in a population of wild Fundulus heteroclitus that has adapted to chronic, extreme PAH pollution. In wild-type fish, CYP1A is highly induced by PAH. In PAH-tolerant fish, CYP1A induction is blunted. Since CYP1A metabolically activates PAH, this memory protects these fish from PAH-mediated cancer. However, PAH-tolerant fish reared in clean water recover CYP1A inducibility, indicating that blunted induction is a non-genetic memory of prior exposure. To explore this possibility, we bred depurated wild fish from PAH-sensitive and -tolerant populations, manually fertilized exposure-naive embryos, and challenged them with PAH. We observed epigenetic control of the reversible memory of generational PAH stress in F1 PAH-tolerant embryos. Specifically, we observed a bivalent domain in the CYP1A promoter enhancer comprising both activating and repressive histone post-translational modifications. Activating modifications, relative to repressive ones, showed greater increases in response to PAH in sensitive embryos, relative to tolerant, consistent with greater gene activation. Also, PAH-tolerant adult fish showed persistent induction of CYP1A long after exposure cessation, which is consistent with defective CYP1A shutoff and recovery to baseline. Since CYP1A expression is inversely correlated with cancer risk, these results indicate that PAH-tolerant fish have epigenetic protection against PAH-induced cancer in early life that degrades in response to continuous gene activation.

SignificanceEpigenetic memory, or the inheritance across cell division within an organism or across generations, of environmental exposure response is a compelling phenomenon with limited understanding of mechanism. Here, we characterized an epigenetic memory at the CYP1A gene in pollution-adapted Fundulus heteroclitus. We found that the CYP1A promoter enhancer contains a bivalent domain, comprising both active and repressive histone modifications, that shows reduced function correlating with reduced gene induction by its pollutant activator. In early life, this memory protects fish against pollution-induced cancer. However, this reduced function carries a cost; adult fish show defective transcriptional recovery of CYP1A, which increases cancer risk later in life. These results provide an initial mechanism for a model epigenetic memory and highlight potential costs.
]]></description>
<dc:creator>Carrothers, S.</dc:creator>
<dc:creator>Trevisan, R.</dc:creator>
<dc:creator>Jayasundara, N.</dc:creator>
<dc:creator>Pelletier, N.</dc:creator>
<dc:creator>Weeks, E.</dc:creator>
<dc:creator>Meyer, J. N.</dc:creator>
<dc:creator>Di Giulio, R. T.</dc:creator>
<dc:creator>Weinhouse, C.</dc:creator>
<dc:date>2024-08-16</dc:date>
<dc:identifier>doi:10.1101/2024.08.14.607951</dc:identifier>
<dc:title><![CDATA[An epigenetic memory at the CYP1A gene in cancer-resistant, pollution-adapted killifish]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-08-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.08.15.608167v1?rss=1">
<title>
<![CDATA[
Traffic Jam activates the Flamenco piRNA cluster locus and the Piwi pathway to ensure transposon silencing and Drosophila fertility. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.08.15.608167v1?rss=1</link>
<description><![CDATA[
Flamenco (Flam) is the most prominent piRNA cluster locus expressed in Drosophila ovarian follicle cells, and it is required for female fertility to silence gypsy/mdg4 transposons. To determine how Flam is regulated, we used promoter-bashing reporter assays in OSS cells to uncover novel enhancer sequences within the first exons of Flam. We confirmed the enhancer sequence relevance in vivo with new Drosophila Flam deletion mutants of these regions that compromised Flam piRNA expression and lowered female fertility from activated transposons. Our proteomic analysis of proteins associated with these enhancer sequences discovered the transcription factor Traffic Jam (TJ). Tj knockdowns in OSS cells caused a decrease in Flam transcripts, Flam piRNAs, and multiple Piwi pathway genes. A TJ ChIP-seq analysis from whole flies and OSS cells confirmed TJ binding exactly at the enhancer that was deleted in the new Flam mutant as well as at multiple Piwi pathway gene enhancers. Interestingly, TJ also bound the Long Terminal Repeats of transposons that had decreased expression after Tj knockdowns in OSS cells. Our study reveals the integral role TJ plays in the on-going arms race between selfish transposons and their suppression by the host Piwi pathway and the Flam piRNA cluster locus.
]]></description>
<dc:creator>Rivera, A.</dc:creator>
<dc:creator>Lee, J.-H. R.</dc:creator>
<dc:creator>Gupta, S.</dc:creator>
<dc:creator>Yang, L.</dc:creator>
<dc:creator>Goel, R. K.</dc:creator>
<dc:creator>Zaia, J.</dc:creator>
<dc:creator>Lau, N. C.</dc:creator>
<dc:date>2024-08-16</dc:date>
<dc:identifier>doi:10.1101/2024.08.15.608167</dc:identifier>
<dc:title><![CDATA[Traffic Jam activates the Flamenco piRNA cluster locus and the Piwi pathway to ensure transposon silencing and Drosophila fertility.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-08-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.08.18.608482v1?rss=1">
<title>
<![CDATA[
Comparative Transcriptomes of Canine and Human Prostate Cancers Identify Mediators of Castration Resistance 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.08.18.608482v1?rss=1</link>
<description><![CDATA[
Prostate cancer continues to be one of the most lethal cancers in men. While androgen-deprivation therapy is initially effective in treating prostate cancer, most cases of advanced prostate cancer eventually progress to castration-resistant prostate cancer (CRPC), which is incurable. Similarly, the most aggressive form of prostatic carcinoma occurs in dogs that have been castrated. To identify molecular similarities between canine prostate cancer and human CRPC, we performed a comparative analysis of gene expression profiles. Through this transcriptomic analysis, we found that prostatic carcinoma in castrated dogs demonstrate an androgen indifferent phenotype, characterized by low androgen receptor and neuroendocrine associated genes. Notably, we identified two genes, ISG15 and AZGP1 that were consistently up- and downregulated, respectively, in both canine prostatic carcinoma and human CRPC. Additionally, we identified several other genes, including GPX3, S100P, and IFITM1, that exhibited similar expression patterns in both species. Protein-protein interaction network analysis demonstrated that these 5 genes were part of a larger network of interferon-induced genes, suggesting that they may act together in signaling pathways that are disrupted in prostate cancer. Accordingly, our findings suggest that the interferon pathway may play a role in the development and progression of CRPC in both dogs and humans and chart a new therapeutic approach.
]]></description>
<dc:creator>Angel, M. R.</dc:creator>
<dc:creator>Seguin, B.</dc:creator>
<dc:creator>Lohr, C. V.</dc:creator>
<dc:creator>Beer, T.</dc:creator>
<dc:creator>Feliciano, J.</dc:creator>
<dc:creator>Ramsey, S. A.</dc:creator>
<dc:creator>Thomas, G. V.</dc:creator>
<dc:date>2024-08-18</dc:date>
<dc:identifier>doi:10.1101/2024.08.18.608482</dc:identifier>
<dc:title><![CDATA[Comparative Transcriptomes of Canine and Human Prostate Cancers Identify Mediators of Castration Resistance]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-08-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.08.14.607971v1?rss=1">
<title>
<![CDATA[
Aquaporin-4 mis-localization slows glymphatic clearance of α-synuclein and promotes α-synuclein pathology and aggregate propagation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.08.14.607971v1?rss=1</link>
<description><![CDATA[
The appearance of misfolded and aggregated proteins is a pathological hallmark of numerous neurodegenerative diseases including Alzheimers disease and Parkinsons disease. Sleep disruption is proposed to contribute to these pathological processes and is a common early feature among neurodegenerative disorders. Synucleinopathies are a subclass of neurodegenerative conditions defined by the presence of -synuclein aggregates, which may not only enhance cell death, but also contribute to disease progression by seeding the formation of additional aggregates in neighboring cells. The mechanisms driving intercellular transmission of aggregates remains unclear. We propose that disruption of sleep-active glymphatic function, caused by loss of precise perivascular AQP4 localization, inhibits -synuclein clearance and facilitates -synuclein propagation and seeding. We examined human post-mortem frontal cortex and found that neocortical -synuclein pathology was associated with AQP4 mis-localization throughout the gray matter. Using a transgenic mouse model lacking the adapter protein -syntrophin, we observed that loss of perivascular AQP4 localization impairs the glymphatic clearance of -synuclein from intersititial to cerebrospinal fluid. Using a mouse model of -synuclein propogation, using pre-formed fibril injection, we observed that loss of perivascular AQP4 localization increased -synuclein aggregates. Our results indicate -synuclein clearance and propagation are mediated by glymphatic function and that AQP4 mis-localization observed in the presence of human synucleinopathy may contribute to the development and propagation of Lewy body pathology in conditions such as Lewy Body Dementia and Parkinsons disease.

SummaryIn a human postmortem case series, we observe that neocortical Lewy body pathology is associated with mis-localization of the astroglial water channel aquaporin-4 (AQP4). In mice, -synuclein is cleared from the brain along perivascular pathways, while loss of perivascular AQP4 localization impairs glymphatic -synuclein clearance to the CSF. Furthermore, loss of perivascular AQP4 localization promotes the development and propagation of -synuclein aggregates.
]]></description>
<dc:creator>Braun, M.</dc:creator>
<dc:creator>Simon, M. J.</dc:creator>
<dc:creator>Jang, J.</dc:creator>
<dc:creator>Sanderson, K.</dc:creator>
<dc:creator>Swierz, J.</dc:creator>
<dc:creator>Sevao, M.</dc:creator>
<dc:creator>Pincus, A. B.</dc:creator>
<dc:creator>Schaser, A. J.</dc:creator>
<dc:creator>Elliott, J. E.</dc:creator>
<dc:creator>Lim, M. M.</dc:creator>
<dc:creator>Unni, V. K.</dc:creator>
<dc:creator>Schindler, A. G.</dc:creator>
<dc:creator>Keene, C. D.</dc:creator>
<dc:creator>Latimer, C. S.</dc:creator>
<dc:creator>Iliff, J.</dc:creator>
<dc:date>2024-08-19</dc:date>
<dc:identifier>doi:10.1101/2024.08.14.607971</dc:identifier>
<dc:title><![CDATA[Aquaporin-4 mis-localization slows glymphatic clearance of α-synuclein and promotes α-synuclein pathology and aggregate propagation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-08-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.08.15.608144v1?rss=1">
<title>
<![CDATA[
Sex differences in nucleus accumbens core circuitry engaged by binge-like ethanol drinking 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.08.15.608144v1?rss=1</link>
<description><![CDATA[
Growing parity in Alcohol Use Disorder (AUD) diagnoses in men and women necessitates consideration of sex as a biological variable. In humans and rodents, the nucleus accumbens core (NAcc) regulates alcohol binge drinking, a risk factor for developing AUD. We labeled NAcc inputs with a viral retrograde tracer and quantified whole-brain c-Fos to determine the regions and NAcc inputs differentially engaged in male and female mice during binge-like ethanol drinking. We found that binge-like ethanol drinking females had 129 brain areas with greater c-Fos than males. Moreover, ethanol engaged more NAcc inputs in binge-like ethanol drinking females (as compared with males), including GABAergic and glutamatergic inputs. Relative to water controls, ethanol increased network modularity and decreased connectivity in both sexes and did so more dramatically in males. These results demonstrate that early-stage binge-like ethanol drinking engages brain regions and NAcc-inputs and alters network dynamics in a sex-specific manner.

O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=86 SRC="FIGDIR/small/608144v1_ufig1.gif" ALT="Figure 1">
View larger version (24K):
org.highwire.dtl.DTLVardef@c47435org.highwire.dtl.DTLVardef@1a97d69org.highwire.dtl.DTLVardef@68b2cborg.highwire.dtl.DTLVardef@1b2453a_HPS_FORMAT_FIGEXP  M_FIG O_FLOATNOGraphical Summary:C_FLOATNO Binge-like ethanol drinking engages more regions and NAcc inputs in female relative to male mice.Left: Comparison of regions with both greater c-Fos expression and c-Fos+GFP colocalization in female relative to male ethanol drinking mice.

Right: NAcc inputs engaged by binge-like ethanol drinking compared to water drinking mice, sex collapsed. Thalamic (TH) regions include left parasubthalamic nucleus, left anteromedial nucleus of the thalamus, left central medial nucleus of the thalamus, left medial group of the dorsal thalamus, left subparafascicular nucleus, left peripeduncular nucleus, and right paraventricular nucleus of the thalamus. EW, Edinger-Westphal nucleus; GU, gustatory areas.

Bottom Middle: NAcc inputs with greater engagement in male than female ethanol drinking mice (left and right main olfactory bulbs (MOB).

Bottom Right: NAcc inputs with greater engagement in female than male ethanol drinking mice. Amygdala (AMY) regions include left anterior amygdalar area and left intercalated amygdalar area. Hippocampal (HPF) regions include right dentate gyrus, right Field CA1, right Field CA2, and right Field CA3. Hypothalamic (HY) regions include left and right lateral hypothalamic area, left and right periventricular hypothalamic nucleus, preoptic part, right dorsomedial nucleus of the hypothalamus, right posterior hypothalamic nucleus, and right ventromedial hypothalamic nucleus. Midbrain (MB) regions include left and right midbrain reticular nucleus, retrorubral area, left and right superior colliculus, motor related, left nucleus of the brachium of the inferior colliculus, left nucleus of the posterior commissure, left olivary pretectal nucleus, left posterior pretectal nucleus, left superior colliculus, sensory related, and right substantia nigra, reticular parts. Pontine (P) regions include left superior central nucleus raphe, left supratrigeminal nucleus, right nucleus raphe pontis, right pontine reticular nucleus, and right superior olivary complex. Thalamic (TH) regions include left and right lateral dorsal nucleus of the thalamus, right dorsal part of the lateral geniculate complex, right lateral posterior nucleus of the thalamus, and right parasubthalamic nucleus. CB, Cerebellum; MA, magnocellular nucleus; SSp-m, primary somatosensory cortex, mouth; Created with BioRender.com. See Table S18 for additional information.

C_FIG
]]></description>
<dc:creator>Chan, A. E.</dc:creator>
<dc:creator>Anderson, J. Q.</dc:creator>
<dc:creator>Grigsby, K. E.</dc:creator>
<dc:creator>Jensen, B. E.</dc:creator>
<dc:creator>Ryabinin, A. E.</dc:creator>
<dc:creator>Ozburn, A. R.</dc:creator>
<dc:date>2024-08-19</dc:date>
<dc:identifier>doi:10.1101/2024.08.15.608144</dc:identifier>
<dc:title><![CDATA[Sex differences in nucleus accumbens core circuitry engaged by binge-like ethanol drinking]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-08-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.08.17.608337v1?rss=1">
<title>
<![CDATA[
Biological sex and BMI influence the longitudinal evolution of adolescent and young adult MRI-visible perivascular spaces 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.08.17.608337v1?rss=1</link>
<description><![CDATA[
Background and PurposeAn association recently emerged between magnetic resonance imaging (MRI)-visible perivascular spaces (MV-PVS) with intracerebral solute clearance and neuroinflammation, in adults. However, it is unknown how MV-PVS change throughout adolescence and what factors influence MV-PVS volume and morphology. This study assesses the temporal evolution of MV-PVS volume in adolescents and young adults, and secondarily evaluates the relationship between MV-PVS, age, sex, and body mass index (BMI).

Materials and MethodsThis analysis included a 783 participant cohort from the longitudinal multicenter National Consortium on Alcohol and Neurodevelopment in Adolescence study that involved up to 6 imaging visits spanning 5 years. Healthy adolescents aged 12-21 years at study entry with at least two MRI scans were included. The primary outcome was mean MV-PVS volume (mm3/white matter cm3).

ResultsOn average, males had greater MV-PVS volume at all ages compared to females. A linear mixed-effect model for MV-PVS volume was performed. Mean BMI and increases in a persons BMI were associated with increases in MV-PVS volume over time. In females only, changes in BMI correlated with MV-PVS volume. One unit increase in BMI above a persons average BMI was associated with a 0.021 mm3/cm3 increase in MV-PVS volume (p<0.001).

ConclusionThis longitudinal study showed sex differences in MV-PVS features during adolescence and young adulthood. Importantly, we report that increases in BMI from a persons mean BMI are associated with increases in MV-PVS volume in females only. These findings suggest a potential link between MV-PVS, sex, and BMI that warrants future study.
]]></description>
<dc:creator>Yamamoto, E. A.</dc:creator>
<dc:creator>Koike, S.</dc:creator>
<dc:creator>Wong, C.</dc:creator>
<dc:creator>Dennis, L.</dc:creator>
<dc:creator>Luther, M. N.</dc:creator>
<dc:creator>Scatena, A.</dc:creator>
<dc:creator>Khambadkone, S.</dc:creator>
<dc:creator>Iliff, J.</dc:creator>
<dc:creator>Lim, M. M.</dc:creator>
<dc:creator>Levendovszky, S. R.</dc:creator>
<dc:creator>Elliott, J.</dc:creator>
<dc:creator>Barisano, G.</dc:creator>
<dc:creator>Muller-Oehring, E.</dc:creator>
<dc:creator>Morales, A. M.</dc:creator>
<dc:creator>Baker, F.</dc:creator>
<dc:creator>Nagel, B.</dc:creator>
<dc:creator>Piantino, J.</dc:creator>
<dc:date>2024-08-19</dc:date>
<dc:identifier>doi:10.1101/2024.08.17.608337</dc:identifier>
<dc:title><![CDATA[Biological sex and BMI influence the longitudinal evolution of adolescent and young adult MRI-visible perivascular spaces]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-08-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.08.19.608555v1?rss=1">
<title>
<![CDATA[
Towards Digital Quantification of Ploidy from Pan-Cancer Digital Pathology Slides using Deep Learning 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.08.19.608555v1?rss=1</link>
<description><![CDATA[
Abnormal DNA ploidy, found in numerous cancers, is increasingly being recognized as a contributor in driving chromosomal instability, genome evolution, and the heterogeneity that fuels cancer cell progression. Furthermore, it has been linked with poor prognosis of cancer patients. While next-generation sequencing can be used to approximate tumor ploidy, it has a high error rate for near-euploid states, a high cost and is time consuming, motivating alternative rapid quantification methods. We introduce PloiViT, a transformer-based model for tumor ploidy quantification that outperforms traditional machine learning models, enabling rapid and cost-effective quantification directly from pathology slides. We trained PloiViT on a dataset of fifteen cancer types from The Cancer Genome Atlas and validated its performance in multiple independent cohorts. Additionally, we explored the impact of self-supervised feature extraction on performance. PloiViT, using self-supervised features, achieved the lowest prediction error in multiple independent cohorts, exhibiting better generalization capabilities. Our findings demonstrate that PloiViT predicts higher ploidy values in aggressive cancer groups and patients with specific mutations, validating PloiViT potential as complementary for ploidy assessment to next-generation sequencing data. To further promote its use, we release our models as a user-friendly inference application and a Python package for easy adoption and use.
]]></description>
<dc:creator>Carrillo-Perez, F.</dc:creator>
<dc:creator>Cramer, E. M.</dc:creator>
<dc:creator>Pizurica, M.</dc:creator>
<dc:creator>Andor, N.</dc:creator>
<dc:creator>Gevaert, O.</dc:creator>
<dc:date>2024-08-20</dc:date>
<dc:identifier>doi:10.1101/2024.08.19.608555</dc:identifier>
<dc:title><![CDATA[Towards Digital Quantification of Ploidy from Pan-Cancer Digital Pathology Slides using Deep Learning]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-08-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.08.21.608907v1?rss=1">
<title>
<![CDATA[
CD4+ but not CD8+ T cells are required for protection against severe guinea pig cytomegalovirus infections 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.08.21.608907v1?rss=1</link>
<description><![CDATA[
Human cytomegalovirus (HCMV) is a ubiquitous herpesvirus and the leading cause of infectious disease related birth defects worldwide. How the immune response modulates the risk of intrauterine transmission of HCMV after maternal infection remains poorly understood. Maternal T cells likely play a critical role in preventing infection at the maternal-fetal interface and limiting spread across the placenta, but concerns exist that immune responses to infection may also cause placental dysfunction and adverse pregnancy outcomes. This study investigated the role of CD4+ and CD8+ T cells in a guinea pig model of primary cytomegalovirus infection. Monoclonal antibodies specific to guinea pig CD4 and CD8 were used to deplete T cells in non- pregnant and in pregnant guinea pigs after mid-gestation. CD4+ T cell depletion increased the severity of illness, caused significantly elevated viral loads, and increased the rate of congenital guinea pig cytomegalovirus (GPCMV) infection relative to animals treated with control antibody. CD8+ T cell depletion was comparably well tolerated and did not significantly affect the weight of infected guinea pigs or viral loads in their blood or tissue. However, significantly more viral genomes and transcripts were detected in the placenta and decidua of CD8+ T cell depleted dams post-infection. This study corroborates earlier findings made in nonhuman primates that maternal CD4+ T cells play a critical role in limiting the severity of primary CMV infection during pregnancy while also revealing that other innate and adaptive immune responses can compensate for an absent CD8+ T cell response in -CD8-treated guinea pigs.

Author SummaryCongenital cytomegalovirus infection is a leading cause of adverse pregnancy outcomes and preventable disability in children. Using guinea pigs, a well-established small animal model of congenital infection and intrauterine development, this study tested how depleting T cells affects the course of primary cytomegalovirus infections. Severe illness and high rates of congenital infection were observed when helper CD4+ T cells were depleted. The depletion of killer CD8+ T cells did not affect the severity of disease or the rate of congenital infection but did increase the amount of virus that was detected in the placenta. A greater understanding how an immune response can prevent the infection of the placenta and developing offspring is needed to inform vaccine and therapeutic development. This study not only describes a new reagent that can be used to study the guinea pig immune system but also sheds new light in how adaptive immunity regulates congenital viral infection.
]]></description>
<dc:creator>Rollman, T. B.</dc:creator>
<dc:creator>Berkebile, Z. W.</dc:creator>
<dc:creator>Hicks, D. M.</dc:creator>
<dc:creator>Hatfield, J. S.</dc:creator>
<dc:creator>Chauhan, P.</dc:creator>
<dc:creator>Pravetoni, M.</dc:creator>
<dc:creator>Schleiss, M. R.</dc:creator>
<dc:creator>Milligan, G. N.</dc:creator>
<dc:creator>Morgan, T. K.</dc:creator>
<dc:creator>Bierle, C. J.</dc:creator>
<dc:date>2024-08-21</dc:date>
<dc:identifier>doi:10.1101/2024.08.21.608907</dc:identifier>
<dc:title><![CDATA[CD4+ but not CD8+ T cells are required for protection against severe guinea pig cytomegalovirus infections]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-08-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.08.28.610193v1?rss=1">
<title>
<![CDATA[
Brain Charts for the Rhesus Macaque Lifespan 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.08.28.610193v1?rss=1</link>
<description><![CDATA[
Recent efforts to chart human brain growth across the lifespan using large-scale MRI data have provided reference standards for human brain development. However, similar models for nonhuman primate (NHP) growth are lacking. The rhesus macaque, a widely used NHP in translational neuroscience due to its similarities in brain anatomy, phylogenetics, cognitive, and social behaviors to humans, serves as an ideal NHP model. This study aimed to create normative growth charts for brain structure across the macaque lifespan, enhancing our understanding of neurodevelopment and aging, and facilitating cross-species translational research. Leveraging data from the PRIMatE Data Exchange (PRIME-DE) and other sources, we aggregated 1,522 MRI scans from 1,024 rhesus macaques. We mapped non-linear developmental trajectories for global and regional brain structural changes in volume, cortical thickness, and surface area over the lifespan. Our findings provided normative charts with centile scores for macaque brain structures and revealed key developmental milestones from prenatal stages to aging, highlighting both species-specific and comparable brain maturation patterns between macaques and humans. The charts offer a valuable resource for future NHP studies, particularly those with small sample sizes. Furthermore, the interactive open resource (https://interspeciesmap.childmind.org) supports cross-species comparisons to advance translational neuroscience research.
]]></description>
<dc:creator>Alldritt, S.</dc:creator>
<dc:creator>Ramirez, J. S. B.</dc:creator>
<dc:creator>Vos de Wael, R.</dc:creator>
<dc:creator>Bethlehem, R.</dc:creator>
<dc:creator>Seidlitz, J.</dc:creator>
<dc:creator>Wang, Z.</dc:creator>
<dc:creator>Nenning, K.-H.</dc:creator>
<dc:creator>Esper, N. B.</dc:creator>
<dc:creator>Smallwood, J.</dc:creator>
<dc:creator>Franco, A. R.</dc:creator>
<dc:creator>Byeon, K.</dc:creator>
<dc:creator>Alexander-Bloch, A.</dc:creator>
<dc:creator>Messinger, A.</dc:creator>
<dc:creator>Chen, A.</dc:creator>
<dc:creator>Thiele, A.</dc:creator>
<dc:creator>Weiss, A. R.</dc:creator>
<dc:creator>Shmuel, A.</dc:creator>
<dc:creator>Fox, A.</dc:creator>
<dc:creator>Tusche, A.</dc:creator>
<dc:creator>Brambrink, A. M.</dc:creator>
<dc:creator>Falchier, A.</dc:creator>
<dc:creator>Hiba, B.</dc:creator>
<dc:creator>Jarraya, B.</dc:creator>
<dc:creator>Jung, B.</dc:creator>
<dc:creator>Butler, B.</dc:creator>
<dc:creator>Russ, B. E.</dc:creator>
<dc:creator>Howell, B. R.</dc:creator>
<dc:creator>Drzewiecki, C. M.</dc:creator>
<dc:creator>Guedj, C.</dc:creator>
<dc:creator>Schwiedrzik, C. M.</dc:creator>
<dc:creator>Amiez, C.</dc:creator>
<dc:creator>Wilson, C. R. E.</dc:creator>
<dc:creator>Klink, C.</dc:creator>
<dc:creator>Kroenke, C.</dc:creator>
<dc:creator>Constantinidis, C.</dc:creator>
<dc:creator>Garin, C. M.</dc:creator>
<dc:creator>Poirier, C.</dc:creator>
<dc:creator>Gale, D. J.</dc:creator>
<dc:creator>Leopold, D. A.</dc:creator>
<dc:creator>Rudko, D. A.</dc:creator>
<dc:creator>Amaral, D. G.</dc:creator>
<dc:creator>Tsao, D.</dc:creator>
<dc:creator>Cook, D. J.</dc:creator>
<dc:creator></dc:creator>
<dc:date>2024-08-30</dc:date>
<dc:identifier>doi:10.1101/2024.08.28.610193</dc:identifier>
<dc:title><![CDATA[Brain Charts for the Rhesus Macaque Lifespan]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-08-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.08.29.610280v1?rss=1">
<title>
<![CDATA[
Centromeric transposable elements and epigenetic status drive karyotypic variation in the eastern hoolock gibbon 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.08.29.610280v1?rss=1</link>
<description><![CDATA[
Great apes have maintained a stable karyotype with few large-scale rearrangements; in contrast, gibbons have undergone a high rate of chromosomal rearrangements coincident with rapid centromere turnover. Here we characterize assembled centromeres in the Eastern hoolock gibbon, Hoolock leuconedys (HLE), finding a diverse group of transposable elements (TEs) that differ from the canonical alpha satellites found across centromeres of other apes. We find that HLE centromeres contain a CpG methylation centromere dip region, providing evidence this epigenetic feature is conserved in the absence of satellite arrays; nevertheless, we report a variety of atypical centromeric features, including protein-coding genes and mismatched replication timing. Further, large structural variations define HLE centromeres and distinguish them from other gibbons. Combined with differentially methylated TEs, topologically associated domain boundaries, and segmental duplications at chromosomal breakpoints, we propose that a "perfect storm" of multiple genomic attributes with propensities for chromosome instability shaped gibbon centromere evolution.
]]></description>
<dc:creator>Hartley, G. A.</dc:creator>
<dc:creator>Okhovat, M.</dc:creator>
<dc:creator>Hoyt, S. J.</dc:creator>
<dc:creator>Fuller, E.</dc:creator>
<dc:creator>Pauloski, N.</dc:creator>
<dc:creator>Alexandre, N.</dc:creator>
<dc:creator>Alexandrov, I. A.</dc:creator>
<dc:creator>Drennan, R. S.</dc:creator>
<dc:creator>Dubocanin, D.</dc:creator>
<dc:creator>Gilbert, D. M.</dc:creator>
<dc:creator>Mao, Y.</dc:creator>
<dc:creator>McCann, C.</dc:creator>
<dc:creator>Neph, S.</dc:creator>
<dc:creator>Ryabov, F.</dc:creator>
<dc:creator>Sasaki, T.</dc:creator>
<dc:creator>Storer, J. M.</dc:creator>
<dc:creator>Svendsen, D.</dc:creator>
<dc:creator>Troy, W.</dc:creator>
<dc:creator>Wells, J.</dc:creator>
<dc:creator>Core, L.</dc:creator>
<dc:creator>Stergachis, A.</dc:creator>
<dc:creator>Carbone, L.</dc:creator>
<dc:creator>O'Neill, R. J.</dc:creator>
<dc:date>2024-08-30</dc:date>
<dc:identifier>doi:10.1101/2024.08.29.610280</dc:identifier>
<dc:title><![CDATA[Centromeric transposable elements and epigenetic status drive karyotypic variation in the eastern hoolock gibbon]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-08-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.08.29.610348v1?rss=1">
<title>
<![CDATA[
Distinct functional domains of Dystroglycan regulate inhibitory synapse formation and maintenance in cerebellar Purkinje cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.08.29.610348v1?rss=1</link>
<description><![CDATA[
Dystroglycan is a cell adhesion molecule that localizes to synapses throughout the nervous system. While Dystroglycan is required to maintain inhibitory synapses from cerebellar molecular layer interneurons (MLIs) onto Purkinje cells (PCs) whether initial synaptogenesis during development is dependent on Dystroglycan has not been examined. We show that conditional deletion of Dystroglycan from Purkinje cells prior to synaptogenesis results in impaired MLI:PC synapse formation and function due to reduced presynaptic inputs and abnormal postsynaptic GABAA receptor clustering. Using genetic manipulations that disrupt glycosylation of Dystroglycan or truncate its cytoplasmic domain, we show that Dystroglycans role in synapse function requires both extracellular and intracellular interactions, whereas synapse formation requires only extracellular interactions. Together, these findings provide molecular insight into the mechanism of inhibitory synapse formation and maintenance in cerebellar cortex.
]]></description>
<dc:creator>Jahncke, J. N.</dc:creator>
<dc:creator>Schnell, E.</dc:creator>
<dc:creator>Wright, K. M.</dc:creator>
<dc:date>2024-08-30</dc:date>
<dc:identifier>doi:10.1101/2024.08.29.610348</dc:identifier>
<dc:title><![CDATA[Distinct functional domains of Dystroglycan regulate inhibitory synapse formation and maintenance in cerebellar Purkinje cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-08-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.08.29.610369v1?rss=1">
<title>
<![CDATA[
Atlas-scale Single-cell DNA Methylation Profiling with sciMETv3 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.08.29.610369v1?rss=1</link>
<description><![CDATA[
Single-cell methods to assess DNA methylation have not yet achieved the same level of cell throughput compared to other modalities. Here, we describe sciMETv3, a combinatorial indexing-based technique that builds on our prior technology, sciMETv2. SciMETv3 achieves nearly a 100-fold improvement in cell throughput by increasing the index space while simultaneously reducing hands-on time and total costs per experiment. To reduce the sequencing burden of the assay, we demonstrate compatibility of sciMETv3 with capture techniques that enrich for regulatory regions, as well as the ability to leverage enzymatic conversion which can yield higher library diversity. We showcase the throughput of sciMETv3 by producing a >140k cell library from human middle frontal gyrus split across four multiplexed individuals using both Illumina and Ultima sequencing instrumentation. This library was prepared over two days by one individual and required no expensive equipment (e.g. a flow sorter, as required by sciMETv2). The same experiment produced an estimated 650k additional cells that were not sequenced, representing the power of sciMETv3 to meet the throughput needs of the most demanding atlas-scale projects. Finally, we demonstrate the compatibility of sciMETv3 with multimodal assays by introducing sciMET+ATAC, which will enable high- throughput exploration of the interplay between two layers of epigenetic regulation within the same cell, as well as the ability to directly integrate single-cell methylation datasets with existing single-cell ATAC-seq.

HighlightsO_LIAtlas-scale production of single-cell DNA methylation libraries in a single experiment
C_LIO_LIProtocols and evaluation using both Illumina and Ultima Genomics sequencing platforms
C_LIO_LICompatibility of sciMETv3 with capture techniques to reduce sequencing burden
C_LIO_LICompatibility of sciMETv3 with enzymatic conversion methods
C_LIO_LIGeneration of an integrated >140,000 cell dataset from human middle frontal gyrus across four individuals
C_LIO_LIAbility to profile both ATAC and genome-wide DNA methylation from the same cells and integration with datasets from each modality
C_LIO_LIA novel implementation of the s3-ATAC technology that leverages a nanowell chip for increased throughput
C_LI

MotivationDNA methylation forms a basal layer of epigenomic regulatory control, shaping the genomic permissiveness of mammalian cells during lineage specification and development. Aberrant DNA methylation has been associated with myriad health conditions ranging from developmental disorders to cancer. The high cell type specificity necessitates analysis at the single-cell level, much like transcription or other epigenomic properties. However, robust and cost-effective techniques to produce atlas-scale datasets have not been realized for DNA methylation. Here, we directly meet this need by introducing sciMETv3, a high-throughput protocol capable of producing hundreds of thousands of single-cell DNA methylation profiles in a single experiment.
]]></description>
<dc:creator>Nichols, R. V.</dc:creator>
<dc:creator>Rylaarsdam, L.</dc:creator>
<dc:creator>O'Connell, B. L.</dc:creator>
<dc:creator>Shipony, Z.</dc:creator>
<dc:creator>Iremadze, N.</dc:creator>
<dc:creator>Acharya, S. N.</dc:creator>
<dc:creator>Adey, A.</dc:creator>
<dc:date>2024-08-30</dc:date>
<dc:identifier>doi:10.1101/2024.08.29.610369</dc:identifier>
<dc:title><![CDATA[Atlas-scale Single-cell DNA Methylation Profiling with sciMETv3]]></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/10.1101/2024.09.04.611325v1?rss=1">
<title>
<![CDATA[
Structure and inhibition mechanisms of Mycobacterium tuberculosis essential transporterefflux protein A 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.09.04.611325v1?rss=1</link>
<description><![CDATA[
A broad chemical genetics screen in Mycobacterium tuberculosis (Mtb) to identify inhibitors of established or previously untapped targets for therapeutic development yielded compounds (BRD-8000.3 and BRD-9327) that inhibit the essential efflux pump EfpA. To understand the mechanisms of inhibition by these compounds, we determined the structures of EfpA with inhibitors bound at 2.7 -3.4 [A] resolution. Our structures reveal different mechanisms of inhibition for the two inhibitors. BRD-8000.3 binds in a tunnel making contact with the lipid bilayer and extending toward the central cavity to displace the fatty acid chain of a lipid molecule bound in the apo structure, suggesting its blocking of an access route for a natural lipidic substrate, in contrast to its uncompetitive mechanism for the small molecule substrate ethidium bromide which likely enters through an alternative tunnel. Meanwhile, BRD-9327 binds in the outer vestibule without complete blockade of the substrate path to the outside, suggesting its possible inhibition of the dynamical motion necessary for "alternate access" to the two different sides of the membrane, as is characteristic of major facilitator superfamily (MFS) transporters. Both inhibitors may have a role in inhibiting the "alternate access" mechanism that could account for the uncompetitive nature of their efflux of some substrates. Our results explain the basis of the synergy of these inhibitors and their potential for combination in a multi drug strategy for anti-tuberculosis therapy. They also potentially point to a possible function for this essential efflux pump as a lipid transporter. The structures provide a foundation for rational modification of these inhibitors to increase potency.
]]></description>
<dc:creator>Khandelwal, N. K.</dc:creator>
<dc:creator>Gupta, M.</dc:creator>
<dc:creator>Gomez, J. E.</dc:creator>
<dc:creator>Barkho, S.</dc:creator>
<dc:creator>Guan, Z.</dc:creator>
<dc:creator>Eng, A. Y.</dc:creator>
<dc:creator>Kawate, T.</dc:creator>
<dc:creator>Balasubramani, S. G.</dc:creator>
<dc:creator>Sali, A.</dc:creator>
<dc:creator>Hung, D. T.</dc:creator>
<dc:creator>Stroud, R. M.</dc:creator>
<dc:date>2024-09-05</dc:date>
<dc:identifier>doi:10.1101/2024.09.04.611325</dc:identifier>
<dc:title><![CDATA[Structure and inhibition mechanisms of Mycobacterium tuberculosis essential transporterefflux protein A]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-09-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.09.05.611490v1?rss=1">
<title>
<![CDATA[
AI-Based Discovery and CryoEM Structural Elucidation of a KATP Channel Pharmacochaperone 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.09.05.611490v1?rss=1</link>
<description><![CDATA[
Pancreatic KATP channel trafficking defects underlie congenital hyperinsulinism (CHI) cases unresponsive to the KATP channel opener diazoxide, the mainstay medical therapy for CHI. Current clinically used KATP channel inhibitors have been shown to act as pharmacochaperones and restore surface expression of trafficking mutants; however, their therapeutic utility for KATP trafficking impaired CHI is hindered by high-affinity binding, which limits functional recovery of rescued channels. Recent structural studies of KATP channels employing cryo-electron microscopy (cryoEM) have revealed a promiscuous pocket where several known KATP pharmacochaperones bind. The structural knowledge provides a framework for discovering KATP channel pharmacochaperones with desired reversible inhibitory effects to permit functional recovery of rescued channels. Using an AI-based virtual screening technology AtomNet(R) followed by functional validation, we identified a novel compound, termed Aekatperone, which exhibits chaperoning effects on KATP channel trafficking mutations. Aekatperone reversibly inhibits KATP channel activity with a half-maximal inhibitory concentration (IC50) [~] 9 M. Mutant channels rescued to the cell surface by Aekatperone showed functional recovery upon washout of the compound. CryoEM structure of KATP bound to Aekatperone revealed distinct binding features compared to known high affinity inhibitor pharmacochaperones. Our findings unveil a KATP pharmacochaperone enabling functional recovery of rescued channels as a promising therapeutic for CHI caused by KATP trafficking defects.
]]></description>
<dc:creator>Elsheikh, A.</dc:creator>
<dc:creator>Driggers, C. M.</dc:creator>
<dc:creator>Truong, H. H.</dc:creator>
<dc:creator>Yang, Z.</dc:creator>
<dc:creator>Allen, J.</dc:creator>
<dc:creator>Henriksen, N.</dc:creator>
<dc:creator>Walczewska-Szewc, K.</dc:creator>
<dc:creator>Shyng, S.-L.</dc:creator>
<dc:date>2024-09-05</dc:date>
<dc:identifier>doi:10.1101/2024.09.05.611490</dc:identifier>
<dc:title><![CDATA[AI-Based Discovery and CryoEM Structural Elucidation of a KATP Channel Pharmacochaperone]]></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/10.1101/2024.09.06.611254v1?rss=1">
<title>
<![CDATA[
Brain-wide mapping of oligodendrocyte organization and oligodendrogenesis across the murine lifespan 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.09.06.611254v1?rss=1</link>
<description><![CDATA[
Insulating sheaths of myelin accelerate neuronal signaling in complex networks of the mammalian brain. In the CNS, myelin sheaths are exclusively produced by oligodendrocytes, which continue to be generated throughout life to change patterns of myelination. However, a brain-wide analysis of oligodendrocyte dynamics across the lifespan has not been performed. We developed a rapid, robust cellular mapping pipeline involving tissue clearing, lightsheet microscopy, atlas alignment, and automated segmentation to define the location of all oligodendrocytes in the mouse brain. This analysis demonstrated the remarkable consistency of oligodendrocyte patterns between hemispheres, individuals, and sexes, and established that oligodendrocyte maps estimate myelin coverage. We trained a vision transformer to identify newly generated oligodendrocytes from millions of mature cells, highlighting age- and region-specific differences in oligodendrogenesis, and revealing areas of enhanced oligodendrocyte resilience and regenerative capacity following demyelination, demonstrating the utility of this pipeline for uncovering brain-wide oligodendrocyte dynamics in health and disease.
]]></description>
<dc:creator>Xu, Y. K. T.</dc:creator>
<dc:creator>Bush, A.</dc:creator>
<dc:creator>Musheyev, E.</dc:creator>
<dc:creator>Kim, A. A.</dc:creator>
<dc:creator>Zhang, S.</dc:creator>
<dc:creator>Eugenin von Bernhardi, J.</dc:creator>
<dc:creator>Sulam, J.</dc:creator>
<dc:creator>Bergles, D. E.</dc:creator>
<dc:date>2024-09-08</dc:date>
<dc:identifier>doi:10.1101/2024.09.06.611254</dc:identifier>
<dc:title><![CDATA[Brain-wide mapping of oligodendrocyte organization and oligodendrogenesis across the murine lifespan]]></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/10.1101/2024.09.07.611840v1?rss=1">
<title>
<![CDATA[
A computationally guided approach to improve expression of VHH binders 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.09.07.611840v1?rss=1</link>
<description><![CDATA[
The variable heavy chain fragments derived from camelid antibodies, called VHHs or nanobodies, have recently shown promise as high-affinity reagents. They offer higher stability compared to conventional antibodies and fragments thereof. Furthermore, their smaller size ([~]15-20 kDa) allows better targeting of molecules localized inside the cell and in crowded environments, like tissues and protein aggregates. Despite these advantages, nanobody clones screened using phage display can suffer from poor soluble expression, which we hypothesized, is due to the presence of hydrophobic hotspots on their surface. In this work, we propose a novel computationally guided workflow for screening and production of nanobody binders for optimized expression. After an initial round of phage display screens against our target (K-Ras), we modeled the lead candidates to generate Spatial Aggregation Propensity (SAP) maps to highlight the hydrophobic hotspots with single amino acid resolution, which were subsequently used to guide mutagenesis of the binders for soluble expression. We followed two approaches to perform point hydrophilic mutations: i) performing point hydrophilic mutations in the hydrophobic hotspots; ii) combining point mutation resulting from a round of random mutagenesis that show favorable SAP scores. Both approaches led a remarkable increase in soluble expression which allowed production and characterization of their binding to their target (K-Ras) on soluble ELISA, and biolayer interferometry. We observed that the latter approach resulted in clones with stronger binding affinity compared to the former approach. Our results emphasize the need to perform a round of random mutagenesis to identify point mutations, which can then be used in an in-silico guided pipeline to identify the right combination of mutations for high soluble expression.
]]></description>
<dc:creator>OZDEMIR, E. S.</dc:creator>
<dc:creator>Tolley, J.</dc:creator>
<dc:creator>Goncalves, F.</dc:creator>
<dc:creator>Gomes, M.</dc:creator>
<dc:creator>Wagnell, E.</dc:creator>
<dc:creator>Branchaud, B.</dc:creator>
<dc:creator>Ranganathan, S.</dc:creator>
<dc:date>2024-09-13</dc:date>
<dc:identifier>doi:10.1101/2024.09.07.611840</dc:identifier>
<dc:title><![CDATA[A computationally guided approach to improve expression of VHH binders]]></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/10.1101/2024.09.12.611703v1?rss=1">
<title>
<![CDATA[
Retinoic acid-mediated homeostatic plasticity drives cell type-specific CP-AMPAR accumulation in nucleus accumbens core and incubation of cocaine craving 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.09.12.611703v1?rss=1</link>
<description><![CDATA[
Incubation of cocaine craving, a translationally relevant model for the persistence of drug craving during abstinence, ultimately depends on strengthening of nucleus accumbens core (NAcc) synapses through synaptic insertion of homomeric GluA1 Ca2+-permeable AMPA receptors (CP-AMPARs). Here we tested the hypothesis that CP-AMPAR upregulation results from a form of homeostatic plasticity, previously characterized in vitro and in other brain regions, that depends on retinoic acid (RA) signaling in dendrites. Under normal conditions, ongoing synaptic transmission maintains intracellular Ca2+ at levels sufficient to suppress RA synthesis. Prolonged blockade of neuronal activity results in disinhibition of RA synthesis, leading to increased GluA1 translation and synaptic insertion of homomeric GluA1 CP-AMPARs. Using slice recordings, we found that increasing RA signaling in NAcc medium spiny neurons (MSN) from drug-naive rats rapidly upregulates CP-AMPARs, and that this pathway is operative only in MSN expressing the D1 dopamine receptor. In MSN recorded from rats that have undergone incubation of craving, this effect of RA is occluded; instead, interruption of RA signaling in the slice normalizes the incubation-associated elevation of synaptic CP-AMPARs. Paralleling this in vitro finding, interruption of RA signaling in the NAcc of  incubated rats normalizes the incubation-associated elevation of cue-induced cocaine seeking. These results suggest that RA signaling becomes tonically active in the NAcc during cocaine withdrawal and, by maintaining elevated CP-AMPAR levels, contributes to the incubation of cocaine craving.
]]></description>
<dc:creator>Hwang, E.-K.</dc:creator>
<dc:creator>Wunsch, A. M.</dc:creator>
<dc:creator>Wolf, M. E.</dc:creator>
<dc:date>2024-09-14</dc:date>
<dc:identifier>doi:10.1101/2024.09.12.611703</dc:identifier>
<dc:title><![CDATA[Retinoic acid-mediated homeostatic plasticity drives cell type-specific CP-AMPAR accumulation in nucleus accumbens core and incubation of cocaine craving]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-09-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.09.12.612675v1?rss=1">
<title>
<![CDATA[
A Comprehensive Proteogenomic and Spatial Analysis of Innate and Acquired Resistance of Metastatic Melanoma to Immune Checkpoint Blockade Therapies 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.09.12.612675v1?rss=1</link>
<description><![CDATA[
While a subset of patients with metastatic melanoma achieves durable responses to immune checkpoint blockade (ICB) therapies, the majority ultimately exhibit either innate or acquired resistance to these treatments. However, the molecular mechanisms underlying resistance to ICB therapies remain elusive and are warranted to elucidate. Here, we comprehensively investigated the tumor and tumor immune microenvironment (TIME) of paired pre- and post-treatment tumor specimens from metastatic melanoma patients who were primary or secondary resistance to anti-CTLA-4 and/or anti-PD-1/PD-L1 therapies. Differentially expressed gene (DEG) analysis and single-sample gene set enrichment analysis (ssGSEA) with transcriptomic data identified cell cycle and c-MYC signaling as pathway-based resistance signatures. And weighted gene co-expression network analysis (WGCNA) revealed the activation of a cross-resistance meta-program involving key signaling pathways related to tumor progression in ICB resistant melanoma. Moreover, spatially-resolved, image-based immune monitoring analysis by using NanoStrings digital spatial profiling (DSP) and Cyclic Immunofluorescence (CyCIF) showed infiltration of suppressive immune cells in the tumor microenvironment of melanoma with resistance to ICB therapies. Our study reveals the molecular mechanisms underlying resistance to ICB therapies in patients with metastatic melanoma by conducting such integrated analyses of multi-dimensional data, and provides rationale for salvage therapies that will potentially overcome resistance to ICB therapies.

Statement of translational relevanceThis study paves the way for the creation of innovative therapeutic strategies, aimed at subverting resistance to immune checkpoint blockade (ICB) therapies in metastatic melanoma patients. By unraveling the specific molecular mechanisms underlying resistance, scientists can design effective alternative treatments that target pathways such as pathways associated with cell cycle dysregulation and c-MYC signaling. Furthermore, through the application of advanced immune monitoring techniques such as NanoString Digital Spatial Profiling (DSP) and Cyclic Immunofluorescence (CyCIF), this study has significantly enriched our understanding of the tumor microenvironment. This enhanced characterization facilitates the discovery of potential biomarkers that may forecast a patients response to ICB treatment. Ultimately, these advancements could potentially refine patient outcomes and foster the development of more personalized cancer treatments in the future.
]]></description>
<dc:creator>wei, s.</dc:creator>
<dc:creator>Du, K.</dc:creator>
<dc:creator>Lan, H.</dc:creator>
<dc:creator>Yang, Z.</dc:creator>
<dc:creator>Deng, Y.</dc:creator>
<dc:creator>Wei, Z.</dc:creator>
<dc:creator>Frederick, D. T.</dc:creator>
<dc:creator>Lee, J.</dc:creator>
<dc:creator>Labrie, M.</dc:creator>
<dc:creator>Tian, T.</dc:creator>
<dc:creator>Moll, T.</dc:creator>
<dc:creator>Chen, Y.</dc:creator>
<dc:creator>Sullivan, R. J.</dc:creator>
<dc:creator>Mills, G. B.</dc:creator>
<dc:creator>Boland, G. M.</dc:creator>
<dc:creator>Flaherty, K.</dc:creator>
<dc:creator>liu, l.</dc:creator>
<dc:creator>Herlyn, M.</dc:creator>
<dc:creator>Zhang, G.</dc:creator>
<dc:date>2024-09-15</dc:date>
<dc:identifier>doi:10.1101/2024.09.12.612675</dc:identifier>
<dc:title><![CDATA[A Comprehensive Proteogenomic and Spatial Analysis of Innate and Acquired Resistance of Metastatic Melanoma to Immune Checkpoint Blockade Therapies]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-09-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.09.13.612869v1?rss=1">
<title>
<![CDATA[
Contribution of amygdala to dynamic model arbitration under uncertainty 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.09.13.612869v1?rss=1</link>
<description><![CDATA[
Intrinsic uncertainty in the reward environment requires the brain to run multiple models simultaneously to predict outcomes based on preceding cues or actions, commonly referred to as stimulus- and action-based learning. Ultimately, the brain also must adopt appropriate choice behavior using reliability of these models. Here, we combined multiple experimental and computational approaches to quantify concurrent learning in monkeys performing tasks with different levels of uncertainty about the model of the environment. By comparing behavior in control monkeys and monkeys with bilateral lesions to the amygdala or ventral striatum, we found evidence for dynamic, competitive interaction between stimulus-based and action-based learning, and for a distinct role of the amygdala. Specifically, we demonstrate that the amygdala adjusts the initial balance between the two learning systems, thereby altering the interaction between arbitration and learning that shapes the time course of both learning and choice behaviors. This novel role of the amygdala can account for existing contradictory observations and provides testable predictions for future studies into circuit-level mechanisms of flexible learning and choice under uncertainty.
]]></description>
<dc:creator>Woo, J. H.</dc:creator>
<dc:creator>Costa, V. D.</dc:creator>
<dc:creator>Taswell, C. A.</dc:creator>
<dc:creator>Rothenhoefer, K. M.</dc:creator>
<dc:creator>Averbeck, B.</dc:creator>
<dc:creator>Soltani, A.</dc:creator>
<dc:date>2024-09-15</dc:date>
<dc:identifier>doi:10.1101/2024.09.13.612869</dc:identifier>
<dc:title><![CDATA[Contribution of amygdala to dynamic model arbitration under uncertainty]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-09-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.09.15.613147v1?rss=1">
<title>
<![CDATA[
Changes in nucleus accumbens core translatome accompanying incubation of cocaine craving 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.09.15.613147v1?rss=1</link>
<description><![CDATA[
In the  incubation of cocaine craving model of relapse, rats exhibit progressive intensification (incubation) of cue-induced craving over several weeks of forced abstinence from cocaine self-administration. The expression of incubated craving depends on plasticity of excitatory synaptic transmission in nucleus accumbens core (NAcC) medium spiny neurons (MSN). Previously, we found that the maintenance of this plasticity and the expression of incubation depends on ongoing protein translation, and the regulation of translation is altered after incubation of cocaine craving. Here we used male and female rats that express Cre recombinase in either dopamine D1 receptor- or adenosine 2a (A2a) receptor-expressing MSN to express a GFP-tagged ribosomal protein in a cell-type specific manner, enabling us to use Translating Ribosome Affinity Purification (TRAP) to isolate actively translating mRNAs from both MSN subtypes for analysis by RNA-seq. We compared rats that self-administered saline or cocaine. Saline rats were assessed on abstinence day (AD) 1, while cocaine rats were assessed on AD1 or AD40-50. For both D1-MSN and A2a-MSN, there were few differentially translated genes between saline and cocaine AD1 groups. In contrast, pronounced differences in the translatome were observed between cocaine rats on AD1 and AD40-50, and this was far more robust in D1-MSN. Notably, all comparisons revealed sex differences in translating mRNAs. Sequencing results were validated by qRT-PCR for several genes of interest. This study, the first to combine TRAP-seq, transgenic rats, and a cocaine self-administration paradigm, identifies translating mRNAs linked to incubation of cocaine craving in D1-MSN and A2a-MSN of the NAcC.
]]></description>
<dc:creator>Kawa, A. B.</dc:creator>
<dc:creator>Hashimoto, J. G.</dc:creator>
<dc:creator>Beutler, M. M.</dc:creator>
<dc:creator>Guizzetti, M.</dc:creator>
<dc:creator>Wolf, M. E.</dc:creator>
<dc:date>2024-09-16</dc:date>
<dc:identifier>doi:10.1101/2024.09.15.613147</dc:identifier>
<dc:title><![CDATA[Changes in nucleus accumbens core translatome accompanying incubation of cocaine craving]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-09-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.09.18.613774v1?rss=1">
<title>
<![CDATA[
A functional screen for ubiquitin regulation identifies an E3 ligase secreted by Pseudomonas aeruginosa 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.09.18.613774v1?rss=1</link>
<description><![CDATA[
Ubiquitin signaling controls many aspects of eukaryotic biology, including targeted protein degradation and immune defense. Remarkably, invading bacterial pathogens have adapted secreted effector proteins that hijack host ubiquitination to gain control over host responses. These ubiquitin-targeted effectors can exhibit, for example, E3 ligase or deubiquitinase activities, often without any sequence or structural homology to eukaryotic ubiquitin regulators. Such convergence in function poses a challenge to the discovery of additional bacterial virulence factors that target ubiquitin. To overcome this, we have developed a workflow to harvest natively secreted bacterial effectors and functionally screen them for ubiquitin regulatory activities. After benchmarking this approach on diverse ligase and deubiquitinase activities from Salmonella Typhimurium, Enteropathogenic Escherichia coli, and Shigella flexneri, we applied it to the identification of a cryptic E3 ligase activity secreted by Pseudomonas aeruginosa. We identified an unreported P. aeruginosa E3 ligase, which we have termed Pseudomonas Ub ligase 1 (PUL-1), that resembles none of the other E3 ligases previously established in or outside of the eukaryotic system. Importantly, in an animal model of P. aeruginosa infection, PUL-1 ligase activity plays an important role in regulating virulence. Thus, our workflow for the functional identification of ubiquitin-targeted effector proteins carries promise for expanding our appreciation of how host ubiquitin regulation contributes to bacterial pathogenesis.
]]></description>
<dc:creator>Roberts, C. G.</dc:creator>
<dc:creator>Kaur, S.</dc:creator>
<dc:creator>Ogden, A. J.</dc:creator>
<dc:creator>Divine, M. E.</dc:creator>
<dc:creator>Warren, G. D.</dc:creator>
<dc:creator>Kang, D.</dc:creator>
<dc:creator>Kirienko, N. V.</dc:creator>
<dc:creator>Geurink, P. P.</dc:creator>
<dc:creator>Mulder, M. P.</dc:creator>
<dc:creator>Nakayasu, E. S.</dc:creator>
<dc:creator>McDermott, J. E.</dc:creator>
<dc:creator>Adkins, J. N.</dc:creator>
<dc:creator>Aballay, A.</dc:creator>
<dc:creator>Pruneda, J. N.</dc:creator>
<dc:date>2024-09-18</dc:date>
<dc:identifier>doi:10.1101/2024.09.18.613774</dc:identifier>
<dc:title><![CDATA[A functional screen for ubiquitin regulation identifies an E3 ligase secreted by Pseudomonas aeruginosa]]></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/10.1101/2024.09.14.613069v1?rss=1">
<title>
<![CDATA[
Melanocortin-4 receptor antagonist TCMCB07 alleviates chemotherapy-induced anorexia and weight loss 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.09.14.613069v1?rss=1</link>
<description><![CDATA[
Cancer patients undergoing chemotherapy often experience anorexia and weight loss that significantly deteriorates overall health, reduces treatment tolerance and quality of life, and worsens oncologic outcomes. There are currently few effective therapeutic options to mitigate these side effects. The central melanocortin system, which plays a pivotal role in regulating appetite and energy homeostasis, presents a logical target for treating anorexia and weight loss. In this preclinical study, we evaluated the efficacy of TCMCB07, a synthetic antagonist of the melanocortin-4 receptor, in mitigating anorexia and weight loss in several rat models of chemotherapy: cisplatin, 5-fluorouracil, cyclophosphamide, vincristine, doxorubicin, and a combination of irinotecan and 5-fluorouracil. Our results indicate that peripheral administration of TCMCB07 improved appetite, stabilized body weight, preserved fat and heart mass, and slightly protected lean mass after multiple cycles of chemotherapy. Furthermore, combining TCMCB07 with a growth differentiation factor 15 antibody enhanced treatment effectiveness. Similar effects from TCMCB07 treatment were observed in a rat tumor model following combination chemotherapy. No significant adverse effects nor increased chemotherapy-related toxicities were observed with TCMCB07 treatment. These findings suggest that peripheral administration of TCMCB07 holds promise as a therapeutic approach for alleviating chemotherapy-induced anorexia and weight loss, potentially benefiting numerous patients undergoing chemotherapy.

Graphical abstract

O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=143 SRC="FIGDIR/small/613069v1_ufig1.gif" ALT="Figure 1">
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]]></description>
<dc:creator>Zhu, X.</dc:creator>
<dc:creator>Potterfield, R.</dc:creator>
<dc:creator>Gruber, K. A.</dc:creator>
<dc:creator>Zhang, E.</dc:creator>
<dc:creator>Newton, S.</dc:creator>
<dc:creator>Norgard, M. A.</dc:creator>
<dc:creator>Levasseur, P. R.</dc:creator>
<dc:creator>Bai, P.</dc:creator>
<dc:creator>Chen, X.</dc:creator>
<dc:creator>Gu, Q.</dc:creator>
<dc:creator>Grossberg, A.</dc:creator>
<dc:creator>Marks, D. L.</dc:creator>
<dc:date>2024-09-19</dc:date>
<dc:identifier>doi:10.1101/2024.09.14.613069</dc:identifier>
<dc:title><![CDATA[Melanocortin-4 receptor antagonist TCMCB07 alleviates chemotherapy-induced anorexia and weight loss]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-09-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.09.20.614142v1?rss=1">
<title>
<![CDATA[
Heart rate variability impairment during sleep in Veterans with REM sleep behavior disorder, traumatic brain injury, and posttraumatic stress disorder: An early potential window into autonomic dysfunction? 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.09.20.614142v1?rss=1</link>
<description><![CDATA[
Individuals with comorbid REM sleep behavior disorder (RBD) and neurotrauma (defined by traumatic brain injury and post-traumatic stress disorder) have an earlier age of RBD symptom onset, increased RBD-related symptom severity and more neurological features indicative of prodromal synucleinopathy compared to RBD only. An early sign of neurodegenerative condition is autonomic dysfunction, which we sought to evaluate by examining heart rate variability during sleep. Participants with overnight polysomnography were recruited from the VA Portland Health Care System. Veterans without neurotrauma or RBD (controls; n=19), with RBD only (RBD, n=14), and with RBD and neurotrauma (RBD+NT, n=19) were evaluated. Eligible 5-minute NREM and REM epochs without apneas/hypopneas, microarousals, and ectopic beats were analyzed for frequency and time domain (e.g. low frequency power, LF; high frequency power, HF; root mean square of successive RR intervals, RMSSD; % of RR intervals that vary [&ge;]50 ms, pNN50) heart rate variability outcomes. Heart rate did not significantly differ between groups in any sleep stage. Time domain and frequency domain variables (e.g., LF power, HF power, RMSSD, and pNN50) were significantly reduced in the RBD and RBD+NT groups compared to controls and RBD only during NREM sleep. There were no group differences detected during REM sleep. These data suggest significant reductions in heart rate variability during NREM sleep in RBD+NT participants, suggesting greater autonomic dysfunction compared to controls or RBD alone. Heart rate variability during sleep may be an early, promising biomarker, yielding mechanistic insight for diagnosis and prognosis of early neurodegeneration in this vulnerable population.

STATEMENT OF SIGNIFICANCEComorbid REM sleep behavior disorder (RBD) and neurotrauma (NT, traumatic brain injury + post-traumatic stress disorder; RBD+NT) is associated with increased neurodegenerative symptom burden and worsened health. Sleep and autonomic function are integrally and bidirectionally related to neurodegenerative processes. In the current study, we sought to determine if early signs of autonomic dysfunction, measured via heart rate variability (HRV), were present during sleep in comorbid RBD+NT compared to RBD only and controls. Our data show reduced time and frequency domain HRV during NREM sleep in RBD+NT Veterans compared to RBD only and controls. These data contribute evidence that participants with RBD and comorbid NT demonstrate significantly worse autonomic dysfunction compared to age/sex matched participants with RBD alone.
]]></description>
<dc:creator>Cunningham, H. A.</dc:creator>
<dc:creator>Dovek, L.</dc:creator>
<dc:creator>Recoder, N.</dc:creator>
<dc:creator>Bryant-Ekstrand, M. D.</dc:creator>
<dc:creator>Ligman, B.</dc:creator>
<dc:creator>Piantino, J.</dc:creator>
<dc:creator>Lim, M. M.</dc:creator>
<dc:creator>Elliott, J. E.</dc:creator>
<dc:date>2024-09-24</dc:date>
<dc:identifier>doi:10.1101/2024.09.20.614142</dc:identifier>
<dc:title><![CDATA[Heart rate variability impairment during sleep in Veterans with REM sleep behavior disorder, traumatic brain injury, and posttraumatic stress disorder: An early potential window into autonomic dysfunction?]]></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/10.1101/2024.09.25.615077v1?rss=1">
<title>
<![CDATA[
Multi-omics identification of extracellular components of the fetal monkey and human neocortex. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.09.25.615077v1?rss=1</link>
<description><![CDATA[
During development, precursor cells are continuously and intimately interacting with their extracellular environment, which guides their ability to generate functional tissues and organs. Much is known about the development of the neocortex in mammals. This information has largely been derived from histological analyses, heterochronic cell transplants, and genetic manipulations in mice, and to a lesser extent from transcriptomic and histological analyses in humans. However, these approaches have not led to a characterization of the extracellular composition of the developing neocortex in any species. Here, using a combination of single-cell transcriptomic analyses from published datasets, and our proteomics and immunohistofluorescence analyses, we provide a more comprehensive and unbiased picture of the early developing fetal neocortex in humans and non-human primates. Our findings provide a starting point for further hypothesis-driven studies on structural and signaling components in the developing cortex that had previously not been identified.
]]></description>
<dc:creator>Vilicich, F.</dc:creator>
<dc:creator>Vettiatil, D.</dc:creator>
<dc:creator>Kattapong Graber, S.</dc:creator>
<dc:creator>Nawsheen, N.</dc:creator>
<dc:creator>Patel, N.</dc:creator>
<dc:creator>Quezada, A.</dc:creator>
<dc:creator>Gurney, E.</dc:creator>
<dc:creator>Smith, E. M.</dc:creator>
<dc:creator>Nelson, H.</dc:creator>
<dc:creator>Pesci, S.</dc:creator>
<dc:creator>Atrio, J.</dc:creator>
<dc:creator>Moreno, N.</dc:creator>
<dc:creator>Jones, A.</dc:creator>
<dc:creator>Murphy, M.</dc:creator>
<dc:creator>Benfield, N.</dc:creator>
<dc:creator>Hennelbold, J.</dc:creator>
<dc:creator>Solansky, N.</dc:creator>
<dc:creator>Lisgo, S.</dc:creator>
<dc:creator>Glass, I.</dc:creator>
<dc:creator>Birth Defects Research Laboratory,</dc:creator>
<dc:creator>Sidoli, S.</dc:creator>
<dc:date>2024-09-26</dc:date>
<dc:identifier>doi:10.1101/2024.09.25.615077</dc:identifier>
<dc:title><![CDATA[Multi-omics identification of extracellular components of the fetal monkey and human neocortex.]]></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/10.1101/2024.09.26.615308v1?rss=1">
<title>
<![CDATA[
Pancreatic cancer cells overexpressing interleukin 6 induce T-cell-mediated tumor clearance and durable anti-tumor immune response 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.09.26.615308v1?rss=1</link>
<description><![CDATA[
Background & AimsTumor immune resistance is recognized as a contributor to low survivorship in pancreatic ductal adenocarcinoma (PDAC). The inflammatory cytokine interleukin-6 (IL-6) promotes polarization of CD4 T cell populations away from immune tolerance, and induces differentiation of cytotoxic CD8 T cells. This work aims to test whether IL-6 could stimulate an anti-tumor response in PDAC

MethodsWe overexpressed IL-6 in multiple KrasG12D/+, Tp53R172H/+, Pdx1-Cre (KPC) cell lines, which were orthotopically implanted in mice (OT-PDACIL6). We followed mouse survival and measured tumor growth, tumor histology, and plasma IL-6 at 5 and 10 days after tumor implantation. We measured tumor immune cell infiltration via flow cytometry and histology. We used antibody-based T cell depletion and secondary tumor implantation rechallenge to test the dependency of the durable immune reaction on T cells. We use lipid nanoparticle (LNP)-based delivery of IL-6 mRNA to the pancreas as an orthogonal approach for testing the effect of elevated IL-6 in the tumor microenvironment on anti-tumor T cell invasion.

ResultsImproved survival occurred in all instances of OT-PDACIL6, with one cell line (KxPxCx) reproducibly resulting in long-term recurrence-free survival. With KxPxCx cells, circulating IL-6 was 100-fold higher in OT-KxPxCxIL6 than in OT-KxPxCxparental mice. Flow cytometry revealed increased T cells and NK cells, and decreased T regulatory cells, and we observed significantly increased lymphoid aggregates in OT-KXPXCXIL6 as compared to OT--KxPxCxparental tumors. Antibody-based CD4+ and CD8+ T cell depletion prevented tumor clearance and completely abolished the survival advantage in OT-KxPxCxIL6 mice. The anti-tumor immune response to OT-KxPxCxIL6 rendered mice immune to re-challenge with OT-KxPxCxparental tumors. LNP delivery of IL-6 to the pancreas elevated systemic IL-6 levels [~]50 fold, lowered tumor burden, and increased anti-tumor T cell phenotypes.

ConclusionsLocally high IL-6 concentrations potently enhance the T cell-mediated anti-tumor response to PDAC.

SYNOPSISInterleukin-6 induces rapid and durable T cell-driven immune clearance of pancreatic ductal adenocarcinoma. The anti-tumor immune microenvironment is hallmarked by increased lymphoid aggregate formation, increased CD4 T cell abundance, and decreased Treg abundance.
]]></description>
<dc:creator>Arneson-Wissink, P. C.</dc:creator>
<dc:creator>Bartlett, A. Q.</dc:creator>
<dc:creator>Mendez, H.</dc:creator>
<dc:creator>Zhu, X.</dc:creator>
<dc:creator>Dickie, J.</dc:creator>
<dc:creator>McWhorter, M.</dc:creator>
<dc:creator>Levasseur, P. R.</dc:creator>
<dc:creator>Diba, P.</dc:creator>
<dc:creator>Byrne, K.</dc:creator>
<dc:creator>Scott, G. D.</dc:creator>
<dc:creator>Eil, R.</dc:creator>
<dc:creator>Grossberg, A.</dc:creator>
<dc:date>2024-09-27</dc:date>
<dc:identifier>doi:10.1101/2024.09.26.615308</dc:identifier>
<dc:title><![CDATA[Pancreatic cancer cells overexpressing interleukin 6 induce T-cell-mediated tumor clearance and durable anti-tumor immune response]]></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/10.1101/2024.09.27.615497v1?rss=1">
<title>
<![CDATA[
Quantifying Cancer Drug Targetability across Biochemical Pathways 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.09.27.615497v1?rss=1</link>
<description><![CDATA[
The evaluation of cancer drug treatments within the complex context of biological networks and pathways is challenging. Drugs can be "promiscuous," targeting multiple genes or pathways, not just the ones of interest. This complexity makes it difficult to assess the full impact of a drug solely based on individual knowledgebases. We make the impact more comprehensible through the characterization of drug targetability in terms of a combination of biochemical pathway data from the Reactome Knowledgebase and drug-target interaction data from the Cancer Targetome. This leads to a graphical representation of drug targetability. We further improve comprehensibility by developing metrics to quantify the graph structures in order to evaluate drug target specificity and navigate the hierarchical nature of biological networks and identify candidate treatment options to increase specificity and minimize off-target effects.. We have developed software that calculates these metrics for drugs in the Cancer Targetome. The software is available for research purposes only at https://github.com/sehagler/drug_targetability.
]]></description>
<dc:creator>Hagler, S.</dc:creator>
<dc:date>2024-10-01</dc:date>
<dc:identifier>doi:10.1101/2024.09.27.615497</dc:identifier>
<dc:title><![CDATA[Quantifying Cancer Drug Targetability across Biochemical Pathways]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-10-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.10.02.616342v1?rss=1">
<title>
<![CDATA[
Autophosphorylation of the oncogenic protein TEL-ABL confers resistance to the allosteric ABL inhibitor asciminib 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.10.02.616342v1?rss=1</link>
<description><![CDATA[
Chromosomal translocations that fuse the ABL1 gene to BCR and TEL cause human leukemias. Oligomerization and the loss of an inhibitory myristoylation modification lead to unregulated kinase activity of the BCR-ABL and TEL-ABL fusion proteins. ATP-competitive ABL inhibitors, such as imatinib and ponatinib, are effective against both fusion proteins. We discovered that asciminib, an allosteric inhibitor of BCR-ABL that binds to the myristoyl binding site in the ABL kinase domain, is >2000-fold less potent against TEL-ABL than BCR-ABL in cell-growth assays. This is surprising because the ABL components of the two fusion proteins, including the asciminib binding sites, have identical sequence. We deleted a short helical segment in the ABL kinase domain that closes over asciminib when it is bound. This deletion results in asciminib resistance in BCR-ABL, but has no effect on TEL-ABL, suggesting that the native autoinhibitory mechanism that asciminib engages in BCR-ABL is disrupted in TEL-ABL. We show, using mammalian cell expression and single-molecule microscopy, that BCR-ABL is mainly dimeric while TEL-ABL forms higher-order oligomers. Oligomerization can promote trans-autophosphorylation of ABL, and we find that a regulatory phosphorylation site in the SH3 domain of ABL (Tyr 89) is highly phosphorylated in TEL-ABL. This phosphorylation is expected to disassemble the autoinhibited conformation of ABL, thereby preventing asciminib binding. We show that TEL-ABL is intrinsically susceptible to inhibition by asciminib, but that increased phosphorylation results in resistance. Our results demonstrate that different ABL fusion proteins can have dramatically different responses to allosteric inhibitors due to differential phosphorylation.

One Sentence SummaryWhen TEL-ABL is phosphorylated, it is insensitive to asciminib. However, when TEL-ABL is dephosphorylated by a phosphatase, asciminib sensitivity is restored.
]]></description>
<dc:creator>Muratcioglu, S.</dc:creator>
<dc:creator>Eide, C. A.</dc:creator>
<dc:creator>Gorday, K.</dc:creator>
<dc:creator>Sumpena, E.</dc:creator>
<dc:creator>Zuo, W.</dc:creator>
<dc:creator>Groves, J. T.</dc:creator>
<dc:creator>Druker, B. J.</dc:creator>
<dc:creator>Kuriyan, J.</dc:creator>
<dc:date>2024-10-02</dc:date>
<dc:identifier>doi:10.1101/2024.10.02.616342</dc:identifier>
<dc:title><![CDATA[Autophosphorylation of the oncogenic protein TEL-ABL confers resistance to the allosteric ABL inhibitor asciminib]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-10-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.10.02.616147v1?rss=1">
<title>
<![CDATA[
Baby Open Brains: An Open-Source Repository of Infant Brain Segmentations 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.10.02.616147v1?rss=1</link>
<description><![CDATA[
Reproducibility of neuroimaging research on infant brain development remains limited due to highly variable protocols and processing approaches. Progress towards reproducible pipelines is limited by a lack of benchmarks such as gold standard brain segmentations. Addressing this core limitation, we constructed the Baby Open Brains (BOBs) Repository, an open source resource comprising manually curated and expert-reviewed infant brain segmentations. Markers and expert reviewers manually segmented anatomical MRI data from 71 infant imaging visits across 51 participants, using both T1w and T2w images per visit. Anatomical images showed dramatic differences in myelination and intensities across the 1 to 9 month age range, emphasizing the need for densely sampled gold standard manual segmentations in these ages. The BOBs repository is publicly available through the Masonic Institute for the Developing Brain (MIDB) Open Data Initiative, which links S3 storage, Datalad for version control, and BrainBox for visualization. This repository represents an open-source paradigm, where new additions and changes can be added, enabling a community-driven resource that will improve over time and extend into new ages and protocols. These manual segmentations and the ongoing repository provide a benchmark for evaluating and improving pipelines dependent upon segmentations in the youngest populations. As such, this repository provides a vitally needed foundation for early-life large-scale studies such as HBCD.
]]></description>
<dc:creator>Feczko, E. J.</dc:creator>
<dc:creator>Stoyell, S. M.</dc:creator>
<dc:creator>Moore, L. A.</dc:creator>
<dc:creator>Alexopoulos, D.</dc:creator>
<dc:creator>Bagonis, M.</dc:creator>
<dc:creator>Barrett, K.</dc:creator>
<dc:creator>Bower, B.</dc:creator>
<dc:creator>Cavender, A.</dc:creator>
<dc:creator>Chamberlain, T. A.</dc:creator>
<dc:creator>Conan, G.</dc:creator>
<dc:creator>Day, T. K.</dc:creator>
<dc:creator>Goradia, D.</dc:creator>
<dc:creator>Graham, A.</dc:creator>
<dc:creator>Heisler-Roman, L.</dc:creator>
<dc:creator>Hendrickson, T. J.</dc:creator>
<dc:creator>Houghton, A.</dc:creator>
<dc:creator>Kardan, O.</dc:creator>
<dc:creator>Kiffmeyer, E. A.</dc:creator>
<dc:creator>Lee, E. G.</dc:creator>
<dc:creator>Lundquist, J. T.</dc:creator>
<dc:creator>Lucena, C.</dc:creator>
<dc:creator>Martin, T.</dc:creator>
<dc:creator>Mummaneni, A.</dc:creator>
<dc:creator>Myricks, M.</dc:creator>
<dc:creator>Narnur, P.</dc:creator>
<dc:creator>Perrone, A. J.</dc:creator>
<dc:creator>Reiners, P.</dc:creator>
<dc:creator>Rueter, A. R.</dc:creator>
<dc:creator>Saw, H.</dc:creator>
<dc:creator>Styner, M.</dc:creator>
<dc:creator>Sung, S.</dc:creator>
<dc:creator>Tiklasky, B.</dc:creator>
<dc:creator>Wisnowski, J. L.</dc:creator>
<dc:creator>Yacoub, E.</dc:creator>
<dc:creator>Zimmermann, B.</dc:creator>
<dc:creator>Smyser, C. D.</dc:creator>
<dc:creator>Rosenberg, M. D.</dc:creator>
<dc:creator>Fair, D. A.</dc:creator>
<dc:creator>Elison, J. T.</dc:creator>
<dc:date>2024-10-03</dc:date>
<dc:identifier>doi:10.1101/2024.10.02.616147</dc:identifier>
<dc:title><![CDATA[Baby Open Brains: An Open-Source Repository of Infant Brain Segmentations]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-10-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.10.03.616354v1?rss=1">
<title>
<![CDATA[
An effective method of measuring nanobody binding kinetics and competition-based epitope mapping using biolayer interferometry 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.10.03.616354v1?rss=1</link>
<description><![CDATA[
Protein-protein interactions (PPI) underpin nearly all biological processes, and understanding the molecular mechanisms governing these interactions is crucial for the progress of biomedical sciences. The emergence of AI-driven computational tools can help reshape the methods in structural biology, however model data often quires empirical validation. The large scale of predictive modeling data will therefore benefit from optimized methodologies for the high-throughput biochemical characterization of PPIs. Biolayer interferometry (BLI) is one of very few approaches that can determine the rate of biomolecular interactions, called kinetics, and of the commonly available kinetic measurement techniques, it is the most suitable for high-throughput experimental designs. Here, we provide step-by-step instructions on how to perform kinetics experiments using BLI. We further describe the basis and execution of competition and epitope binning experiments, which are particularly useful for antibody and nanobody screening applications. The procedure requires 3 hours to complete and is suitable for users with minimal experience with biochemical techniques.
]]></description>
<dc:creator>Bates, T. A.</dc:creator>
<dc:creator>Gurmessa, S. K.</dc:creator>
<dc:creator>Reyes-Weinstein, J. B.</dc:creator>
<dc:creator>Trank-Greene, M.</dc:creator>
<dc:creator>Wrynla, X. H.</dc:creator>
<dc:creator>Anastas, A.</dc:creator>
<dc:creator>Anley, T. W.</dc:creator>
<dc:creator>Hinchliff, A.</dc:creator>
<dc:creator>Shinde, U.</dc:creator>
<dc:creator>Burke, J. E.</dc:creator>
<dc:creator>Tafesse, F. G.</dc:creator>
<dc:date>2024-10-04</dc:date>
<dc:identifier>doi:10.1101/2024.10.03.616354</dc:identifier>
<dc:title><![CDATA[An effective method of measuring nanobody binding kinetics and competition-based epitope mapping using biolayer interferometry]]></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/10.1101/2024.10.03.616430v1?rss=1">
<title>
<![CDATA[
Disruption of riboflavin biosynthesis in mycobacteria establishes 5-amino-6-D-ribitylaminouracil (5-A-RU) as key precursor of MAIT cell agonists 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.10.03.616430v1?rss=1</link>
<description><![CDATA[
Mucosal-associated invariant T (MAIT) cells exhibit an intrinsic ability to recognize and respond to microbial infections. The semi-invariant antigen recognition receptor of MAIT cells specifically detects the non-polymorphic antigen-presenting molecule, major histocompatibility complex class I-related protein 1 (MR1), which primarily binds riboflavin-derived metabolites of microbial origin. To further interrogate the dependence of these antigens on riboflavin biosynthesis in mycobacteria, we deleted individual genes in the riboflavin biosynthesis pathways in Mycobacterium smegmatis (Msm) and Mycobacterium tuberculosis (Mtb) and evaluated the impact thereof on MAIT cell activation. Blocking the early steps of the pathway by deletion of RibA2 or RibG profoundly reduced, but did not completely ablate, MAIT cell activation by Msm or Mtb, whereas deletion of RibC, which catalyzes the last step in the pathway, had no significant effect. Interestingly, deletion of RibH specifically enhanced MAIT cell recognition of Mtb whereas loss of lumazine synthase (RibH) activity had no impact on MAIT cell activation by Msm. MAIT cell activation by Msm was likewise unaffected by blocking the production of the MAIT cell antagonist, Fo (by inhibiting its conversion from the riboflavin pathway intermediate, 5-amino-6-D-ribitylaminouracil (5-A-RU), through the deletion of fbiC). Together, these results confirm a central role for 5-AR-U in generating mycobacterial MR1 ligands and reveal similarities and differences between Msm and Mtb in terms of the impact of riboflavin pathway disruption on MAIT cell activation.

Author summaryMucosal-associated invariant T (MAIT) cells are an abundant population of innate-like T-cells which respond to microbial infections. These specialized cells recognize the MR1 molecule, which presents microbial metabolites derived from riboflavin (vitamin B2) biosynthesis. These cells are enriched in the airways and in some cases reduced in the peripheral blood of tuberculosis (TB) infected individuals suggestive of a role in the early response to infection by Mycobacterium tuberculosis. In this study, we investigated the effect of deleting individual genes in the riboflavin biosynthesis pathway on MAIT cell activation by Mycobacterium tuberculosis or Mycobacterium smegmatis. Our findings revealed that disrupting early stages in the pathway profoundly reduced but did not eliminate MAIT cell activation by both mycobacterial species. However, blocking the penultimate step in the pathway, catalyzed by the enzyme, lumazine synthase, led specifically to increased MAIT cell recognition of M. tuberculosis. Our results confirm the pivotal role of the riboflavin pathway intermediate, 5-A-RU, in generating mycobacterial ligands that serve as MAIT cell agonists. By enhancing our understanding of how MAIT cells recognize mycobacterial infections, the results of this study could inform strategies for the development of vaccines and/or immunotherapies for TB.
]]></description>
<dc:creator>Chengalroyen, M. D.</dc:creator>
<dc:creator>Oketade, N.</dc:creator>
<dc:creator>Worley, A.</dc:creator>
<dc:creator>Lucas, M.</dc:creator>
<dc:creator>Ramirez, L. N.</dc:creator>
<dc:creator>Raphela, M. L.</dc:creator>
<dc:creator>Swarbrick, G.</dc:creator>
<dc:creator>Warner, D. F.</dc:creator>
<dc:creator>Lewinsohn, D.</dc:creator>
<dc:creator>Mehaffy, C.</dc:creator>
<dc:creator>Adams, E. J.</dc:creator>
<dc:creator>Hildebrand, W.</dc:creator>
<dc:creator>Dobos, K. M.</dc:creator>
<dc:creator>Mizrahi, V.</dc:creator>
<dc:creator>Lewinsohn, D.</dc:creator>
<dc:date>2024-10-04</dc:date>
<dc:identifier>doi:10.1101/2024.10.03.616430</dc:identifier>
<dc:title><![CDATA[Disruption of riboflavin biosynthesis in mycobacteria establishes 5-amino-6-D-ribitylaminouracil (5-A-RU) as key precursor of MAIT cell agonists]]></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/10.1101/2024.10.13.618056v1?rss=1">
<title>
<![CDATA[
Cryo-electron tomography reveals coupled flavivirus replication, budding and maturation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.10.13.618056v1?rss=1</link>
<description><![CDATA[
Flaviviruses replicate their genomes in replication organelles (ROs) formed as bud-like invaginations on the endoplasmic reticulum (ER) membrane, which also functions as the site for virion assembly. While this localization is well established, it is not known to what extent viral membrane remodeling, genome replication, virion assembly, and maturation are coordinated. Here, we imaged tick-borne flavivirus replication in human cells using cryo-electron tomography. We find that the RO membrane bud is shaped by a combination of a curvature-establishing coat and the pressure from intraluminal template RNA. A protein complex at the RO base extends to an adjacent membrane, where immature virions bud. Naturally occurring furin site variants determine whether virions mature in the immediate vicinity of ROs. We further visualize replication in mouse brain tissue by cryo-electron tomography. Taken together, these findings reveal a close spatial coupling of flavivirus genome replication, budding, and maturation.
]]></description>
<dc:creator>Dahmane, S.</dc:creator>
<dc:creator>Schexnaydre, E.</dc:creator>
<dc:creator>Zhang, J.</dc:creator>
<dc:creator>Rosendal, E.</dc:creator>
<dc:creator>Chotiwan, N.</dc:creator>
<dc:creator>Kumari Singh, B.</dc:creator>
<dc:creator>Yau, W.-L.</dc:creator>
<dc:creator>Lundmark, R.</dc:creator>
<dc:creator>Barad, B.</dc:creator>
<dc:creator>Grotjahn, D. A.</dc:creator>
<dc:creator>Liese, S.</dc:creator>
<dc:creator>Carlson, A.</dc:creator>
<dc:creator>Overby, A.</dc:creator>
<dc:creator>Carlson, L.-A.</dc:creator>
<dc:date>2024-10-13</dc:date>
<dc:identifier>doi:10.1101/2024.10.13.618056</dc:identifier>
<dc:title><![CDATA[Cryo-electron tomography reveals coupled flavivirus replication, budding and maturation]]></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/10.1101/2024.10.14.618249v1?rss=1">
<title>
<![CDATA[
In vitro inhibition of the CFTR ion channel in the cervix thickens cervical mucus 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.10.14.618249v1?rss=1</link>
<description><![CDATA[
Cervical mucus changes throughout the menstrual cycle in response to hormonal fluctuations, regulating access of sperm and pathogens to the reproductive tract. CFTR is an anion channel that plays a critical role in mediating epithelial mucus secretions. Primary endocervical cells obtained from rhesus macaques Macaca mulatta were cultured using conditional reprogramming and treated with vehicle controls or CFTR inhibitors. In order to measure changes in hydration and viscosity of secreted mucus, we adapted two airway mucus assays, airway surface liquid and particle-tracking microrheology, for our endocervical culture system. Endocervical cells treated with CFTR inhibitors demonstrated dehydrated, thicker mucus secretions compared to controls in both assay outputs. Our studies suggest that CFTR may be an important mediator of fertility changes and provide experimental evidence for the infertility phenotype seen in women with cystic fibrosis. Additionally, assays developed in these studies provide new endpoints for assessing cervical mucus changes in vitro.

Summary SentenceInhibition of CFTR, a key epithelial ion channel in primary endocervical cells leads to dehydrated, thickened mucus production in vitro
]]></description>
<dc:creator>Looney, R. J.</dc:creator>
<dc:creator>Roberts, M.</dc:creator>
<dc:creator>Markovetz, M.</dc:creator>
<dc:creator>Godiah, R.</dc:creator>
<dc:creator>Yao, S.</dc:creator>
<dc:creator>Golgotiu, K.</dc:creator>
<dc:creator>Wei, S.</dc:creator>
<dc:creator>Cellucci, C.</dc:creator>
<dc:creator>Han, L.</dc:creator>
<dc:date>2024-10-15</dc:date>
<dc:identifier>doi:10.1101/2024.10.14.618249</dc:identifier>
<dc:title><![CDATA[In vitro inhibition of the CFTR ion channel in the cervix thickens cervical mucus]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-10-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.10.15.618515v1?rss=1">
<title>
<![CDATA[
FBXW7 regulates MYRF levels to control myelin capacity and homeostasis in the adult CNS 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.10.15.618515v1?rss=1</link>
<description><![CDATA[
Myelin, along with the oligodendrocytes (OLs) that produce it, is essential for proper central nervous system (CNS) function in vertebrates. Although the accurate targeting of myelin to axons and its maintenance are critical for CNS performance, the molecular pathways that regulate these processes remain poorly understood. Through a combination of zebrafish genetics, mouse models, and primary OL cultures, we found FBXW7, a recognition subunit of an E3 ubiquitin ligase complex, is a regulator of adult myelination in the CNS. Loss of Fbxw7 in myelinating OLs resulted in increased myelin sheath lengths with no change in myelin thickness. As the animals aged, they developed progressive abnormalities including myelin outfolds, disrupted paranodal organization, and ectopic ensheathment of neuronal cell bodies with myelin. Through biochemical studies we found that FBXW7 directly binds and degrades the N-terminal of Myelin Regulatory Factor (N-MYRF), to control the balance between oligodendrocyte myelin growth and homeostasis.
]]></description>
<dc:creator>Collins, H. Y.</dc:creator>
<dc:creator>Doan, R. A.</dc:creator>
<dc:creator>Li, J.</dc:creator>
<dc:creator>Early, J. E.</dc:creator>
<dc:creator>Madden, M. E.</dc:creator>
<dc:creator>Simkins, T.</dc:creator>
<dc:creator>Lyons, D. A.</dc:creator>
<dc:creator>Monk, K. R.</dc:creator>
<dc:creator>Emery, B.</dc:creator>
<dc:date>2024-10-18</dc:date>
<dc:identifier>doi:10.1101/2024.10.15.618515</dc:identifier>
<dc:title><![CDATA[FBXW7 regulates MYRF levels to control myelin capacity and homeostasis in the adult CNS]]></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/10.1101/2024.10.20.619319v1?rss=1">
<title>
<![CDATA[
Understanding the development of enzalutamide resistance based on a functional single-cell approach 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.10.20.619319v1?rss=1</link>
<description><![CDATA[
Most metastatic prostate cancers (PCa) initially depend on androgen for survival and proliferation. Thus, anti-androgen or castration therapies are the mainstay treatment. Although effective at first, androgen-dependent PCa (ADPC) universally develops therapy resistance, thereby evolving to the incurable disease, called castration resistant PCa (CRPC). Currently, mechanisms underlying the emergence of CRPC from ADPC are largely unclear. We used single-cell RNA-sequencing (scRNA-Seq) to determine how a therapy-naive ADPC cell line - LNCaP responds to the anti-androgen drug, enzalutamide. We found that most cells expressed the drug-target androgen receptor (AR+), while a small subpopulation ([~]12%) expressed low or no AR (ARlow/-). Gene set enrichment analysis (GSEA) revealed that AR+ and ARlow/- cells were enriched with significantly different gene expressions and signaling pathways. Unexpectedly, ARlow/- cells displayed robust transcriptional response, including upregulations of genes and pathways involved in clinical CRPC. Next, we isolate ARlow/- and AR+ cells from the LNCaP cell line, and functionally confirmed the enzalutamide resistant phenotype of ARlow/- cells in vitro and in xenograft models in vivo. Finally, to explore a therapeutic option for ARlow/- cells, we found that ARlow/- cells expressed low levels of NAD+ biosynthesis genes, notably NAPRT, indicating a possible vulnerability to inhibitors blocking NAD+ synthesis. Indeed, treating ARlow/- cells with NAD+ synthesis inhibitors, FK866 and OT-82, significantly inhibited the survival and proliferation of ARlow/- cells, thus suggesting a possible novel therapeutic option for ADT and enzalutamide resistant PCa.

SUMMARYSingle-cell RNA-Sequencing reveals heterogeneities of tumor cell populations. In most cases, however, the functional significance of the observed heterogeneity is not tested. In this study, we first identified a possible therapy-resistant prostate cancer cell subpopulation with scRNA-Seq, then confirmed the resistant phenotype with single cell and colony - based cloning and functional testing. In addition, we also identified a therapeutic vulnerability of the resistant cells.
]]></description>
<dc:creator>Xue, C.</dc:creator>
<dc:creator>Ko, H.-K.</dc:creator>
<dc:creator>Shi, K.</dc:creator>
<dc:creator>Pittsenbarger, J.</dc:creator>
<dc:creator>Dao, L. V.</dc:creator>
<dc:creator>Shi, K.</dc:creator>
<dc:creator>Libmann, M.</dc:creator>
<dc:creator>Geng, H.</dc:creator>
<dc:creator>Qian, D. Z.</dc:creator>
<dc:date>2024-10-22</dc:date>
<dc:identifier>doi:10.1101/2024.10.20.619319</dc:identifier>
<dc:title><![CDATA[Understanding the development of enzalutamide resistance based on a functional single-cell approach]]></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/10.1101/2024.10.23.619844v1?rss=1">
<title>
<![CDATA[
AI-readiness for Biomedical Data: Bridge2AI Recommendations 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.10.23.619844v1?rss=1</link>
<description><![CDATA[
Biomedical research is rapidly adopting artificial intelligence (AI). Yet the inherent complexity of biomedical data preparation requires implementing actionable, robust criteria for ethical and explainable AI (XAI) at the "pre-model" stage, encompassing data acquisition, detailed transformations, and ethical governance. Simple conformance to FAIR (Findable, Accessible, Interoperable, Reusable) Principles is insufficient.

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

This framework sets a benchmark for preparing ethical, reusable datasets in biomedical AI and provides standardized methods for reliable pre-model data evaluation.
]]></description>
<dc:creator>Clark, T.</dc:creator>
<dc:creator>Caufield, H.</dc:creator>
<dc:creator>Mohan, J. A.</dc:creator>
<dc:creator>Al Manir, S.</dc:creator>
<dc:creator>Amorim, E.</dc:creator>
<dc:creator>Eddy, J.</dc:creator>
<dc:creator>Gim, N.</dc:creator>
<dc:creator>Gow, B.</dc:creator>
<dc:creator>Goar, W.</dc:creator>
<dc:creator>Haendel, M.</dc:creator>
<dc:creator>Hansen, J. N.</dc:creator>
<dc:creator>Harris, N.</dc:creator>
<dc:creator>Hermjakob, H.</dc:creator>
<dc:creator>McWeeney, S. K.</dc:creator>
<dc:creator>Nebeker, C.</dc:creator>
<dc:creator>Nikolov, M.</dc:creator>
<dc:creator>Shaffer, J.</dc:creator>
<dc:creator>Sheffield, N.</dc:creator>
<dc:creator>Sheynkman, G.</dc:creator>
<dc:creator>Stevenson, J.</dc:creator>
<dc:creator>Mungall, C.</dc:creator>
<dc:creator>Chen, J. Y.</dc:creator>
<dc:creator>Wagner, A.</dc:creator>
<dc:creator>Kong, S. W.</dc:creator>
<dc:creator>Ghosh, S. S.</dc:creator>
<dc:creator>Patel, B.</dc:creator>
<dc:creator>Williams, A.</dc:creator>
<dc:creator>Munoz-Torres, M. C.</dc:creator>
<dc:date>2024-10-25</dc:date>
<dc:identifier>doi:10.1101/2024.10.23.619844</dc:identifier>
<dc:title><![CDATA[AI-readiness for Biomedical Data: Bridge2AI Recommendations]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-10-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.10.27.620502v1?rss=1">
<title>
<![CDATA[
Transcriptional profiles of murine oligodendrocyte precursor cells across the lifespan 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.10.27.620502v1?rss=1</link>
<description><![CDATA[
Oligodendrocyte progenitor cells (OPCs) are highly dynamic, widely distributed glial cells of the central nervous system (CNS) that are responsible for generating myelinating oligodendrocytes during development. By also generating new oligodendrocytes in the adult CNS, OPCs allow formation of new myelin sheaths in response to environmental and behavioral changes and play a crucial role in regenerating myelin following demyelination (remyelination). However, the rates of OPC proliferation and differentiation decline dramatically with aging, which may impair homeostasis, remyelination, and adaptive myelination during learning. To determine how aging influences OPCs, we generated a novel transgenic mouse line that expresses membrane-anchored EGFP under the endogenous promoter/enhancer of Matrilin-4 (Matn4-mEGFP) and performed high-throughput single-cell RNA sequencing, providing enhanced resolution of transcriptional changes during key transitions from quiescence to proliferation and differentiation across the lifespan. Comparative analysis of OPCs isolated from mice aged 30 to 720 days, revealed that aging induces distinct inflammatory transcriptomic changes in OPCs in different states, including enhanced activation of HIF-1 and Wnt pathways. Inhibition of these pathways in acutely isolated OPCs from aged animals restored their ability to differentiate, suggesting that this enhanced signaling may contribute to the decreased regenerative potential of OPCs with aging. This Matn4-mEGFP mouse line and single-cell mRNA datasets of cortical OPCs across ages help to define the molecular changes guiding their behavior in various physiological and pathological contexts.
]]></description>
<dc:creator>Heo, D.</dc:creator>
<dc:creator>Kim, A. A.</dc:creator>
<dc:creator>Neumann, B.</dc:creator>
<dc:creator>Doze, V. N.</dc:creator>
<dc:creator>Xu, Y. K. T.</dc:creator>
<dc:creator>Mironova, Y. A.</dc:creator>
<dc:creator>Slosberg, J.</dc:creator>
<dc:creator>Goff, L. A.</dc:creator>
<dc:creator>Franklin, R. J. M.</dc:creator>
<dc:creator>Bergles, D. E.</dc:creator>
<dc:date>2024-10-28</dc:date>
<dc:identifier>doi:10.1101/2024.10.27.620502</dc:identifier>
<dc:title><![CDATA[Transcriptional profiles of murine oligodendrocyte precursor cells across the lifespan]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-10-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.10.28.620713v1?rss=1">
<title>
<![CDATA[
Reductions of Grin2a in adolescent dopamine neurons confers aberrant salience and related psychosis phenotype 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.10.28.620713v1?rss=1</link>
<description><![CDATA[
Psychosis is a hallmark of schizophrenia. It involves a collection of symptoms that are typically associated with disrupted dopamine signaling and emerges during adolescence or early adulthood. Most schizophrenia-associated genes, however, involve glutamatergic or other ubiquitous targets that do not explain the latent expression of psychosis or dopaminergic abnormalities. Here, we describe an etiologically relevant model for the adolescent onset of dopamine-related dysfunction in schizophrenia. We focused on GRIN2A, the gene encoding the GluN2A subunit of the NMDA receptor, as both the common loss-of-function variants and the rare missense variants in this gene are risk factors for schizophrenia. We find that GluN2A levels distinctly decline in dopamine neuron-containing regions throughout adolescence while remaining stable in other regions. This suggested that adolescent dopamine neurons may be particularly vulnerable to further reductions in GluN2A caused by a damaging variant of GRIN2A. Consistent with this idea, we find that selective knockout of Grin2a in adolescent rat dopamine neurons results in a psychosis-relevant behavioral phenotype. This manipulation also reduced dopamine release in response to unexpected outcomes in young adults, reflective of prediction error signaling abnormalities observed in the clinical population. These data provide mechanistic insight into how GRIN2A mutations may contribute to the delayed onset of dopamine-related symptoms and provide a model for identifying course altering treatments for schizophrenia.
]]></description>
<dc:creator>Kielhold, M. L.</dc:creator>
<dc:creator>Jacobs, D. S.</dc:creator>
<dc:creator>Pacheco, A. T.</dc:creator>
<dc:creator>Lebowitz, J. J.</dc:creator>
<dc:creator>Williams, J. T.</dc:creator>
<dc:creator>Langdon, A. J.</dc:creator>
<dc:creator>Zweifel, L.</dc:creator>
<dc:creator>Moghaddam, B.</dc:creator>
<dc:date>2024-10-28</dc:date>
<dc:identifier>doi:10.1101/2024.10.28.620713</dc:identifier>
<dc:title><![CDATA[Reductions of Grin2a in adolescent dopamine neurons confers aberrant salience and related psychosis phenotype]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-10-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.10.29.620922v1?rss=1">
<title>
<![CDATA[
Drosophila with-no-lysine and frayed Act in Parallel to dSarm to Drive Neurodegeneration Following dNmnat Depletion 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.10.29.620922v1?rss=1</link>
<description><![CDATA[
How neurons to sense when they are terminally dysfunctional and activate neurodegeneration remains poorly defined. The pro-degenerative NAD+ hydrolase dSarm/SARM1 can act as a metabolic sensor by detecting pathological changes in NAD+/NMN and subsequently induce catastrophic axon degeneration. Here we show Drosophila with-no-lysine kinase (dWnk), which can directly sense Cl-, K+ and osmotic pressure, is required for neurodegeneration induced by depletion of the NAD+ biosynthetic enzyme dNmnat. dWnk functions in parallel to dSarm and acts through the downstream kinase Frayed to promote axon degeneration and neuronal cell death. dWnk and dSarm ultimately converge on the BTB-Back domain molecule Axundead (Axed) to execute neurodegeneration. Our work argues that neurons use direct sensors of both metabolism (dSarm/SARM1) and ionic/osmotic status (dWnk) to evaluate cellular health and, when dysfunctional, promote neurodegeneration though a common axon death signaling molecule, Axundead.
]]></description>
<dc:creator>Avetisyan, A.</dc:creator>
<dc:creator>Barria, R.</dc:creator>
<dc:creator>Sheehan, A.</dc:creator>
<dc:creator>Freeman, M. R.</dc:creator>
<dc:date>2024-10-31</dc:date>
<dc:identifier>doi:10.1101/2024.10.29.620922</dc:identifier>
<dc:title><![CDATA[Drosophila with-no-lysine and frayed Act in Parallel to dSarm to Drive Neurodegeneration Following dNmnat Depletion]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-10-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.10.31.621388v1?rss=1">
<title>
<![CDATA[
Thalamic integration of basal ganglia and cerebellar circuits during motor learning 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.10.31.621388v1?rss=1</link>
<description><![CDATA[
The ability to control movement and learn new motor skills is one of the fundamental functions of the brain. The basal ganglia (BG) and the cerebellum (CB) are two key brain regions involved in controlling movement, and neuronal plasticity within these two regions is crucial for acquiring new motor skills. However, how these regions interact to produce a cohesive unified motor output remains elusive. Here, we discovered that a subset of neurons in the motor thalamus receive converging synaptic inputs from both BG and CB. By performing multi-site fiber photometry in mice learning motor tasks, we found that motor thalamus neurons integrate BG and CB signals and show distinct movement-related activity. Lastly, we found a critical role of these thalamic neurons and their BG and CB inputs in motor learning and control. These results identify the thalamic convergence of BG and CB and its crucial role in integrating movement signals.

HighlightsO_LIIndividual neurons in motor thalamus receive converging synaptic input from SNr and DCN projections.
C_LIO_LIThalamic neurons with SNr and DCN input are concentrated at the border between VM and VAL thalamic nuclei.
C_LIO_LIThalamic neurons functionally integrate SNr and DCN activity and adapt with motor learning.
C_LIO_LIThalamic neurons and their inputs from SNr and DCN are critical for learning and executing motor tasks.
C_LI
]]></description>
<dc:creator>Roth, R. H.</dc:creator>
<dc:creator>Muniak, M. A.</dc:creator>
<dc:creator>Huang, C. J.</dc:creator>
<dc:creator>Hwang, F.-J.</dc:creator>
<dc:creator>Sun, Y.</dc:creator>
<dc:creator>Min, C.</dc:creator>
<dc:creator>Mao, T.</dc:creator>
<dc:creator>Ding, J. B.</dc:creator>
<dc:date>2024-11-01</dc:date>
<dc:identifier>doi:10.1101/2024.10.31.621388</dc:identifier>
<dc:title><![CDATA[Thalamic integration of basal ganglia and cerebellar circuits during motor learning]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-11-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.10.30.621184v1?rss=1">
<title>
<![CDATA[
Single-cell, spatial, and fate-mapping analyses uncover niche dependent diversity of cochlear myeloid cells. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.10.30.621184v1?rss=1</link>
<description><![CDATA[
Recent advances in fate mapping and single-cell technologies have revealed how the dynamics and function of tissue-resident macrophages are shaped by their environment. However, macrophages in sensory organs such as the cochlea where the central nervous system and peripheral nervous system meet remain understudied. Combining single-cell transcriptomics, fate mapping, and parabiosis experiments, we show that five types of myeloid cells including three tissue-resident macrophage subpopulations, coexist in the mouse cochlea. The three macrophage subsets showed different potential functions in relationship with their specific topography across cochlear compartments. Further analysis revealed that they were partially derived from yolk sac progenitors during development, while in adulthood, most cochlear macrophages were long-term resident. Finally, we showed that cochlear macrophage morphology and density changed during aging. Our findings show that cochlea is a microenvironment with a unique heterogeneity of macrophages in terms of gene expression, spatial distribution, ontogeny, and function.
]]></description>
<dc:creator>Chiot, A.</dc:creator>
<dc:creator>Felgner, M. J.</dc:creator>
<dc:creator>Brownell, D.</dc:creator>
<dc:creator>Rott, K. H.</dc:creator>
<dc:creator>Bogachuk, A.</dc:creator>
<dc:creator>Rosmus, D.-D.</dc:creator>
<dc:creator>Masuda, T.</dc:creator>
<dc:creator>Ching, A.</dc:creator>
<dc:creator>Atkinson, P. J.</dc:creator>
<dc:creator>Prinz, M.</dc:creator>
<dc:creator>Sachs, K.</dc:creator>
<dc:creator>Cheng, A. G.</dc:creator>
<dc:creator>Wieghofer, P.</dc:creator>
<dc:creator>Ajami, B.</dc:creator>
<dc:date>2024-11-02</dc:date>
<dc:identifier>doi:10.1101/2024.10.30.621184</dc:identifier>
<dc:title><![CDATA[Single-cell, spatial, and fate-mapping analyses uncover niche dependent diversity of cochlear myeloid cells.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-11-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.11.02.621556v1?rss=1">
<title>
<![CDATA[
A high-throughput protein tagging toolkit that retains endogenous UTRs for studying gene regulation in Kinetoplastids. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.11.02.621556v1?rss=1</link>
<description><![CDATA[
Kinetoplastid parasites cause diseases that threaten human and animal health. To survive transitions between vertebrate hosts and insect vectors, these parasites rely on precise regulation of gene expression to adapt to environmental changes. Since gene regulation in Kinetoplastids is primarily post-transcriptional, developing efficient genetic tools for modifying genes at their endogenous loci while preserving regulatory mRNA elements is crucial for studying their complex biology.

We present a CRISPR/Cas9-based tagging system that preserves untranslated regulatory elements and uses a viral 2A peptide from Thosea asigna to generate two separate proteins from a single transcript: a drug-selectable marker and a tagged protein of interest. This dual-function design maintains native control elements, allowing discrimination between regulation of transcript abundance, translational efficiency, and post-translational events.

We validate the system by tagging six Trypanosoma brucei proteins and demonstrate: (i) high-efficiency positive selection and separation of drug-selectable marker and target protein, (ii) preservation of regulatory responses to environmental cues like heat shock and iron availability, and (iii) maintenance of stage-specific regulation during developmental transitions. This versatile toolkit is applicable to all kinetoplastids amenable to CRISPR/Cas9 editing, providing a powerful reverse genetic tool for studying post-transcriptional regulation and protein function in organisms where post-transcriptional control is dominant.
]]></description>
<dc:creator>Carbajo, C. G.</dc:creator>
<dc:creator>Han, X.</dc:creator>
<dc:creator>Savur, B.</dc:creator>
<dc:creator>Upadhyaya, A.</dc:creator>
<dc:creator>Taha, F.</dc:creator>
<dc:creator>Tinti, M.</dc:creator>
<dc:creator>Wheeler, R. J.</dc:creator>
<dc:creator>Yates, P. A.</dc:creator>
<dc:creator>Tiengwe, C.</dc:creator>
<dc:date>2024-11-03</dc:date>
<dc:identifier>doi:10.1101/2024.11.02.621556</dc:identifier>
<dc:title><![CDATA[A high-throughput protein tagging toolkit that retains endogenous UTRs for studying gene regulation in Kinetoplastids.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-11-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.11.02.621577v1?rss=1">
<title>
<![CDATA[
SloR-SRE binding to the S. mutans mntH promoter is cooperative. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.11.02.621577v1?rss=1</link>
<description><![CDATA[
Streptococcus mutans is a commensal member of the plaque microbiome. It is especially prevalent when dietary sugars are available for S. mutans fermentation, generating acid byproducts that lower plaque pH and foster tooth decay. S. mutans can survive in the transient conditions of the mouth, in part because it can regulate the uptake of manganese and iron during periods of feast when metal ions are available, and famine when they are limiting. S. mutans depends on a 25kDa metalloregulatory protein, called SloR, to modulate uptake of these cations across the bacterial cell surface. When bound to manganese, SloR, binds to palindromic recognition elements in the promoter of the sloABC genes that encode the major manganese transporter in S. mutans. Reports in the literature describ MntH, an ancillary manganese transporter in S. mutans, that is also subject to SloR control. In the present study, we performed expression profiling experiments that reveal coordinate regulation of the sloABC and mntH genes at the level of transcription. In addition, we describe a role for the mntH gene product that is redundant with that of the sloABC-encoded metal ion uptake machinery. The results of DNA binding studies support direct SloR binding to the mntH promoter region which, like that at the sloABC promoter, harbors three palindromic recognition elements to which SloR binds cooperatively to repress downstream transcription. These findings expand our understanding of the SloR metalloregulome and elucidate SloR-DNA binding that is essential for S. mutans metal ion homeostasis and fitness in the oral cavity.
]]></description>
<dc:creator>Ziogas, M.</dc:creator>
<dc:creator>Drummond, I. Y.</dc:creator>
<dc:creator>Todorovic, I.</dc:creator>
<dc:creator>Kraczkowsky, K. N.</dc:creator>
<dc:creator>Zhang, H.</dc:creator>
<dc:creator>Wu, H.</dc:creator>
<dc:creator>Spatafora, G.</dc:creator>
<dc:date>2024-11-03</dc:date>
<dc:identifier>doi:10.1101/2024.11.02.621577</dc:identifier>
<dc:title><![CDATA[SloR-SRE binding to the S. mutans mntH promoter is cooperative.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-11-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.11.04.621595v1?rss=1">
<title>
<![CDATA[
Multi-omics analysis in mouse primary cortical neurons reveals complex positive and negative biological interactions between constituent compounds in Centella asiatica 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.11.04.621595v1?rss=1</link>
<description><![CDATA[
BackgroundA water extract of the Ayurvedic plant Centella asiatica (CAW) improves cognitive function in mouse models of aging and Alzheimers disease, and affects dendritic arborization, mitochondrial activity and oxidative stress in mouse primary neurons. Triterpenes (TT) and caffeoylquinic acids (CQA) are constituents associated with these bioactivities of CAW although little is known about how interactions between these compounds contribute to the plants therapeutic benefit.

MethodsMouse primary cortical neurons were treated with CAW, or equivalent concentrations of four TT combined, eight CQA combined, or these twelve compounds combined (TTCQA). Treatment effects on the cell transcriptome (18,491 genes) and metabolome (192 metabolites) relative to vehicle control were evaluated using RNAseq and metabolomic analyses respectively.

ResultsExtensive differentially expressed genes (DEGs) were seen with all treatments, as well as evidence of interactions between compounds. Notably many DEGs seen with TT treatment were not observed in the TTCQA condition, possibly suggesting CQA reduced the effects of TT. Moreover, additional gene activity seen with CAW as compared to TTCQA indicate the presence of additional compounds in CAW that further modulate TTCQA interactions. Weighted Gene Correlation Network Analysis (WGCNA) identified 4 gene co-expression modules altered by treatments that were associated with extracellular matrix organization, fatty acid metabolism, cellular response to stress and stimuli, and immune function. Compound interaction patterns were seen at the eigengene level in these modules. Interestingly, in metabolomics analysis, the TTCQA treatment saw the highest number of changes in individual metabolites (20), followed by CQA (15), then TT (8) and finally CAW (3). WGCNA analysis found two metabolomics modules with significant eigenmetabolite differences for TT and CQA, and possible compound interactions at this level.

ConclusionsFour gene expression modules and two metabolite modules were altered by the four types of treatments applied. This methodology demonstrated the existence of both negative and positive interactions between TT, CQA and additional compounds found in CAW on the transcriptome and metabolome of mouse primary cortical neurons.
]]></description>
<dc:creator>Chamberlin, S. R.</dc:creator>
<dc:creator>Zweig, J.</dc:creator>
<dc:creator>Neff, C.</dc:creator>
<dc:creator>Marney, L.</dc:creator>
<dc:creator>Choi, J.</dc:creator>
<dc:creator>Yang, L.</dc:creator>
<dc:creator>Maier, C.</dc:creator>
<dc:creator>Soumyanath, A.</dc:creator>
<dc:creator>McWeeney, S.</dc:creator>
<dc:creator>Gray, N. E.</dc:creator>
<dc:date>2024-11-04</dc:date>
<dc:identifier>doi:10.1101/2024.11.04.621595</dc:identifier>
<dc:title><![CDATA[Multi-omics analysis in mouse primary cortical neurons reveals complex positive and negative biological interactions between constituent compounds in Centella asiatica]]></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/10.1101/2024.11.06.621895v1?rss=1">
<title>
<![CDATA[
Cocaine, via ΔFosB, remodels gene expression and excitability in ventral hippocampus 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.11.06.621895v1?rss=1</link>
<description><![CDATA[
Ventral hippocampus (vHPC) CA1 pyramidal neurons send glutamatergic projections to nucleus accumbens (NAc), and this vHPC-NAc circuit mediates cocaine seeking and reward, but it is unclear whether vHPC-NAc neuron properties are modulated by cocaine exposure to drive subsequent behavior. The immediate early gene transcription factor {Delta}FosB is induced throughout the brain by cocaine and is critical for cocaine seeking, but its function in vHPC-NAc neurons is not understood. We now show that circuit-specific knockout of {Delta}FosB in vHPC-NAc neurons impaired cocaine reward expression and forced abstinence-induced seeking. We also found that vHPC-NAc excitability was decreased by experimenter-administered repeated cocaine and cocaine self-administration, and this cocaine-induced excitability decrease was mediated by {Delta}FosB expression. To uncover the mechanism of this change in circuit function, we used circuit-specific translating ribosome affinity purification (TRAP) to assess cocaine-induced, {Delta}FosB-dependent changes in gene expression in vHPC-NAc. We found that cocaine causes a {Delta}FosB-dependent increase in the expression of calreticulin, an ER-resident calcium-buffering protein. Calreticulin expression mediated vHPC-NAc excitability and was necessary for cocaine reward. These findings uncover a novel, non-canonical mechanism by which cocaine increases calreticulin in vHPC leading to decreased vHPC-NAc excitability and drives cocaine seeking and reward.
]]></description>
<dc:creator>Eagle, A. L.</dc:creator>
<dc:creator>Doyle, M. A.</dc:creator>
<dc:creator>Sugimoto, C.</dc:creator>
<dc:creator>Dykstra, M. M.</dc:creator>
<dc:creator>Kuhn, H. M.</dc:creator>
<dc:creator>Murray, B. R.</dc:creator>
<dc:creator>Bastle, R. M.</dc:creator>
<dc:creator>He, J.</dc:creator>
<dc:creator>Maze, I.</dc:creator>
<dc:creator>Mazei-Robison, M.</dc:creator>
<dc:creator>Robison, A. J.</dc:creator>
<dc:date>2024-11-07</dc:date>
<dc:identifier>doi:10.1101/2024.11.06.621895</dc:identifier>
<dc:title><![CDATA[Cocaine, via ΔFosB, remodels gene expression and excitability in ventral hippocampus]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-11-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.11.06.622325v1?rss=1">
<title>
<![CDATA[
TIA1 Mediates Divergent Inflammatory Responses to Tauopathy in Microglia and Macrophages 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.11.06.622325v1?rss=1</link>
<description><![CDATA[
The RNA binding protein TIA1 is known to regulate stress responses. Here we show that TIA1 plays a much broader role in inflammatory cells, being required for the microglial sensome. We crossed TIA1 cKO mice (using a CX3CR1 driven cre element) to PS19 MAPT P301S tauopathy mice. The peripheral macrophages of TIA1 cKO mice exhibited a hyper-inflammatory phenotype with increased cytokine signaling, as expected. Surprisingly, the brains of these mice showed striking reductions in inflammation, including decreases in microglial inflammatory cytokines (TNF and IL-1{beta}) and sensome markers (CLEC7A, TREM2, ITGAX); these reductions were accompanied by corresponding decreases in tau pathology. Analysis of the brain TIA1 protein interactome identified brain selective TIA1 protein mediated pathways, including strong interactions with the microglial protein C1q, which directs pruning of dystrophic neurons. These results uncover a previously unknown regulatory role for TIA1 in microglial activation in the context of neurodegenerative disease and highlights the divergent regulation of two mononuclear phagocytic lineages: microglia and macrophages.
]]></description>
<dc:creator>Webber, C. J.</dc:creator>
<dc:creator>Van Der Spek, S.</dc:creator>
<dc:creator>Cruz, A. L. F.</dc:creator>
<dc:creator>Puri, S.</dc:creator>
<dc:creator>Zhang, C.</dc:creator>
<dc:creator>Aw, J. T. M.</dc:creator>
<dc:creator>Papadimitriou, G.-Z.</dc:creator>
<dc:creator>Roberts, R.</dc:creator>
<dc:creator>Jiang, K.</dc:creator>
<dc:creator>Tran, T. N.</dc:creator>
<dc:creator>Zhang, L.</dc:creator>
<dc:creator>Taylor, A. K.</dc:creator>
<dc:creator>Wang, Z.</dc:creator>
<dc:creator>Porter, J.</dc:creator>
<dc:creator>Sotiropoulos, I.</dc:creator>
<dc:creator>Emili, A.</dc:creator>
<dc:creator>Silva, J. M.</dc:creator>
<dc:creator>Li, H.</dc:creator>
<dc:creator>Wolozin, B.</dc:creator>
<dc:date>2024-11-07</dc:date>
<dc:identifier>doi:10.1101/2024.11.06.622325</dc:identifier>
<dc:title><![CDATA[TIA1 Mediates Divergent Inflammatory Responses to Tauopathy in Microglia and Macrophages]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-11-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.11.07.622384v1?rss=1">
<title>
<![CDATA[
Convolutional neural network models describe the encoding subspace of local circuits in auditory cortex 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.11.07.622384v1?rss=1</link>
<description><![CDATA[
Convolutional neural networks (CNNs) provide powerful models of neural sensory encoding, but their complexity makes it difficult to discern computations that support their performance. To address this limitation, we developed a linear-nonlinear subspace model that identifies the most informative sensory dimensions captured by a CNN. A CNN was trained on single-neuron data recorded from auditory cortex of ferrets during presentation of a large natural sound set. Each neurons linear tuning subspace was computed by applying dimensionality reduction to the gradient of CNN output relative to input. Subspace projections were combined nonlinearly to predict neural activity. The resulting model was functionally equivalent to the CNN. Analysis of trained models showed that responses of local neural populations sparsely tiled a shared stimulus subspace. Encoding properties also differed between cell types and layers, reflecting their position in the cortical circuit. More generally, these results establish a framework for interpreting deep learning-based encoding models.

Significance statementAuditory cortex mediates the representation and discrimination of complex sound features. Many models have been proposed for cortical sound encoding, varying in their generality, interpretability, and ease of fitting. It has been difficult to determine if and what different functional properties are captured by different models. This study shows that two families of encoding models, convolutional neural networks (CNNs) and tuning subspace models, account for the same functional properties, providing an important analytical link between accurate models that are easy to fit (CNNs) and models that are straightforward to interpret (tuning subspace).
]]></description>
<dc:creator>Wingert, J. C.</dc:creator>
<dc:creator>Parida, S.</dc:creator>
<dc:creator>Norman-Haignere, S. V.</dc:creator>
<dc:creator>David, S. V.</dc:creator>
<dc:date>2024-11-08</dc:date>
<dc:identifier>doi:10.1101/2024.11.07.622384</dc:identifier>
<dc:title><![CDATA[Convolutional neural network models describe the encoding subspace of local circuits in auditory cortex]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-11-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.11.12.623260v1?rss=1">
<title>
<![CDATA[
Proteomic characterization of Mycobacterium tuberculosis subjected to carbon starvation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.11.12.623260v1?rss=1</link>
<description><![CDATA[
Mycobacterium tuberculosis (Mtb) is the causative agent of tuberculosis (TB), the leading cause of infectious-disease related deaths worldwide. TB infections present as a spectrum from active to latent disease. In the human host, Mtb faces hostile environments, such as nutrient deprivation, hypoxia, and low pH. Under these conditions, Mtb can enter a dormant, but viable, state characterized by a lack of cell replication and increased resistance to antibiotics. These dormant Mtb pose a major challenge to curing infections and eradicating TB globally. In the current study, we subjected Mtb to carbon starvation (CS), a culture condition that induces growth stasis and mimics nutrient-starved conditions associated with dormancy in vivo. We provide a detailed analysis of the proteome in CS compared to replicating samples. We observed extensive proteomic reprogramming, with 36% of identified proteins significantly altered in CS. Many enzymes involved in oxidative phosphorylation and lipid metabolism were retained or upregulated in CS. The cell wall biosynthetic machinery was present in CS, although numerous changes in the abundance of peptidoglycan, arabinogalactan, and mycolic acid biosynthetic enzymes likely result in pronounced remodeling of the cell wall. Many clinically approved anti-TB drugs target cell wall biosynthesis, and we found that these enzymes were largely retained in CS. Lastly, we compared our results to those of other dormancy models and propose that CS produces a physiologically-distinct state of stasis compared to hypoxia in Mtb.
]]></description>
<dc:creator>Devlin, K. L.</dc:creator>
<dc:creator>Leach, D. T.</dc:creator>
<dc:creator>Stratton, K. G.</dc:creator>
<dc:creator>Lamichhane, G.</dc:creator>
<dc:creator>Lin, V.</dc:creator>
<dc:creator>Beatty, K. E.</dc:creator>
<dc:date>2024-11-12</dc:date>
<dc:identifier>doi:10.1101/2024.11.12.623260</dc:identifier>
<dc:title><![CDATA[Proteomic characterization of Mycobacterium tuberculosis subjected to carbon starvation]]></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/10.1101/2024.11.13.623052v1?rss=1">
<title>
<![CDATA[
A genome-wide ATLAS of liver chromatin architecture reveals that sex dictates diet-induced nucleosome dynamics 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.11.13.623052v1?rss=1</link>
<description><![CDATA[
The three-dimensional organization of the genome plays an important role in cellular function. Alterations between open and closed chromatin states contributes to DNA binding, collaborative transcriptional activities and informs post-transcriptional processing. The liver orchestrates systemic metabolic control and has the ability to mount a rapid adaptive response to environmental challenges. We interrogated the chromatin architecture in liver under different dietary cues. Using ATAC-seq, we mapped over 120,000 nucleosome peaks, revealing a remarkably preserved hepatic chromatin landscape across feeding conditions. Stringent analysis of nucleosome rearrangements in response to diet revealed that sex is the dominant factor segregating changes in chromatin accessibility. A lipid-rich diet led to a more accessible chromatin confirmation at promoter regions in female mice along with enrichment of promoter binding CCAAT-binding domain proteins. Male liver exhibited stronger binding for nutrient sensing nuclear receptors. Integrative analysis with gene expression corroborated a role for chromatin states in informing functional differences in metabolic traits. We distinguished the impact of gonadal sex and chromosomal sex as determinants of chromatin modulation by diet using the Four Core Genotypes mouse model. Our data provide mechanistic evidence underlying the regulation for the critical sex-dimorphic GWAS gene, Pnpla3. In summary, we provide a comprehensive epigenetic resource in murine liver that uncovers the complexity of chromatin dynamics in response to diet and sex.

HighlightsATAC-Seq, RNA-Seq, and FCG model-integrated analysis unravel sex differences in chromatin accessibility and transcriptome responses to dietary challenges.
Lipid-rich diet led to sex-biased chromatin confirmation at promoter regions.
Gonadal sex emerged as the most prevalent determinant of the sex bias hepatic chromatin modulation by lipid-rich diets.
The critical sex-dimorphic GWAS gene Pnpla3 is suppressed by testosterone, which underlies hepatic differences in expression between the sexes.
]]></description>
<dc:creator>ZHANG, Z.</dc:creator>
<dc:creator>Su, V.</dc:creator>
<dc:creator>Wiese, C. B.</dc:creator>
<dc:creator>Cheng, L.</dc:creator>
<dc:creator>Wang, D.</dc:creator>
<dc:creator>Cui, Y.</dc:creator>
<dc:creator>Kim, J.</dc:creator>
<dc:creator>Wu, X.</dc:creator>
<dc:creator>Tran, P. H.</dc:creator>
<dc:creator>Zhou, Z.</dc:creator>
<dc:creator>Casero, D.</dc:creator>
<dc:creator>Li, W.</dc:creator>
<dc:creator>Hevener, A. L.</dc:creator>
<dc:creator>Reue, K.</dc:creator>
<dc:creator>Sallam, T.</dc:creator>
<dc:date>2024-11-15</dc:date>
<dc:identifier>doi:10.1101/2024.11.13.623052</dc:identifier>
<dc:title><![CDATA[A genome-wide ATLAS of liver chromatin architecture reveals that sex dictates diet-induced nucleosome dynamics]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-11-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.11.21.624171v1?rss=1">
<title>
<![CDATA[
Ezh2 Delays Activation of Differentiation Genes During Normal Cerebellar Granule Neuron Development and in Medulloblastoma 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.11.21.624171v1?rss=1</link>
<description><![CDATA[
Medulloblastoma (MB) is the most common malignant brain tumour in children. The Sonic Hedgehog (SHH)-medulloblastoma subtype arises from the cerebellar granule neuron lineage. Terminally differentiated neurons are incapable of undergoing further cell division, so an effective treatment for this tumour could be to force neuronal differentiation. Differentiation therapy provides a potential alternative for patients with medulloblastoma who harbor mutations that impair cell death pathways (TP53), which is associated a with high mortality. To this end, our goal was to explore epigenetic regulation of cerebellar granule neuron differentiation in medulloblastoma cells. Key regulators were discovered using chromatin immunoprecipitation with high-throughput sequencing. DNA-bound protein and chromatin protein modifications were investigated across all genes. We discovered that Ezh2-mediated tri-methylation of the H3 histone (H3K27me3), occurred on more than half of the 787 genes whose transcription normally increases as granule neurons terminally differentiate. Conditional knockout of Ezh2 led to early initiation of differentiation in granule neuron precursors (GNPs), but only after cell cycle exit had occurred. Similarly, in MB cells, neuronal differentiation could be induced by preventing H3K27me3 modifications using an Ezh2 inhibitor (UNC1999), but only when UNC1999 was combined with forced cell cycle exit driven by a CDK4/6 inhibitor (Palbociclib). Ezh2 emerges as a powerful restraint upon post-mitotic differentiation during normal GNP development and combination of Ezh2 inhibition with cell cycle exit leads to MB cell differentiation.
]]></description>
<dc:creator>Purzner, J.</dc:creator>
<dc:creator>Brown, A. S.</dc:creator>
<dc:creator>Purzner, T.</dc:creator>
<dc:creator>Ellis, L.</dc:creator>
<dc:creator>Broski, S.</dc:creator>
<dc:creator>Litzenburger, U.</dc:creator>
<dc:creator>Andrews, K.</dc:creator>
<dc:creator>Sharma, A.</dc:creator>
<dc:creator>Wang, X.</dc:creator>
<dc:creator>Taylor, M. D.</dc:creator>
<dc:creator>Cho, Y.-J.</dc:creator>
<dc:creator>Fuller, M. T.</dc:creator>
<dc:creator>Scott, M. P.</dc:creator>
<dc:date>2024-11-21</dc:date>
<dc:identifier>doi:10.1101/2024.11.21.624171</dc:identifier>
<dc:title><![CDATA[Ezh2 Delays Activation of Differentiation Genes During Normal Cerebellar Granule Neuron Development and in Medulloblastoma]]></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/10.1101/2024.11.22.624784v1?rss=1">
<title>
<![CDATA[
Unraveling SARS-CoV-2 Host-Response Heterogeneity through Longitudinal Molecular Subtyping 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.11.22.624784v1?rss=1</link>
<description><![CDATA[
Hospitalized COVID-19 patients exhibit diverse immune responses during acute infection, which are associated with a wide range of clinical outcomes. However, understanding these immune heterogeneities and their links to various clinical complications, especially long COVID, remains a challenge. In this study, we performed unsupervised subtyping of longitudinal multi-omics immunophenotyping in over 1,000 hospitalized patients, identifying two critical subtypes linked to mortality or mechanical ventilation with prolonged hospital stay and three severe subtypes associated with timely acute recovery. We confirmed that unresolved systemic inflammation and T-cell dysfunctions were hallmarks of increased severity and further distinguished patients with similar acute respiratory severity by their distinct immune profiles, which correlated with differences in demographic and clinical complications. Notably, one critical subtype (SubF) was uniquely characterized by early excessive inflammation, insufficient anticoagulation, and fatty acid dysregulation, alongside higher incidences of hematologic, cardiac, and renal complications, and an elevated risk of long COVID. Among the severe subtypes, significant differences in viral clearance and early antiviral responses were observed, with one subtype (SubC) showing strong early T-cell cytotoxicity but a poor humoral response, slower viral clearance, and greater risks of chronic organ dysfunction and long COVID. These findings provide crucial insights into the complex and context-dependent nature of COVID-19 immune responses, highlighting the importance of personalized therapeutic strategies to improve both acute and long-term outcomes.
]]></description>
<dc:creator>Wang, K.</dc:creator>
<dc:creator>Nie, Y.</dc:creator>
<dc:creator>Maguire, C.</dc:creator>
<dc:creator>Syphurs, C.</dc:creator>
<dc:creator>Sheen, H.</dc:creator>
<dc:creator>Karoly, M.</dc:creator>
<dc:creator>Lapp, L.</dc:creator>
<dc:creator>Gygi, J. P.</dc:creator>
<dc:creator>Jayavelu, N. D.</dc:creator>
<dc:creator>Patel, R. K.</dc:creator>
<dc:creator>Hoch, A.</dc:creator>
<dc:creator>IMPACC Network,</dc:creator>
<dc:creator>Corry, D.</dc:creator>
<dc:creator>Kheradmand, F.</dc:creator>
<dc:creator>McComsey, G. A.</dc:creator>
<dc:creator>Fernandez-Sesma, A.</dc:creator>
<dc:creator>Simon, V.</dc:creator>
<dc:creator>Metcalf, J. P.</dc:creator>
<dc:creator>Agudelo Higuita, N. I.</dc:creator>
<dc:creator>Messer, W. B.</dc:creator>
<dc:creator>David, M. M.</dc:creator>
<dc:creator>Nadeau, K. C.</dc:creator>
<dc:creator>Kraft, M.</dc:creator>
<dc:creator>Bime, C.</dc:creator>
<dc:creator>Schaenman, J.</dc:creator>
<dc:creator>Erle, D.</dc:creator>
<dc:creator>Calfee, C. S.</dc:creator>
<dc:creator>Atkinson, M. A.</dc:creator>
<dc:creator>Brackenridge, S. C.</dc:creator>
<dc:creator>Hafler, D. A.</dc:creator>
<dc:creator>Shaw, A. C.</dc:creator>
<dc:creator>Rahman, A.</dc:creator>
<dc:creator>Hough, C. L.</dc:creator>
<dc:creator>Geng, L. N.</dc:creator>
<dc:creator>Ozonoff, A.</dc:creator>
<dc:creator>Haddad, E. K.</dc:creator>
<dc:creator>Reed, E. F.</dc:creator>
<dc:creator>Bakel, H. v.</dc:creator>
<dc:creator>Kim-Schulze, S. H.</dc:creator>
<dc:creator>Krammer, F.</dc:creator>
<dc:creator>Wilson, M.</dc:creator>
<dc:creator>Eckalbar, W.</dc:creator>
<dc:creator>Bosinger,</dc:creator>
<dc:date>2024-11-25</dc:date>
<dc:identifier>doi:10.1101/2024.11.22.624784</dc:identifier>
<dc:title><![CDATA[Unraveling SARS-CoV-2 Host-Response Heterogeneity through Longitudinal Molecular Subtyping]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-11-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.11.25.625083v1?rss=1">
<title>
<![CDATA[
Targeting the menopause transition with metformin improves breast cancer outcomes, but discontinuation has deleterious effects on metabolic health: Findings from a preclinical model of postmenopausal breast cancer. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.11.25.625083v1?rss=1</link>
<description><![CDATA[
BackgroundWomen with obesity and/or type-II-diabetes have an increased breast cancer risk, increased metastasis, and poorer prognosis, especially after menopause. In a rat model of high-fat-diet and menopause-induced weight gain, we previously reported that treatment with the anti-diabetic drug metformin for 8-weeks after ovariectomy (OVX; modeling menopause) reduced growth of existing mammary tumors and inhibited new tumor formation. This identified the menopause transition as a potential window-of-opportunity for interventions to decrease obesity-associated breast cancer incidence and disease progression. Here, we extend these findings to determine if limiting metformin to the peak window of OVX-induced weight gain would have similar anti-cancer effects.

FindingsMetformin during the first four weeks following OVX is critical to reducing tumor burden, as rats treated with metformin early (weeks0-4-postOVX) had reduced tumor burden. Conversely, initiating metformin later in the postOVX period (weeks 4-8postOVX) did not reduce cancer burden. Despite improved tumor outcomes, metformin withdrawal after the early postOVX time had detrimental metabolic effects, including weight gain and increased adiposity, insulin, IGF1, and HOMA-IR, which correlate with increased cancer risk.

ConclusionsThese data reveal early-postmenopause as a critical window when metformin decreases progression of existing disease and highlights the importance of maintaining treatment to prevent metabolic dysregulation, which could promote secondary tumors/metastasis. These findings also help explain the disconnect between epidemiological studies reporting anticancer benefits of metformin and more recent clinical trials that failed to see similar efficacy, potentially due to issues of timing and/or inclusion of women outside the early postmenopausal window and/or without underlying metabolic dysfunction.
]]></description>
<dc:creator>Corleto, K. A.</dc:creator>
<dc:creator>Schedin, P.</dc:creator>
<dc:creator>Kotta, A. S.</dc:creator>
<dc:creator>Strandmo, J. L.</dc:creator>
<dc:creator>Foster, S. M.</dc:creator>
<dc:creator>Lammoglia, N.</dc:creator>
<dc:creator>Karmakar, M.</dc:creator>
<dc:creator>Carroll, R. J.</dc:creator>
<dc:creator>MacLean, P. S.</dc:creator>
<dc:creator>Giles, E. D.</dc:creator>
<dc:date>2024-11-26</dc:date>
<dc:identifier>doi:10.1101/2024.11.25.625083</dc:identifier>
<dc:title><![CDATA[Targeting the menopause transition with metformin improves breast cancer outcomes, but discontinuation has deleterious effects on metabolic health: Findings from a preclinical model of postmenopausal breast cancer.]]></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/10.1101/2024.11.26.625543v1?rss=1">
<title>
<![CDATA[
Systemic exposure to COVID-19 virus-like particles modulates firing patterns of cortical neurons in the living mouse brain 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.11.26.625543v1?rss=1</link>
<description><![CDATA[
Severe Acute Respiratory Syndrome Corona Virus 2 (SARS-CoV-2) causes a systemic infection that affects the central nervous system. We used virus-like particles (VLPs) to explore how exposure to the SARS-CoV-2 proteins affects brain activity patterns in wild-type (WT) mice and in mice that express the wild-type human tau protein (htau mice). VLP exposure elicited dose-dependent changes in corticosterone and distinct chemokine levels. Longitudinal two-photon microscopy recordings of primary somatosensory and motor cortex neurons that express the jGCaMP7s calcium sensor tracked modifications of neuronal activity patterns following exposure to VLPs. There was a substantial short-term increase in stimulus-evoked activity metrics in both WT and htau VLP-injected mice, while htau mice showed also increased spontaneous activity metrics and increase activity in the vehicle-injected group. Over the following weeks, activity metrics in WT mice subsided, but remained above baseline levels. For htau mice, activity metrics either remain elevated or decreased to lower levels than baseline. Overall, our data suggest that exposure to the SARS-CoV-2 VLPs leads to strong short-term disruption of cortical activity patterns in mice with long-term residual effects. The htau mice, which have a more vulnerable genetic background, exhibited more severe pathobiology that may lead to more adverse outcomes.
]]></description>
<dc:creator>Das, A.</dc:creator>
<dc:creator>Icardi, J.</dc:creator>
<dc:creator>Borovicka, J.</dc:creator>
<dc:creator>Holden, S.</dc:creator>
<dc:creator>Harrison, H. F.</dc:creator>
<dc:creator>Hirsch, A. J.</dc:creator>
<dc:creator>Raber, J.</dc:creator>
<dc:creator>Dana, H.</dc:creator>
<dc:date>2024-11-27</dc:date>
<dc:identifier>doi:10.1101/2024.11.26.625543</dc:identifier>
<dc:title><![CDATA[Systemic exposure to COVID-19 virus-like particles modulates firing patterns of cortical neurons in the living mouse brain]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-11-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.11.28.625832v1?rss=1">
<title>
<![CDATA[
The TLR7/8 agonist INI-4001 enhances the immunogenicity of a Powassan virus-like-particle vaccine 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.11.28.625832v1?rss=1</link>
<description><![CDATA[
Powassan virus (POWV) is a pathogenic tick-borne flavivirus that causes fatal neuroinvasive disease in humans. There are currently no approved therapies or vaccines for POWV infection. Here, we develop a POW virus-like-particle (POW-VLP) based vaccine adjuvanted with the novel synthetic Toll-like receptor 7/8 agonist INI-4001. We demonstrate that INI-4001 outperforms both alum and the Toll-like receptor 4 agonist INI-2002 in enhancing the immunogenicity of a dose-sparing POW-VLP vaccine in mice. INI-4001 increases the magnitude and breadth of the antibody response as measured by whole-virus ELISA, induces neutralizing antibodies measured by FRNT, reduces viral burden in the brain of infected mice measured by RT qPCR, and confers 100% protection from lethal challenge with both lineages of POWV. We show that the antibody response induced by INI-4001 is more durable than standard alum, and 80% of mice remain protected from lethal challenge 9-months post-vaccination. Lastly, we show that the protection elicited by INI-4001 adjuvanted POW-VLP vaccine is unaffected by either CD4+ or CD8+ T cell depletion and can be passively transferred to unvaccinated mice indicating that protection is mediated through humoral immunity. This study highlights the utility of novel synthetic adjuvants in VLP-based vaccines.

Author summaryPowassan virus (POWV) is an emerging pathogenic tick-borne flavivirus for which there is no vaccine. Current tick-borne flavivirus vaccines are less than ideal and use formalin-inactivated virus adjuvanted with alum. These vaccines require thorough inactivation of the antigen and frequent boosting to maintain immunity. In this study, we describe the development of a POWV vaccine using Powassan virus-like-particles (POW-VLPs) adjuvanted with either of two novel Toll-like receptor (TLR) agonists, the TLR4 agonist INI-2002 or the TLR7/8 agonist INI-4001. We show that INI-4001 enhances the antibody response, reduces POWV neuroinvasion, and elicits full protection from lethal POWV infection in mice prime-boost vaccinated with low doses of POW-VLP. We further show that this protection is mediated by a humoral immune response which is both broader and more durable than a POW-VLP vaccine formulated with alum. These findings demonstrate the effectiveness of the novel synthetic TLR7/8 agonist INI-4001 as an adjuvant for low-dose VLP-based vaccines and the ability of this vaccine platform to improve upon current tick-borne flavivirus vaccine methodology.
]]></description>
<dc:creator>Crawford, M. W.</dc:creator>
<dc:creator>Abdelwahab, W. M.</dc:creator>
<dc:creator>Siram, K.</dc:creator>
<dc:creator>Parkins, C. J.</dc:creator>
<dc:creator>Harrison, H. F.</dc:creator>
<dc:creator>Osman, S. R.</dc:creator>
<dc:creator>Schweitzer, D.</dc:creator>
<dc:creator>Evans, J. T.</dc:creator>
<dc:creator>Burkhart, D. J.</dc:creator>
<dc:creator>Pinto, A. K.</dc:creator>
<dc:creator>Brien, J. D.</dc:creator>
<dc:creator>Smith, J. L.</dc:creator>
<dc:creator>Hirsch, A. J.</dc:creator>
<dc:date>2024-12-03</dc:date>
<dc:identifier>doi:10.1101/2024.11.28.625832</dc:identifier>
<dc:title><![CDATA[The TLR7/8 agonist INI-4001 enhances the immunogenicity of a Powassan virus-like-particle vaccine]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-12-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.12.03.626649v1?rss=1">
<title>
<![CDATA[
Multinucleated giant cells are hallmarks of ovarian aging with unique immune and degradation-associated molecular signatures 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.12.03.626649v1?rss=1</link>
<description><![CDATA[
The ovary is one of the first organs to exhibit signs of aging, characterized by reduced tissue function, chronic inflammation, and fibrosis. Multinucleated giant cells (MNGCs), formed by macrophage fusion, typically occur in chronic immune pathologies, including infectious and non-infectious granulomas and the foreign body response1, but are also observed in the aging ovary2-4. The function and consequence of ovarian MNGCs remain unknown as their biological activity is highly context-dependent, and their large size has limited their isolation and analysis through technologies such as single-cell RNA sequencing. In this study, we define ovarian MNGCs through a deep analysis of their presence across age and species using advanced imaging technologies as well as their unique transcriptome using laser capture microdissection. MNGCs form complex interconnected networks that increase with age in both mouse and nonhuman primate ovaries. MNGCs are characterized by high Gpnmb expression, a putative marker of ovarian and non-ovarian MNGCs5,6. Pathway analysis highlighted functions in apoptotic cell clearance, lipid metabolism, proteolysis, immune processes, and increased oxidative phosphorylation and antioxidant activity. Thus, MNGCs have signatures related to degradative processes, immune function, and high metabolic activity. These processes were enriched in MNGCs compared to primary ovarian macrophages, suggesting discrete functionality. MNGCs express CD4 and colocalize with T-cells, which were enriched in regions of MNGCs, indicative of a close interaction between these immune cell types. These findings implicate MNGCs in modulation of the ovarian immune landscape during aging given their high penetrance and unique molecular signature that supports degradative and immune functions.
]]></description>
<dc:creator>Converse, A.</dc:creator>
<dc:creator>Perry, M. J.</dc:creator>
<dc:creator>Dipali, S. S.</dc:creator>
<dc:creator>Isola, J. V.</dc:creator>
<dc:creator>Kelley, E. B.</dc:creator>
<dc:creator>Varberg, J. M.</dc:creator>
<dc:creator>Zelinkski, M. B.</dc:creator>
<dc:creator>Gerton, J. L.</dc:creator>
<dc:creator>Stout, M.</dc:creator>
<dc:creator>Pritchard, M. T.</dc:creator>
<dc:creator>Duncan, F.</dc:creator>
<dc:date>2024-12-04</dc:date>
<dc:identifier>doi:10.1101/2024.12.03.626649</dc:identifier>
<dc:title><![CDATA[Multinucleated giant cells are hallmarks of ovarian aging with unique immune and degradation-associated molecular signatures]]></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/10.1101/2024.12.01.626246v1?rss=1">
<title>
<![CDATA[
A negative binomial latent factor model for paired microbiome sequencing data 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.12.01.626246v1?rss=1</link>
<description><![CDATA[
MotivationMicrobiome compositional data are often collected from several body sites and exhibit dependency among them. Analyzing microbial compositions from different sites jointly allows for effective borrowing of information by exploiting the underlying cross-site correlation, which can lead to more effective statistical analysis, especially when the sample size at one or both sites is limited. To this end, we introduce a joint model for microbiome compositions at two (or more) sites within the same subjects. Our model incorporates (i) latent factors shared across two body sites to explain the common subject effects and to serve as the source of correlation between the two sites; and (ii) mixtures of latent factors to allow heterogeneity among the samples in their level of cross-site association. The model is illustrated with synthetic data and we apply it in a case study involving samples of the urinary and vaginal microbiome collected from women.

ResultsSimulation studies show how common subject effects influence regression analysis results; a stronger association between two sites in the data causes a greater degree of bias in the analysis. The model with latent factors mitigates the bias present in the model without latent factors, whereas the two models perform comparably for the data set without paired associations. In a case study involving samples collected from a study on the female urogenital microbiome with aging (e.g., the UMICRO study), our model leads to the detection of covariate associations of the vaginal and urinary microbiome composition that are otherwise not statistically significant under a similar regression model applied to the two sites separately. Our model also enables prediction of the microbial abundance at one site based on observations from another site. We also consider a model extension that allows the clustering of subjects (samples) and cluster-specific levels of paired association. Under the extended modeling framework, the clusters can be classified according to their association strengths.
]]></description>
<dc:creator>Kim, H.</dc:creator>
<dc:creator>Siddiqui, N. Y.</dc:creator>
<dc:creator>Karstens, L.</dc:creator>
<dc:creator>Ma, L.</dc:creator>
<dc:date>2024-12-05</dc:date>
<dc:identifier>doi:10.1101/2024.12.01.626246</dc:identifier>
<dc:title><![CDATA[A negative binomial latent factor model for paired microbiome sequencing data]]></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/10.1101/2024.12.01.626256v1?rss=1">
<title>
<![CDATA[
Alpha-synuclein knockout impairs melanoma development and alters DNA damage repair in the TG3 mouse model in a sex-dependent manner 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.12.01.626256v1?rss=1</link>
<description><![CDATA[
Strong evidence suggests links between Parkinsons Disease (PD) and melanoma, as studies have found that people with PD are at an increased risk of developing melanoma and those with melanoma are at increased risk of developing PD. Although these clinical associations are well-established, the cellular and molecular pathways linking these diseases are poorly understood. Recent studies have found a previously unrecognized role for the neurodegeneration-associated protein alpha-synuclein (Syn) in melanoma; the overexpression of Syn promotes melanoma cell proliferation and metastasis. However, to our knowledge, no studies have investigated the role of Syn in in vivo melanoma models outside of a xenograft paradigm. Our study created and characterized Snca knockout in the spontaneously developing melanoma TG3 mouse line, TG3+/+Snca-/-. We show that Syn loss-of-function significantly delays melanoma onset and slows tumor growth in vivo. Furthermore, decreased tumor volume is correlated with a decreased DNA damage signature and increased apoptotic markers, indicating a role for Syn in modulating the DNA damage response (DDR) pathway. Overall, our study provides evidence that targeting Syn and its role in modulating the DDR and melanomagenesis could serve as a promising new therapeutic target.
]]></description>
<dc:creator>Arnold, M. R.</dc:creator>
<dc:creator>Chen, S.</dc:creator>
<dc:creator>Unni, V. K.</dc:creator>
<dc:date>2024-12-05</dc:date>
<dc:identifier>doi:10.1101/2024.12.01.626256</dc:identifier>
<dc:title><![CDATA[Alpha-synuclein knockout impairs melanoma development and alters DNA damage repair in the TG3 mouse model in a sex-dependent manner]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-12-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.12.02.626426v1?rss=1">
<title>
<![CDATA[
IFNγ-dependent metabolic reprogramming restrains an immature, pro-metastatic lymphatic state in melanoma 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.12.02.626426v1?rss=1</link>
<description><![CDATA[
Lymphatic vessels play a crucial role in activating anti-tumor immune surveillance but also contribute to metastasis and systemic tumor progression. Whether distinct lymphatic phenotypes exist that govern the switch between immunity and metastasis remains unclear. Here we reveal that cytotoxic immunity normalizes lymphatic function and uncouples immune and metastatic potential. We find that in mice and humans, intratumoral lymphatic vessel density negatively correlates with productive cytotoxic immune responses and identify IFN{gamma} as an intrinsic inhibitor of lymphangiogenesis. Specific deletion of the Ifngr1 in lymphatic endothelial cells (LECs) greatly expanded the intratumoral lymphatic network and drove the emergence of a tip-like endothelial state, promoting lymph node metastasis but not dendritic cell migration. IFN{gamma} inhibits oxidative phosphorylation, which is required for proliferation and acquisition of the pathologic transcriptional state. Our data indicate that IFN{gamma} induces a phenotypic switch in tumor-associated lymphatic vessels to reinforce canonical immune surveillance and block metastasis.
]]></description>
<dc:creator>Karakousi, T.</dc:creator>
<dc:creator>Cristaldi, V.</dc:creator>
<dc:creator>Lopes de Oliveira, M. L.</dc:creator>
<dc:creator>Medeiros Geraldo, L. H.</dc:creator>
<dc:creator>Gonzalez-Robles, T.</dc:creator>
<dc:creator>da Silva, G.</dc:creator>
<dc:creator>Breazeale, A. P.</dc:creator>
<dc:creator>Encarnacion Rosado, J.</dc:creator>
<dc:creator>Pozniak, J.</dc:creator>
<dc:creator>Kimmelman, A.</dc:creator>
<dc:creator>Ruggles, K.</dc:creator>
<dc:creator>Marine, J. C.</dc:creator>
<dc:creator>Chandel, N. S.</dc:creator>
<dc:creator>Lund, A. W.</dc:creator>
<dc:date>2024-12-05</dc:date>
<dc:identifier>doi:10.1101/2024.12.02.626426</dc:identifier>
<dc:title><![CDATA[IFNγ-dependent metabolic reprogramming restrains an immature, pro-metastatic lymphatic state in melanoma]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-12-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.12.03.626640v1?rss=1">
<title>
<![CDATA[
Mechanism of small heat shock protein client sequestration and induced polydispersity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.12.03.626640v1?rss=1</link>
<description><![CDATA[
Small heat shock proteins (sHSPs) act as first responders during cellular stress by recognizing and sequestering destabilized proteins (clients), preventing their aggregation and facilitating downstream refolding or degradation1-3. This chaperone function is critically important to proteostasis, conserved across all kingdoms of life, and associated with various protein misfolding diseases in humans4,5. Mechanistic insights into how sHSPs sequester destabilized clients have been limited due to the extreme molecular plasticity and client-induced polydispersity of sHSP/client complexes6-8. Here, we present high-resolution cryo-EM structures of the sHSP from Methanocaldococcus jannaschii (mjHSP16.5) in both the apo-state and in an ensemble of client-bound states. The ensemble not only reveals key molecular mechanisms by which sHSPs respond to and sequester client proteins, but also provides insights into the cooperative nature of chaperone-client interactions. Engagement with destabilized client induces a polarization of stability across the mjHSP16.5 scaffold, proposed to facilitate higher-order assembly and enhance client sequestration capacity. Some higher-order sHSP oligomers appear to form through simple insertion of dimeric subunits into new geometrical features, while other higher-order states suggest multiple sHSP/client assembly pathways. Together, these results provide long-sought insights into the chaperone function of sHSPs and highlight the relationship between polydispersity and client sequestration under stress conditions.
]]></description>
<dc:creator>Miller, A. P.</dc:creator>
<dc:creator>Reichow, S. L.</dc:creator>
<dc:date>2024-12-06</dc:date>
<dc:identifier>doi:10.1101/2024.12.03.626640</dc:identifier>
<dc:title><![CDATA[Mechanism of small heat shock protein client sequestration and induced polydispersity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-12-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.12.06.626879v1?rss=1">
<title>
<![CDATA[
UniFORM: Towards Universal Immunofluorescence Normalization for Multiplex Tissue Imaging 
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</title>
<link>https://biorxiv.org/content/10.1101/2024.12.06.626879v1?rss=1</link>
<description><![CDATA[
Multiplexed tissue imaging (MTI) technologies enable high-dimensional spatial analysis of tumor microenvironments but face challenges with technical variability in staining intensities. Existing normalization methods, including Z-score, ComBat, and MxNorm, often fail to account for the heterogeneous, right-skewed expression patterns of MTI data, compromising signal alignment and downstream analyses. We present UniFORM, a non-parametric, Python-based pipeline that uses an automated rigid landmark functional data registration approach for normalizing both feature- and pixel-level MTI data. Designed specifically for the distributional characteristics of MTI datasets, UniFORM operates without prior distributional assumptions and performs robustly regardless of distribution modality, including both unimodal and bimodal patterns. It removes technical variation by aligning the biologically invariant component of the signal, typically the negative (non-expressing) population, while preserving biologically meaningful variation in the positive population, thereby maintaining tissue-specific expression patterns essential for downstream analysis. Benchmarking across three distinct MTI platform datasets demonstrates that UniFORM outperforms existing methods in mitigating batch effects while maintaining biological signal fidelity. This is evidenced by improved marker distribution alignment and positive population preservation, enhanced kBET and Silhouette scores, and improved downstream analyses such as UMAP visualizations and Leiden clustering. UniFORM also introduces a novel guided fine-tuning option for complex and heterogeneous datasets. Although optimized for fluorescence-based platforms, UniFORM provides a scalable and robust solution for MTI data normalization, enabling accurate and biologically meaningful interpretations.
]]></description>
<dc:creator>Wang, K.</dc:creator>
<dc:creator>Ait-Ahmad, K.</dc:creator>
<dc:creator>Kupp, S.</dc:creator>
<dc:creator>Sims, Z.</dc:creator>
<dc:creator>Cramer, E.</dc:creator>
<dc:creator>Sayar, Z.</dc:creator>
<dc:creator>Yu, J.</dc:creator>
<dc:creator>Wong, M. H.</dc:creator>
<dc:creator>Mills, G. B.</dc:creator>
<dc:creator>Eksi, S. E.</dc:creator>
<dc:creator>Chang, Y. H.</dc:creator>
<dc:date>2024-12-10</dc:date>
<dc:identifier>doi:10.1101/2024.12.06.626879</dc:identifier>
<dc:title><![CDATA[UniFORM: Towards Universal Immunofluorescence Normalization for Multiplex Tissue Imaging]]></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/10.1101/2024.12.10.627859v1?rss=1">
<title>
<![CDATA[
Recognizing amino acid sidechains in a medium resolution cryo-electron density map 
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</title>
<link>https://biorxiv.org/content/10.1101/2024.12.10.627859v1?rss=1</link>
<description><![CDATA[
Building an accurate atomic structure model of a protein into a cryo-electron microscopy (cryo-EM) map at worse than 3 [A] resolution is difficult. To facilitate this task, we devised a method for assigning the amino acid residue sequence to the backbone fragments traced in an input cryo-EM map (EMSequenceFinder). EMSequenceFinder relies on a Bayesian scoring function for ranking 20 standard amino acid residue types at a given backbone position, based on the fit to a density map, map resolution, and secondary structure propensity. The fit to a density is quantified by a convolutional neural network that was trained on [~]5.56 million amino acid residue densities extracted from cryo-EM maps at 3-10 [A] resolution and corresponding atomic structure models deposited in the Electron Microscopy Data Bank (EMDB). We benchmarked EMSequenceFinder by predicting the sequences of 58,044 distinct L-helix and {beta}-strand fragments, given the fragment backbone coordinates fitted in their density maps. EMSequenceFinder identifies the correct sequence as the best-scoring sequence in 77.8% of these cases. We also assessed EMSequenceFinder on separate datasets of cryo-EM maps at resolutions from 4 to 6 L. The accuracy of EMSequenceFinder (63.5%) was better than that of three tested state-of-the-art methods, including findMysequence (45%), ModelAngelo (27%), and sequence_from_map in Phenix (12.9%). We further illustrate EMSequenceFinder by threading the SARS-CoV-2 NSP2 sequence into eight cryo-EM maps at resolutions from 3.7 to 7.0 [A]. EMSequenceFinder is implemented in our open-source Integrative Modeling Platform (IMP) program. Thus, it is expected to be helpful for integrative structure modeling based on a cryo-EM map and other information, such as models of protein complex components and chemical crosslinks between them. EMSequenceFinder is available as part of our open source IMP distribution at https://integrativemodeling.org/.
]]></description>
<dc:creator>Mondal, D.</dc:creator>
<dc:creator>Kumar, V.</dc:creator>
<dc:creator>Satler, T.</dc:creator>
<dc:creator>Ramachandran, R.</dc:creator>
<dc:creator>Saltzberg, D.</dc:creator>
<dc:creator>Chemmama, I.</dc:creator>
<dc:creator>Pilla, K. B.</dc:creator>
<dc:creator>Echeverria, I.</dc:creator>
<dc:creator>Webb, B. M.</dc:creator>
<dc:creator>Gupta, M.</dc:creator>
<dc:creator>Verba, K. A.</dc:creator>
<dc:creator>Sali, A.</dc:creator>
<dc:date>2024-12-12</dc:date>
<dc:identifier>doi:10.1101/2024.12.10.627859</dc:identifier>
<dc:title><![CDATA[Recognizing amino acid sidechains in a medium resolution cryo-electron density map]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-12-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.12.11.628012v1?rss=1">
<title>
<![CDATA[
Conformational plasticity of human acid-sensing ion channel 1a 
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</title>
<link>https://biorxiv.org/content/10.1101/2024.12.11.628012v1?rss=1</link>
<description><![CDATA[
Acid-sensing ion channels (ASICs) are typically activated by acidic environments and contribute to nociception and synaptic plasticity. ASIC1a is the most abundant subunit in the central nervous system and forms homomeric channels permeable to Na+ and Ca2+, making it a compelling therapeutic target for acidotic pathologies including stroke and traumatic brain injury. However, a complete conformational library of human ASIC1a in its various functional states has yet to be described. Using cryo-EM, we obtained hASIC1a structures across a pH range between 8.5 and 5.7, as well as in the presence of a toxin agonist and a gating-modulating mutation. We identify six major conformations that establish linear transmembrane helices to be associated with an open state, delineate mechanistic differences between proton and toxin activation, and demonstrate that desensitization leads to unexpected conformational diversity in the transmembrane domain. Together, they provide a three-dimensional rationalization of decades of structure-function studies on ASIC.
]]></description>
<dc:creator>Cahill, J.</dc:creator>
<dc:creator>Hartfield, K. A.</dc:creator>
<dc:creator>Heusser, S. A.</dc:creator>
<dc:creator>Poulsen, M. H.</dc:creator>
<dc:creator>Yoshioka, C.</dc:creator>
<dc:creator>Pless, S. A.</dc:creator>
<dc:creator>Baconguis, I.</dc:creator>
<dc:date>2024-12-12</dc:date>
<dc:identifier>doi:10.1101/2024.12.11.628012</dc:identifier>
<dc:title><![CDATA[Conformational plasticity of human acid-sensing ion channel 1a]]></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/10.1101/2024.12.18.629013v1?rss=1">
<title>
<![CDATA[
Synaptic facilitation enhances the reliability and precision of high frequency neurotransmission 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.12.18.629013v1?rss=1</link>
<description><![CDATA[
The small and tortuous volume of synaptic clefts limits the diffusion of Ca2+ ions during high frequency spiking. Extracellular Ca2+ levels ([Ca2+]o) of 0.8 mM or lower have been measured or calculated for different synapses. Here, we recorded evoked postsynaptic potentials (EPSP) and action potentials (AP) from young adult male and female mouse auditory brainstem principal neurons to investigate the relationship between neurotransmission reliability, stimulation frequency and [Ca2+]o. In 0.8 mM [Ca2+]o, we observed AP failures during afferent fiber stimulation at 100 Hz. Surprisingly, AP failures, EPSP-AP latency and jitter were all greatly reduced when stimulation frequency was increased to 500 Hz. Analysis of the EPSP/AP waveform revealed marked facilitation at 500 Hz that was not present at 100 Hz. Raising [Ca2+]o to 1.2 mM or 2.0 mM reduced or eliminated facilitation and, in these conditions, stimulation at 500 Hz increased the number of AP failures. In 0.8 mM [Ca2+]o, afferent fiber stimulation over a broad range of frequencies from 10-1000 Hz produced three different types of spiking responses: Type I cells exhibited band-pass filtering, with best response at approximately 500 Hz, Type II cells exhibited low-pass filtering above 600 Hz, and Type III cells exhibited shallow band-pass filtering centered at approximately 300 Hz. To predict AP success or failure, we built a model based on three factors: size of the EPSP, membrane potential immediately prior to the synaptic event and the number of preceding failures. We conclude that synaptic facilitation can contribute positively to the maintenance of reliable and precise high frequency neurotransmission in the auditory brainstem.

Significance StatementFacilitation of evoked postsynaptic currents is a common feature of synapses. The strength of facilitation and its role in reaching spike threshold depends on intrinsic properties of the synapse, stimulation frequency, and extracellular Ca2+ concentration ([Ca2+]o). Physiological levels of [Ca2+]o can vary from 0.8 to 1.2 mM depending on synaptic activity. In the mouse Medial Nucleus of the Trapezoid Body, synaptic facilitation is readily observable in slice experiments using relatively low (0.8 mM) [Ca2+]o, but is partially or completely obscured by short-term synaptic depression when [Ca2+]o is high (1.2 or 2.0 mM). Here we show that facilitation can rescue the reliability of high-frequency (500 Hz) action potential firing in low [Ca2+]o, resulting in band-pass filter transmission.
]]></description>
<dc:creator>Bernal-Correa, A. M.</dc:creator>
<dc:creator>Dagostin, A.</dc:creator>
<dc:creator>von Gersdorff, H.</dc:creator>
<dc:creator>Kushmerick, C.</dc:creator>
<dc:date>2024-12-18</dc:date>
<dc:identifier>doi:10.1101/2024.12.18.629013</dc:identifier>
<dc:title><![CDATA[Synaptic facilitation enhances the reliability and precision of high frequency neurotransmission]]></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/10.1101/2024.12.18.628944v1?rss=1">
<title>
<![CDATA[
Statins inhibit onco-dimerization of the 4Ig isoform of B7-H3 
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</title>
<link>https://biorxiv.org/content/10.1101/2024.12.18.628944v1?rss=1</link>
<description><![CDATA[
B7-H3 (CD276), a member of the B7-family of immune checkpoint proteins, has been shown to have immunological and non-immunological effects promoting tumorigenesis [1, 2] and expression correlates with poor prognosis for many solid tumors, including cervical, ovarian and breast cancers [3-6]. We recently identified a tumor-cell autochthonous tumorigenic role for dimerization of the 4Ig isoform of B7-H3 (4Ig-B7-H3) [7], where 4Ig-B7-H3 dimerization in cis activated tumor-intrinsic cellular proliferation and tumorigenesis pathways, providing a novel opportunity for therapeutic intervention. Herein, a live cell split-luciferase complementation strategy was used to visualize 4Ig-B7-H3 homodimerization in a high-throughput small molecule screen (HTS) to identify modulators of this protein-protein interaction (PPI). Notably, the HTS identified several compounds that converged on lipid metabolism (including HMG-CoA reductase inhibitors, also known as statins) as significant inhibitors of 4Ig-B7-H3 dimerization (p < 0.01). In vitro and in vivo murine studies provided evidence that statin-mediated disruption of 4Ig-B7-H3 dimerization was associated with anti-tumor effects. Statin-mediated anti-cancer efficacy was selective for B7-H3-expressing tumors and retrospective analysis of clinical tumor specimens supported the hypothesis that concurrent statin use enhanced clinical outcomes for patients in a B7-H3 restricted manner. Thus, disruption of 4Ig-B7-H3 dimerization provides an unanticipated molecular mechanism linking statin use in cancer therapy and prevention with immune checkpoint.
]]></description>
<dc:creator>Sutton, M. N.</dc:creator>
<dc:creator>Glazer, S. E.</dc:creator>
<dc:creator>Al Zaki, A.</dc:creator>
<dc:creator>Napoli, A.</dc:creator>
<dc:creator>Yang, P.</dc:creator>
<dc:creator>Bhosale, P.</dc:creator>
<dc:creator>Liu, J.</dc:creator>
<dc:creator>Gammon, S. T.</dc:creator>
<dc:creator>Piwnica-Worms, D.</dc:creator>
<dc:date>2024-12-20</dc:date>
<dc:identifier>doi:10.1101/2024.12.18.628944</dc:identifier>
<dc:title><![CDATA[Statins inhibit onco-dimerization of the 4Ig isoform of B7-H3]]></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/10.1101/2024.12.18.629070v1?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/10.1101/2024.12.18.629263v1?rss=1">
<title>
<![CDATA[
Host and rabies virus gene expression is shaped by human brain cell type and reveals a preexisting pro-viral transcriptional state in astrocytes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.12.18.629263v1?rss=1</link>
<description><![CDATA[
How virus-host cell interactions and innate immune antagonism shape neurotropic infection dynamics across diverse brain cell types are largely unknown. To "unmask" and study how innate immune inhibition affects cell type-specific transcriptional regulation of the human and viral genome, we performed single-cell RNA sequencing of human brain cell co-cultures, comparing an isolate of rabies virus (RABV) to its mutant incapable of antagonizing interferon- and NF-{kappa}B-dependent responses. RABV gene expression was shaped by host cell type. RABV induced small-scale, cell-type conserved transcriptional programs that likely support infection by 1) hijacking negative transcriptional feedback of pro-viral factors while 2) reducing anti-viral RNAs. Unexpectedly, disinhibited innate immune signaling increased RABV transcription, most strikingly in an infection-independent "pro-viral" astrocyte subpopulation. Further analysis suggested that "pro-viral"-like astrocytes are a rare subtype in the human brain and are primed to protect the brain during viral infection in concert with interferon-sensitive microglia recalcitrant to infection.
]]></description>
<dc:creator>Feige, L.</dc:creator>
<dc:creator>Young, K.</dc:creator>
<dc:creator>Cerapio, J. P.</dc:creator>
<dc:creator>Kozaki, T.</dc:creator>
<dc:creator>Kergoat, L.</dc:creator>
<dc:creator>Libri, V.</dc:creator>
<dc:creator>Ginhoux, F.</dc:creator>
<dc:creator>Hasan, M.</dc:creator>
<dc:creator>Ben Ameur, L.</dc:creator>
<dc:creator>Chin, G.</dc:creator>
<dc:creator>Goode, Z.</dc:creator>
<dc:creator>Bourhy, H.</dc:creator>
<dc:creator>Saunders, A.</dc:creator>
<dc:date>2024-12-20</dc:date>
<dc:identifier>doi:10.1101/2024.12.18.629263</dc:identifier>
<dc:title><![CDATA[Host and rabies virus gene expression is shaped by human brain cell type and reveals a preexisting pro-viral transcriptional state in astrocytes]]></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/10.1101/2024.12.19.629517v1?rss=1">
<title>
<![CDATA[
DNAJC13 localization to endosomes is opposed by its J domain and its disordered C-terminal tail 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.12.19.629517v1?rss=1</link>
<description><![CDATA[
Endosomes are a central sorting hub for membrane cargos. DNAJC13/RME-8 plays a critical role in endosomal trafficking by regulating the endosomal recycling or degradative pathways. DNAJC13 localizes to endosomes through its N-terminal Plekstrin Homology (PH)-like domain, which directly binds endosomal phosphoinositol-3-phosphate (PI(3)P). However, little is known about how DNAJC13 localization is regulated. Here, we show that two regions within DNAJC13, its J domain and disordered C-terminal tail, act as negative regulators of its PH-like domain. Using a structure-function approach combined with quantitative proteomics, we mapped these control points to a conserved YLT motif in the C-terminal tail as well as the catalytic HPD triad in its J domain. Mutation of either motif enhanced DNAJC13 endosomal localization in cells and increased binding to PI(3)P in vitro. Further, these effects required the N-terminal PH-like domain. We show that, similar to other PI(3)P binding domains, the N-terminal PH-like domain binds PI(3)P weakly in isolation and requires oligomerization for efficient PI(3)P binding and endosomal localization. Together, these results demonstrate that interaction between DNAJC13 and PI(3)P serves as a molecular control point for regulating DNAJC13 localization to endosomes.

Significance StatementO_LIDNAJC13 controls endosomal sorting by regulating proteins which mediate the endosomal recycling and degradative subdomains.
C_LIO_LIHere we show that subcellular localization of DNAJC13 is regulated through the coordinated action of three of its domains: the PH-like domain which has low affinity for PI(3)P, the J domain, and a YLT motif in its disordered C-terminus.
C_LIO_LIThis study defines a novel mechanism by which DNAJC13 function is regulated.
C_LI
]]></description>
<dc:creator>Adoff, H.</dc:creator>
<dc:creator>Novy, B.</dc:creator>
<dc:creator>Holland, E.</dc:creator>
<dc:creator>Lobingier, B.</dc:creator>
<dc:date>2024-12-20</dc:date>
<dc:identifier>doi:10.1101/2024.12.19.629517</dc:identifier>
<dc:title><![CDATA[DNAJC13 localization to endosomes is opposed by its J domain and its disordered C-terminal tail]]></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/10.1101/2024.12.20.629740v1?rss=1">
<title>
<![CDATA[
Anti-Cancer Immune Priming with Beta-Radioligand Therapy and Isoform-Selective Targeting of 4Ig-B7-H3 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.12.20.629740v1?rss=1</link>
<description><![CDATA[
Radioligand therapy (RLT), a re-emerging oncologic strategy using molecularly-targeted therapeutic radioisotopes, clinically reduces tumor burden and enhances survival for select patients with otherwise unresponsive advanced prostate cancer and neuroendocrine tumors. Developing new approaches to next generation targets and a better understanding of systemic immune effects could broaden the impact of RLT. Aside from contributions to immune checkpoint, B7-H3 (CD276) is an attractive oncologic target because of its widespread and high differential expression across a variety of solid tumors compared to normal tissues. However, B7-H3 has two isoforms: a 4Ig-B7-H3 isoform, the dominant transmembrane protein expressed on tumors and tumor immune microenvironments (TIME), and a 2Ig-B7-H3 isoform, a soluble ectodomain protein, representing a circulating, and in the context of RLT, significant shed (decoy) antigen. To enhance tumor-specific binding and circumvent confounding soluble 2Ig-B7-H3, a novel IgG2a monoclonal antibody (MIL33B) was generated with high affinity for 4Ig-B7-H3 (72 picomolar) and 8- to 18-fold selectivity over soluble 2Ig-B7-H3. Live cell fluorescence microscopy using AF594-labeled MIL33B demonstrated strong membranous localization and target specificity. PET-CT imaging with 89Zr-labeled MIL33B confirmed robust tumor-selective target binding in vivo in murine xenograft (HeLa cervical) and syngeneic tumor models (4T1 breast, B16F10 melanoma, and CT26 colorectal) expressing human 4Ig-B7-H3. As a single dose beta-emitting systemic RLT therapeutic, 90Y-labeled MIL33B (100 Ci) produced 53% long-term survival in a 4Ig-B7-H3-dependent manner in an otherwise fatal established CT26 colorectal tumor model. Immunologic analysis showed that 90Y-MIL33B RLT functioned as an immune priming event, engaging downstream CD8+ T-cell activation and inducing immunological memory in vivo, thus illustrating the potential of systemic beta-RLT to target both primary and metastatic sites. Thus, MIL33B showcases a strategy to selectively target 4Ig-B7-H3 for beta-RLT, warranting further investigation as an immune priming tactic alone or in combination for cancer therapy.

Statement of significanceModest antigen expression levels, even if target tissue-selective, combined with ectodomain shedding (soluble decoy antigens) can generally hinder targeted diagnostic and therapeutic strategies, but are especially challenging for radioligand therapy, PET imaging, and in vivo diagnostics wherein high specific activity radioisotopes necessitate use of low masses of biocarrier. Binding, absorption and non-specific tissue deposition of radiolabeled biocarriers by decoy antigens can significantly misdirect systemic radiation, reducing therapeutic efficacy. An antibody development process with a focus aimed at on-target affinity for folded proteins on live cells resulted in a novel picomolar affinity antibody selectively targeting membranous 4Ig-B7-H3 over soluble decoy 2Ig-B7-H3. This antibody shows promise as a transformative systemic beta-radioligand therapy platform for immune priming applications in oncology, and potentially in cardiology, rheumatology, and autoimmunity.
]]></description>
<dc:creator>Glazer, S. E.</dc:creator>
<dc:creator>Sutton, M. N.</dc:creator>
<dc:creator>Yang, P.</dc:creator>
<dc:creator>Pisaneschi, F.</dc:creator>
<dc:creator>Sebastian, M. M.</dc:creator>
<dc:creator>Gammon, S. T.</dc:creator>
<dc:creator>Piwnica-Worms, D.</dc:creator>
<dc:date>2024-12-22</dc:date>
<dc:identifier>doi:10.1101/2024.12.20.629740</dc:identifier>
<dc:title><![CDATA[Anti-Cancer Immune Priming with Beta-Radioligand Therapy and Isoform-Selective Targeting of 4Ig-B7-H3]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-12-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.12.26.630255v1?rss=1">
<title>
<![CDATA[
Generation of a transgenic P. cynomolgi parasite expressing P. vivax circumsporozoite protein for testing pre-erythrocytic malaria vaccines in non-human primates 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.12.26.630255v1?rss=1</link>
<description><![CDATA[
Malaria, caused by infection with Plasmodium parasites, exacts a heavy toll worldwide. There are two licensed vaccines for malaria as well as two monoclonal antibodies that have shown promising efficacy in field trials. Both vaccines and monoclonals target the major surface protein (circumsporozoite protein, CSP) of Plasmodium falciparum. Yet Pf is only one of the four major species of Plasmodium that infects humans. Plasmodium vivax is the second leading cause of malaria but Pv vaccine and monoclonal development lags far behind P. falciparum owing to the lack of basic preclinical tools such as in vitro culture or mouse models that replicate the key biological features of P. vivax. Notably among these features is the ability to form dormant liver stages (hypnozoites) that reactivate and drive the majority of P. vivax malaria burden.

Plasmodium cynomolgi is a simian parasite which is genotypically and phenotypically very close to P. vivax, can infect common research non-human primates and replicates many features of Pv including relapsing hypnozoites. Recently, a strain of Pc has been adapted to in vitro culture allowing parasite transgenesis. Here, we created a transgenic P. cynomolgi parasite in which the endogenous Pc CSP has been replaced with Pv CSP with the goal of enabling preclinical study of anti-Pv CSP interventions to protect against primary and relapse infections. We show that the in vitro-generated transgenic Pc[PvCSP] parasite expresses both serotypes of Pv CSP and retains full functionality in vivo including the ability to transmit to laboratory-reared Anopheles mosquitos and cause relapsing infection in rhesus macaques. To our knowledge, this is the first gene replacement in a relapsing Plasmodium species. This work can directly enable in vivo development of anti-Pv CSP interventions and provide a blueprint for the study of relapsing malaria through reverse genetics.
]]></description>
<dc:creator>Aleshnick, M.</dc:creator>
<dc:creator>Hegde, S.</dc:creator>
<dc:creator>Jennison, C.</dc:creator>
<dc:creator>Mikolajczak, S.</dc:creator>
<dc:creator>Vaughan, A. M.</dc:creator>
<dc:creator>Haumpy, D.</dc:creator>
<dc:creator>Straimer, J.</dc:creator>
<dc:creator>Wilder, B.</dc:creator>
<dc:date>2024-12-26</dc:date>
<dc:identifier>doi:10.1101/2024.12.26.630255</dc:identifier>
<dc:title><![CDATA[Generation of a transgenic P. cynomolgi parasite expressing P. vivax circumsporozoite protein for testing pre-erythrocytic malaria vaccines in non-human primates]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-12-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.12.27.630549v1?rss=1">
<title>
<![CDATA[
The RNA-binding protein HuR impairs adipose tissue anabolism in pancreatic cancer cachexia 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.12.27.630549v1?rss=1</link>
<description><![CDATA[
BackgroundCachexia is defined by chronic loss of fat and muscle, is a frequent complication of pancreatic ductal adenocarcinoma (PDAC), and negatively impacts patient outcomes. Nutritional supplementation cannot fully reverse tissue wasting, and the mechanisms underlying this phenotype are unclear. This work aims to define the relative contributions of catabolism and anabolism to adipose wasting in PDAC-bearing mice. Human antigen R (HuR) is an RNA-binding protein recently shown to suppress adipogenesis. We hypothesize that fat wasting results from a loss of adipose anabolism driven by increased HuR activity in adipocytes of PDAC-bearing mice.

MethodsAdult C57BL/6J mice received orthotopic PDAC cell (KrasG12D; p53R172H/+; Pdx1-cre) (PDAC) or PBS (sham) injections. Mice exhibiting moderate cachexia (9 days after injection) were fasted for 24h, or fasted 24h and refed 24h before euthanasia. A separate cohort of PDAC mice were treated with an established HuR inhibitor (KH-3, 100 mg/kg) and subjected to the fast/refeed paradigm. We analyzed body mass, gross fat pad mass, and adipose tissue mRNA expression. We quantified lipolytic rate as the normalized quantity of glycerol released from 3T3-L1 adipocytes in vitro, and gonadal fat pads (gWAT) ex vivo.

Results3T3-L1 adipocytes treated with PDAC cell conditioned media (CM) had lower expression of lipolysis and lipogenesis genes than control cells, and did not display elevated lipolysis as measured by liberated glycerol. PDAC gWAT cultured ex vivo displayed decreased lipolysis compared to sham gWAT (-54.7%). PDAC and sham mice lost equivalent fat mass after a 24h fast, however, PDAC mice could not restore inguinal fat pads (iWAT) (-40.5%) or gWAT (-31.8%) mass after refeeding. RNAseq revealed 572 differentially expressed genes in gWAT from PDAC compared to sham mice. Downregulated genes (n=126) were associated with adipogenesis (adj p=0.05), and expression of adipogenesis master regulators Pparg and Cebpa were reduced in gWAT from PDAC mice. Immunohistochemistry revealed increased HuR staining in gWAT (+74.9%) and iWAT (+41.2%) from PDAC mice. Inhibiting HuR binding restored lipogenesis in refed animals with a concomitant increase in iWAT mass (+131.7%).

ConclusionsOur work highlights deficient adipose anabolism as a driver of reduced lipid content in 3T3-L1 adipocytes treated with PDAC conditioned media and PDAC mice. The small molecule KH-3, which disrupts HuR binding, restored adipose anabolism in PDAC mice. This highlights HuR as a potentially targetable regulatory node for adipose anabolism in cancer cachexia.
]]></description>
<dc:creator>Arneson-Wissink, P. C.</dc:creator>
<dc:creator>Pelz, K.</dc:creator>
<dc:creator>Worley, B.</dc:creator>
<dc:creator>Mendez, H.</dc:creator>
<dc:creator>Pham, P.</dc:creator>
<dc:creator>McCarthy, G.</dc:creator>
<dc:creator>Chitsazan, A.</dc:creator>
<dc:creator>Brody, J. R.</dc:creator>
<dc:creator>Grossberg, A.</dc:creator>
<dc:date>2024-12-28</dc:date>
<dc:identifier>doi:10.1101/2024.12.27.630549</dc:identifier>
<dc:title><![CDATA[The RNA-binding protein HuR impairs adipose tissue anabolism in pancreatic cancer cachexia]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-12-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.12.30.630700v1?rss=1">
<title>
<![CDATA[
Rapid clonal selection within early hematopoietic cell compartments presages outcome to ivosidenib combination therapy 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.12.30.630700v1?rss=1</link>
<description><![CDATA[
Acquired resistance to targeted non-intensive therapies is common in myeloid malignancies. Yet, key questions remain as to how rapidly resistant clones are selected by treatment and in which hematopoietic cell compartments clonal selection occurs. To address this gap, we studied clonal responses to ivosidenib + venetoclax {+/-} azacitidine combination therapy in 8 patients with IDH1-mutant myeloid malignancy. Whilst all 8 patients initially responded to treatment, 6 relapsed and 2 remained in sustained remission for > 4 years. To study longitudinal clonal dynamics through hematopoietic differentiation, we performed high-sensitivity single-cell genotyping in index-sorted sequential patient samples. In all patients who relapsed, therapy-resistant clones were selected rapidly, within 1-3 treatment cycles, at times when hematopoiesis was still largely sustained by either normal or pre-leukemic cells. Selection of therapy-resistant clones preceded overt treatment failure by months or even years. Relapse was associated either with clones harboring newly-detected myeloid driver mutations or expansion of minor pre-existing clones that had reduced fitness prior to treatment. In both cases, resistant clones were selected within immature cell populations previously shown to contain leukemic stem cell (LSC) potential, preceding malignant expansion of these compartments by immunophenotyping. In contrast, in both patients remaining in remission, leukemic clones were eradicated and rapidly replaced by clonal and wild-type hematopoiesis. These observations suggest that, in patients treated with non-intensive ivosidenib combination therapy, rapid clonal selection occurs in populations with LSC potential, where failure to eliminate either genetically evolved or persistent leukemic clones ultimately leads to relapse.

Key points- Rapid selection of leukemic clones occurs within small populations with LSC potential, months or years prior to relapse.

- Rapid eradication of leukemic clones leads to sustained remission in the context of ivosidenib combination therapy.
]]></description>
<dc:creator>Turkalj, S.</dc:creator>
<dc:creator>Stoilova, B.</dc:creator>
<dc:creator>Groom, A. J.</dc:creator>
<dc:creator>Radtke, F. A.</dc:creator>
<dc:creator>Mecklenbrauck, R.</dc:creator>
<dc:creator>Jakobsen, N. A.</dc:creator>
<dc:creator>Lachowiez, C. A.</dc:creator>
<dc:creator>Metzner, M.</dc:creator>
<dc:creator>Usukhbayar, B.</dc:creator>
<dc:creator>Salazar, M. A.</dc:creator>
<dc:creator>Zeng, Z.</dc:creator>
<dc:creator>Loghavi, S.</dc:creator>
<dc:creator>Marvin-Peek, J.</dc:creator>
<dc:creator>Koerber, V.</dc:creator>
<dc:creator>Ravandi, F.</dc:creator>
<dc:creator>Issa, G.</dc:creator>
<dc:creator>Kadia, T.</dc:creator>
<dc:creator>Symeonido, V.</dc:creator>
<dc:creator>de Groot, A. P.</dc:creator>
<dc:creator>Kantarjian, H.</dc:creator>
<dc:creator>Takahashi, K.</dc:creator>
<dc:creator>Konopleva, M.</dc:creator>
<dc:creator>DiNardo, C. D.</dc:creator>
<dc:creator>Vyas, P.</dc:creator>
<dc:date>2024-12-31</dc:date>
<dc:identifier>doi:10.1101/2024.12.30.630700</dc:identifier>
<dc:title><![CDATA[Rapid clonal selection within early hematopoietic cell compartments presages outcome to ivosidenib combination therapy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-12-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2024.12.30.630781v1?rss=1">
<title>
<![CDATA[
Analysis of mouse lens morphological and proteomic abnormalities following depletion of βB3-crystallin 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2024.12.30.630781v1?rss=1</link>
<description><![CDATA[
Crystallin proteins serve as both essential structural and as well as protective components of the ocular lens and are required for the transparency and light refraction properties of the organ. The mouse lens crystallin proteome is represented by A-, B-, {beta}A1-, {beta}A2-, {beta}A3-, {beta}A4-, {beta}B1-, {beta}B2-, {beta}B3-, {gamma}A-, {gamma}B-, {gamma}C-, {gamma}D-, {gamma}E, {gamma}F-, {gamma}N-, and {gamma}S-crystallin proteins encoded by 16 genes. Their mutations are responsible for lens opacification and early onset cataract formation. While many cataract-causing missense and nonsense mutations are known for these proteins, including the human CRYBB3 gene, the mammalian loss-of function model of the Crybb3 gene remains to be established. Herein, we generated the first mouse model via deletion of the Crybb3 promoter that abolished expression of the {beta}B3-crystallin. Histological analysis of lens morphology using newborn {beta}B3-crystallin-deficient lenses revealed disrupted lens morphology with early-onset phenotypic variability. In-depth lens proteomics at four time points (newborn, 3-weeks, 6-weeks, and 3-months) showed both down- and up-regulation of various proteins, with the highest divergence from control mice observed in 3-months lenses. Apart from the {beta}B3-crystallin, another protein Smarcc1/Baf155 was down-regulated in all four samples. In addition, downregulation of Hspe1, Pdlim1, Ast/Got, Lsm7, Ddx23, and Acad11 was found in three time points. Finally, we show that the {beta}B3-crystallin promoter region, which contains multiple binding sites for the transcription factors AP-2, c-Jun, c-Maf, Etv5, and Pax6 is activated by FGF2 in primary lens cell culture experiments. Together, these studies establish the mouse Crybb3 loss-of-function model and its disrupted crystallin and non-crystallin proteomes.
]]></description>
<dc:creator>Rayee, D.</dc:creator>
<dc:creator>Wilmarth, P. A.</dc:creator>
<dc:creator>VanSlyke, J. K.</dc:creator>
<dc:creator>Zientek, K.</dc:creator>
<dc:creator>Reddy, A. P.</dc:creator>
<dc:creator>Musil, L. S.</dc:creator>
<dc:creator>David, L. L.</dc:creator>
<dc:creator>Cvekl, A.</dc:creator>
<dc:date>2024-12-31</dc:date>
<dc:identifier>doi:10.1101/2024.12.30.630781</dc:identifier>
<dc:title><![CDATA[Analysis of mouse lens morphological and proteomic abnormalities following depletion of βB3-crystallin]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-12-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.01.07.631771v1?rss=1">
<title>
<![CDATA[
Noise-induced hearing loss enhances Ca2+-dependent spontaneous bursting activity in lateralcochlear efferents 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.01.07.631771v1?rss=1</link>
<description><![CDATA[
Exposure to loud and/or prolonged noise damages cochlear hair cells and triggers downstream changes in synaptic and electrical activity in multiple brain regions, resulting in hearing loss and altered speech comprehension. It remains unclear however whether or not noise exposure also compromises the cochlear efferent system, a feedback pathway in the brain that fine-tunes hearing sensitivity in the cochlea. We examined the effects of noise-induced hearing loss on the spontaneous action potential (AP) firing pattern in mouse lateral olivocochlear (LOC) neurons. This spontaneous firing exhibits a characteristic burst pattern dependent on Ca2+ channels, and we therefore also examined the effects of noise-induced hearing loss on the function of these and other ion channels. The burst pattern was sustained by an interaction between inactivating Ca2+ currents contributed largely by L-type channels, and steady outward currents mediated by Ba2+-sensitive inwardly-rectifying and two-pore domain K+ channels. One week following exposure to loud broadband noise, hearing thresholds were significantly elevated, and the duration of AP bursts was increased, likely as a result of an enhanced Ca2+ current. Additional effects of noise-induced hearing loss included alteration of Ca2+-dependent inactivation of Ca2+ currents and a small elevation of outward K+ currents. We propose that noise-induced hearing loss enhances efferent activity and may thus amplify the release of neurotransmitters and neuromodulators (i.e., neuropeptides), potentially altering sensory coding within the damaged cochlea.

Significance StatementAlthough the effects of noise-induced hearing loss on the auditory afferent system have been extensively studied, little is known about its impact on the auditory efferent system, which modulates hearing sensitivity via feedback from the brain. Additionally, while Ca2+ channels are related to numerous neurological diseases, their involvement in auditory disorders is underexplored. This study bridges these gaps by examining Ca2+ channel-driven spontaneous burst firing in lateral olivocochlear (LOC) neurons, the most numerous auditory efferent neurons. Noise-induced hearing loss differentially affects Ca2+ channel subtypes by increasing high-voltage activated currents that further prolong burst firing and suggesting altered intracellular Ca2+ signaling. These significant changes in LOC firing behavior may profoundly impact their downstream targets in the cochlea.
]]></description>
<dc:creator>Hong, H.</dc:creator>
<dc:creator>Trussell, L. O.</dc:creator>
<dc:date>2025-01-08</dc:date>
<dc:identifier>doi:10.1101/2025.01.07.631771</dc:identifier>
<dc:title><![CDATA[Noise-induced hearing loss enhances Ca2+-dependent spontaneous bursting activity in lateralcochlear efferents]]></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/10.1101/2025.01.10.632348v1?rss=1">
<title>
<![CDATA[
Developmental axon diameter growth of central nervous system axons does not depend on ensheathment or myelination by oligodendrocytes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.01.10.632348v1?rss=1</link>
<description><![CDATA[
Myelination facilitates the rapid conduction of action potentials along axons. In the central nervous system (CNS), myelinated axons vary over 100-fold in diameter, with conduction speed scaling linearly with increasing diameter. Axon diameter and myelination are closely interlinked, with axon diameter exerting a strong influence on myelination. Conversely, myelinating Schwann cells in the peripheral nervous system can both positively and negatively affect axon diameter. However, whether axon diameter is regulated by CNS oligodendrocytes is less clear. Here, we investigated CNS axon diameter growth in the absence of myelin using mouse (Mbpshi/shi and Myrf conditional knockout) and zebrafish (olig2 morpholino) models. We find that neither the ensheathment of axons, nor the formation of compact myelin are required for CNS axons to achieve appropriate and diverse diameters. This indicates that developmental CNS axon diameter growth is independent of myelination, and shows that myelinating cells of CNS and PNS differentially influence axonal morphology.
]]></description>
<dc:creator>Bin, J. M.</dc:creator>
<dc:creator>Emberley, K.</dc:creator>
<dc:creator>Buscham, T. J.</dc:creator>
<dc:creator>Eichel-Vogel, M. A.</dc:creator>
<dc:creator>Doan, R. A.</dc:creator>
<dc:creator>Steyer, A. M.</dc:creator>
<dc:creator>Nolan, M. F.</dc:creator>
<dc:creator>Möbius, W.</dc:creator>
<dc:creator>Monk, K. R.</dc:creator>
<dc:creator>Werner, H. B.</dc:creator>
<dc:creator>Emery, B.</dc:creator>
<dc:creator>Lyons, D. A.</dc:creator>
<dc:date>2025-01-10</dc:date>
<dc:identifier>doi:10.1101/2025.01.10.632348</dc:identifier>
<dc:title><![CDATA[Developmental axon diameter growth of central nervous system axons does not depend on ensheathment or myelination by oligodendrocytes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-01-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.01.10.632481v1?rss=1">
<title>
<![CDATA[
The bile acid-sensitive ion channel is gated by Ca2+-dependent conformational changes in the transmembrane domain 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.01.10.632481v1?rss=1</link>
<description><![CDATA[
The bile acid-sensitive ion channel (BASIC) is the least understood member of the mammalian epithelial Na+ channel/degenerin (ENaC/DEG) superfamily of ion channels, which are involved in a variety of physiological processes. While some members of this superfamily, including BASIC, are inhibited by extracellular Ca2+ (Ca2+), the molecular mechanism underlying Ca2+ modulation remains unclear. Here, by determining the structure of human BASIC in the presence and absence of Ca2+ using single particle cryo-electron microscopy (cryo-EM), we reveal Ca2+-dependent conformational changes in the transmembrane domain and {beta}-linkers. Electrophysiological experiments further show that a glutamate residue in the extracellular vestibule of the pore underpins the Ca2+-binding site, whose occupancy determines the conformation of the pore and therefore ion flow through the channel. These results reveal the molecular principles governing gating of BASIC and its regulation by Ca2+ ions, demonstrating that Ca2+ ions modulate BASIC function via changes in protein conformation rather than solely from pore-block, as proposed for other members of this superfamily.
]]></description>
<dc:creator>Freitas, M. M.</dc:creator>
<dc:creator>Gouaux, E.</dc:creator>
<dc:date>2025-01-12</dc:date>
<dc:identifier>doi:10.1101/2025.01.10.632481</dc:identifier>
<dc:title><![CDATA[The bile acid-sensitive ion channel is gated by Ca2+-dependent conformational changes in the transmembrane domain]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-01-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.01.11.632510v1?rss=1">
<title>
<![CDATA[
Cholesterol deficiency directs autophagy-dependent secretion of extracellular vesicles 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.01.11.632510v1?rss=1</link>
<description><![CDATA[
Extracellular vesicle (EV) secretion is an important, though not fully understood, intercellular communication process. Lipid metabolism has been shown to regulate EV activity, though the impact of specific lipid classes is unclear. Through analysis of small EVs (sEVs), we observe aberrant increases in sEV release within genetic models of cholesterol biosynthesis disorders, where cellular cholesterol is diminished. Inhibition of cholesterol synthesis at multiple synthetic steps mimics genetic models in terms of cholesterol reduction and sEVs secreted. Further analyses of sEVs from cholesterol-depleted cells revealed structural deficits and altered surface marker expression, though these sEVs were also more easily internalized by recipient cells. Transmission electron microscopy of cells with impaired cholesterol biosynthesis demonstrated multivesicular and multilamellar structures potentially associated with autophagic defects. We further found autophagic vesicles being redirected toward late endosomes at the expense of autophagolysosomes. Through CRISPR-mediated inhibition of autophagosome formation, we mechanistically determined that release of sEVs after cholesterol depletion is autophagy dependent. We conclude that cholesterol imbalance initiates autophagosome-dependent secretion of sEVs, which may have pathological relevance in diseases of cholesterol disequilibrium.
]]></description>
<dc:creator>Yaeger, J. D.</dc:creator>
<dc:creator>Sengupta, S.</dc:creator>
<dc:creator>Walz, A. L.</dc:creator>
<dc:creator>Morita, M.</dc:creator>
<dc:creator>Morgan, T. K.</dc:creator>
<dc:creator>Vermeer, P. D.</dc:creator>
<dc:creator>Francis, K. R.</dc:creator>
<dc:date>2025-01-12</dc:date>
<dc:identifier>doi:10.1101/2025.01.11.632510</dc:identifier>
<dc:title><![CDATA[Cholesterol deficiency directs autophagy-dependent secretion of extracellular vesicles]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-01-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.01.13.632726v1?rss=1">
<title>
<![CDATA[
A Cross-Species Atlas of the Dorsal Vagal Complex Reveals Neural Mediators of Cagrilintide's Effects on Energy Balance 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.01.13.632726v1?rss=1</link>
<description><![CDATA[
Amylin analogs, including potential anti-obesity therapies like cagrilintide, act on neurons in the brainstem dorsal vagal complex (DVC) that express calcitonin receptors (CALCR). These receptors, often combined with receptor activity-modifying proteins (RAMPs), mediate the suppression of food intake and body weight. To understand the molecular and neural mechanisms of cagrilintide action, we used single-nucleus RNA sequencing to define 89 cell populations across the rat, mouse, and non-human primate caudal brainstem. We then integrated spatial profiling to reveal neuron distribution in the rat DVC. Furthermore, we compared the acute and long-term transcriptional responses to cagrilintide across DVC neurons of rats, which exhibit strong cagrilintide responsiveness, and mice, which respond poorly to cagrilintide over the long term. We found that cagrilintide promoted long-term transcriptional changes, including increased prolactin releasing hormone (Prlh) expression, in the nucleus of the solitary tract (NTS) Calcr/Prlh cells in rats, but not in mice, suggesting the importance of NTS Calcr/Prlh cells for sustained weight loss. Indeed, activating rat area postrema Calcr cells briefly reduced food intake but failed to decrease food intake or body weight over the long term. Overall, these results not only provide a cross-species and spatial atlas of DVC cell populations but also define the molecular and neural mediators of acute and long-term cagrilintide action.
]]></description>
<dc:creator>Ludwig, M. Q.</dc:creator>
<dc:creator>Coester, B.</dc:creator>
<dc:creator>Gordian, D.</dc:creator>
<dc:creator>Hassan, S.</dc:creator>
<dc:creator>Tomlinson, A. J.</dc:creator>
<dc:creator>Toure, M. H.</dc:creator>
<dc:creator>Christensen, O. P.</dc:creator>
<dc:creator>Moltke-Prehn, A.</dc:creator>
<dc:creator>Brown, J. M.</dc:creator>
<dc:creator>Rausch, D. M.</dc:creator>
<dc:creator>Gowda, A.</dc:creator>
<dc:creator>Wu, I.</dc:creator>
<dc:creator>Kernodle, S.</dc:creator>
<dc:creator>Dong, V.</dc:creator>
<dc:creator>Ayensu-Mensah, M.</dc:creator>
<dc:creator>Sabatini, P. V.</dc:creator>
<dc:creator>Shin, J. H.</dc:creator>
<dc:creator>Kirigiti, M.</dc:creator>
<dc:creator>Egerod, K. L.</dc:creator>
<dc:creator>Le Foll, C.</dc:creator>
<dc:creator>Lundh, S.</dc:creator>
<dc:creator>Gerstenberg, M. K.</dc:creator>
<dc:creator>Lutz, T. A.</dc:creator>
<dc:creator>Kievit, P.</dc:creator>
<dc:creator>Secher, A.</dc:creator>
<dc:creator>Raun, K.</dc:creator>
<dc:creator>Myers, M. G.</dc:creator>
<dc:creator>Pers, T. H.</dc:creator>
<dc:date>2025-01-15</dc:date>
<dc:identifier>doi:10.1101/2025.01.13.632726</dc:identifier>
<dc:title><![CDATA[A Cross-Species Atlas of the Dorsal Vagal Complex Reveals Neural Mediators of Cagrilintide's Effects on Energy Balance]]></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/10.1101/2025.01.16.633282v1?rss=1">
<title>
<![CDATA[
Distinct cell types along thick ascending limb express pathways for monovalent and divalent cation transport 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.01.16.633282v1?rss=1</link>
<description><![CDATA[
Kidney thick ascending limb cells reabsorb sodium, potassium, calcium, and magnesium and contribute to urinary concentration. These cells are typically viewed as of a single type that recycles potassium across the apical membrane and generates a lumen-positive transepithelial voltage driving calcium and magnesium reabsorption, although variability in potassium channel expression has been reported. Additionally, recent transcriptomic analyses suggest that different cell types exist along this segment, but classifications have varied and have not led to a new consensus model. We used immunolocalization, electrophysiology and enriched single nucleus RNA-Seq to identify thick ascending limb cell types in rat, mouse and human. We identified three major TAL cell types defined by expression of potassium channels and claudins. One has apical potassium channels, low basolateral potassium conductance, and is bordered by a sodium-permeable claudin. A second lacks apical potassium channels, has high basolateral potassium conductance and is bordered by calcium- and magnesium-permeable claudins. A third type also lacks apical potassium channels and has a high basolateral potassium conductance, but these cells are ringed by sodium-permeable claudins. The recognition of diverse cell types resolves longstanding questions about how solute transport can be modulated selectively and how disruption of these cells leads to human disease.
]]></description>
<dc:creator>Bahena-Lopez, J.</dc:creator>
<dc:creator>Demirci, H.</dc:creator>
<dc:creator>Smorodchenko, A.</dc:creator>
<dc:creator>Su, X.-T.</dc:creator>
<dc:creator>Nelson, J.</dc:creator>
<dc:creator>Yang, C.-L.</dc:creator>
<dc:creator>Wang, W.-H.</dc:creator>
<dc:creator>Curry, J.</dc:creator>
<dc:creator>Duan, X.-P.</dc:creator>
<dc:creator>Sharkovska, Y.</dc:creator>
<dc:creator>Liu, R.</dc:creator>
<dc:creator>Yilmaz, D. E.</dc:creator>
<dc:creator>Quintanova, C.</dc:creator>
<dc:creator>Emberly, K.</dc:creator>
<dc:creator>Emery, B.</dc:creator>
<dc:creator>Himmerkus, N.</dc:creator>
<dc:creator>Bleich, M.</dc:creator>
<dc:creator>Bachmann, S.</dc:creator>
<dc:creator>Ellison, D. H.</dc:creator>
<dc:date>2025-01-21</dc:date>
<dc:identifier>doi:10.1101/2025.01.16.633282</dc:identifier>
<dc:title><![CDATA[Distinct cell types along thick ascending limb express pathways for monovalent and divalent cation transport]]></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/10.1101/2025.01.16.633406v1?rss=1">
<title>
<![CDATA[
A nonhuman primate model mirrors human congenital cytomegalovirus infection and reveals a spectrum of vertical transmission outcomes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.01.16.633406v1?rss=1</link>
<description><![CDATA[
Congenital cytomegalovirus (cCMV) is the leading infectious cause of birth defects worldwide, yet immune determinants of protection to inform design of a maternal vaccine remain elusive. Here, we characterized the outcome of primary rhesus CMV (RhCMV) infection during pregnancy in an immune competent nonhuman primate (NHP) model. RhCMV DNA was detected in amniotic fluid and/or fetal tissues in six of 12 (50% placental transmission) CMV-naive rhesus macaque dams inoculated intravenously with RhCMV in early second trimester gestation. Widespread tissue dissemination dominated by one of two inoculated RhCMV strains was present in one fetus (8.3% cCMV disease). Placental RhCMV transmission was associated with elevated fetal and maternal plasma TNF-alpha and reduced maternal brain-derived neurotrophic factor and IL-10 levels. CMV exposure during pregnancy had a broad impact on the placenta and fetus even in the absence of congenital infection as evidenced by RhCMV infection at the maternal-fetal interface in all 12 dams, along with significantly reduced placental efficiency and fetal growth metrics compared to gestation-matched control pregnancies. This NHP model recapitulates key aspects of human cCMV and provides new insight into barriers and biomarkers of successful vertical transmission.

One sentence summary: The nonhuman primate model mirrors the epidemiology of human congenital CMV (cCMV) after primary infection and reveals its transmission bottlenecks.
]]></description>
<dc:creator>Manuel, T. D.</dc:creator>
<dc:creator>Mostrom, M. J.</dc:creator>
<dc:creator>Crooks, C.</dc:creator>
<dc:creator>Davalos, A.</dc:creator>
<dc:creator>Barfield, R.</dc:creator>
<dc:creator>Scheef, E. A.</dc:creator>
<dc:creator>Kendall, S.</dc:creator>
<dc:creator>Midkiff, C. C.</dc:creator>
<dc:creator>Sprehe, L. M.</dc:creator>
<dc:creator>Trexler, M.</dc:creator>
<dc:creator>Boquet, F.</dc:creator>
<dc:creator>Shroyer, M.</dc:creator>
<dc:creator>Danner, V.</dc:creator>
<dc:creator>Doyle-Meyers, L.</dc:creator>
<dc:creator>Mirza, A.</dc:creator>
<dc:creator>Lammi, S.</dc:creator>
<dc:creator>Otero, C. E.</dc:creator>
<dc:creator>Lee, M.</dc:creator>
<dc:creator>Rogers, L.</dc:creator>
<dc:creator>Kowalik, T.</dc:creator>
<dc:creator>Chan, C.</dc:creator>
<dc:creator>Permar, S. R.</dc:creator>
<dc:creator>Blair, R. V.</dc:creator>
<dc:creator>Frueh, K.</dc:creator>
<dc:creator>Malouli, D.</dc:creator>
<dc:creator>Granek, J. A.</dc:creator>
<dc:creator>Owzar, K.</dc:creator>
<dc:creator>Kaur, A.</dc:creator>
<dc:date>2025-01-21</dc:date>
<dc:identifier>doi:10.1101/2025.01.16.633406</dc:identifier>
<dc:title><![CDATA[A nonhuman primate model mirrors human congenital cytomegalovirus infection and reveals a spectrum of vertical transmission outcomes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-01-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.01.20.633926v1?rss=1">
<title>
<![CDATA[
Combinations comprising dual β-lactams and a β-lactamase inhibitor achieve optimal synergistic inhibition of Mycobacterium abscessus growth 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.01.20.633926v1?rss=1</link>
<description><![CDATA[
The historical model, which posits that {beta}-lactams inhibit bacterial growth while {beta}-lactamase inhibitors (BLIs) merely protect {beta}-lactams from enzymatic degradation, fails to fully explain their activity against Mycobacterium abscessus (Mab). This study demonstrates that synergistic effects extend beyond the traditional one {beta}-lactam+one BLI paradigm, refuting the oversimplified mechanistic framework. First, {beta}-lactam-based BLIs such as clavulanic acid, sulbactam, and tazobactam exhibit intrinsic antibacterial activity against Mab. These agents synergized not only with {beta}-lactams but also with one another, undermining their historical classification as mere {beta}-lactamase inhibitors. The data indicate that their activity is not limited to inhibiting {beta}-lactamases but extends to directly targeting critical bacterial processes. Second, dual {beta}-lactam combinations exhibit synergism against Mab even in the absence of BLIs. For example, despite being rapidly hydrolyzed by the native {beta}-lactamase BlaMab, amoxicillin demonstrates strong synergism with {beta}-lactams such as imipenem or ceftaroline. This suggests that the second {beta}-lactam either acts as a functional BLI surrogate or targets complementary pathways. Supporting this, experiments using penicillin- and carbapenem-based probes revealed that {beta}-lactams bind to multiple Mab proteins simultaneously, reinforcing the idea that their synergy arises from targeting complementary essential proteins. Finally, triple combinations comprising dual {beta}-lactam and one BLI, such as amoxicillin + ceftaroline + avibactam, achieved very high synergy, underscoring the complementary roles of dual {beta}-lactams and BLIs. The evidence in this study necessitates a revised model that can more accurately explain the activities of {beta}-lactams and BLIs and underscores the potential for optimizing {beta}-lactam/BLI regimens against Mab.

IMPORTANCEThis research challenges old assumptions about how antibiotics fight bacteria, particularly Mycobacterium abscessus (Mab), a tough-to-treat infection. Traditionally, {beta}-lactam antibiotics were thought to stop bacterial growth, while {beta}-lactamase inhibitors (BLIs) just protected them from breakdown. However, this study reveals that BLIs like clavulanic acid can work together with another BLI or {beta}-lactam antibiotics for stronger effects. Surprisingly, even combinations comprising two BLIs can be highly effective, showing they target multiple critical bacterial processes simultaneously. Triple combinations--two {beta}-lactams and one BLI--proved especially powerful. These findings overturn outdated ideas, offering a smarter way to use these drugs to combat difficult infections and save lives.
]]></description>
<dc:creator>Rimal, B.</dc:creator>
<dc:creator>Xie, Y.</dc:creator>
<dc:creator>Panthi, C. M.</dc:creator>
<dc:creator>Devlin, K. L.</dc:creator>
<dc:creator>Beatty, K. E.</dc:creator>
<dc:creator>Lamichhane, G.</dc:creator>
<dc:date>2025-01-21</dc:date>
<dc:identifier>doi:10.1101/2025.01.20.633926</dc:identifier>
<dc:title><![CDATA[Combinations comprising dual β-lactams and a β-lactamase inhibitor achieve optimal synergistic inhibition of Mycobacterium abscessus growth]]></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/10.1101/2025.01.17.633673v1?rss=1">
<title>
<![CDATA[
Neoplastic immune mimicry is a generalizable phenomenon in breast cancer and epithelial CD69 enables early tumor progression 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.01.17.633673v1?rss=1</link>
<description><![CDATA[
Dedifferentiation programs are commonly enacted during breast cancer progression to enhance tumor cell fitness. Increased cellular plasticity within the neoplastic compartment of tumors correlates with disease aggressiveness, often culminating in greater resistance to cytotoxic therapies or augmented metastatic potential. Here we report that subpopulations of dedifferentiated neoplastic breast epithelial cells express canonical leukocyte cell surface receptor proteins and have thus named this cellular program "immune mimicry." We document neoplastic cells engaging in immune mimicry within public human breast tumor single-cell RNA-seq datasets, histopathological breast tumor specimens, breast cancer cell lines, as well as in murine transgenic and cell line-derived mammary cancer models. Immune-mimicked neoplastic cells harbor hallmarks of dedifferentiation and are enriched in treatment-resistant and high-grade breast tumors. We corroborated these observations in aggressive breast cancer cell lines where anti-proliferative cytotoxic chemotherapies drove epithelial cells toward immune mimicry. Moreover, in subsequent proof-of-concept studies, we demonstrate that expression of the CD69 leukocyte activation protein by neoplastic cells confers a proliferative advantage that facilitates early tumor growth and therefore conclude that neoplastic breast epithelial cells upregulating leukocyte surface receptors potentiate malignancy. Moving forward, neoplastic immune mimicry should be evaluated for prognostic utility in additional breast cancer cohorts to determine its potential for patient stratification. Future research should evaluate correlates with distal metastases, progression-free survival, overall survival, and therapeutic response/resistance.

Statement of SignificanceNeoplastic breast epithelial cells express surface receptors canonically attributed to leukocytes and are associated with therapy resistance and aggressive tumor behavior.
]]></description>
<dc:creator>Berens, E. B.</dc:creator>
<dc:creator>Khou, S.</dc:creator>
<dc:creator>Huang, E.</dc:creator>
<dc:creator>Hoffman, A.</dc:creator>
<dc:creator>Johnson, B.</dc:creator>
<dc:creator>Kirchberger, N.</dc:creator>
<dc:creator>Sivagnanam, S.</dc:creator>
<dc:creator>Calistri, N. L.</dc:creator>
<dc:creator>Derrick, D.</dc:creator>
<dc:creator>Liby, T. A.</dc:creator>
<dc:creator>McLean, I. C.</dc:creator>
<dc:creator>Alanizi, A. A.</dc:creator>
<dc:creator>Ozmen, F.</dc:creator>
<dc:creator>Ozmen, T. Y.</dc:creator>
<dc:creator>Mills, G. B.</dc:creator>
<dc:creator>Hwang, E. S.</dc:creator>
<dc:creator>Schedin, P. J.</dc:creator>
<dc:creator>Gonzalez, H.</dc:creator>
<dc:creator>Werb, Z.</dc:creator>
<dc:creator>Heiser, L. M.</dc:creator>
<dc:creator>Coussens, L. M.</dc:creator>
<dc:date>2025-01-22</dc:date>
<dc:identifier>doi:10.1101/2025.01.17.633673</dc:identifier>
<dc:title><![CDATA[Neoplastic immune mimicry is a generalizable phenomenon in breast cancer and epithelial CD69 enables early tumor progression]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-01-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.01.23.634593v1?rss=1">
<title>
<![CDATA[
Inferring single-cell dynamics of a molecular reporter from unlabeled live-cell light microscopy data analyzed with delay embedding 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.01.23.634593v1?rss=1</link>
<description><![CDATA[
Quantification of the temporal sequence of molecular behavior in live individual cells holds promise for improving causal and mechanistic models of cell biology. In recent years, different methods for inferring molecular labeling from microscopy data have been developed, especially in the context of "virtual pathology", but less effort has been directed to the context of single-cell dynamics and live-cell imaging. We demonstrate that phase-contrast live-cell imaging of MCF10A cells, without labeling data, is predictive of dynamical, single-cell behavior of the cell-cycle reporter Human DNA Helicase B (HDHB) - in particular, of the nuclear vs. cytoplasmic localization of this fluorescent reporter of cyclin-dependent kinase activity. Prediction quality improves substantially when temporal sequences of images are combined in a "delay embedding" framework. When different featurizations of the imaging data are examined, we find that features derived from a variational auto-encoder (VAE) outperform "classical" image features derived from shape and texture. We find the best performance, with Pearson R[~] 0.9 on test data, using VAE features augmented by categorical predictions, all within the delay-embedding framework - in comparison to R [~] 0.5 based on "ordinary" regression with VAE features.
]]></description>
<dc:creator>Singh, J.</dc:creator>
<dc:creator>Copperman, J.</dc:creator>
<dc:creator>Heiser, L. M.</dc:creator>
<dc:creator>Zuckerman, D. M.</dc:creator>
<dc:date>2025-01-24</dc:date>
<dc:identifier>doi:10.1101/2025.01.23.634593</dc:identifier>
<dc:title><![CDATA[Inferring single-cell dynamics of a molecular reporter from unlabeled live-cell light microscopy data analyzed with delay embedding]]></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/10.1101/2025.01.24.634817v1?rss=1">
<title>
<![CDATA[
HMG box-containing protein 1 (HBP1) prevents pancreatic injury in experimental pancreatitis but accelerates pancreatic neoplasia progression 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.01.24.634817v1?rss=1</link>
<description><![CDATA[
Background & AimsPancreatitis is an inflammatory disease of the exocrine pancreas and a known risk factor for pancreatic ductal adenocarcinoma (PDAC). Previously, we identified HMG- box transcription factor 1 (HBP1) as a potential master transcription factor (TF) in the early progression of PDAC, with its expression associated with poor patient survival, underscoring its significance in pancreatic disease. However, the functional role of HBP1 in the onset and progression of acute pancreatitis (AP) remains unknown.

MethodsWe examined HBP1 expression in human pancreatitis samples and a cerulein-induced AP mouse model. Pancreatic-specific conditional HBP1 knockout mice, with or without an oncogenic Kras mutation, were generated and compared to their littermate controls. Spatial transcriptomics and multiplexed protein assays, histological analysis, and immunostaining were utilized to characterize pathological changes. Findings from mouse models were validated using inducible HBP1-overexpressing human pancreatic ductal epithelial cells.

ResultsHBP1 was upregulated in pancreatic exocrine cells in human chronic pancreatitis and mouse acute pancreatitis, with its expression in human chronic pancreatitis correlating with cancer presence. Pancreatic HBP1 ablation disrupted acinar homeostasis by impairing autophagic flux and exacerbating inflammation following injury. In the presence of oncogenic KRAS, HBP1 ablation delayed the formation of pancreatic intraepithelial neoplasia (PanIN), the precursor to PDAC, and slowed its progression to higher-grade lesions.

ConclusionsHBP1 upregulation in pancreatitis mitigates pancreatic inflammatory injury; however, in the presence of oncogenic KRAS, it facilitates PanIN progression. Thus, HBP1 serves as a critical regulator in both pancreatitis and early pancreatic neoplasia, representing a potential therapeutic target for intervening pancreatitis and PanIN progression.
]]></description>
<dc:creator>lee, S.-W.</dc:creator>
<dc:creator>Ekstrom, T.</dc:creator>
<dc:creator>Manalo, E.</dc:creator>
<dc:creator>Ye, S.</dc:creator>
<dc:creator>Berry, M.</dc:creator>
<dc:creator>Grygoryev, D.</dc:creator>
<dc:creator>Szczepaniak, M.</dc:creator>
<dc:creator>Marmolejo, C. O.</dc:creator>
<dc:creator>Haverlack, S.</dc:creator>
<dc:creator>Lee, J.</dc:creator>
<dc:creator>Shah, V. M.</dc:creator>
<dc:creator>Keith, D.</dc:creator>
<dc:creator>Muschler, J. L.</dc:creator>
<dc:creator>Chin, K.</dc:creator>
<dc:creator>Sears, R. C.</dc:creator>
<dc:creator>Weisberg, S. P.</dc:creator>
<dc:creator>Morgan, T. K.</dc:creator>
<dc:creator>kim, j.</dc:creator>
<dc:date>2025-01-27</dc:date>
<dc:identifier>doi:10.1101/2025.01.24.634817</dc:identifier>
<dc:title><![CDATA[HMG box-containing protein 1 (HBP1) prevents pancreatic injury in experimental pancreatitis but accelerates pancreatic neoplasia progression]]></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/10.1101/2025.02.01.634535v1?rss=1">
<title>
<![CDATA[
A spatially resolved single cell proteomic atlas of Small Bowel Adenocarcinoma 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.02.01.634535v1?rss=1</link>
<description><![CDATA[
Small bowel adenocarcinoma (SBA) is a rare malignancy marked with a poor prognosis. The cellular and proteomic heterogeneity within the tumor immune microenvironment (TIME) of SBA is a likely driver of prognosis, disease progression and response to therapy. We have addressed a major gap in knowledge of the TIME in SBA using highly multiplexed, protein imaging of the SBA tumor-immune ecosystem generating a comprehensive, single-cell level, spatial, proteomic atlas of TIME in > 600,000 cells from 136 tumor and matched normal samples from clinically and genomically annotated SBA patients (N=37). The SBA TIME Atlas informs on spatial distribution and interactions of tumor-intrinsic processes, diverse immune cell types, immune checkpoints, and vascularization. Enrichment of proliferating epithelial tissue, stem cells, and likely pro-tumor immune signatures in the tumor niche is contrasted by the representation of naive and early-effector T-cells in the epithelial compartments of adjacent normal tissues. Epithelial-stem-immune cell spatial architectures within tumor and matched-normal niches were strongly associated with patient survival, suggesting malignancy is driven by spatial architecture beyond the tumor microenvironment. The blueprints of therapeutically actionable immune checkpoints at the interface between epithelial and microenvironmental T-cells as well as macrophages have established a guideline for precision immunotherapies tailored to the TIME composition in SBA. We expect that this SBA atlas will contribute to a deeper understanding of the immune contexture in this rare disease as well as other gastrointestinal cancers and help guide future precision immune-oncology strategies.
]]></description>
<dc:creator>Dereli, Z.</dc:creator>
<dc:creator>Bozorgui, B.</dc:creator>
<dc:creator>Sanchez, M.</dc:creator>
<dc:creator>Hornstein, N. J.</dc:creator>
<dc:creator>Thibault, G.</dc:creator>
<dc:creator>Wang, H.</dc:creator>
<dc:creator>Mills, G. B.</dc:creator>
<dc:creator>Weinstein, J. N.</dc:creator>
<dc:creator>Overman, M. J.</dc:creator>
<dc:creator>Korkut, A.</dc:creator>
<dc:date>2025-02-04</dc:date>
<dc:identifier>doi:10.1101/2025.02.01.634535</dc:identifier>
<dc:title><![CDATA[A spatially resolved single cell proteomic atlas of Small Bowel Adenocarcinoma]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-02-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.02.01.636070v1?rss=1">
<title>
<![CDATA[
Centella asiatica improves sleep quality and quantity in aged mice 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.02.01.636070v1?rss=1</link>
<description><![CDATA[
Age-related sleep disruption is common in older adults. Not only does the total amount of time spent in sleep decline, but the number of arousals during sleep increases with age. As sleep is important for both memory consolidation and to prevent neurodegenerative pathology, this decline in sleep and/or sleep consolidation may underlie age-related cognitive decline and dementias. Furthermore, treatment of sleep disruption can improve quality of life. However, few interventions have successfully reversed age-related sleep decline. Extracts from the plant Centella asiatica have demonstrated neuroprotective effects in human, rodent, and fly models of aging and neurodegenerative diseases, and is a promising intervention for dementias, yet little is known about how these extracts affect sleep patterns. Here, we administered Centella asiatica water extract (CAW) dosed or control chow to male and female C57BL6/J mice aged 18 months. Effects on sleep composition were determined using electrodes that recorded EEG and EMG signals. We found that CAW dosed chow (1000 mg/kg/day) increased REM sleep time in aged male mice and decreased the number of arousals during sleep observed in aged females, compared to age- and sex-matched controls. We conclude that CAW administered in food has a moderate, sex-dependent effect on sleep quantity and quality.

Statement of SignificanceSleep declines with age and may underline age-related cognitive changes. However, few interventions have successfully reversed age-related sleep and cognitive decline. This study found that botanical extract from the plant Centella asiatica increased total REM sleep time in aged male mice, and decreased sleep fragmentation in aged female mice, compared to age- and sex-matched controls. Whether these moderate, sex-dependent effect sizes on sleep in aged mice are impactful enough to affect cognition, quality of life, and/or neurodegenerative pathology could be explored in future studies.
]]></description>
<dc:creator>Dovek, L.</dc:creator>
<dc:creator>Tinsley, C. E.</dc:creator>
<dc:creator>Gutowsky, K.</dc:creator>
<dc:creator>McDaniel, K.</dc:creator>
<dc:creator>Potter, Z.</dc:creator>
<dc:creator>Ruffins, M.</dc:creator>
<dc:creator>Milman, N.</dc:creator>
<dc:creator>Wong, C.</dc:creator>
<dc:creator>Soumyanath, A.</dc:creator>
<dc:creator>Gray, N. E.</dc:creator>
<dc:creator>Lim, M.</dc:creator>
<dc:date>2025-02-06</dc:date>
<dc:identifier>doi:10.1101/2025.02.01.636070</dc:identifier>
<dc:title><![CDATA[Centella asiatica improves sleep quality and quantity in aged mice]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-02-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.02.07.632847v1?rss=1">
<title>
<![CDATA[
mRNA stability factor HuR promotes immune evasion in pancreatic ductal adenocarcinoma 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.02.07.632847v1?rss=1</link>
<description><![CDATA[
The tumor microenvironment (TME) of pancreatic ductal adenocarcinoma (PDAC) is characterized by a limited infiltration of tumor-specific T cells and anti-tumor T cell activity. Extracellular factors in the PDAC TME have been widely reported to mediate immune suppression, but the contribution from tumor-intrinsic factors is not well understood. The RNA-binding protein, HuR (ELAVL1), is enriched in PDAC and negatively correlates with T cell infiltration. In an immunocompetent Kras-p53-Cre (KPC) orthotopic model of PDAC, we found that genetic disruption of HuR impaired tumor growth due to a novel role of HuR inducing T-cell suppression. Importantly, we found that HuR depletion in tumors enhanced both T cell number and activation states and diminished myeloid phenotypes by comprehensive spatial profiling of the PDAC TME. Mechanistically, HuR mediated the stabilization of mTOR pathway transcripts, and inhibition of mTOR activity rescued the impaired function of local T cells. Translating these findings, we demonstrated that HuR depletion sensitized PDAC tumors to immune checkpoint blockade, while isogenic, wildtype tumors are resistant. For the first time, we show that HuR facilitates tumor immune suppression in PDAC by inhibiting T cell infiltration and function and implicate targeting HuR as a potential therapeutic strategy in combination with immunotherapy.

SIGNIFICANCEThis study identified a novel mechanism that HuR supports pancreatic tumor growth by restricting T cell infiltration, promoting immune evasion. Our work supports targeting strategies against HuR in PDAC with the goal of enhancing PDAC sensitivity to immune-based cancer therapies, such as checkpoint blockade and T cell transfer.
]]></description>
<dc:creator>Guo, Y.</dc:creator>
<dc:creator>Finan, J. M.</dc:creator>
<dc:creator>Bartlett, A. Q.</dc:creator>
<dc:creator>Sivagnanam, S.</dc:creator>
<dc:creator>Blise, K. E.</dc:creator>
<dc:creator>Kirchberger, N.</dc:creator>
<dc:creator>Betre, K.</dc:creator>
<dc:creator>Hawthorne, K.</dc:creator>
<dc:creator>Chen, C.</dc:creator>
<dc:creator>Grossberg, A.</dc:creator>
<dc:creator>Xia, Z.</dc:creator>
<dc:creator>Coussens, L. M.</dc:creator>
<dc:creator>Sears, R. C.</dc:creator>
<dc:creator>Brody, J. R.</dc:creator>
<dc:creator>Eil, R.</dc:creator>
<dc:date>2025-02-07</dc:date>
<dc:identifier>doi:10.1101/2025.02.07.632847</dc:identifier>
<dc:title><![CDATA[mRNA stability factor HuR promotes immune evasion in pancreatic ductal adenocarcinoma]]></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/10.1101/2025.02.06.636959v1?rss=1">
<title>
<![CDATA[
Cerebellar climbing fibers convey perceptual choice during decision-making 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.02.06.636959v1?rss=1</link>
<description><![CDATA[
Cerebellar climbing fibers are thought to signal reward prediction errors in non-motor functions. By imaging postsynaptic responses of climbing fibers onto mouse Purkinje cell dendrites during auditory discrimination, we found that climbing fibers in crus I can encode cue identities or perceptual choices. These responses were reshaped by reversal learning. Optogenetic perturbation of climbing fiber activity impaired discrimination. These results suggest a feedforward role of climbing fibers in perceptual decision-making.
]]></description>
<dc:creator>Yiu, T. L.</dc:creator>
<dc:creator>Wilson, E. A.</dc:creator>
<dc:creator>Yahiro, T.</dc:creator>
<dc:creator>Ma, L.</dc:creator>
<dc:creator>Qin, M.</dc:creator>
<dc:creator>Zhong, H.</dc:creator>
<dc:date>2025-02-08</dc:date>
<dc:identifier>doi:10.1101/2025.02.06.636959</dc:identifier>
<dc:title><![CDATA[Cerebellar climbing fibers convey perceptual choice during decision-making]]></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/10.1101/2025.02.09.637183v1?rss=1">
<title>
<![CDATA[
IFNγ regulates MR1 transcription and antigen presentation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.02.09.637183v1?rss=1</link>
<description><![CDATA[
Antigen presentation molecules play key roles in activating T cell immunity. Multiple complementary pathways are known to regulate classical MHC-I molecules at transcriptional, translational, and post-translational levels. Intracellular trafficking mechanisms dictating post-transcriptional regulation of MR1, the MHC Class I-like molecule which restricts MAIT cells, have been an area of focus; however, little is known about MR1 transcriptional regulation. We demonstrate that, similar to classical MHC-I, interferons regulate MR1 transcription. Treatment of airway epithelial cells (AEC) with recombinant IFN{beta} or IFN{gamma} variably increased MR1 transcripts, while only IFN{gamma} significantly increased surface MR1 expression and enhanced antigen presentation to MAIT cells. The MR1 promoter contains binding motifs for interferon regulatory factor 1 (IRF1), an important MHC-I transcription factor. IRF1 knockout reduced IFN{gamma}-stimulated MR1 transcription, surface expression, and antigen presentation. Conversely, knockout of Nod-like Receptor family CARD domain containing 5 (NLRC5), a critical component of IFN{gamma}-induced MHC-I transcription, did not significantly impact MR1 expression. These findings were corroborated in primary human AEC treated with IFN{gamma}. In co-culture experiments, MAIT cells incubated with Streptococcus pneumoniae-infected primary AEC produced sufficient IFN{gamma} to stimulate upregulation of MR1 expression. Our data support a model where IFN{gamma} from activated MAIT cells or another source stimulates IRF1-dependent MR1 expression and antigen presentation, leading to greater MAIT cell activation. A robust MR1-dependent MAIT cell response may be beneficial for early infection responses, allowing minimal antigen stimulus to generate greater proinflammatory activity.
]]></description>
<dc:creator>Huber, M. E.</dc:creator>
<dc:creator>Larson, E. A.</dc:creator>
<dc:creator>Lust, T. N.</dc:creator>
<dc:creator>Heisler, C. M.</dc:creator>
<dc:creator>Harriff, M. J.</dc:creator>
<dc:date>2025-02-10</dc:date>
<dc:identifier>doi:10.1101/2025.02.09.637183</dc:identifier>
<dc:title><![CDATA[IFNγ regulates MR1 transcription and antigen presentation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-02-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.02.10.637549v1?rss=1">
<title>
<![CDATA[
Astrocyte gap junctions and Kir channels contribute to K+ buffering and regulate neuronal excitability 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.02.10.637549v1?rss=1</link>
<description><![CDATA[
ABSRACTAstrocytes are connected in a functional syncytium via gap junctions, which is thought to contribute to maintenance of extracellular K+ homeostasis. The prevailing hypothesis is that K+ released during neuronal firing is taken up by astrocytes via Kir channels and then distributed among neighboring astrocytes via gap junctions. Previous reports examining the role of Kir channels and gap junctions have shown both hyperexcitability and depression when each mechanism is blocked. Here, we tested the effect of blocking Kir channels and gap junctions, both independently and simultaneously, on field activity of cortical slices in response to a 3 s, 20 Hz stimulation train. Independently blocking either Kir channels or gap junctions increased the amplitude of the first fEPSC (field excitatory post-synaptic current) in response to a stimulation train, followed by suppression of fEPSCs during sustained stimulation. Surprisingly, blocking both gap junctions and Kir channels enhanced the suppression of neuronal activity, resulting in a [~]75% decrease in fiber volley (pre-synaptic action potentials) amplitude in the first response, followed by a fast and strong suppression of sustained fEPSCs. Our results demonstrate that blocking Kir channels and gap junctions can increase the excitability of neurons when firing is sparse, but suppression results when the firing frequency is increased to cortical physiological ranges. This suggest that K+ buffering via Kir and gap junctions, likely mediated by astrocytes, together play a critical role in maintaining neuronal excitability, particularly during sustained activity.
]]></description>
<dc:creator>Bojovic, D.</dc:creator>
<dc:creator>Dagostin, A.</dc:creator>
<dc:creator>Sullivan, S.</dc:creator>
<dc:creator>von Gersdorff, H.</dc:creator>
<dc:creator>Mishra, A.</dc:creator>
<dc:date>2025-02-11</dc:date>
<dc:identifier>doi:10.1101/2025.02.10.637549</dc:identifier>
<dc:title><![CDATA[Astrocyte gap junctions and Kir channels contribute to K+ buffering and regulate neuronal excitability]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-02-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.02.11.637423v1?rss=1">
<title>
<![CDATA[
Virus-against-virus dominant-negative interference strategy targeting a viral CC chemokine prevents cytomegalovirus-related neurodevelopmental pathogenesis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.02.11.637423v1?rss=1</link>
<description><![CDATA[
BackgroundCongenital cytomegalovirus (CMV) infections are one leading cause of human neurodevelopmental disorders. Increasing evidence for the pathogenic involvement of brain immune alterations was obtained in the recent years. Host and virus-encoded chemokines might play important roles in CMV-related neuropathogenesis by regulating leukocyte trafficking and microglia recruitment in the CMV-infected brains, and by interfering with key neurodevelopmental steps. In a rat model of CMV infection of the fetal brain in utero that leads to detrimental neurologic and other severe phenotypes postnatally, we reported on the early alteration of microglia and on the infiltration of the infected brains by lymphoid and myeloid cells. Particularly, expression of the r129 gene encoding the viral chemokine RCK3 was detected as early as 24h post-infection; together with the previously reported chemotaxis properties exerted by RCK3 on lymphocytes and on macrophages in vitro, this suggested that RCK3 might be involved in the brain immune cell alterations seen in the CMV-infected developing brains, and in the related neuropathogenesis.

MethodsInfection of the rat fetal brain was done by intracerebroventricular injections in utero of either rat CMV encoding wild-type (wt) RCK3 (RCMV-wt), or a mutant CMV counterpart encoding RCK3 with a deletion in its chemokine domain (RCMV-r129{Delta}NT). As RCMV-r129{Delta}NT had shown dominant-negative effects on the chemotaxis properties of RCK3 in vitro, simultaneous and successive co-infection rescue assays were also performed. The detrimental postnatal phenotypes in vivo and the epileptiform activity ex vivo usually detected after infection of the rat developing brain with RCMV-wt were monitored in the RCMV-r129{Delta}NT condition and in co-infection assays.

ResultsIn sharp contrast with RCMV-wt whose infection of the fetal brain led to decreased postnatal survival, impaired sensorimotor development, hindlimb hyperextension and epileptic seizures in neonatal pups, RCMV-r129{Delta}NT infection was not associated with any severe postnatal phenotype in vivo. Consistently, the epileptiform activity recorded in most neocortical slices from RCMV-wt-infected pups was not detected in any slice from RCMV-r129{Delta}NT-infected pups. Simultaneous co-infection assays led to dramatic prevention against the postnatal phenotypes in vivo and the altered network activity ex vivo, revealing a dose-dependent rescuing effect exerted by RCMV-r129{Delta}NT on RCMV-wt. Importantly, successful rescue was also obtained when the mutant RCMV-r129{Delta}NT was inoculated in the fetal brains either before or after infection with RCMV-wt.

SignificanceOur data demonstrate the crucial neuropathogenic role of RCK3 CMV chemokine in vivo. The dramatic success and apparent safety of the dominant-negative RCK3 rescue assays in vivo provide a proof-of-principle for the beneficial use of a virus-against-virus approach against CMV-related pathogenesis.
]]></description>
<dc:creator>Bauer, S.</dc:creator>
<dc:creator>Tarhini, S.</dc:creator>
<dc:creator>Buhler, E.</dc:creator>
<dc:creator>Saha, S.</dc:creator>
<dc:creator>Stamminger, T.</dc:creator>
<dc:creator>Streblow, D. N.</dc:creator>
<dc:creator>Burnashev, N.</dc:creator>
<dc:creator>Luche, H.</dc:creator>
<dc:creator>Szepetowski, P.</dc:creator>
<dc:date>2025-02-12</dc:date>
<dc:identifier>doi:10.1101/2025.02.11.637423</dc:identifier>
<dc:title><![CDATA[Virus-against-virus dominant-negative interference strategy targeting a viral CC chemokine prevents cytomegalovirus-related neurodevelopmental pathogenesis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-02-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.02.12.637926v1?rss=1">
<title>
<![CDATA[
Identification of a multi-omics factor predictive of long COVID in the IMPACC study 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.02.12.637926v1?rss=1</link>
<description><![CDATA[
Following SARS-CoV-2 infection, [~]10-35% of COVID-19 patients experience long COVID (LC), in which often debilitating symptoms persist for at least three months. Elucidating the biologic underpinnings of LC could identify therapeutic opportunities. We utilized machine learning methods on biologic analytes and patient reported outcome surveys provided over 12 months after hospital discharge from >500 hospitalized COVID-19 patients in the IMPACC cohort to identify a multi-omics "recovery factor". IMPACC participants who experienced LC had lower recovery factor scores compared to participants without LC. Biologic characterization revealed increased levels of plasma proteins associated with inflammation, elevated transcriptional signatures of heme metabolism, and decreased androgenic steroids in LC patients. The recovery factor was also associated with altered circulating immune cell frequencies. Notably, recovery factor scores were predictive of LC occurrence in patients as early as hospital admission, irrespective of acute disease severity. Thus, the recovery factor identifies patients at risk of LC early after SARS-CoV-2 infection and reveals LC biomarkers and potential treatment targets.
]]></description>
<dc:creator>Gabernet, G.</dc:creator>
<dc:creator>Maciuch, J.</dc:creator>
<dc:creator>Gygi, J. P.</dc:creator>
<dc:creator>Moore, J. F.</dc:creator>
<dc:creator>Hoch, A.</dc:creator>
<dc:creator>Syphurs, C.</dc:creator>
<dc:creator>Chu, T.</dc:creator>
<dc:creator>Jayavelu, N. D.</dc:creator>
<dc:creator>Corry, D. B.</dc:creator>
<dc:creator>Kheradmand, F.</dc:creator>
<dc:creator>Baden, L. R.</dc:creator>
<dc:creator>Sekaly, R.-P.</dc:creator>
<dc:creator>McComsey, G. A.</dc:creator>
<dc:creator>Haddad, E. K.</dc:creator>
<dc:creator>Cairns, C. B.</dc:creator>
<dc:creator>Rouphael, N.</dc:creator>
<dc:creator>Fernandez-Sesma, A.</dc:creator>
<dc:creator>Simon, V.</dc:creator>
<dc:creator>Metcalf, J. P.</dc:creator>
<dc:creator>Agudelo Higuita, N. I.</dc:creator>
<dc:creator>Hough, C. L.</dc:creator>
<dc:creator>Messer, W. B.</dc:creator>
<dc:creator>Davis, M. M.</dc:creator>
<dc:creator>Nadeau, K. C.</dc:creator>
<dc:creator>Pulendran, B.</dc:creator>
<dc:creator>Kraft, M.</dc:creator>
<dc:creator>Bime, C.</dc:creator>
<dc:creator>Reed, E. F.</dc:creator>
<dc:creator>Schaenman, J.</dc:creator>
<dc:creator>Erle, D. J.</dc:creator>
<dc:creator>Calfee, C. S.</dc:creator>
<dc:creator>Atkinson, M. A.</dc:creator>
<dc:creator>Brackenridge, S. C.</dc:creator>
<dc:creator>Melamed, E.</dc:creator>
<dc:creator>Shaw, A. C.</dc:creator>
<dc:creator>Hafler, D. A.</dc:creator>
<dc:creator>Ozonoff, A.</dc:creator>
<dc:creator>Bosinger, S. E.</dc:creator>
<dc:creator>Eckalbar, W.</dc:creator>
<dc:creator>Maecker, H. T.</dc:creator>
<dc:creator>Kim-Schulze, S.</dc:creator>
<dc:creator></dc:creator>
<dc:date>2025-02-14</dc:date>
<dc:identifier>doi:10.1101/2025.02.12.637926</dc:identifier>
<dc:title><![CDATA[Identification of a multi-omics factor predictive of long COVID in the IMPACC study]]></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/10.1101/2025.02.12.637953v1?rss=1">
<title>
<![CDATA[
Reversible lipid mediated pH-gating of connexin-46/50 by cryo-EM 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.02.12.637953v1?rss=1</link>
<description><![CDATA[
Gap junctions, formed by connexin proteins, establish direct electrical and metabolic coupling between cells, enabling coordinated tissue responses. These channels universally respond to intracellular pH changes, closing under acidic conditions to limit the spread of cytotoxic signals during cellular stress, such as ischemia. Using cryo-electron microscopy (cryo-EM), we uncover insights into the structural mechanism of pH-gating in native lens connexin-46/50 (Cx46/50) gap junctions. Mild acidification drives lipid infiltration into the channel pore, displacing the N-terminal (NT) domain and stabilizing pore closure. Lipid involvement is both essential and fully reversible, with structural transitions involving an ensemble of gated-states formed through non-cooperative NT domain movement as well as minor populations of a distinct destabilized open-state. These findings provide molecular insights into pH-gating dynamics, illustrating how structural changes may regulate gap junction function under cellular stress and linking Cx46/50 dysregulation to age-related cataract formation.
]]></description>
<dc:creator>Jarodsky, J. M.</dc:creator>
<dc:creator>Myers, J. B.</dc:creator>
<dc:creator>Reichow, S. L.</dc:creator>
<dc:date>2025-02-14</dc:date>
<dc:identifier>doi:10.1101/2025.02.12.637953</dc:identifier>
<dc:title><![CDATA[Reversible lipid mediated pH-gating of connexin-46/50 by cryo-EM]]></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/10.1101/2025.02.12.637955v1?rss=1">
<title>
<![CDATA[
Calcium induced N-terminal gating and pore collapse in connexin-46/50 gap junctions 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.02.12.637955v1?rss=1</link>
<description><![CDATA[
Gap junctions facilitate electrical and metabolic coupling essential for tissue function. Under ischemic conditions (e.g., heart attack or stroke), elevated intracellular calcium (Ca2+) levels uncouple these cell-to-cell communication pathways to protect healthy cells from cytotoxic signals. Using single-particle cryo-EM, we elucidate details of the Ca2+-induced gating mechanism of native connexin-46/50 (Cx46/50) gap junctions. The resolved structures reveal Ca2+ binding sites within the channel pore that alter the chemical environment of the permeation pathway and induce diverse occluded and gated states through N-terminal domain remodeling. Moreover, subunit rearrangements lead to pore collapse, enabling steric blockade by the N-terminal domains, reminiscent of the "iris model" of gating proposed over four decades ago. These findings unify and expand key elements of previous gating models, providing mechanistic insights into how Ca2+ signaling regulates gap junction uncoupling and broader implications for understanding cell stress responses and tissue protection.
]]></description>
<dc:creator>Flores, J. A.</dc:creator>
<dc:creator>O'Neill, S. E.</dc:creator>
<dc:creator>Jarodsky, J. M.</dc:creator>
<dc:creator>Reichow, S. L.</dc:creator>
<dc:date>2025-02-14</dc:date>
<dc:identifier>doi:10.1101/2025.02.12.637955</dc:identifier>
<dc:title><![CDATA[Calcium induced N-terminal gating and pore collapse in connexin-46/50 gap junctions]]></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/10.1101/2025.02.12.637899v1?rss=1">
<title>
<![CDATA[
UB-MBX-46 is a potent and selective antagonist of the human P2X7 receptor developed by structure-based drug design 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.02.12.637899v1?rss=1</link>
<description><![CDATA[
The P2X7 receptor is an ATP-gated ion channel that activates inflammatory pathways involved in diseases such as cancer, atherosclerosis, and neurodegeneration. However, despite the potential benefits of blocking overactive signaling, no P2X7 receptor antagonists have been approved for clinical use. Interspecies variation among existing antagonists has proven challenging, in part due to the dearth of molecular information on different receptor orthologs. Here, to identify distinct molecular features in the human receptor, we determine high-resolution cryo-EM structures of the full-length wild-type human P2X7 receptor in apo closed and ATP-bound open state conformations and draw comparisons with new and existing structures of other orthologs. We also report a cryo-EM structure of the human receptor in complex with an adamantane-based inhibitor, which we leverage, in conjunction with functional data and molecular dynamics simulations, to design a potent and selective antagonist with a unique polycyclic scaffold. Functional and structural analysis reveal how this optimized ligand, UB-MBX-46, interacts with the classical allosteric pocket of the human P2X7 receptor with picomolar potency and high selectivity, revealing its significant therapeutic potential.
]]></description>
<dc:creator>Oken, A. C.</dc:creator>
<dc:creator>Turcu, A. L.</dc:creator>
<dc:creator>Tzortzini, E.</dc:creator>
<dc:creator>Georgiou, K.</dc:creator>
<dc:creator>Nagel, J.</dc:creator>
<dc:creator>Barniol-Xicota, M.</dc:creator>
<dc:creator>Kim, G.-R.</dc:creator>
<dc:creator>Lee, S.-D.</dc:creator>
<dc:creator>Nicke, A.</dc:creator>
<dc:creator>Kim, Y.-C.</dc:creator>
<dc:creator>Müller, C. E.</dc:creator>
<dc:creator>Kolocouris, A.</dc:creator>
<dc:creator>Vazquez, S.</dc:creator>
<dc:creator>Mansoor, S. E.</dc:creator>
<dc:date>2025-02-16</dc:date>
<dc:identifier>doi:10.1101/2025.02.12.637899</dc:identifier>
<dc:title><![CDATA[UB-MBX-46 is a potent and selective antagonist of the human P2X7 receptor developed by structure-based drug design]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-02-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.02.18.638792v1?rss=1">
<title>
<![CDATA[
Effect of obesity on the acute response to SARS-CoV-2 infection and development of post-acute sequelae of COVID-19 (PASC) in nonhuman primates 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.02.18.638792v1?rss=1</link>
<description><![CDATA[
Long-term adverse consequences of SARS-CoV-2 infection, termed "long COVID" or post-acute sequelae of COVID (PASC), are a major component of overall COVID-19 disease burden. Prior obesity and metabolic disease increase the severity of acute disease, but SARS-CoV-2 infection also contributes to the development of new-onset metabolic disease. Since the COVID pandemic occurred in the context of the global obesity epidemic, an important question is the extent to which pre-existing obesity modifies long-term responses to SARS-CoV-2 infection. We utilized a nonhuman primate model to compare the effects of infection with the SARS-CoV-2 delta variant in lean and obese/insulin-resistant adult male rhesus macaques over a 6-month time course. While some longitudinal responses to SARS-CoV-2 infection, including overall viral dynamics, SARS-CoV-2-specific IgG induction, cytokine profiles, and tissue persistence of viral RNA, did not appreciably differ between lean and obese animals, other responses, including neutralizing Ab dynamics, lung pathology, body weight, degree of insulin sensitivity, adipocytokine profiles, body temperature, and nighttime activity levels were significantly different in lean versus obese animals. Furthermore, several parameters in lean animals were altered following SARS-CoV-2 infection to resemble those in obese animals. Notably, persistent changes in multiple parameters were present in most animals, suggesting that PASC may be more prevalent than estimated from self-reported symptoms in human studies.
]]></description>
<dc:creator>Sauter, K. A.</dc:creator>
<dc:creator>Webb, G. M.</dc:creator>
<dc:creator>Bader, L.</dc:creator>
<dc:creator>Kreklywich, C. N.</dc:creator>
<dc:creator>Takahashi, D. L.</dc:creator>
<dc:creator>Zaro, C.</dc:creator>
<dc:creator>McGuire, C. M.</dc:creator>
<dc:creator>Lewis, A. D.</dc:creator>
<dc:creator>Colgin, L. M. A.</dc:creator>
<dc:creator>Kirigiti, M. A.</dc:creator>
<dc:creator>Blomenkamp, H.</dc:creator>
<dc:creator>Pessoa, C.</dc:creator>
<dc:creator>Humkey, M.</dc:creator>
<dc:creator>Hulahan, J.</dc:creator>
<dc:creator>Sleeman, M.</dc:creator>
<dc:creator>Zweig, R. C.</dc:creator>
<dc:creator>Thomas, S.</dc:creator>
<dc:creator>Thomas, A.</dc:creator>
<dc:creator>Gao, L.</dc:creator>
<dc:creator>Hirsch, A. J.</dc:creator>
<dc:creator>Levy, M.</dc:creator>
<dc:creator>Cherry, S. R.</dc:creator>
<dc:creator>Kahn, S. E.</dc:creator>
<dc:creator>Slifka, M. K.</dc:creator>
<dc:creator>Sacha, J. B.</dc:creator>
<dc:creator>Kievit, P.</dc:creator>
<dc:creator>Roberts, C.</dc:creator>
<dc:date>2025-02-19</dc:date>
<dc:identifier>doi:10.1101/2025.02.18.638792</dc:identifier>
<dc:title><![CDATA[Effect of obesity on the acute response to SARS-CoV-2 infection and development of post-acute sequelae of COVID-19 (PASC) in nonhuman primates]]></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/10.1101/2025.02.20.639336v1?rss=1">
<title>
<![CDATA[
Epigenetic and Transcriptomic Alterations Precede Amyloidosis in the Hippocampus of the Alzheimer's Disease AppNL-G-F Knock in Mouse Model 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.02.20.639336v1?rss=1</link>
<description><![CDATA[
Detecting and understanding the early stages of Alzheimers disease (AD) is essential for uncovering initial mechanisms of neuropathology and devising effective interventions. In this study, we leveraged the humanized AppNL-G-F mouse which exhibits early-onset amyloid pathology with a predictable timeline, to investigate molecular changes in the hippocampus and blood before the onset of severe neuropathology and independent of aging. Employing a multi-omics approach, we identified alterations in chromatin accessibility, gene expression, and DNA methylation associated with early amyloidosis. Chromatin accessibility changes were prominent in excitatory neurons during early pathology, with a later shift to inhibitory neurons, potentially reflecting compensatory mechanisms to mitigate excitatory neuron dysregulation. Despite broadly comparable hippocampal cell composition, transcriptomic comparisons between wild-type and AppNL-G-F mice revealed major gene expression differences, particularly in pathways related to mitochondrial function and protein biosynthesis, preceding severe amyloid plaque deposition. In later stages, upregulation of immune and neuroinflammatory pathways was observed, aligning with established neuroinflammatory processes in AD. Additionally, we identified extensive DNA methylation differences in both the blood and hippocampus of AppNL-G-Fmice during early and late stages of pathology. Many differentially methylated regions in the blood, even at early pathology stages, were associated with cis-regulatory elements in the brain and were located near differentially expressed genes in the hippocampus. These regions were enriched in pathways associated with brain function, including neuron development and synaptic processes, highlighting a connection between blood methylation patterns and brain activity. This finding suggests the potential use of blood DNA methylation as a biomarker for the early detection of amyloidosis. Notably, we identified five candidate biomarker genes, including Rbfox1 and Camta1, with epigenetic dysregulation detectable in both the brain and blood prior to severe amyloid accumulation. Our study, leveraging a unique AD mouse model and a multi-omics approach, highlights epigenetic signatures of AD before the onset of clinical symptoms, providing a foundation for future research into early diagnosis and therapeutic strategies, as well as potential blood biomarkers.
]]></description>
<dc:creator>Okhovat, M.</dc:creator>
<dc:creator>Layman, C.</dc:creator>
<dc:creator>Davis, B. A.</dc:creator>
<dc:creator>Pederson, A.</dc:creator>
<dc:creator>O'Niel, A.</dc:creator>
<dc:creator>Holden, S.</dc:creator>
<dc:creator>Kessler, K.</dc:creator>
<dc:creator>Acharya, S.</dc:creator>
<dc:creator>Wheeler, K. J.</dc:creator>
<dc:creator>Nevonen, K. A.</dc:creator>
<dc:creator>Herrera, J.</dc:creator>
<dc:creator>Ward, S.</dc:creator>
<dc:creator>Vigh-Conrad, K.</dc:creator>
<dc:creator>Adey, A.</dc:creator>
<dc:creator>Raber, J.</dc:creator>
<dc:creator>Carbone, L.</dc:creator>
<dc:date>2025-02-21</dc:date>
<dc:identifier>doi:10.1101/2025.02.20.639336</dc:identifier>
<dc:title><![CDATA[Epigenetic and Transcriptomic Alterations Precede Amyloidosis in the Hippocampus of the Alzheimer's Disease AppNL-G-F Knock in Mouse Model]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-02-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.02.24.639678v1?rss=1">
<title>
<![CDATA[
Enhancement of Therapeutic Transgene Insertion for Murine Phenylketonuria 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.02.24.639678v1?rss=1</link>
<description><![CDATA[
Low in vivo transgene integration frequency limits the therapeutic efficacy of homology-directed repair (HDR)-mediated gene insertion as a treatment for Mendelian disorders. This study demonstrates improved efficacy of HDR-mediated gene insertion for the treatment of murine phenylalanine hydroxylase (PAH) deficiency, a model of human phenylketonuria (PKU), through pharmacologic inhibition of competing DNA repair pathways. Targeted integration of a Pah-expressing transgene into the hepatocytes of neonatal mice was enhanced with vanillin, a potent inhibitor of non-homologous end joining (NHEJ). This was further improved following combination of vanillin and novobiocin, an inhibitor of microhomology-mediated end joining (MMEJ). Combined NHEJ and MMEJ inhibition yielded PAH-expressing transgene insertions in approximately 10% of targeted alleles and was associated with a 70.6% decrease in serum phenylalanine. Demonstrating that pharmacologic inhibition of DNA repair pathways that compete with HDR can significantly enhance HDR-mediated transgene insertion in vivo.
]]></description>
<dc:creator>Martinez, M.</dc:creator>
<dc:creator>Richards, D. Y.</dc:creator>
<dc:creator>Winn, S. R.</dc:creator>
<dc:creator>Baris, A. M.</dc:creator>
<dc:creator>Vonada, A.</dc:creator>
<dc:creator>Dudley, S.</dc:creator>
<dc:creator>Harper, L.</dc:creator>
<dc:creator>Harding, C. O.</dc:creator>
<dc:date>2025-02-28</dc:date>
<dc:identifier>doi:10.1101/2025.02.24.639678</dc:identifier>
<dc:title><![CDATA[Enhancement of Therapeutic Transgene Insertion for Murine Phenylketonuria]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-02-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.02.25.640225v1?rss=1">
<title>
<![CDATA[
Comprehensive identification of β-lactam antibiotic polypharmacology in Mycobacterium tuberculosis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.02.25.640225v1?rss=1</link>
<description><![CDATA[
Infections with Mycobacterium tuberculosis (Mtb) cause tuberculosis (TB), which requires at least six months of treatment with multiple antibiotics. There is emergent interest in using {beta}-lactam antibiotics to improve treatment outcomes for patients. These drugs target cell wall biosynthesis, but a comprehensive list of enzymes inhibited by {beta}-lactams in Mtb is lacking. In the current study, we sought to identify and characterize Mtb enzymes inhibited by {beta}-lactam antibiotics using physiological conditions representative of both acute and chronic TB disease. We used new activity-based probes based on the {beta}-lactam antibiotic meropenem due to its approval by the World Health Organization for TB treatment. Activity-based probes label enzymes based on both substrate specificity and catalytic mechanism, enabling precise identification of drug targets. We identified previously undiscovered targets of meropenem in addition to known cell wall biosynthetic enzymes. We validated {beta}-lactam binding and hydrolysis for six newly identified targets: Rv1723, Rv2257c, Rv0309, DapE (Rv1202), MurI (Rv1338), and LipD (Rv1923). Our results demonstrate that there are at least 30 enzymes in Mtb vulnerable to inhibition by meropenem. This is many more {beta}-lactam targets than historically described, suggesting that efficacy in Mtb is a direct result of polypharmacology.
]]></description>
<dc:creator>Devlin, K. L.</dc:creator>
<dc:creator>Hutchinson, E.</dc:creator>
<dc:creator>Dearing, H. N.</dc:creator>
<dc:creator>Levine, S. R.</dc:creator>
<dc:creator>Reid, D. J.</dc:creator>
<dc:creator>Leach, D. T.</dc:creator>
<dc:creator>Griggs, L. H.</dc:creator>
<dc:creator>Lomas, G. X.</dc:creator>
<dc:creator>Gorham, L. J.</dc:creator>
<dc:creator>Wright, A. T.</dc:creator>
<dc:creator>Lamichhane, G.</dc:creator>
<dc:creator>Lin, V.</dc:creator>
<dc:creator>Beatty, K. E.</dc:creator>
<dc:date>2025-02-28</dc:date>
<dc:identifier>doi:10.1101/2025.02.25.640225</dc:identifier>
<dc:title><![CDATA[Comprehensive identification of β-lactam antibiotic polypharmacology in Mycobacterium tuberculosis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-02-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.03.04.641060v1?rss=1">
<title>
<![CDATA[
Identification of Direct-acting nsP2 Helicase Inhibitors with Anti-alphaviral Activity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.03.04.641060v1?rss=1</link>
<description><![CDATA[
Alphaviruses are mosquito-borne RNA viruses that pose a significant public health threat, with no FDA-approved antiviral therapeutics available. The non-structural protein 2 helicase (nsP2hel) is an enzyme involved in unwinding dsRNA essential for alphavirus replication. This study reports the discovery and optimization of first-in-class oxaspiropiperidine inhibitors targeting nsP2hel. Structure-activity relationship (SAR) studies identified potent cyclic sulfonamide analogs with nanomolar antiviral activity against chikungunya virus (CHIKV). Biochemical analyses of nsP2hel ATPase and RNA unwindase activities showed these compounds act by a non-competitive mode suggesting that they are allosteric inhibitors. Viral resistance mutations mapped to nsP2hel and a fluorine-labeled analog exhibited direct binding to the protein by 19F NMR. The lead inhibitor, 2o, demonstrated broad-spectrum antialphaviral activity, reducing titers of CHIKV, Mayaro virus (MAYV), and Venezuelan equine encephalitis virus (VEEV). These findings support nsP2hel as a viable target for development of broad-spectrum direct-acting antialphaviral drugs.

Graphical Abstract

O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=108 SRC="FIGDIR/small/641060v1_ufig1.gif" ALT="Figure 1">
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org.highwire.dtl.DTLVardef@856df5org.highwire.dtl.DTLVardef@1f6225borg.highwire.dtl.DTLVardef@4997d5org.highwire.dtl.DTLVardef@18f42f2_HPS_FORMAT_FIGEXP  M_FIG C_FIG
]]></description>
<dc:creator>Bose, M. R.</dc:creator>
<dc:creator>Sears, J. D.</dc:creator>
<dc:creator>Talbot, K. M.</dc:creator>
<dc:creator>Su, Y.-W. N.</dc:creator>
<dc:creator>Houliston, S.</dc:creator>
<dc:creator>Hossain, M. A.</dc:creator>
<dc:creator>Davis-Gilbert, Z. W.</dc:creator>
<dc:creator>Zhao, C.</dc:creator>
<dc:creator>Oh, H. J.</dc:creator>
<dc:creator>Brown, P. J.</dc:creator>
<dc:creator>Sanders, M. K.</dc:creator>
<dc:creator>Moorman, S. R.</dc:creator>
<dc:creator>Ojha, D.</dc:creator>
<dc:creator>Burdick, J. E.</dc:creator>
<dc:creator>Law, I.</dc:creator>
<dc:creator>Morales, N. L.</dc:creator>
<dc:creator>Martinez, S. A.</dc:creator>
<dc:creator>Loppnau, P.</dc:creator>
<dc:creator>Perez, J. G.</dc:creator>
<dc:creator>Drobish, A. M.</dc:creator>
<dc:creator>Morrison, T. E.</dc:creator>
<dc:creator>Streblow, Z. J.</dc:creator>
<dc:creator>Streblow, D. N.</dc:creator>
<dc:creator>Arrowsmith, C. H.</dc:creator>
<dc:creator>Vargason, A.</dc:creator>
<dc:creator>Counago, R. M.</dc:creator>
<dc:creator>Halabelian, L.</dc:creator>
<dc:creator>Arnold, J. J.</dc:creator>
<dc:creator>Cameron, C. E.</dc:creator>
<dc:creator>Moorman, N. J.</dc:creator>
<dc:creator>Heise, M. T.</dc:creator>
<dc:creator>Willson, T. M.</dc:creator>
<dc:date>2025-03-10</dc:date>
<dc:identifier>doi:10.1101/2025.03.04.641060</dc:identifier>
<dc:title><![CDATA[Identification of Direct-acting nsP2 Helicase Inhibitors with Anti-alphaviral Activity]]></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/10.1101/2025.03.10.642481v1?rss=1">
<title>
<![CDATA[
Analysis of factors that regulate HIV-1 fusion in reverse 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.03.10.642481v1?rss=1</link>
<description><![CDATA[
Based on observations that HIV-1 envelope (Env) proteins on the surfaces of cells have the capacity to fuse with neighboring cells or enveloped viruses that express CD4 receptors and CXCR4 co-receptors, we tested factors that affect the capacities of lentiviral vectors pseudotyped with CD4 and CXCR4 variants to infect Env-expressing cells. The process, which we refer to as fusion in reverse, involves the binding and activation of cellular Env proteins to fuse membranes with lentiviruses carrying CD4 and CXCR4 proteins. We have found that infection via fusion in reverse depends on cell surface Env levels, is inhibitable by an HIV-1-specific fusion inhibitor, and preferentially requires lentiviral pseudotyping with a glycosylphosphatidylinositol (GPI) anchored CD4 variant, and a cytoplasmic tail-truncated CXCR4 protein. We have demonstrated that latently HIV-1-infected cells can be specifically infected using this mechanism, and that activation of latently infected cells increases infection efficiency. The fusion in reverse approach allowed us to characterize how alteration of CD4 plus CXCR4 lipid membranes affected Env protein activities. In particular, we found that perturbation of membrane cholesterol levels did not affect Env activity. In contrast, viruses assembled in cells deficient for long chain sphingolipids showed increased infectivities, while viruses that incorporated a lipid scramblase were non-infectious. Our results yield new insights as to factors that influence envelope protein functions.
]]></description>
<dc:creator>Alfadhli, A.</dc:creator>
<dc:creator>Barklis, R. L.</dc:creator>
<dc:creator>Tafesse, F.</dc:creator>
<dc:creator>Barklis, E.</dc:creator>
<dc:date>2025-03-13</dc:date>
<dc:identifier>doi:10.1101/2025.03.10.642481</dc:identifier>
<dc:title><![CDATA[Analysis of factors that regulate HIV-1 fusion in reverse]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-03-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.03.17.643805v1?rss=1">
<title>
<![CDATA[
Human Neonatal MR1T Cells Have Diverse TCR Usage, are Less Cytotoxic and are Unable to Respond to Many Common Childhood Pathogens. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.03.17.643805v1?rss=1</link>
<description><![CDATA[
Neonatal sepsis is a leading cause of childhood mortality. Understanding immune cell development can inform strategies to combat this. MR1-restricted T (MR1T) cells can be defined by their recognition of small molecules derived from microbes, self, and drug and drug-like molecules, presented by the MHC class 1-related molecule (MR1). In healthy adults, the majority of MR1T cells express an invariant -chain; TRAV1-2/TRAJ33/12/20 and are referred to as mucosal-associated invariant T (MAIT) cells. Neonatal MR1T cells isolated from cord blood (CB) demonstrate more diversity in MR1T TCR usage, with the majority of MR1-5-OP-RU-tetramer(+) cells being TRAV1-2(-). To better understand this diversity, we performed single-cell-RNA-seq/TCR-seq (scRNA-seq/scTCR-seq) on MR1-5-OP-RU-tetramer(+) cells from CB (n=5) and adult participants (n=5). CB-derived MR1T cells demonstrate a less cytotoxic/pro-inflammatory phenotype, and a more diverse TCR repertoire. A panel of CB and adult MAIT and TRAV1-2(-) MR1T cell clones were generated, and CB-derived clones were unable to recognize several common riboflavin-producing childhood pathogens (S. aureus, S. pneumoniae, M. tuberculosis). Biochemical and structural investigation of one CB MAIT TCR (CB964 A2; TRAV1-2/TRBV6-2) showed a reduction in binding affinity toward the canonical MR1-antigen, 5-OP-RU, compared to adult MAIT TCRs that correlated with differences in {beta}-chain contribution in the TCR-MR1 interface. Overall, this data shows that CB MAIT and TRAV1-2(-) MR1T cells, express a diverse TCR repertoire, a more restricted childhood pathogen recognition profile and diminished cytotoxic and pro-inflammatory capacity. Understanding this diversity, along with the functional ability of TRAV1-2(-) MR1T cells, could provide insight into increased neonatal susceptibility to infections.
]]></description>
<dc:creator>Kain, D.</dc:creator>
<dc:creator>Abdelhady, W.</dc:creator>
<dc:creator>McElfresh, G.</dc:creator>
<dc:creator>Cansler, M.</dc:creator>
<dc:creator>Swarbrick, G.</dc:creator>
<dc:creator>Poa, K.</dc:creator>
<dc:creator>McNeice, C.</dc:creator>
<dc:creator>Boggy, G.</dc:creator>
<dc:creator>Rott, K.</dc:creator>
<dc:creator>Null, M.</dc:creator>
<dc:creator>Lewinsohn, D.</dc:creator>
<dc:creator>Rossjohn, J.</dc:creator>
<dc:creator>Bimber, B. N.</dc:creator>
<dc:creator>Lewinsohn, D.</dc:creator>
<dc:date>2025-03-20</dc:date>
<dc:identifier>doi:10.1101/2025.03.17.643805</dc:identifier>
<dc:title><![CDATA[Human Neonatal MR1T Cells Have Diverse TCR Usage, are Less Cytotoxic and are Unable to Respond to Many Common Childhood Pathogens.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-03-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.03.19.644227v1?rss=1">
<title>
<![CDATA[
MYC Serine 62 phosphorylation promotes its binding to DNA double strand breaks to facilitate repair and cell survival under genotoxic stress. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.03.19.644227v1?rss=1</link>
<description><![CDATA[
Genomic instability is a hallmark of cancer, driving oncogenic mutations that enhance tumor aggressiveness and drug resistance. MYC, a master transcription factor that is deregulated in nearly all human tumors, paradoxically induces replication stress and associated DNA damage while also increasing expression of DNA repair factors and mediating resistance to DNA-damaging therapies. Emerging evidence supports a non-transcriptional role for MYC in preserving genomic integrity at sites of active transcription and protecting stalled replication forks under stress. Understanding how MYCs genotoxic and genoprotective functions diverge may reveal new therapeutic strategies for MYC-driven cancers. Here, we identify a non-canonical role of MYC in DNA damage response (DDR) through its direct association with DNA breaks. We show that phosphorylation at serine 62 (pS62-MYC) is crucial for the efficient recruitment of MYC to damage sites, its interaction with repair factors BRCA1 and RAD51, and effective DNA repair to support cell survival under stress. Mass spectrometry analysis with MYC-BioID2 during replication stress reveals a shift in MYCs interactome, maintaining DDR associations while losing transcriptional regulators. These findings establish pS62-MYC as a key regulator of genomic stability and a potential therapeutic target in cancers.
]]></description>
<dc:creator>Cohn, G. M.</dc:creator>
<dc:creator>Daniel, C.</dc:creator>
<dc:creator>Eng, J.</dc:creator>
<dc:creator>Sun, X.-X.</dc:creator>
<dc:creator>Pelz, C.</dc:creator>
<dc:creator>Chin, K.</dc:creator>
<dc:creator>Smith, A.</dc:creator>
<dc:creator>Lopez, C. D.</dc:creator>
<dc:creator>Brody, J. R.</dc:creator>
<dc:creator>DAI, M.</dc:creator>
<dc:creator>Sears, R. C.</dc:creator>
<dc:date>2025-03-20</dc:date>
<dc:identifier>doi:10.1101/2025.03.19.644227</dc:identifier>
<dc:title><![CDATA[MYC Serine 62 phosphorylation promotes its binding to DNA double strand breaks to facilitate repair and cell survival under genotoxic stress.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-03-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.03.20.644395v1?rss=1">
<title>
<![CDATA[
Combining a rhesus cytomegalovirus/SIV vaccine with neutralizing antibody to protect against SIV challenge in rhesus macaques 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.03.20.644395v1?rss=1</link>
<description><![CDATA[
A vaccine is widely regarded as necessary for the control of the HIV pandemic and eventual eradication of AIDS. Neutralizing antibodies and MHC-E-restricted CD8+ T cells have both been shown capable of vaccine protection against the simian counterpart of HIV, SIV, in rhesus macaques. Here we provide preliminary evidence that combining these orthogonal antiviral mechanisms can provide increased protection against SIV challenge such that replication arrest observed following vaccination with a rhesus cytomegalovirus (RhCMV/SIV)-based vaccine was enhanced in the presence of passively administered incompletely protective levels of neutralizing antibody. The report invites studies involving larger cohorts of macaques and alternate routes of providing neutralizing antibody.
]]></description>
<dc:creator>Coppola, J.</dc:creator>
<dc:creator>Parren, M.</dc:creator>
<dc:creator>Bastidas, R.</dc:creator>
<dc:creator>Saye-Francisco, K.</dc:creator>
<dc:creator>Malvin, J.</dc:creator>
<dc:creator>Jardine, J. G.</dc:creator>
<dc:creator>Gilbride, R. M.</dc:creator>
<dc:creator>Ojha, S.</dc:creator>
<dc:creator>Feltham, S.</dc:creator>
<dc:creator>Morrow, D.</dc:creator>
<dc:creator>Barber-Axthelm, A.</dc:creator>
<dc:creator>Bochart, R.</dc:creator>
<dc:creator>Fast, R.</dc:creator>
<dc:creator>Oswald, K.</dc:creator>
<dc:creator>Shoemaker, R.</dc:creator>
<dc:creator>Lifson, J. D.</dc:creator>
<dc:creator>Picker, L. J.</dc:creator>
<dc:creator>Burton, D. R.</dc:creator>
<dc:creator>Hansen, S. G.</dc:creator>
<dc:date>2025-03-25</dc:date>
<dc:identifier>doi:10.1101/2025.03.20.644395</dc:identifier>
<dc:title><![CDATA[Combining a rhesus cytomegalovirus/SIV vaccine with neutralizing antibody to protect against SIV challenge in rhesus macaques]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-03-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.03.21.644674v1?rss=1">
<title>
<![CDATA[
The Photoswitchable Cannabinoid Azo-HU308 Enables Optical Control of Ca2+ Signaling in Pancreatic β-Cells via a Non CB2 TRPC Channel Mechanism 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.03.21.644674v1?rss=1</link>
<description><![CDATA[
Background and PurposeCa2+ plays a critical role in regulating insulin secretion from pancreatic {beta}-cells, a process modulated by various cell surface receptors, including cannabinoid receptors (CBRs). However, our understanding of cannabinoid signaling in {beta}-cells is complicated by the intricate pharmacology of cannabinoid ligands and their inherent hydrophobicity, which hinders precise control of receptor activation. This study aims to investigate the effects of the light-activatable CB2 receptor agonist, azo-HU308, on {beta}-cell Ca2+ dynamics.

Experimental ApproachWe employ fluorescent Ca2+ imaging in INS-1 832/13 (INS-1) {beta}-cells to measure Ca2+ transients induced by azo-HU308 and photoactivation with UV-A light. We then apply a pharmacological screen using a various CBR and TRP channel antagonists to determine the mechanism by which azo-HU308 enables optical control of {beta}-cell Ca2+ levels.

Key ResultsWe observed that azo-HU308 triggers a robust increase in intracellular Ca2+ when isomerized to the cis-form with UV-A light. The effect was repeatable over multiple cycles of irradiation and gradually desensitized on each sequential UV-light pulse. A pharmacological screen determined that the Ca2+ increase was not due to CB2 receptor activation and Ca2+ release from intracellular stores, but rather influx of extracellular Ca2+ through TRPC channels.

Conclusions and Implicationsazo-HU308 enables robust, repeatable stimulation of Ca2+ in INS-1 pancreatic {beta}-cells when triggered by UV-A light. This study presents a novel tool to optically control {beta}-cell Ca2+ dynamics, and sheds light on a new mechanism by which synthetic cannabinoids affect Ca2+ signaling through non-GPCR targets.
]]></description>
<dc:creator>Viray, A. E.</dc:creator>
<dc:creator>Frank, J. A.</dc:creator>
<dc:date>2025-03-25</dc:date>
<dc:identifier>doi:10.1101/2025.03.21.644674</dc:identifier>
<dc:title><![CDATA[The Photoswitchable Cannabinoid Azo-HU308 Enables Optical Control of Ca2+ Signaling in Pancreatic β-Cells via a Non CB2 TRPC Channel Mechanism]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-03-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.03.24.644849v1?rss=1">
<title>
<![CDATA[
Dissecting the steps in early Simian Immunodeficiency Virus dissemination following mucosal and intravenous infection of rhesus macaques 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.03.24.644849v1?rss=1</link>
<description><![CDATA[
In cases of HIV transmission, the typical delay from exposure to detectable viremia is approximately one week. This delay from exposure to viremia suggests that during initial expansion of virus from a limited number of founder lineages, there exists a period of low infected cell population. It is during this period of low infected cell population that the virus may be more vulnerable to clearance via primed immune responses targeting infected cells (e.g., antibody-dependent cellular cytotoxicity (ADCC) or CD8+ T cell killing/suppression). Potential future prophylactic harnessing of these immune mechanisms for early virally infected cell clearance will rely on an understanding of the earliest stages of viral replication and dissemination. The factors that dictate the rate of early viral spread, termed the  dissemination bottleneck could include target-cell-mediated effects, the anatomical microenvironment, or the organ of the first infected cells. In this study, we use the barcoded Simian Immunodeficiency Virus (SIV) infection model to assess the contribution of various anatomical and cellular mechanisms to the SIV dissemination bottleneck. Viral, cellular, and anatomically-mediated heterogeneity in viral replication each introduce a degree of variability into the early phases of viral spread, and when multiple founder lineages are present, this variability in early growth results in a large distribution in lineage sizes. Therefore, we use a comparison of viral lineage size variability across multiple experimental SIV infection models to examine the relative contribution to the overall dissemination bottleneck of viral-mediated stochasticity of cellular infection (e.g., integration site), infected cell phenotype (e.g., activation state), anatomical variability, and initial viral spread within the genital tract. We estimate that inherent heterogeneity in viral production by infected cells corresponds to 23 to 44% of the dissemination bottleneck, but the majority (56 to 77%) arises from anatomical heterogeneity (presumably heterogeneity in how conducive local microenvironments are to viral replication).

Author SummaryA brief window exists immediately following HIV transmission where low initial levels of virus and infected cells may be susceptible to immune clearance. A better understanding of the bottlenecks encountered by the virus during this window is necessary when designing therapies to clear HIV during this early period. We used a barcoded Simian Immunodeficiency Virus (SIV) infection model to track the early dissemination of multiple viral lineages after mucosal and intravenous inoculation. We observed up to 105-fold differences in lineage size between transmitted barcodes within a single animal two weeks after infection, suggesting very different trajectories of virial growth. By comparing lineage size diversity after mucosal and intravenous transmission and in vitro replication, we determined the contribution of anatomical and cellular mechanisms to the relative growth of different clonotypes. Although we expected that the processes of local dissemination may lead to greater lineage size diversity after mucosal transmission, this was not the case, and we saw no difference compared to intravenous transmission. We found that around a quarter of the diversity in clonotype size could be attributed to early cellular infection events, with the remainder likely attributable to differences in clonotype specific establishment and dissemination in vivo.
]]></description>
<dc:creator>Docken, S. S.</dc:creator>
<dc:creator>Macairan, A.</dc:creator>
<dc:creator>Schlub, T. E.</dc:creator>
<dc:creator>Fennessey, C. M.</dc:creator>
<dc:creator>Varco-Merth, B.</dc:creator>
<dc:creator>Picker, L. J.</dc:creator>
<dc:creator>Okoye, A. A.</dc:creator>
<dc:creator>Immonen, T. T.</dc:creator>
<dc:creator>Cromer, D.</dc:creator>
<dc:creator>Keele, B. F.</dc:creator>
<dc:creator>Davenport, M. P.</dc:creator>
<dc:date>2025-03-28</dc:date>
<dc:identifier>doi:10.1101/2025.03.24.644849</dc:identifier>
<dc:title><![CDATA[Dissecting the steps in early Simian Immunodeficiency Virus dissemination following mucosal and intravenous infection of rhesus macaques]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-03-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.03.31.645839v1?rss=1">
<title>
<![CDATA[
Protocol for Extracting Circulating Cell-Free DNA from Murine Saliva: Insights into Oral and Systemic Disease Research 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.03.31.645839v1?rss=1</link>
<description><![CDATA[
Circulating cell-free DNA (cfDNA) consists of small fragments of extracellular DNA from mammalian and bacterial cells found in bodily fluids such as blood and saliva, and it has been strongly recognized as a critical biomarker for various disease diagnoses, prognoses, and therapeutic monitoring. In this study, we present a reproducible protocol for efficiently isolating cfDNA from murine saliva using an innovative swabbing method in conjunction with the QIAamp MinElute ccfDNA Mini Kit. The quantification of isolated cfDNA is detected by a Qubit Fluorometer. Moreover, qualification assessment is conducted through BioAnalyzer analysis. This protocol facilitates research on saliva-derived cfDNA in the context of oral and systemic diseases in murine models.

Graphical Abstract

O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=200 SRC="FIGDIR/small/645839v1_ufig1.gif" ALT="Figure 1">
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]]></description>
<dc:creator>Elamin, I.</dc:creator>
<dc:creator>Rao, M. S.</dc:creator>
<dc:creator>Figliozzi, R. W.</dc:creator>
<dc:creator>Maahs, J. C.</dc:creator>
<dc:creator>Balish, M.</dc:creator>
<dc:creator>Hsia, S. V.</dc:creator>
<dc:creator>Fugolin, A. P. P.</dc:creator>
<dc:creator>Fan, J.</dc:creator>
<dc:date>2025-04-02</dc:date>
<dc:identifier>doi:10.1101/2025.03.31.645839</dc:identifier>
<dc:title><![CDATA[Protocol for Extracting Circulating Cell-Free DNA from Murine Saliva: Insights into Oral and Systemic Disease Research]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-04-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.04.02.646827v1?rss=1">
<title>
<![CDATA[
Complex carbohydrates catabolism capacity of bladder microbiota inhabiting healthy and overactive female bladders 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.04.02.646827v1?rss=1</link>
<description><![CDATA[
Overactive bladder syndrome (OAB) is a poorly understood symptom complex that affects 40% of females over the age of 40, with clinical features including urinary urgency and incontinence. In addition to inflammation, oxidative stress, nerve damage and reduced blood flow, alterations in the urinary microbiome (urobiome), specifically in bladder bacterial diversity, have been reported to be associated with OAB. Bladder bacteria are members of the urobiome along with viruses, archaea, fungi, and protozoans. The urobiome metabolism, particularly in relationship to host complex sugars (glycans), has been investigated recently in terms of glycosaminoglycan (GAG) utilization. Nevertheless, other urinary free oligosaccharides (FOS) have not yet been explored in both OAB and urobiome contexts. Similarly, a comprehensive search of microbial genes involved in host glycan metabolism in the bladder of adult females with or without OAB has not yet been reported. In this study, we investigated urinary FOS by mass spectrometry in women without OAB (asymptomatic controls), with OAB without incontinence (dry OAB), or with OAB with incontinence (wet OAB or urgency urinary incontinence, UUI). We also questioned the ability of commensal bladder bacteria to digest these FOS and other glycans, using bioinformatic tools to query publicly available bladder genomes isolated from affected and unaffected adult females to identify genes that encode polysaccharide lyases (PL) and glycoside hydrolases (GH). Our results show that FOS are present in a similar level in affected and unaffected controls with a few exceptions: ten FOS were found to differ between the OAB dry groups and either the control (four) or UUI (six) groups. Our results indicate that bladder microbiota from adult females both with and without OAB have the genetic capacity to digest host glycans and dietary sugars with subtle differences. Bladder bacteria isolated from females with OAB possess more GH/PL genes for host mucins, whereas bladder bacteria from controls possess more GH/PL genes for GAG digestion. In the control group, specifically, the genus Streptococcus possessed genes for the PL8 and GH88 enzymes, known to be involved in host GAG digestion. These novel bioinformatic data can enable future biochemical exploration of the urobiomes metabolism toward specific host glycans, such as GAGs, mucins O-glycans and N-glycans.
]]></description>
<dc:creator>Gourdine, J.-P.</dc:creator>
<dc:creator>Pacentine, I. V.</dc:creator>
<dc:creator>Barstad, A.</dc:creator>
<dc:creator>Dahl, E. M.</dc:creator>
<dc:creator>Gregory, W. T.</dc:creator>
<dc:creator>Wolfe, A. J.</dc:creator>
<dc:creator>Sysoeva, T. A.</dc:creator>
<dc:creator>Karstens, L.</dc:creator>
<dc:date>2025-04-04</dc:date>
<dc:identifier>doi:10.1101/2025.04.02.646827</dc:identifier>
<dc:title><![CDATA[Complex carbohydrates catabolism capacity of bladder microbiota inhabiting healthy and overactive female bladders]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-04-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.04.03.647095v1?rss=1">
<title>
<![CDATA[
Integrative Analysis of EGF, OSM, and TGFB Signaling Pathways Reveals Synergistic Mechanisms Driving Cell Motility Through CXCR2 Chemotactic Signaling and CREB Activation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.04.03.647095v1?rss=1</link>
<description><![CDATA[
The microenvironment surrounding cells plays a critical role in determining cellular phenotype. Key components of the microenvironment include the diverse milieu of ligands and cytokines bind cell surface receptors to initiate changes in molecular programs. While the responses to extracellular signals have been extensively studied in isolation, little is known about the effects of combinations of signals on phenotypic and transcriptional responses. In this study, we used a coordinated approach to systematically investigate the combinatorial effects of the cytokines Oncostatin M (OSM) and Transforming Growth Factor Beta 1 (TGFB), and the growth factor Epidermal Growth Factor (EGF) on MCF10A mammary epithelial cells. Quantitative analysis of live-cell imaging data revealed a complex array of phenotypic responses after ligand treatment, including changes in proliferation, motility, cell clustering, and cytoplasmic size. We observed that all ligand combinations produce emergent phenotypic responses distinct from the maximal effects of individual ligands, indicating induction of new molecular programs. Companion RNA sequencing studies revealed a synergistic upregulation of a small but specific transcriptional program, including genes involved in cell migration, epithelial differentiation, and chemotactic signaling. Notably, these included chemokines such as CXCL3, CXCL5, and PPBP, which are known drivers of epithelial proliferation and migration. Additionally, transcription factor enrichment analyses and Reverse Phase Protein Array (RPPA) studies highlighted distinct changes in pathway utilization and transcription factor activity following combination treatment, including enhanced activation of MAP kinase and CREB signaling, compared to treatment with either agent alone. Using partial least squares regression, we identified robust transcriptional signatures associated with quantitative cellular phenotypes. We validated these signatures in independent datasets, confirming that they generalize across cellular contexts. Finally, an in-depth functional analysis of cell motility with RNA interference and pathway inhibition revealed that synergistic upregulation of CXCR2 signaling, mediated by CREB transcription factor activation, contributes to increases in cell motility across ligand conditions. These findings underscore the importance of combinatorial signaling in reprogramming epithelial phenotypes and reveal potential therapeutic targets for disrupting synergistic pathways in disease contexts such as cancer progression. Together, this study provides a framework for understanding how complex ligand interactions shape phenotypic and molecular landscapes.
]]></description>
<dc:creator>McLean, I. C.</dc:creator>
<dc:creator>Gross, S.</dc:creator>
<dc:creator>Copperman, J.</dc:creator>
<dc:creator>Derrick, D.</dc:creator>
<dc:creator>Heiser, L.</dc:creator>
<dc:date>2025-04-04</dc:date>
<dc:identifier>doi:10.1101/2025.04.03.647095</dc:identifier>
<dc:title><![CDATA[Integrative Analysis of EGF, OSM, and TGFB Signaling Pathways Reveals Synergistic Mechanisms Driving Cell Motility Through CXCR2 Chemotactic Signaling and CREB Activation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-04-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.04.06.647399v1?rss=1">
<title>
<![CDATA[
Incubation of oxycodone craving is associated with CP-AMPAR upregulation in D1 and D2 receptor-expressing medium spiny neurons in nucleus accumbens core and shell 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.04.06.647399v1?rss=1</link>
<description><![CDATA[
A major problem in treating opioid use disorder is persistence of craving after protracted abstinence. This has been modeled in rodents using the incubation of craving model, in which cue-induced drug seeking increases over the first weeks of abstinence from drug self-administration and then remains high for an extended period. Incubation has been reported for several opioids, including oxycodone, but little is known about underlying synaptic plasticity. In contrast, it is well established that incubation of cocaine and methamphetamine craving depends on strengthening of glutamate synapses in the nucleus accumbens (NAc) through incorporation of calcium-permeable AMPARs (CP-AMPARs). CP-AMPARs have higher conductance than the calcium-impermeable AMPARs that mediate NAc excitatory transmission in drug-naive animals, as well as other distinct properties. Here we examined AMPAR transmission in medium spiny neurons (MSN) of NAc core and shell subregions in rats during forced abstinence from extended-access oxycodone self-administration. In early abstinence (prior to incubation), CP-AMPAR levels were low. After 17-33 days of abstinence (when incubation is stably plateaued), CP-AMPAR levels were significantly elevated in both subregions. These results explain the prior demonstration that infusion of a selective CP-AMPAR antagonist into NAc core or shell subregions prevents expression of oxycodone incubation. Then, using transgenic rats, we found CP-AMPAR upregulation on both D1 and D2 receptor-expressing MSN, which contrasts with selective upregulation on D1 MSN after cocaine and methamphetamine incubation. Overall, our results demonstrate a common role for CP-AMPAR upregulation in psychostimulant and oxycodone incubation, albeit with differences in MSN subtype-specificity.
]]></description>
<dc:creator>Mount, K. A.</dc:creator>
<dc:creator>Kuhn, H. M.</dc:creator>
<dc:creator>Hwang, E.-K.</dc:creator>
<dc:creator>Beutler, M. M.</dc:creator>
<dc:creator>Wolf, M. E.</dc:creator>
<dc:date>2025-04-06</dc:date>
<dc:identifier>doi:10.1101/2025.04.06.647399</dc:identifier>
<dc:title><![CDATA[Incubation of oxycodone craving is associated with CP-AMPAR upregulation in D1 and D2 receptor-expressing medium spiny neurons in nucleus accumbens core and shell]]></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/10.1101/2025.04.01.646698v1?rss=1">
<title>
<![CDATA[
Hepatic Yap1 activates systemic catabolism and muscle loss during organ repair: evidence for a liver-derived common mechanism with cancer cachexia. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.04.01.646698v1?rss=1</link>
<description><![CDATA[
Recovery from critical injury concomitant with restoration of functional organ mass invokes a systemic catabolic response leading to muscle and fat loss, known as cachexia. We interrogated this process using mouse models of organ repair, including liver regeneration after hepatectomy, Yap1-mediated hepatomegaly, and pneumonectomy. Both hepatectomy and Yap1 induced systemic catabolism. Muscle and adipose wasting scaled with degree of liver growth, with 10-25% reduction of muscle mass and 50-80% of fat mass. In contrast, non-regenerative lung injury did not induce tissue wasting. Liver growth elevated resting energy expenditure. Tracer studies demonstrated redistribution of muscle-derived cholesterol and amino acids to regenerating liver. Gene expression changes in livers and muscles showed high concordance between the liver growth models and cancer cachexia models, including pancreatic adenocarcinoma cachexia. We propose that cachexia is a normal and essential reparative process in organ repair and regeneration and further, that cancer cachexia is a pathological exacerbation of an adaptive process mediated by activation of Yap1 in the liver.
]]></description>
<dc:creator>HAMIDI, T.</dc:creator>
<dc:creator>Yang, Y.</dc:creator>
<dc:creator>Robinson, T. P.</dc:creator>
<dc:creator>Kronbergs, A.</dc:creator>
<dc:creator>Au, E.</dc:creator>
<dc:creator>Rupert, J. E.</dc:creator>
<dc:creator>Zhong, X.</dc:creator>
<dc:creator>Lal, S.</dc:creator>
<dc:creator>Liang, T.</dc:creator>
<dc:creator>Robling, A. G.</dc:creator>
<dc:creator>Hoggatt, A.</dc:creator>
<dc:creator>Carvalho, R.</dc:creator>
<dc:creator>Cury, S. S.</dc:creator>
<dc:creator>Wells, C.</dc:creator>
<dc:creator>Zimmers, T.</dc:creator>
<dc:creator>Koniaris, L. G.</dc:creator>
<dc:date>2025-04-07</dc:date>
<dc:identifier>doi:10.1101/2025.04.01.646698</dc:identifier>
<dc:title><![CDATA[Hepatic Yap1 activates systemic catabolism and muscle loss during organ repair: evidence for a liver-derived common mechanism with cancer cachexia.]]></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/10.1101/2025.04.02.646829v1?rss=1">
<title>
<![CDATA[
Genome-wide analysis reveals genes mediating resistance to paraquat neurodegeneration in Drosophila 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.04.02.646829v1?rss=1</link>
<description><![CDATA[
Parkinsons disease (PD) is thought to develop through a complex interplay of genetic and environmental factors. Epidemiological studies have linked exposure to certain pesticides such as paraquat with elevated PD risk, although how a persons genetic makeup influences disease risk upon exposure remains unknown. Here, we used a genome-wide approach to uncover genes that play a role in resistance to paraquat-induced dopaminergic neurodegeneration in Drosophila. We developed a paraquat exposure model displaying delayed-onset dopaminergic (DA) neurodegeneration to recapitulate this aspect of human disease. We reveal that genetic background is a strong determinant of paraquat-induced DA neurodegeneration susceptibility across a series of nearly 200 fly strains called the Drosophila genetic reference panel (DGRP). Through unbiased genome-wide analysis and follow-up validation, we identify two candidate paraquat resistance genes, luna and CG32264. In gene-level studies, decreased expression of luna or CG32264 is associated with paraquat-induced DA neuron loss while overexpression of either gene prevents neurodegeneration in vivo. The mammalian ortholog of CG32264 is Phactr2, which has previously been linked to human idiopathic PD risk in several populations. Hence, our results reveal genes regulating paraquat-induced DA neuron loss that intersect with human PD risk variants, supporting the potential relevance of our findings to PD and underscoring a role for gene-environment interactions in pesticide-related DA neurodegeneration.

ARTICLE SUMMARYParaquat is a widely used herbicide linked to increased PD risk and to dopaminergic neurodegeneration in animal studies. Gene-environment interactions likely influence whether an individual exposed to paraquat eventually manifests PD and presents a major opportunity to yield insight into PD genetics. We developed a paraquat-induced neurodegeneration model in Drosophila, applied this model to nearly 200 fly strains belonging to the Drosophila Genetic Reference Panel and used a genome-wide association approach to identify candidate modifier genes of paraquat-induced dopamine neuron loss which we subsequently validated through RNAi and overexpression functional testing. Through this approach, we reveal two novel paraquat resistance genes, luna and CG32264. Strikingly, the putative mammalian ortholog of CG32264 (Phactr2) was previously linked to human PD, supporting the potential relevance of our findings to human disease.
]]></description>
<dc:creator>Villalobos-Cantor, S.</dc:creator>
<dc:creator>Arreola-Bustos, A.</dc:creator>
<dc:creator>Martin, I.</dc:creator>
<dc:date>2025-04-08</dc:date>
<dc:identifier>doi:10.1101/2025.04.02.646829</dc:identifier>
<dc:title><![CDATA[Genome-wide analysis reveals genes mediating resistance to paraquat neurodegeneration in Drosophila]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-04-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.04.11.648464v1?rss=1">
<title>
<![CDATA[
CODAvision: best practices and a user-friendly interface for rapid, customizable segmentation of medical images 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.04.11.648464v1?rss=1</link>
<description><![CDATA[
Image-based machine learning tools have emerged as powerful resources for analyzing medical images, with deep learning-based semantic segmentation commonly utilized to enable spatial quantification of structures in images. However, customization and training of segmentation algorithms requires advanced programming skills and intricate workflows, limiting their accessibility to many investigators. Here, we present a protocol and software for automatic segmentation of medical images guided by a graphical user interface (GUI) using the CODAvision algorithm. This workflow simplifies the process of semantic segmentation of microanatomical structures by enabling users to train highly customizable deep learning models without extensive coding expertise. The protocol outlines best practices for creating robust training datasets, configuring model parameters, and optimizing performance across diverse biomedical image modalities.

CODAvision enhances the usability of the CODA algorithm (Nature Methods, 2022) by streamlining parameter configuration, model training, and performance evaluation, automatically generating quantitative results and comprehensive reports. We expand beyond the original implementation of CODA to serial histology by demonstrating robust performance across numerous medical image modalities and diverse biological questions. We provide sample results in data types including histology, magnetic resonance imaging (MRI), and computed tomography (CT). We demonstrate the diverse use of this tool in applications including quantification of metastatic burden in in vivo models and deconvolution of spot-based spatial transcriptomics datasets. This protocol is designed for researchers with interest in rapid design of highly customizable semantic segmentation algorithms and a basic understanding of programming and anatomy.
]]></description>
<dc:creator>Matos-Romero, V.</dc:creator>
<dc:creator>Gomez-Becerril, J.</dc:creator>
<dc:creator>Forjaz, A.</dc:creator>
<dc:creator>Dequiedt, L.</dc:creator>
<dc:creator>Newton, T.</dc:creator>
<dc:creator>Joshi, S.</dc:creator>
<dc:creator>Shen, Y.</dc:creator>
<dc:creator>Hanna, E.</dc:creator>
<dc:creator>Nair, P. R.</dc:creator>
<dc:creator>Sivasubramanian, A.</dc:creator>
<dc:creator>Wang, J.</dc:creator>
<dc:creator>Lasse-Opsahl, E. L.</dc:creator>
<dc:creator>Bell, A. T. F.</dc:creator>
<dc:creator>Czum, J.</dc:creator>
<dc:creator>Steenbergen, C.</dc:creator>
<dc:creator>Dai, D.-F.</dc:creator>
<dc:creator>Wood, L. D.</dc:creator>
<dc:creator>Kagohara, L. T.</dc:creator>
<dc:creator>Fertig, E. J.</dc:creator>
<dc:creator>Pasca di Magliano, M.</dc:creator>
<dc:creator>Shatzel, J.</dc:creator>
<dc:creator>McCarty, O. J.</dc:creator>
<dc:creator>Lo, J. O.</dc:creator>
<dc:creator>Rosenberg, A.</dc:creator>
<dc:creator>Hruban, R. H.</dc:creator>
<dc:creator>Munoz-Barrutia, A.</dc:creator>
<dc:creator>Wirtz, D.</dc:creator>
<dc:creator>Kiemen, A. L.</dc:creator>
<dc:date>2025-04-14</dc:date>
<dc:identifier>doi:10.1101/2025.04.11.648464</dc:identifier>
<dc:title><![CDATA[CODAvision: best practices and a user-friendly interface for rapid, customizable segmentation of medical images]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-04-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.04.10.648051v1?rss=1">
<title>
<![CDATA[
Submicron-Sampling of Living Cells by Macrophages 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.04.10.648051v1?rss=1</link>
<description><![CDATA[
SUMMARY PARAGRAPHAn effective immune system must sample and appreciate healthy-self identity to prevent autoimmunity and to contrast to pathogenic insults1-3. Self-proteins are presented to T cells in the thymus during immune cell development2,3, and must be presented throughout the body to both maintain regulatory T cell populations4-6 and provide a tonic signal to maintain conventional T cells over time7-9. The ready observations of continuous apoptosis in some organs together with the ingestion of that material by myeloid populations has led to a conventional understanding of ongoing cell-death as a major source of self-antigens10, complemented in some situations by uptake of free-floating cell-derived vesicles. Here, we used a series of companion imaging and vesicular labeling technologies to reveal an alternate process undertaken by macrophages that results in non-destructive and direct sampling of living cells. The process requires cell-cell contact, does not require caspase activation, and takes place via a trogocytosis-like stretching of the target cell into the macrophage, leading to the generation of submicron-sized vesicles containing cytoplasm. Using a high-dimensional flow-based method for labeling vesicles ingested under this versus other conditions, we find that live-sampled material is distinctly processed, is poorly subject to fusion with lysosomes, and produces ensuing differential effects on the presentation of those to CD4 versus CD8 T cells. Disrupting this trafficking by redirecting antigen to the lysosome significantly reduced the associated macrophage-mediated priming of CD8 T cells. This demonstrates an important and substantial sampling of living cells by the immune system, with clear consequences for maintaining the border of immunity.
]]></description>
<dc:creator>Fan, A. C.</dc:creator>
<dc:creator>Thota, R.</dc:creator>
<dc:creator>Serwas, N.</dc:creator>
<dc:creator>Marchuk, K.</dc:creator>
<dc:creator>Ruhland, M.</dc:creator>
<dc:creator>Edwards, A.</dc:creator>
<dc:creator>Krummel, M. F.</dc:creator>
<dc:date>2025-04-16</dc:date>
<dc:identifier>doi:10.1101/2025.04.10.648051</dc:identifier>
<dc:title><![CDATA[Submicron-Sampling of Living Cells by Macrophages]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-04-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.04.12.648490v1?rss=1">
<title>
<![CDATA[
KDM5B expression is suppressed by MYC in a negative feedback loop to promote cell survival. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.04.12.648490v1?rss=1</link>
<description><![CDATA[
Both c-MYC and KDM5B are global regulators of gene transcription, cell identity, and master transcription factors in hematopoietic malignancies. In this study we link these two critical factors in a negative feedback loop that controls apoptosis and cell survival. We find the MYC specifically downregulates KDM5B, thereby inhibiting its transcriptional repressive functions. Loss of KDM5B abrogates the cell death response to MYC withdrawal in MYC-dependent cells, indicating KDM5B mediates cell death. We find that KDM5B regulates the MYC network, and specifically demethylates the MYC locus. In summary we have discovered a negative feedback loop between MYC and KDM5B. The MYC-dependent suppression of KDM5B leads to a global increase in H3K4me3 methylation, transcriptional activity, and increase cell survival and tumor progression.
]]></description>
<dc:creator>Chatterjee, T.</dc:creator>
<dc:creator>Beffert, E.</dc:creator>
<dc:creator>Dewson, G. S.</dc:creator>
<dc:creator>Sullivan, D. K.</dc:creator>
<dc:creator>Anchang, B.</dc:creator>
<dc:creator>Sears, R. C.</dc:creator>
<dc:creator>Liefwalker, D.</dc:creator>
<dc:date>2025-04-18</dc:date>
<dc:identifier>doi:10.1101/2025.04.12.648490</dc:identifier>
<dc:title><![CDATA[KDM5B expression is suppressed by MYC in a negative feedback loop to promote cell survival.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-04-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.04.15.648971v1?rss=1">
<title>
<![CDATA[
Maternal obesity programs cardiac remodeling in offspring via epigenetic, metabolic, and immune dysregulations. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.04.15.648971v1?rss=1</link>
<description><![CDATA[
Maternal obesity during pregnancy significantly increases the offsprings risk of later-life cardiovascular disease. This study investigated cardiometabolic perturbations by utilizing a mouse model of maternal high-fat diet (HFD)-induced obesity that recapitulates metabolic abnormalities observed in humans. We report that offspring of HFD-fed mothers (Off-HFD) exhibit a progression of obesity, hypertension, dyslipidemia, and metabolic inflexibility when compared with offspring of regular diet-fed mothers (Off-RD). Deeper investigation of cardiac function further identified significant functional, metabolic, and immune perturbations in adult offspring of mothers on HFD. Specifically, Off-HFD mice presented progressing cardiac hypertrophy with reduced ejection fraction, increased accumulation of fibrotic tissue, mitochondrial dysfunction, and altered immune complexity including increases in cardiac resident and infiltrated macrophages, and decreases in CD4+ and CD8+ T-cell subpopulations. While these alterations may not be immediately catastrophic, they likely predispose the offspring to heightened sensitivity to nutritional, psychological, or environmental stressors. Analysis of DNA methylation in the hearts of newly-weaned offspring from RD and HFD mothers revealed numerous differentially methylated CpGs and regions within genes associated with cardiac development, hypertrophy, mitochondrial function, and immune response. Thus, our study shows epigenetic remodeling early in development, which is likely responsible for the cardiovascular dysregulation observed in adult life. These findings uncover potential windows of opportunity for preventive therapy and early therapeutic interventions.
]]></description>
<dc:creator>Philips, E. A.</dc:creator>
<dc:creator>Alharithi, Y.</dc:creator>
<dc:creator>Wilson, T. D.</dc:creator>
<dc:creator>Broberg, C.</dc:creator>
<dc:creator>Davis, B. A.</dc:creator>
<dc:creator>Koch, S.</dc:creator>
<dc:creator>Carbone, L.</dc:creator>
<dc:creator>Rubinstein, J.</dc:creator>
<dc:creator>Gurley, S. B.</dc:creator>
<dc:creator>Rugonyi, S.</dc:creator>
<dc:creator>Kumar, S.</dc:creator>
<dc:creator>Maloyan, A.</dc:creator>
<dc:date>2025-04-19</dc:date>
<dc:identifier>doi:10.1101/2025.04.15.648971</dc:identifier>
<dc:title><![CDATA[Maternal obesity programs cardiac remodeling in offspring via epigenetic, metabolic, and immune dysregulations.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-04-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.04.30.651574v1?rss=1">
<title>
<![CDATA[
Surface morphometrics reveals local membrane thickness variation in organellar subcompartments 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.04.30.651574v1?rss=1</link>
<description><![CDATA[
Lipid bilayers form the basis of organellar architecture, structure, and compartmentalization in the cell. Decades of biophysical, biochemical, and imaging studies on purified or in vitro reconstituted liposomes have shown that variations in lipid composition influence the physical properties of membranes, such as thickness and curvature. However, similar studies characterizing these membrane properties within the native cellular context have remained technically challenging. Recent advancements in cellular cryo-electron tomography (cryo-ET) imaging enable high-resolution, three-dimensional views of native organellar membrane architecture preserved in near-native conditions. We previously developed a  Surface Morphometrics pipeline that generates surface mesh reconstructions to model and quantify cellular membrane ultrastructure from cryo-ET data. Here, we expand this pipeline to measure the distance between the phospholipid head groups (PHG) of the membrane bilayer as a readout of membrane thickness. Using this approach, we demonstrate thickness variations both within and between distinct organellar membranes. We also demonstrate that membrane thickness positively correlates with other features, such as membrane curvedness. Further, we show that subcompartments of the mitochondrial inner membrane exhibit varying membrane thicknesses that are independent of network morphology (i.e., fragmented versus elongated networks). Finally, we demonstrate that large membrane-associated macromolecular complexes exhibit distinct density profiles that correlate with local variations in membrane thickness. Overall, our updated Surface Morphometrics pipeline provides a framework for investigating how changes in membrane composition in various cellular and disease contexts affect organelle ultrastructure and function.
]]></description>
<dc:creator>Medina, M.</dc:creator>
<dc:creator>Chang, Y.-T.</dc:creator>
<dc:creator>Rahmani, H.</dc:creator>
<dc:creator>Fuentes, D.</dc:creator>
<dc:creator>Barad, B. A.</dc:creator>
<dc:creator>Grotjahn, D.</dc:creator>
<dc:date>2025-05-01</dc:date>
<dc:identifier>doi:10.1101/2025.04.30.651574</dc:identifier>
<dc:title><![CDATA[Surface morphometrics reveals local membrane thickness variation in organellar subcompartments]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-05-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.05.02.651747v1?rss=1">
<title>
<![CDATA[
Differential cell signaling testing for cell-cell communication inference from single-cell data by dominoSignal 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.05.02.651747v1?rss=1</link>
<description><![CDATA[
Algorithms for ligand-receptor network inference have emerged as commonly used tools to estimate cell-cell communication from reference single-cell data. Many studies employ these algorithms to compare signaling between conditions and lack methods to statistically identify signals that are significantly different. We previously developed the cell communication inference algorithm Domino, which considers ligand and receptor gene expression in association with downstream transcription factor activity scoring. We developed the dominoSignal software to innovate upon Domino and extend its functionality to test statistically differential cellular signaling. This new functionality includes compilation of active signals as linkages from multiple subjects in a single-cell data set and testing condition-dependent signaling linkage. The software is applicable for analysis of single-cell data sets with multiple subjects as biological replicates as well as with bootstrapped replicates from data sets with few or pooled subjects. We use simulation studies to benchmark the number of subjects in compared groups and cells within an annotated cell type sufficient to accurately identify differential linkages. We demonstrate the application of the Differential Cell Signaling Test (DCST) in the dominoSignal software to investigate consequences of cancer cell phenotypes and immunotherapy on cell-cell communication in tumor microenvironments. These applications in cancer studies demonstrate the ability of differential cell signaling analysis to infer changes to cell communication networks from therapeutic or experimental perturbations, which is broadly applicable across biological systems.
]]></description>
<dc:creator>Mitchell, J. T.</dc:creator>
<dc:creator>Stapleton, O.</dc:creator>
<dc:creator>Krishnan, K.</dc:creator>
<dc:creator>Nagaraj, S.</dc:creator>
<dc:creator>Lvovs, D.</dc:creator>
<dc:creator>Cherry, C.</dc:creator>
<dc:creator>Poissonnier, A.</dc:creator>
<dc:creator>Horton, W.</dc:creator>
<dc:creator>Adey, A.</dc:creator>
<dc:creator>Rao, V.</dc:creator>
<dc:creator>Huff, A.</dc:creator>
<dc:creator>Zimmerman, J. W.</dc:creator>
<dc:creator>Kagohara, L. T.</dc:creator>
<dc:creator>Zaidi, N.</dc:creator>
<dc:creator>Coussens, L. M.</dc:creator>
<dc:creator>Jaffee, E. M.</dc:creator>
<dc:creator>Elisseeff, J. H.</dc:creator>
<dc:creator>Fertig, E. J.</dc:creator>
<dc:date>2025-05-03</dc:date>
<dc:identifier>doi:10.1101/2025.05.02.651747</dc:identifier>
<dc:title><![CDATA[Differential cell signaling testing for cell-cell communication inference from single-cell data by dominoSignal]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-05-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.05.01.651668v1?rss=1">
<title>
<![CDATA[
Liver biopsies obtained throughout SIV infection reveal evolving interferon stimulated protein expression within distinct monocyte/macrophage subsets 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.05.01.651668v1?rss=1</link>
<description><![CDATA[
ABSTRACTLiver dysfunction is more common and more severe among people with HIV than in the general population. Our previous transcriptomic analysis identified an increased hepatic type-1 interferon (IFN-1) response in simian immunodeficiency virus (SIV) infected rhesus macaques, including pronounced upregulation of the IFN-1 stimulated gene product MX1. Here, we interrogated the role of different cell types in the IFN-1 response, focusing on macrophage subsets. During the acute phase of SIV infection, the expression of MX1 in liver biopsies was elevated in the resident macrophage Kupffer cells (KCs, CD163+ CD206+) as well as liver sinusoidal endothelial cells (LSECs, CD163- CD206+). Chronic infection was associated with increased MX1 expression both within KCs and the recruited monocyte-derived macrophages (MdMs, CD163+ CD206-) although the KCs were associated with the highest MX1 expression levels. We explored factors in the liver potentially associated with the macrophage IFN-1 response: 1) SIV DNA load, 2) bacterial DNA load (assessed via SIV DNA and 16S bacterial DNA qPCR, respectively), and 3) acute-phase hepatocyte lipid accumulation (steatosis). Both SIV and bacteria were elevated during chronic infection and directly correlated with MdM frequency. KC frequency did not correlate with SIV load but inversely correlated with bacterial load and also correlated directly with hepatocyte microvesicular steatosis in acute infection. Hepatocytes expressed variable levels of MX1 during acute and chronic infection. Overall, our data identify MdMs, LSECs and KCs as contributors to the heightened expression of MX1 in the liver during SIV infection. KCs express the highest levels of MX1 and therefore represent a potential target to reduce liver inflammation in people with HIV.

AUTHOR SUMMARYLiver disease disproportionately affects people with HIV (PWH). However, liver biopsies are generally not indicated in humans, making the immune mechanisms underlying HIV-driven liver dysfunction difficult to study. Using a model of HIV infection in which rhesus macaques are infected with SIV, we previously found a strong upregulation of type 1 interferon (IFN-1) signaling as well as an increased number of CD68+ monocyte/macrophage cells in the liver during chronic infection. Here we used longitudinally collected liver biopsies from SIV-infected macaques to describe the kinetics of the IFN-1 response, focusing on MX1, which was a top differentially expressed gene in our previous transcriptomic screen. We also defined two populations of liver macrophages - resident Kupffer cells and recruited monocyte-derived macrophages - to identify which cells produce MX1, what stimuli the cells respond to, and where the cells accumulate within the liver. Our results indicate that Kupffer cells are the major producers of MX1 during acute and chronic infection and that their frequency in the liver correlates with lipid accumulation (steatosis) during acute infection. These data provide insights into how Kupffer cells may promote inflammation and fibrosis and identify Kupffer cells as an important target to limit liver disease in PWH.
]]></description>
<dc:creator>Derby, N.</dc:creator>
<dc:creator>Johnson, B. I.</dc:creator>
<dc:creator>Biswas, S.</dc:creator>
<dc:creator>Fischer, M.</dc:creator>
<dc:creator>Fancher, K. A.</dc:creator>
<dc:creator>Luevano-Santos, C.</dc:creator>
<dc:creator>Yusova, S.</dc:creator>
<dc:creator>Meyer, K. A.</dc:creator>
<dc:creator>Abraham, Y. M.</dc:creator>
<dc:creator>Lutz, S. S.</dc:creator>
<dc:creator>Fisher, C.</dc:creator>
<dc:creator>Pacheco, M. C.</dc:creator>
<dc:creator>Smedley, J. V.</dc:creator>
<dc:creator>Burwitz, B. J.</dc:creator>
<dc:creator>Sodora, D. L.</dc:creator>
<dc:date>2025-05-07</dc:date>
<dc:identifier>doi:10.1101/2025.05.01.651668</dc:identifier>
<dc:title><![CDATA[Liver biopsies obtained throughout SIV infection reveal evolving interferon stimulated protein expression within distinct monocyte/macrophage subsets]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-05-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.05.02.651934v1?rss=1">
<title>
<![CDATA[
Modeling human retinal ganglion cell axonal outgrowth, development, and pathology using pluripotent stem cell-based microfluidic platforms 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.05.02.651934v1?rss=1</link>
<description><![CDATA[
Retinal ganglion cells (RGCs) are highly compartmentalized cells, with long axons serving as the sole connection between the eye and the brain. RGC degeneration in injury and/or disease also occurs in a compartmentalized manner, with distinct injury responses in axonal and somatodendritic compartments. Thus, the goal of this study was to establish a novel microfluidic-based platform for the analysis of RGC compartmentalization in health and disease states. Human pluripotent stem cell (hPSC)-derived RGCs were seeded into microfluidics, enabling the recruitment and isolation of axons apart from the somatodendritic compartment. Initial studies explored axonal outgrowth and compartmentalization of axons and dendrites. We then compared the differential response of RGCs differentiated from hPSCs carrying the OPTN(E50K) glaucoma mutation with isogenic control RGCs in their respective axonal and somatodendritic compartments, followed by analysis of axonal transport. Further, we explored the axonal transcriptome via RNA-seq, focusing on disease-related axonal differences. Finally, we established models to uniquely orient astrocytes along the axonal compartment combined with modulation of astrocyte reactivity as a pathological feature of neurodegeneration. Overall, RGC culture within microfluidic chips allowed enhanced cell growth and maturation, including long-distance axonal projections and proper compartmentalization, while patient-specific RGCs exhibited axonal outgrowth deficits as well as decreased rate of axonal transport. Finally, the induction of astrocyte reactivity uniquely along the proximal region of RGC axons led to the onset of neurodegenerative phenotypes in RGCs. These results represent the first study to effectively recapitulate the highly compartmentalized properties of hPSC-derived RGCs in healthy and disease states, providing a more physiologically relevant in vitro model for neuronal development and degeneration.
]]></description>
<dc:creator>Gomes, C.</dc:creator>
<dc:creator>Huang, K.-C.</dc:creator>
<dc:creator>Lavekar, S. S.</dc:creator>
<dc:creator>Harkin, J.</dc:creator>
<dc:creator>Prosser, C. G.</dc:creator>
<dc:creator>Fang, Y.</dc:creator>
<dc:creator>Kallem, C.</dc:creator>
<dc:creator>Oblak, A.</dc:creator>
<dc:creator>Zhang, C.</dc:creator>
<dc:creator>Meyer, J. S.</dc:creator>
<dc:date>2025-05-08</dc:date>
<dc:identifier>doi:10.1101/2025.05.02.651934</dc:identifier>
<dc:title><![CDATA[Modeling human retinal ganglion cell axonal outgrowth, development, and pathology using pluripotent stem cell-based microfluidic platforms]]></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/10.1101/2025.05.05.652238v1?rss=1">
<title>
<![CDATA[
Multiomic profiling of hypoxic glioblastoma stem cells reveals expansion of subpopulations with distinct epigenetic and CNV profiles 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.05.05.652238v1?rss=1</link>
<description><![CDATA[
Glioblastoma is characterized by extensive intratumoral heterogeneity driven by dynamic mi-croenvironmental cues such as hypoxia. While transcriptional and epigenetic variability have been separately linked to hypoxia responses, the integrated impact of hypoxia on gene regulation and clonal architecture in glioblastoma stem cells (GSCs) remains poorly defined. We applied singlenucleus multi-omics--integrating RNA-seq and ATAC-seq--to patient-derived GSCs cultured under normoxic or hypoxic conditions. This enabled simultaneous profiling of gene expression and chromatin accessibility within the same cells. Transcription factor (TF) regulatory networks were inferred using Dictys, while RNA-chromatin dynamics were modeled with MultiVelo. Clonal structure and copy number variations (CNVs) were resolved at single-cell resolution using RIDDLER on snATAC-seq data. Hypoxia induced the emergence of four distinct GSC subpopulations with unique transcriptomic and epigenetic profiles enriched for mesenchymal, angiogenic, and proliferative signatures. Regulatory network modeling revealed novel hypoxia-associated TFs--SP2, CREM, and ETV3--that modulate downstream oncogenic pathways. Trajectory analysis uncovered hypoxia-driven reversals in RNA-chromatin coupling, revealing dysregulated future transcriptional states of key genes such as MMP16 and SVIL. CNV profiling identified 13 clonal substructures, with specific clones (e.g., 5, 6, 9) selectively enriched under hypoxia and harboring distinct chromosomal alterations. These results demonstrate coordinated remodeling of GSC gene regulation and clonal fitness in response to hypoxic stress. Our findings reveal that hypoxia drives concurrent epigenetic, transcriptomic, and clonal selection in glioblastoma stem cells. This integrated model of hypoxia-induced plasticity provides mechanistic insights into tumor adaptation and identifies novel regulators that may serve as targets for therapeutic intervention in the hypoxic niche of glioblastoma.
]]></description>
<dc:creator>Corseri, A.</dc:creator>
<dc:creator>Moore, T. W.</dc:creator>
<dc:creator>Szczepanski, N.</dc:creator>
<dc:creator>Hwang, H.</dc:creator>
<dc:creator>Zdon, A.</dc:creator>
<dc:creator>Yardimci, G.</dc:creator>
<dc:creator>Tapinos, N.</dc:creator>
<dc:date>2025-05-09</dc:date>
<dc:identifier>doi:10.1101/2025.05.05.652238</dc:identifier>
<dc:title><![CDATA[Multiomic profiling of hypoxic glioblastoma stem cells reveals expansion of subpopulations with distinct epigenetic and CNV profiles]]></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/10.1101/2025.05.07.648613v1?rss=1">
<title>
<![CDATA[
Transfer learning framework via Bayesian group factor analysis incorporating feature-wise dependencies 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.05.07.648613v1?rss=1</link>
<description><![CDATA[
Transfer learning considers distinct but related tasks defined over heterogeneous domains and aims to improve generalization and performance through knowledge transfer between tasks. This approach can be especially advantageous in biomedical contexts with insufficient labeled training data, where joint learning across domains can enable inference in otherwise underpowered datasets. High-dimensional biomedical data is characterized with redundancy, rendering non-linear dependencies among features. Existing models often fail to leverage such feature dependencies during inference, limiting their ability to model complex biological systems. We present a Bayesian group factor analysis transfer learning framework that supports multitask, multi-modal learning. Our approach learns a shared latent space within each domain, simultaneously across multiple domains, and uses a feature-wise prior to model complex relationships. We evaluate our framework using controlled synthetic data experiments and four disjoint patient cancer datasets from acute myeloid leukemia and neuroblastoma. We show that our method improves drug response prediction and more readily recapitulates consensus biomarkers of drug response. Similarly, our approach improves tumor purity prediction and identifies a robust gene signature associated with it. Our framework is scalable, interpretable, and adaptable across target phenotypes, offering a robust solution for a wide range of heterogeneous multi-omics problems.
]]></description>
<dc:creator>Thirumalaisamy, D.</dc:creator>
<dc:creator>Black, N.</dc:creator>
<dc:creator>Gonen, M.</dc:creator>
<dc:creator>Nikolova, O.</dc:creator>
<dc:date>2025-05-10</dc:date>
<dc:identifier>doi:10.1101/2025.05.07.648613</dc:identifier>
<dc:title><![CDATA[Transfer learning framework via Bayesian group factor analysis incorporating feature-wise dependencies]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-05-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.05.07.652786v1?rss=1">
<title>
<![CDATA[
Mitochondrial Translation Inhibition Uncovers a Critical Metabolic-Epigenetic Interface in Renal Cell Carcinoma 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.05.07.652786v1?rss=1</link>
<description><![CDATA[
Background/ObjectivesRenal cell carcinoma (RCC) exhibits distinctive metabolic vulnerabilities that may be therapeutically targeted. This study investigates how tigecycline, an FDA-approved antibiotic that inhibits mitochondrial translation, affects RCC cells and explores potential combinatorial approaches to enhance its efficacy.

MethodsWe employed comprehensive metabolomic profiling, subcellular proteomics, and functional assays to characterize the effects of tigecycline on RCC cell lines, patient-derived organoids, and xenograft models. The synergistic potential of tigecycline with the histone deacetylase inhibitor entinostat was evaluated using combination index analysis.

ResultsTigecycline selectively inhibited mitochondrial translation in RCC cells, reducing mitochondrially-encoded proteins while sparing nuclear-encoded components, profoundly disrupting mitochondrial bioenergetics and reducing tumor growth in xenograft models. Subcellular proteomic analyses revealed that tigecycline treatment triggered significant accumulation of multiple histone variants concurrent with cell cycle arrest. Based on this discovery, combined treatment with tigecycline and entinostat demonstrated remarkable synergism across RCC cell lines and patient derived organoids.

ConclusionsOur findings identify a promising therapeutic opportunity by targeting the crosstalk between mitochondrial function and epigenetic homeostasis in RCC, with potential for rapid clinical translation given the established pharmacological profiles of both agents.
]]></description>
<dc:creator>Eckenstein, K.</dc:creator>
<dc:creator>Cengiz, B.</dc:creator>
<dc:creator>Chang, M.</dc:creator>
<dc:creator>Cartier, J. M.</dc:creator>
<dc:creator>Flory, M. R.</dc:creator>
<dc:creator>Thomas, G. V.</dc:creator>
<dc:date>2025-05-10</dc:date>
<dc:identifier>doi:10.1101/2025.05.07.652786</dc:identifier>
<dc:title><![CDATA[Mitochondrial Translation Inhibition Uncovers a Critical Metabolic-Epigenetic Interface in Renal Cell Carcinoma]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-05-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.05.06.652505v1?rss=1">
<title>
<![CDATA[
Exploring the Dynamics of Follicle Development and Hormone Synthesis: The Role of Oxygen Tension in Rhesus Macaque Follicle Culture 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.05.06.652505v1?rss=1</link>
<description><![CDATA[
In vitro culture of cryopreserved ovarian follicles has the potential to extend fertility options for young women seeking to preserve ovarian tissue prior to undergoing cancer treatments. Successful implementation of this strategy has been elusive and likely requires a more complete understanding of the microenvironment of the developing ovarian follicles, including ovarian oxygen concentrations. The oxygen tension within the reproductive tract plays a crucial role in follicular development and oocyte maturation. While in vitro culture systems often use atmospheric oxygen (21%), the native environment in vivo is significantly lower, ranging from 1.5% to 8.7%. This study aimed to investigate the effects of reduced oxygen tensions (3% and 5%) on follicle survival, growth, antrum formation, and hormone production in cultured secondary follicles from rhesus macaques (macaca mulata). A total of 300 follicles were isolated from 7 animals and cultured under three oxygen conditions: 3%, 5%, and 21% O2. Follicle survival and antrum formation were assessed weekly by microscopy and Kaplan-Meier survival analysis, while growth dynamics and hormone levels (estradiol, progesterone, AMH, and inhibin B) were monitored throughout the culture period.

Results demonstrated that follicles cultured at 3% and 5% oxygen exhibited significantly higher survival rates and antrum formation compared to those cultured at 21% O2. No significant differences in survival were observed between the 3% and 5% oxygen groups. Growth dynamics revealed distinct patterns, with both low oxygen groups promoting more robust and sustained follicle growth, while atmospheric oxygen led to rapid degeneration. Hormonal analysis showed that follicles in 21% O2 had elevated early hormone production but exhibited reduced long-term viability. In contrast, 3% and 5% oxygen delayed hormone production, reflecting a more stable and sustained follicular environment.

These findings underscore the importance of low oxygen tensions in mimicking the physiological conditions of the reproductive tract, improving follicular development, and supporting optimal hormonal function in vitro. This study suggests that further reducing oxygen levels to 3% may offer additional advantages for long-term follicle viability and function in reproductive technologies.

Graphical Abstract

O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=71 SRC="FIGDIR/small/652505v1_ufig1.gif" ALT="Figure 1">
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org.highwire.dtl.DTLVardef@11fcb62org.highwire.dtl.DTLVardef@61c0beorg.highwire.dtl.DTLVardef@1b5328forg.highwire.dtl.DTLVardef@10a7538_HPS_FORMAT_FIGEXP  M_FIG C_FIG
]]></description>
<dc:creator>Wang, K.</dc:creator>
<dc:creator>Wolf, S.</dc:creator>
<dc:creator>Zelinski, M.</dc:creator>
<dc:creator>Krieg, A.</dc:creator>
<dc:date>2025-05-11</dc:date>
<dc:identifier>doi:10.1101/2025.05.06.652505</dc:identifier>
<dc:title><![CDATA[Exploring the Dynamics of Follicle Development and Hormone Synthesis: The Role of Oxygen Tension in Rhesus Macaque Follicle Culture]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-05-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.05.09.653157v1?rss=1">
<title>
<![CDATA[
Predicting cochlear synaptopathy in mice with varying degrees of outer hair cell dysfunction using auditory evoked potentials 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.05.09.653157v1?rss=1</link>
<description><![CDATA[
Human temporal bones suggest a steady decline of cochlear synapses with age and greater synapse loss in adults with a history of military or occupational noise exposure. However, there is currently no validated method of diagnosing this type of cochlear deafferentation in living humans. Animal models indicate that cochlear synaptopathy is associated with reduced auditory brainstem response (ABR) wave 1 amplitude and envelope following response (EFR) magnitude for a sinusoidally amplitude modulated (SAM) tone. However, translating the SAM EFR to humans is complicated because it is difficult to obtain this measurement in humans using the same modulation frequency that showed the strongest relationship with synaptopathy in mice (1000 Hz). Computational modeling suggests that EFR magnitude measured with a rectangular amplitude modulated (RAM) tone may be a more sensitive measure of synaptopathy than the SAM EFR. In addition, because synaptopathy likely co-occurs with outer hair cell dysfunction, a diagnostic assay for synaptopathy needs to be robust even when auditory thresholds are abnormal. This study compared the relative ability of the ABR, SAM EFR, and RAM EFR to predict synapse numbers in mice with a large range of auditory thresholds and degrees of synaptopathy. The results indicate that the RAM EFR modulated at 1000 Hz is the single best predictor of synapse number when there is a broad loss of synapses across frequency, while combining RAM EFR and ABR further improves synapse prediction. In contrast, focal synaptopathy is best predicted by ABR wave 1 amplitude.

Significance StatementThis study assessed the relative ability of two auditory evoked potentials to identify cochlear synaptopathy, a type of cochlear deafferentation that occurs with age and noise exposure, in mice. Performance of these measures in the presence of outer hair cell (OHC) damage was also evaluated because synaptopathy is expected to often co-occur with OHC dysfunction. Concrete recommendations of measurements to use for non-invasive diagnosis of synaptopathy in humans are provided. This represents a significant advance toward diagnosis of a condition that is thought to have a high prevalence in humans. The ability to identify individuals with cochlear synaptopathy is vital for furthering our understanding of how this auditory deficit impairs auditory perception and the future development of treatment options.
]]></description>
<dc:creator>Buran, B. N.</dc:creator>
<dc:creator>Elkins, S.</dc:creator>
<dc:creator>He, W.</dc:creator>
<dc:creator>Bramhall, N. F.</dc:creator>
<dc:date>2025-05-12</dc:date>
<dc:identifier>doi:10.1101/2025.05.09.653157</dc:identifier>
<dc:title><![CDATA[Predicting cochlear synaptopathy in mice with varying degrees of outer hair cell dysfunction using auditory evoked potentials]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-05-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.05.13.651470v1?rss=1">
<title>
<![CDATA[
Serine auxotrophy is a targetable vulnerability driven by PSAT1 suppression in AML 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.05.13.651470v1?rss=1</link>
<description><![CDATA[
Serine metabolism is of growing biologic and therapeutic interest in cancer. Upregulation of the serine synthesis pathway (SSP) can fuel tumor growth, and cancers with this phenotype are often sensitive to SSP inhibitors. In parallel, dietary restriction of serine and glycine (SG) can suppress some cancers, but the determinants of sensitivity to this approach are poorly understood. This is especially true in acute myeloid leukemia (AML), where serine metabolism has been less explored. We report that a subset of human AML cell lines and primary samples are completely dependent on external serine, known as serine auxotrophy. These leukemias consistently suppressed the SSP enzyme PSAT1, failed to synthesize serine, responded to SG restriction in vivo, and were rescued by restoring PSAT1. We also found that AML with an SF3B1 K700E mutation showed additional dependence on the SSP enzyme PHGDH, that SG restriction synergized with venetoclax in serine auxotrophic AML, and that MECOM rearrangement was strongly associated with PSAT1 suppression and serine auxotrophy. These findings define a metabolically distinct AML subtype and nominate it for targeting by SG restriction.
]]></description>
<dc:creator>Sinanidis, I.</dc:creator>
<dc:creator>Tsakiroglou, P.</dc:creator>
<dc:creator>Dubner, B.</dc:creator>
<dc:creator>Foertsch, R.</dc:creator>
<dc:creator>Gondek, J.</dc:creator>
<dc:creator>Choi, I.</dc:creator>
<dc:creator>Paun, B.</dc:creator>
<dc:creator>Zhao, L.</dc:creator>
<dc:creator>Ghiaur, G.</dc:creator>
<dc:creator>Dalton, W. B.</dc:creator>
<dc:date>2025-05-14</dc:date>
<dc:identifier>doi:10.1101/2025.05.13.651470</dc:identifier>
<dc:title><![CDATA[Serine auxotrophy is a targetable vulnerability driven by PSAT1 suppression in AML]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-05-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.05.13.653850v1?rss=1">
<title>
<![CDATA[
PIN1 Drives Cellular Plasticity and Immune Modulation in Chronic Pancreatitis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.05.13.653850v1?rss=1</link>
<description><![CDATA[
Background and AimsChronic pancreatitis (CP) is characterized by inflammation, fibrosis, and acinar-to-ductal metaplasia (ADM). PIN1, known to drive oncogenic signaling and cellular plasticity in cancer, has an unexplored role in CP. This study investigates PIN1s expression and function in CP pathogenesis using human tissues and mouse models.

MethodsPIN1 expression was assessed in human CP tissue microarrays (TMAs) via immunohistochemistry (IHC) and cyclic immunofluorescence (CyCIF). Acute and chronic pancreatitis were induced in wild-type (WT) and PIN1 knockout (PIN1KO) mice using caerulein. Disease progression was monitored histologically, and immune profiling was conducted using flow cytometry. Pharmacological inhibition was performed using a small molecule PIN1 inhibitor-Sulfopin, and effects were evaluated by histology, qPCR, and cytokine analysis. Single-cell RNA sequencing (scRNA-seq) was performed on pancreatic tissues to perform pathway analysis and intercellular communication.

ResultsPIN1 expression was elevated in human CP tissues, correlating with disease severity and ADM. In mice, both acute and chronic pancreatitis increased PIN1 expression, but only in our chronic PIN1KO mice displayed reduced pancreatic injury, fibrosis, ADM, and modulated immune infiltration. Pharmacological PIN1 inhibition mimicked the protective effects of genetic knockout, dampening inflammatory pathways. scRNA-seq revealed that PIN1 inhibition altered the intercellular communication networks between epithelial, immune, and stromal cells.

ConclusionPIN1 drives cellular plasticity, immune modulation, and disease progression in CP. Targeting PIN1 may offer a therapeutic strategy to mitigate CP.

Graphical Abstract

O_FIG O_LINKSMALLFIG WIDTH=184 HEIGHT=200 SRC="FIGDIR/small/653850v1_ufig1.gif" ALT="Figure 1">
View larger version (36K):
org.highwire.dtl.DTLVardef@67f9fcorg.highwire.dtl.DTLVardef@4d4c1forg.highwire.dtl.DTLVardef@c099d7org.highwire.dtl.DTLVardef@b41aa5_HPS_FORMAT_FIGEXP  M_FIG C_FIG Created in BioRender. Shah, V. (2025) https://BioRender.com/undefined
]]></description>
<dc:creator>Shah, V. M.</dc:creator>
<dc:creator>Bartlett, A.</dc:creator>
<dc:creator>Chen, C.</dc:creator>
<dc:creator>Matter, A.</dc:creator>
<dc:creator>Smith, A.</dc:creator>
<dc:creator>Zimny, H.</dc:creator>
<dc:creator>Wang, X.</dc:creator>
<dc:creator>Daniel, C.</dc:creator>
<dc:creator>Eng, J.</dc:creator>
<dc:creator>Youm, I.</dc:creator>
<dc:creator>Tsuda, M.</dc:creator>
<dc:creator>Calistri, N.</dc:creator>
<dc:creator>Reyer, M.</dc:creator>
<dc:creator>Morgan, T. K.</dc:creator>
<dc:creator>Dewson, G.</dc:creator>
<dc:creator>Waugh, T.</dc:creator>
<dc:creator>Keith, D.</dc:creator>
<dc:creator>Haverlack, S.</dc:creator>
<dc:creator>Xia, Z.</dc:creator>
<dc:creator>Chin, K.</dc:creator>
<dc:creator>Sheppard, B.</dc:creator>
<dc:creator>Brody, J.</dc:creator>
<dc:creator>Sears, R. C.</dc:creator>
<dc:date>2025-05-14</dc:date>
<dc:identifier>doi:10.1101/2025.05.13.653850</dc:identifier>
<dc:title><![CDATA[PIN1 Drives Cellular Plasticity and Immune Modulation in Chronic Pancreatitis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-05-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.05.09.653135v1?rss=1">
<title>
<![CDATA[
Unreliable homeostatic action potential broadening in cultured dissociated neurons 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.05.09.653135v1?rss=1</link>
<description><![CDATA[
Homeostatic plasticity preserves neuronal activity against perturbations. Recently, somatic action potential broadening was proposed as a key homeostatic adaptation to chronic inactivity in neocortical neurons. Since action potential shape critically controls calcium entry and neuronal function, broadening provides an attractive homeostatic feedback mechanism to regulate activity. Here, we report that chronic inactivity induced by sodium channel block does not broaden action potentials in neocortical neurons under a wide range of conditions. In contrast, action potentials were broadened in CA3 neurons of organotypic hippocampal cultures by chronic sodium channel block and in hippocampal dissociated cultures by chronic synaptic block. Mechanistically, BK-type potassium channels were proposed to underly inactivity-induced action potential broadening. However, BK channels did not affect action potential duration in our recordings. Our results indicate that action potential broadening can occur in specific neurons and conditions but is not a general mechanism of homeostatic plasticity in cultured neurons.
]]></description>
<dc:creator>Ritzau-Jost, A.</dc:creator>
<dc:creator>Rajayer, S.</dc:creator>
<dc:creator>Nerlich, J.</dc:creator>
<dc:creator>Maciag, F.</dc:creator>
<dc:creator>John, A.</dc:creator>
<dc:creator>Russier, M.</dc:creator>
<dc:creator>Gonzalez Sabater, V.</dc:creator>
<dc:creator>Steiger, L.</dc:creator>
<dc:creator>Coq, J.-O.</dc:creator>
<dc:creator>Eilers, J.</dc:creator>
<dc:creator>Engelhardt, M.</dc:creator>
<dc:creator>Burrone, J.</dc:creator>
<dc:creator>Debanne, D.</dc:creator>
<dc:creator>Heine, M.</dc:creator>
<dc:creator>Smith, S. M.</dc:creator>
<dc:creator>Hallermann, S.</dc:creator>
<dc:date>2025-05-15</dc:date>
<dc:identifier>doi:10.1101/2025.05.09.653135</dc:identifier>
<dc:title><![CDATA[Unreliable homeostatic action potential broadening in cultured dissociated neurons]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-05-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.05.11.653351v1?rss=1">
<title>
<![CDATA[
Isoform-Specific Roles of NTRK2 in Pulmonary Vascular Regeneration 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.05.11.653351v1?rss=1</link>
<description><![CDATA[
Bronchopulmonary dysplasia (BPD) is a chronic lung disease in premature infants with no curative therapy, characterized by impaired alveologenesis and capillary formation. However, the molecular mechanisms underlying endothelial dysfunction, a key driver of BPD pathogenesis in human, remain poorly understood. Here, through multiomic profiling of vascular endothelial cells isolated from control and BPD patient lungs, we uncovered an expansion of general capillary endothelial cells (gCap) with aberrant expression of the neurotrophic receptor tyrosine kinase 2 (NTRK2) in BPD. Importantly, we identified a pathological NTRK2 isoform switch that dictates the regenerative capacity of gCap cells. Full-length NTRK2 (NTRK2-FL) promoted gCap regeneration in response to hyperoxic injury, whereas RBFOX2-mediated splicing of NTRK2-FL into a truncated isoform (NTRK2-T1) contributed to maladaptive responses and irreversible alveolar simplification in severe BPD cases. Restoring NTRK2-FL using lipid nanoparticle-delivered mRNA promoted angiogenesis and reversed alveolar simplification in vessel organoids and BPD-like mice. These findings identified NTRK2 isoform imbalance as a key driver of endothelial dysfunction and support isoform-specific RNA therapy as a promising strategy for vascular regeneration and repair.

HIGHLIGHTS* Multiomic and spatial profiling reveal abnormal gCap subtype in human BPD lungs

* NTRK2 isoform switch dictates endothelial regeneration in response to hyperoxic injury

* RBFOX2 mediates splicing from NTRK2-FL to NTRK2-T1, driving irreversible alveolar damage

* NTRK2-FL mRNA therapy restores vascular regeneration after injury
]]></description>
<dc:creator>Tan, C.</dc:creator>
<dc:creator>Liu, Z.</dc:creator>
<dc:creator>Mao, X.</dc:creator>
<dc:creator>Zhang, Y.</dc:creator>
<dc:creator>Li, X.</dc:creator>
<dc:creator>Pek, N.</dc:creator>
<dc:creator>Fu, H.</dc:creator>
<dc:creator>Liu, Y.</dc:creator>
<dc:creator>Kalinichenko, V. V.</dc:creator>
<dc:creator>Pryhuber, G. S.</dc:creator>
<dc:creator>Lu, R.</dc:creator>
<dc:creator>Lai, L.</dc:creator>
<dc:creator>Miao, Y.</dc:creator>
<dc:creator>Guo, M.</dc:creator>
<dc:creator>Gu, M.</dc:creator>
<dc:date>2025-05-15</dc:date>
<dc:identifier>doi:10.1101/2025.05.11.653351</dc:identifier>
<dc:title><![CDATA[Isoform-Specific Roles of NTRK2 in Pulmonary Vascular Regeneration]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-05-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.05.11.653360v1?rss=1">
<title>
<![CDATA[
A family of E3 ligases extend K11 polyubiquitin on sites of MARUbylation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.05.11.653360v1?rss=1</link>
<description><![CDATA[
Ubiquitin (Ub) cooperation with other post-translational modifications provides a tiered opportunity for protein regulation. Small modifications to Ub such as phosphorylation, acetylation, or ADP-ribosylation have varying impacts on signaling. The Deltex family of E3 ligases was previously implicated in the ubiquitylation of ADP-ribose (ADPr) and ADPr-containing macromolecules. Our previous work found ester-linked mono-ADPr ubiquitylation (MARUbylation) on PARP7 and PARP10 in cells and that this mark is extended with K11 polyUb. We previously screened for E3 ligases that interact with PARP7 through three different approaches and identified six candidates, including the Deltex family member DTX2. One of these hits, RNF114, interacts with various other PARPs, leading us to hypothesize that RNF114 binds to sites of MARUbylation and extends K11 polyUb. Here, we show that DTX2 generates the initial MARUbe on PARP7 in cells, which depends on PARP7 catalytic activity. The MARUbe on PARP7 is extended with K11 polyUb by RNF114. To investigate the mechanism of RNF114 reader/writer function, we developed a click chemistry-inspired chemoenzymatic approach to create a novel fluorescent Ub-ADPr probe for studying its interaction with RNF114. Strikingly, we found that RNF114 has a weak affinity for ADPr and Ub separately but explicitly recognizes the linkage between Ub and ADPr present in MARUbylated species. We used AlphaFold3 modeling to examine the mechanisms of Ub-ADPr recognition and K11-linked polyUb extension by RNF114. We identified a tandem Di19-UIM module in RNF114 as a MARUbe-binding domain (MUBD), thus providing a reader function that interfaces with K11-specific writer activity. Finally, we described a small family of MUBD-containing E3 ligases that demonstrate preference for Ub-ADPr, which we call MARUbe-Targeted Ligases (MUTLs).
]]></description>
<dc:creator>Lacoursiere, R. E.</dc:creator>
<dc:creator>Upadhyaya, K.</dc:creator>
<dc:creator>Kaur Sidhu, J.</dc:creator>
<dc:creator>Bejan, D. S.</dc:creator>
<dc:creator>Rodriguez Siordia, I.</dc:creator>
<dc:creator>Cohen, M. S.</dc:creator>
<dc:creator>Pruneda, J. N.</dc:creator>
<dc:date>2025-05-15</dc:date>
<dc:identifier>doi:10.1101/2025.05.11.653360</dc:identifier>
<dc:title><![CDATA[A family of E3 ligases extend K11 polyubiquitin on sites of MARUbylation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-05-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.05.14.653900v1?rss=1">
<title>
<![CDATA[
Aggregating multimodal cancer data across unaligned embedding spaces maintains tumor of origin signal 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.05.14.653900v1?rss=1</link>
<description><![CDATA[
AI based embeddings offer the possibilities of encoding complex biological data into low dimensional spaces, called embedding spaces, that maintain the relationships between entities. There is an open question about the compatibility of embedding spaces that are created without any coordination. It has been assumed that signals in these unaligned embedding spaces would be destroyed if vectors were aggregated into summed values. We trained embedding models across different data modalities and tested aggregating the values together to test this assumption. Our research shows that signal from unaligned embedded values is conserved and able to still be used for learning tasks, such as data modality and tumor of origin recognition.
]]></description>
<dc:creator>Kirchgaessner, R.</dc:creator>
<dc:creator>Keutler, K.</dc:creator>
<dc:creator>Sivakumar, L.</dc:creator>
<dc:creator>Song, X.</dc:creator>
<dc:creator>Ellrott, K.</dc:creator>
<dc:date>2025-05-18</dc:date>
<dc:identifier>doi:10.1101/2025.05.14.653900</dc:identifier>
<dc:title><![CDATA[Aggregating multimodal cancer data across unaligned embedding spaces maintains tumor of origin signal]]></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/10.1101/2025.05.14.654109v1?rss=1">
<title>
<![CDATA[
Mutations outside the MR1 antigen binding groove differentially inhibit presentation of exogenous antigens 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.05.14.654109v1?rss=1</link>
<description><![CDATA[
The antigen presenting molecule MHC class I-related protein 1 (MR1) binds small molecule metabolites derived from microbial riboflavin biosynthetic pathways and presents them at the cell surface for surveillance by MR1-restricted mucosal-associated invariant T cells (MAIT cells). MR1 ligands can originate in the extracellular space or in endosomal compartments that contain microbial pathogens. Distinct, complementary antigen processing and presentation pathways enable MR1 to survey diverse intracellular locations and present both exogenous and intracellular antigens. Here, we generated a panel of BEAS-2B MR1 KO cells reconstituted with MR1 proteins mutated at amino acids 9 - 16. The mutated MR1 molecules differentially translocated to the cell surface in response to 6-formylpterin and differed in their ability to present mycobacterial antigens to MAIT cell clones. While they barely presented Mycobacterium smegmatis supernatant and other exogenous MAIT cell antigens, their ability to present antigens derived from mycobacterial infection and a 5-A-RU prodrug requiring endosomal processing remained largely intact. Protein co-immunoprecipitation and mass spectrometry-based proteomic analysis showed that mutated MR1 differentially associated with calnexin and {beta}2-microglobulin (B2M). Knock-down of B2M in cells over-expressing MR1 phenocopied the loss of exogenous antigen presentation but did not impact presentation of intracellular antigens. Thus, the MR1-mediated presentation of exogenous antigen appears to be limited by binding to B2M whereas the lower sensitivity to B2M deficiency implies that MAIT cell activation via the endosomal antigen presentation pathway may be limited by the availability of MR1 itself.
]]></description>
<dc:creator>Kulicke, C. A.</dc:creator>
<dc:creator>Lemon, C.</dc:creator>
<dc:creator>Krawic, J. R.</dc:creator>
<dc:creator>Ramirez, L. M. N.</dc:creator>
<dc:creator>Kim, S.-J.</dc:creator>
<dc:creator>Narayanan, G. A.</dc:creator>
<dc:creator>Tafesse, F. G.</dc:creator>
<dc:creator>Hildebrand, W. H.</dc:creator>
<dc:creator>Dobos, K. M.</dc:creator>
<dc:creator>Lewinsohn, D. M.</dc:creator>
<dc:date>2025-05-19</dc:date>
<dc:identifier>doi:10.1101/2025.05.14.654109</dc:identifier>
<dc:title><![CDATA[Mutations outside the MR1 antigen binding groove differentially inhibit presentation of exogenous antigens]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-05-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.05.18.654760v1?rss=1">
<title>
<![CDATA[
Identification and Rescue of Congenital Hyperinsulinism-Associated ABCC8 Mutations that Impair KATP Channel Trafficking 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.05.18.654760v1?rss=1</link>
<description><![CDATA[
ATP-sensitive potassium (KATP) channels composed of Kir6.2 and sulfonylurea receptor 1 (SUR1) couple glucose metabolism with insulin secretion in pancreatic {beta}-cells and are vital to glucose homeostasis. Loss-of-function mutations in SUR1 and Kir6.2, encoded by ABCC8 and KCNJ11, respectively are the commonest causes of severe persistent hypoglycemia in infants and children seen in the rare disease congenital hyperinsulinism (HI). The N-terminal transmembrane domain, TMD0, and the linker immediately C-terminal to TMD0, L0, of SUR1 (TMD0/L0) forms direct contact with Kir6.2 in KATP channels. Mutations in SUR1-TMD0 often impair KATP channel trafficking to the plasma membrane, causing severe disease unresponsive to treatment by the KATP activator diazoxide; however, surface expression and function of many such mutant channels can be rescued by reversible KATP inhibitor pharmacochaperones. Here, we identified seven new SUR1 missense mutations in TMD0/L0 from HI patients unresponsive to diazoxide and investigated their effects on KATP channel expression, function, and response to pharmacochaperones. All seven mutations, N32K, Y124F, P133R, W143R, L171P, G228D, and Y230C, reduced channel function in Rb+ efflux assays. Further characterization by immunoblotting, immunostaining and electrophysiology revealed that Y124F primarily causes defective channel gating, while the others impair channel trafficking to different extents. The trafficking mutations showed varied response to surface expression and function rescue by the reversible KATP inhibitor pharmacochaperones, tolbutamide and Aekatperone. The study underscores the critical role of SUR1-TMD0/L0 in KATP expression and gating. It further highlights the importance of detailed biochemical and functional studies of mutant channels in understanding their pathogenic roles and response to potential pharmacological therapies.
]]></description>
<dc:creator>ElSheikh, A.</dc:creator>
<dc:creator>Kuo, Y.-Y.</dc:creator>
<dc:creator>E. Boodhansingh, K.</dc:creator>
<dc:creator>Yang, Z.</dc:creator>
<dc:creator>A. Stanley, C.</dc:creator>
<dc:creator>De Leon, D. D.</dc:creator>
<dc:creator>Shyng, S.-L.</dc:creator>
<dc:date>2025-05-19</dc:date>
<dc:identifier>doi:10.1101/2025.05.18.654760</dc:identifier>
<dc:title><![CDATA[Identification and Rescue of Congenital Hyperinsulinism-Associated ABCC8 Mutations that Impair KATP Channel Trafficking]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-05-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.05.15.654371v1?rss=1">
<title>
<![CDATA[
Distinct age- and pathology-dependent epitranscriptome and translational dysfunction in a tauopathy mouse model 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.05.15.654371v1?rss=1</link>
<description><![CDATA[
Chronic neurodegenerative diseases, such as tauopathies, cause major metabolic changes in the brain affecting gene expression at both the transcriptional and translational levels. Our understanding of how regulation of translation changes with disease has focused on mRNA and its translational regulatory factors, RNA binding proteins, and microRNAs, despite clear evidence for translational and post-translational dysfunction in ADRD and tauopathies. The neurobiology of tRNA has only recently begun to be studied, but the impact of chronic neurodegenerative diseases on tRNA biology and translational dysfunction is largely unknown. We have previously shown that the tRNA pool and tRNA modifications behave as a system to regulate the cellular stress response by undergoing stress-specific reprogramming and causing selective translation of mRNAs from codon-biased stress response genes. Here we tested this stress-induced tRNA reprogramming and codon-biased translation system in the response to mutant tau expression by performing mass spectrometric quantification of [~]8500 proteins and 49 tRNA modifications, AQRNA-seq analysis of 222 cytosolic and mitochondrial tRNAs and other small RNAs, and informatic analysis of codon usage patterns in >23,000 protein-coding genes. Analysis of these datasets revealed that aging and tauopathy elicit major adaptation of the tRNA transcriptome and epitranscriptome as well as corresponding evidence of a program of translation of families of codon-biased genes for aging and disease responsive proteins. The mitochondrial tRNA transcriptome showed a strong response to aging and disease with 21 of the 22 mt-tRNAs showing age and disease-linked increases in expression, accompanied by mitochondria-specific modifications such as ms2i6A and f5C. Surprisingly, there were few significant changes in the 203 cytosolic tRNA isodecoders. However, a 10-fold increase tRNA isodecoder tRNA-Arg-TCT-5-1 was accompanied by increased translation of proteins encoded by genes highly enriched in its AGA cognate codon. These changes in tRNA biology are mirrored by strongly biased use of synonymous codons among the most highly upregulated and downregulated proteins in the P301S MAPT mice. Taken together these findings suggest the aging and disease brain produces an integrated response for translational control that is highly integrated with changes in tRNA biology.
]]></description>
<dc:creator>Sun, G.</dc:creator>
<dc:creator>Lin, W.</dc:creator>
<dc:creator>Chen, R.</dc:creator>
<dc:creator>Jiang, L.</dc:creator>
<dc:creator>DeMott, M.</dc:creator>
<dc:creator>McShane, A.</dc:creator>
<dc:creator>Chan, M. C. K.</dc:creator>
<dc:creator>Ehrbar, D.</dc:creator>
<dc:creator>Zhang, X.</dc:creator>
<dc:creator>Begley, T.</dc:creator>
<dc:creator>Emili, A.</dc:creator>
<dc:creator>Wolozin, B.</dc:creator>
<dc:creator>Dedon, P.</dc:creator>
<dc:date>2025-05-20</dc:date>
<dc:identifier>doi:10.1101/2025.05.15.654371</dc:identifier>
<dc:title><![CDATA[Distinct age- and pathology-dependent epitranscriptome and translational dysfunction in a tauopathy mouse model]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-05-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.05.16.654579v1?rss=1">
<title>
<![CDATA[
Deep-learning-enabled multi-omics analyses for prediction of future metastasis in cancer 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.05.16.654579v1?rss=1</link>
<description><![CDATA[
Metastasis remains the leading cause of cancer-related mortality, yet predicting future metastasis is a major clinical challenge due to the lack of validated biomarkers and effective assessment methods. Here, we present EmitGCL, a deep-learning framework that accurately predicts future metastasis and its corresponding biomarkers. Based on a comprehensive benchmarking comparison, EmitGCL outperformed other computational tools across six cancer types from seven cohorts of patients with superior sensitivity and specificity. It captured occult metastatic cells in a patient with a lymph node-negative breast cancer, who was declared to have no evidence of disease by conventional imaging methods but was later confirmed to have a metastatic disease. Notably, EmitGCL identified HSP90AA1 and HSP90AB1 as predictable biomarkers for future breast cancer metastasis, which was validated across five independent cohorts of patients (n=420). Furthermore, we demonstrated YY1 transcription factor as a key driver of breast cancer metastasis which was validated through in-silico and CRISPR-based migration assays, suggesting that YY1 is a potential therapeutic target for deterring metastasis.
]]></description>
<dc:creator>Ma, Q.</dc:creator>
<dc:creator>Wang, X.</dc:creator>
<dc:creator>Duan, M.</dc:creator>
<dc:creator>Su, P.-L.</dc:creator>
<dc:creator>Li, J.</dc:creator>
<dc:creator>Krull, J.</dc:creator>
<dc:creator>Jin, J.</dc:creator>
<dc:creator>Chen, H.</dc:creator>
<dc:creator>Sun, Y.</dc:creator>
<dc:creator>Wu, W.</dc:creator>
<dc:creator>He, K.</dc:creator>
<dc:creator>Carpenter, R.</dc:creator>
<dc:creator>Zhang, C.</dc:creator>
<dc:creator>Cao, S.</dc:creator>
<dc:creator>Xu, D.</dc:creator>
<dc:creator>Wang, G.</dc:creator>
<dc:creator>Li, L.</dc:creator>
<dc:creator>Xin, G.</dc:creator>
<dc:creator>Carbone, D. P.</dc:creator>
<dc:creator>Li, Z.</dc:creator>
<dc:date>2025-05-22</dc:date>
<dc:identifier>doi:10.1101/2025.05.16.654579</dc:identifier>
<dc:title><![CDATA[Deep-learning-enabled multi-omics analyses for prediction of future metastasis in cancer]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-05-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.05.18.654742v1?rss=1">
<title>
<![CDATA[
Cervical Mucus Viscoelasticity and Sperm Velocity are Correlated and Concentration-dependent In Vitro 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.05.18.654742v1?rss=1</link>
<description><![CDATA[
BackgroundMucus in the endocervix serves as fertility gatekeeper in the reproductive tract through hormonally regulated changes in biophysical properties. Cervical mucus can thicken to prevent ascension of sperm into the upper reproductive tract or thin to permit fertilization. Current reproductive studies of mucus viscoelastic properties rely on subjective visual appraisal of cervical mucus changes. Our goal was to use particle tracking microrheology (PTMR) to objectively assess cervical mucus viscoelastic properties and associate these measurements with in vitro measures of sperm velocity.

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

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

ConclusionsPTMR and sperm tracking in mucus provide quantitative measure of viscoelastic mucus changes. PTMR is potentially a method for quantitatively assessing fertility potential in the cervix that could be applied to both infertility and contraceptives studies.
]]></description>
<dc:creator>Markovetz, M. R.</dc:creator>
<dc:creator>Wei, S.</dc:creator>
<dc:creator>Celluci, C.</dc:creator>
<dc:creator>Roberts, M.</dc:creator>
<dc:creator>Han, L.</dc:creator>
<dc:date>2025-05-23</dc:date>
<dc:identifier>doi:10.1101/2025.05.18.654742</dc:identifier>
<dc:title><![CDATA[Cervical Mucus Viscoelasticity and Sperm Velocity are Correlated and Concentration-dependent In Vitro]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-05-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.05.20.654990v1?rss=1">
<title>
<![CDATA[
A New Class of Precision Therapeutics that Inhibit Prostate Cancer Mediated Bone Destruction 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.05.20.654990v1?rss=1</link>
<description><![CDATA[
Cancer mediated bone destruction remains a common cause of morbidity and mortality. Here we couple an inhibitor of cell movement to bone trophic bisphosphonates, forming heterobifunctional compounds termed Dual Acting Bone Defender agents (DABDs). DABDs inhibit prostate cancer cell motility and osteoclast mediated bone destruction in vitro. Our lead compound, BTE-EN1, inhibits circulating human prostate cancer cells from forming bone destructive lesions in mice and prolongs life in a dose-dependent fashion. In mice with established metastatic lesions, BTE-EN1 significantly decreases bone destruction by over 60% compared to the control. Mechanistically, BTE-EN1 binds hydroxyapatite, inhibits prenylation of Rap1A, induces apoptosis in osteoclasts, and inhibits Raf1 activation and formation of bone resorptive cavities in osteoclasts, thus retaining the function of its constituent chemicals. BTE-EN1 is not toxic to mice at doses 4,000-fold over those required for efficacy, exhibits anti-cell motility activity across other bone-destructive cancer cell types, and does not interfere with standard-of-care hormone therapy or chemotherapy. BTE-EN1 is a new class of highly active therapeutics that inhibit cancer mediated bone destruction and holds high potential for translation into humans.
]]></description>
<dc:creator>Qiao, F.</dc:creator>
<dc:creator>Gordon, R.</dc:creator>
<dc:creator>Pattanayak, A.</dc:creator>
<dc:creator>Li, W.</dc:creator>
<dc:creator>Knibbe-Hollinger, J.</dc:creator>
<dc:creator>O'Neill, K.</dc:creator>
<dc:creator>Chen, W.</dc:creator>
<dc:creator>Mukthapuram, P. R.</dc:creator>
<dc:creator>Natarajan, A.</dc:creator>
<dc:creator>Bergan, R.</dc:creator>
<dc:date>2025-05-24</dc:date>
<dc:identifier>doi:10.1101/2025.05.20.654990</dc:identifier>
<dc:title><![CDATA[A New Class of Precision Therapeutics that Inhibit Prostate Cancer Mediated Bone Destruction]]></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/10.1101/2025.05.21.655323v1?rss=1">
<title>
<![CDATA[
Molecular mechanism of substrate transport by human peroxisomal ABCD3 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.05.21.655323v1?rss=1</link>
<description><![CDATA[
Peroxisomes are eukaryotic oxidative organelles involved in numerous metabolic functions that include fatty acid oxidation, bile acid synthesis, and detoxification of reactive oxygen species. ATP-binding cassette transporters of the D subfamily (ABCD1-3) mediate the import of CoA thioesters of fatty acids into the peroxisome. ABCD3, the most abundant of these transporters in the peroxisomal membrane, facilitates the transport of a broad spectrum of substrates including branched-chain fatty acids, very long-chain fatty acids, bile salt intermediates, and dicarboxylic acids. Mutations in ABCD3 are associated with defects in congenital bile acid synthesis and variants of Zellweger syndrome. The structural and functional details of the human ABCD3 transporter remain unclear, despite its significance. In this study, we report the cryogenic sample electron microscopy (cryo-EM) structures of full-length human ABCD3 in its apo state and bound to one of the physiological substrates (phytanoyl-CoA) at resolutions of 3.33 [A] and 3.13 [A], respectively. Our biochemical assays reveal that substrate binding induces ATPase activity in ABCD3, suggesting a substrate-dependent conformational change. Structural comparison of the apo and substrate bound states demonstrate that the substrate interaction brings nucleotide-binding domains closer, providing a mechanistic basis of substrate induced ATPase activity. These findings offer critical insights into the transport mechanism of ABCD3 and lay a structural foundation for understanding its role in peroxisomal metabolite import and related diseases.

Significance StatementPeroxisomes are involved in essential cellular metabolic processes that include fatty acid oxidation, bile acid synthesis, and detoxification. The ABCD subfamily of membrane transporter proteins transport fatty acyl-CoA molecules from the cytosol into peroxisomes. Among the ABCD subfamily members, ABCD3 transports branched chain fatty acids and a range of other important metabolites. Dysfunction in ABCD3 has been associated with several inherited and acquired diseases. Using cryo-EM, we determined high-resolution structures of full-length human ABCD3 in both unbound and phytanoyl-CoA-bound forms. We show that substrate binding stimulates ATP hydrolysis activity by apposition of the two nucleotide binding domains of ABCD3, adding structural insight into the mechanism of action of ABCD3 and its role in peroxisomal function and disease.
]]></description>
<dc:creator>Gupta, M.</dc:creator>
<dc:creator>Khandelwal, N. K.</dc:creator>
<dc:creator>Seka, D. J.</dc:creator>
<dc:creator>Balasubramani, S. G.</dc:creator>
<dc:creator>Dickinson, M. S.</dc:creator>
<dc:creator>Myasnikov, A.</dc:creator>
<dc:creator>Echeverria, I.</dc:creator>
<dc:creator>Stroud, R. M.</dc:creator>
<dc:date>2025-05-27</dc:date>
<dc:identifier>doi:10.1101/2025.05.21.655323</dc:identifier>
<dc:title><![CDATA[Molecular mechanism of substrate transport by human peroxisomal ABCD3]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-05-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.05.22.655365v1?rss=1">
<title>
<![CDATA[
Disabling PSGL-1 abrogates immune suppression and resistance to PD-1 blockade in pancreatic cancer 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.05.22.655365v1?rss=1</link>
<description><![CDATA[
Pancreatic ductal adenocarcinoma (PDAC) is a lethal cancer for which there is a critical need to identify novel therapeutic targets. Herein we define PSGL-1 as a checkpoint inhibitor using a syngeneic orthotopic model of PDAC. As with PDAC patients, CD8+ T cells within murine PDAC tumors expressed high levels of PSGL-1. PSGL-1-/- mice displayed striking T cell-dependent control of primary tumors and lung metastases. Extensive spatial remodeling within PDAC tumors occurred in PSGL-1-/- mice with a dramatic loss of proliferating tumor cells and an increase in CD8+ T cell engagement of antigen-presenting cells. The prominent CD8+ T cell infiltrates included subsets of pre-exhausted T cells retaining hallmarks of stemness and multifunctional effector capacity. These changes enabled a near complete response of PDAC to therapeutic PD-1 blockade. Our findings identify PSGL-1 as a key regulator of anti-tumor immunity in PDAC, highlighting its potential as a therapeutic target to limit CD8+ T cell exhaustion and enhance immunotherapy response.

SummaryHope et al describe a pivotal function of PSGL-1 in CD8+ T cell responses to pancreatic ductal adenocarcinoma. Genetic deletion of PSGL-1 elicits tumor control by increasing T cell infiltration and maintaining functional subsets, thereby promoting sensitivity to PD-1 blockade.
]]></description>
<dc:creator>Hope, J. L.</dc:creator>
<dc:creator>Zhang, Y.</dc:creator>
<dc:creator>Hetrick, H. A. F.</dc:creator>
<dc:creator>Sanchez-Hernandez, E. S.</dc:creator>
<dc:creator>Silvestri, B.</dc:creator>
<dc:creator>Smith, B. J.</dc:creator>
<dc:creator>Nakil, S. H.</dc:creator>
<dc:creator>Roy, S.</dc:creator>
<dc:creator>Lin, M.</dc:creator>
<dc:creator>Palete, A. B.</dc:creator>
<dc:creator>Maganti, S.</dc:creator>
<dc:creator>Ling, L.</dc:creator>
<dc:creator>Otero, D. C.</dc:creator>
<dc:creator>Byrne, K.</dc:creator>
<dc:creator>Romano, G.</dc:creator>
<dc:creator>Wang, Y. X.</dc:creator>
<dc:creator>Commisso, C.</dc:creator>
<dc:creator>Bradley, L. M.</dc:creator>
<dc:date>2025-05-27</dc:date>
<dc:identifier>doi:10.1101/2025.05.22.655365</dc:identifier>
<dc:title><![CDATA[Disabling PSGL-1 abrogates immune suppression and resistance to PD-1 blockade in pancreatic cancer]]></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/10.1101/2025.05.29.656943v1?rss=1">
<title>
<![CDATA[
MicroCT enables simultaneous longitudinal tracking of murine pancreatic cancer progression and cachexia 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.05.29.656943v1?rss=1</link>
<description><![CDATA[
Preclinical models of pancreatic ductal adenocarcinoma (PDAC) can greatly benefit from non-invasive imaging for evaluating disease progression and therapeutic response. Imaging approaches that can accurately and simultaneously track primary tumor growth, metastatic dissemination, and host cachexia over time are lacking. Here, we report an optimized dual-contrast micro-computed tomography (microCT) protocol for longitudinal imaging in orthotopic murine models of PDAC. This method enables high-resolution, volumetric quantification of orthotopic primary tumors, liver and lung metastases, and paraspinal skeletal muscle, providing a dynamic view of both tumor and host physiology. MicroCT primary tumor measurements strongly correlated with endpoint tumor weights and outperformed 2D ultrasound in early detection and volumetric accuracy, particularly for small or irregularly shaped tumors. This platform revealed heterogeneous metastatic kinetics across PDAC models and uncovered early, heterogeneous onset of skeletal muscle wasting, a hallmark of cancer cachexia. Notably, this protocol mimics clinical CT surveillance by enabling opportunistic cachexia assessment from tumor imaging datasets and offers substantial advantages over destructive endpoint analyses. Further, microCT radiation had no effect on our model endpoints. By capturing the temporal dynamics of tumor progression and host response, dual contrast microCT serves as a powerful translational platform for preclinical PDAC research and therapeutic testing.

SignificanceDual-contrast microCT provides high-resolution, whole-body, non-invasive imaging in orthotopic murine PDAC models, enabling simultaneous tracking of tumor growth, metastasis, and skeletal muscle wasting-offering a clinically relevant, translational imaging platform.
]]></description>
<dc:creator>Pelz, K. R.</dc:creator>
<dc:creator>Jimenez, P.</dc:creator>
<dc:creator>Daniel, C. J.</dc:creator>
<dc:creator>Newton, S. D.</dc:creator>
<dc:creator>Cunningham, M.</dc:creator>
<dc:creator>Sears, R. C.</dc:creator>
<dc:creator>Worth, P. J.</dc:creator>
<dc:creator>Brody, J. R.</dc:creator>
<dc:creator>Zimmers, T. A.</dc:creator>
<dc:date>2025-06-01</dc:date>
<dc:identifier>doi:10.1101/2025.05.29.656943</dc:identifier>
<dc:title><![CDATA[MicroCT enables simultaneous longitudinal tracking of murine pancreatic cancer progression and cachexia]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-06-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.06.01.657212v1?rss=1">
<title>
<![CDATA[
Synthetic Serum Markers Enable Noninvasive Monitoring of Gene Expression in Primate Brains 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.06.01.657212v1?rss=1</link>
<description><![CDATA[
We demonstrate a noninvasive approach to measure transgene expression in the brain of nonhuman primates using blood tests with engineered reporters called Released Markers of Activity (RMAs). RMAs can exit the brain and enter the bloodstream via reverse-transcytosis across the blood-brain barrier. We demonstrate that these reporters can be used to repeatedly monitor expression of multiple transgenes in cortical and subcortical brain regions simultaneously over a period of three months. RMAs are also sensitive enough to detect circuit-specific Cre-dependent AAV expression. Through this study, the RMA platform provides a cost-efficient, noninvasive tool for neuroscience study of large animals, enabling sensitive, multiplexed, and repeatable measurements of gene expression in the brain with a blood test.
]]></description>
<dc:creator>Lee, S.</dc:creator>
<dc:creator>Romac, M.</dc:creator>
<dc:creator>Romac, M. D.</dc:creator>
<dc:creator>Chernov, M.</dc:creator>
<dc:creator>Li, H.</dc:creator>
<dc:creator>Raisley, E.</dc:creator>
<dc:creator>Szablowski, J. O.</dc:creator>
<dc:creator>Costa, V. D.</dc:creator>
<dc:date>2025-06-02</dc:date>
<dc:identifier>doi:10.1101/2025.06.01.657212</dc:identifier>
<dc:title><![CDATA[Synthetic Serum Markers Enable Noninvasive Monitoring of Gene Expression in Primate Brains]]></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/10.1101/2025.06.08.658506v1?rss=1">
<title>
<![CDATA[
PIVOT: an open-source tool for multi-omic spatial data registration 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.06.08.658506v1?rss=1</link>
<description><![CDATA[
Advances in spatial profiling have resulted in the generation of multi-omic atlases that span biological scales. In general, multiple workflows are required for image registration, coordinate registration, and spot deconvolution to integrate modalities. To improve the throughput of registration of multi-omic cohorts, we introduce PIVOT, a user-friendly and open-source interface for streamlined nonlinear registration. We demonstrate PIVOTs strengths through registration of three multi-omic datasets, and show comparison of its performance to existing workflows.
]]></description>
<dc:creator>Forjaz, A.</dc:creator>
<dc:creator>Romero, V. M.</dc:creator>
<dc:creator>Reucroft, I.</dc:creator>
<dc:creator>Eminizer, M.</dc:creator>
<dc:creator>Kramer, D.</dc:creator>
<dc:creator>Higuera, D.</dc:creator>
<dc:creator>Mojdeganlou, H.</dc:creator>
<dc:creator>Guerrero, P. A.</dc:creator>
<dc:creator>Min, J.</dc:creator>
<dc:creator>Wetzel, M.</dc:creator>
<dc:creator>Lvovs, D.</dc:creator>
<dc:creator>Valentin, A.</dc:creator>
<dc:creator>Shin, S. M.</dc:creator>
<dc:creator>Xuan, Y.</dc:creator>
<dc:creator>Sears, R. C.</dc:creator>
<dc:creator>Chin, K.</dc:creator>
<dc:creator>Maitra, A.</dc:creator>
<dc:creator>Fertig, E. J.</dc:creator>
<dc:creator>Ho, W. J.</dc:creator>
<dc:creator>Kagohara, L. T.</dc:creator>
<dc:creator>Wood, L. D.</dc:creator>
<dc:creator>Wirtz, D.</dc:creator>
<dc:creator>Sidiropoulos, D. N.</dc:creator>
<dc:creator>Kiemen, A. L.</dc:creator>
<dc:date>2025-06-09</dc:date>
<dc:identifier>doi:10.1101/2025.06.08.658506</dc:identifier>
<dc:title><![CDATA[PIVOT: an open-source tool for multi-omic spatial data registration]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-06-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.06.13.659575v1?rss=1">
<title>
<![CDATA[
Multi-omics profiling reveals atypical sugar utilization and identifies a key membrane composition regulator in Streptococcus pneumoniae 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.06.13.659575v1?rss=1</link>
<description><![CDATA[
The human body comprises many different microenvironments, each with their own challenges for microorganisms to overcome in order to survive, and possibly cause infection. The human pathogen Streptococcus pneumoniae is notoriously flexible in this regard, and can adapt to a wide range of host niches, including the nasopharynx, lungs, and cerebrospinal fluid. However, the molecular and genetic underpinnings of this ability remain largely obscure. In this work, using infection-mimicking growth conditions we demonstrate that niche adaptation imposes genome-wide changes on multiple levels, including gene essentiality, expression and membrane lipid composition. In general, we show that gene expression and fitness profiling couple orthogonal sets of genes to environmental stimuli. For instance, N-acetylglucosamine (GlcNAc) import (manLMN) and catabolism (nagAB) genes were required for growth on this sugar, but not differentially expressed in its presence, whereas other amino sugar metabolism pathways were upregulated, but not essential. Surprisingly, we found that pneumococci do not necessarily prefer glucose over GlcNAc and that uptake of GlcNAc in absence of subsequent catabolism was toxic. Moreover, we identified a previously overlooked fatty acid saturation regulator, FasR, controlling membrane composition, rendering it important during heat stress. A fundamental understanding of how genes contribute to bacterial niche adaptation, including nutrient availability or temperature fluctuations, is crucial for understanding successful antibiotic therapy and vaccination strategies and the development of novel anti-infectives.
]]></description>
<dc:creator>de Bakker, V.</dc:creator>
<dc:creator>Liu, X.</dc:creator>
<dc:creator>Tang, J.</dc:creator>
<dc:creator>Barbisan, M.</dc:creator>
<dc:creator>Baker, J. L.</dc:creator>
<dc:creator>Veening, J.-W.</dc:creator>
<dc:date>2025-06-14</dc:date>
<dc:identifier>doi:10.1101/2025.06.13.659575</dc:identifier>
<dc:title><![CDATA[Multi-omics profiling reveals atypical sugar utilization and identifies a key membrane composition regulator in Streptococcus pneumoniae]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-06-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.06.11.659204v1?rss=1">
<title>
<![CDATA[
Selective inhibition of MR1-restricted T cell activation by a novel MR1-targeting nanobody 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.06.11.659204v1?rss=1</link>
<description><![CDATA[
MR1 is a non-polymorphic, ubiquitously expressed, MHC class I-like antigen-presenting molecule that presents small-molecule metabolites to T cells. Studies have shown that MR1 plays a role in microbial infection, inflammation, and tumor immunity. The antigens it presents include metabolites of microbial and self-origin as well as small-molecule drugs and form stable complexes with MR1 that are displayed on the cell surface to activate T cells. However, unlike classical MHC I and II molecules, the fundamental biology of MR1 remains poorly understood, particularly the mechanisms governing antigen loading and intracellular trafficking. This knowledge gap is largely due to the lack of molecular tools available to precisely manipulate MR1 function. In this study, we describe a high-affinity (1.6 nM KD) anti-MR1 nanobody, MR1Nb1. We characterize the binding of this nanobody including affinity by ELISA and kinetics by BLI. Crucially, we map the binding epitope of MR1Nb1 on MR1 by HDX-MS, providing key insights into the mechanism through which it blocks MR1T cell activation. In functional assays MR1Nb1 effectively and specifically blocks MAIT cell activation by cells infected with M. tuberculosis or treated with M. smegmatis supernatant. This nanobody represents a unique and versatile tool for the field, as it can be produced inexpensively and expressed intracellularly within antigen-presenting cells. Hence, our study provides a powerful new molecular probe for dissecting the mechanistic underpinnings of MR1 biology and uncover its broader roles in immunity.
]]></description>
<dc:creator>Bates, T. A.</dc:creator>
<dc:creator>Kulicke, C. A.</dc:creator>
<dc:creator>Gurmessa, S.</dc:creator>
<dc:creator>Kim, S.-J.</dc:creator>
<dc:creator>Suresh, S.</dc:creator>
<dc:creator>Weinstein, J. R.</dc:creator>
<dc:creator>Hinchliff, A.</dc:creator>
<dc:creator>Burke, J. E.</dc:creator>
<dc:creator>Lewinsohn, D.</dc:creator>
<dc:creator>Tafesse, F. G.</dc:creator>
<dc:date>2025-06-17</dc:date>
<dc:identifier>doi:10.1101/2025.06.11.659204</dc:identifier>
<dc:title><![CDATA[Selective inhibition of MR1-restricted T cell activation by a novel MR1-targeting nanobody]]></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/10.1101/2025.06.16.659918v1?rss=1">
<title>
<![CDATA[
Specific GPCRs Elicit Unique Extracellular Vesicle MiRNA Array Signatures: An Exploratory Study 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.06.16.659918v1?rss=1</link>
<description><![CDATA[
3.All cells secrete extracellular vesicles (EVs) containing nucleic acid cargo, including microRNAs (miRNAs), that regulate the function of receiving cells. G protein-coupled receptors (GPCRs) affect intracellular function via multiple signaling cascades. However, the mechanisms of GPCR intercellular signaling through EV miRNA activity are unknown. Human U2 osteosarcoma cells expressing native GPCRs were used to selectively stimulate distinct G protein signaling cascades (Gi, Gq, G12/13, and {beta}-arrestin) by members of specific receptor subclasses including the adenosine receptor A1 (ADORA1), the histamine receptor H1 (HRH1), the frizzled class receptor 4 (FZD4), and the atypical chemokine receptor 3 (ACKR3), respectively. We hypothesized that stimulation of specific classes of GPCRs would cause the release of EVs containing miRNAs with receptor-specific up- or down-regulated expression, affecting unique pathological downstream signaling cascades. Receptor-specific agonists dose-dependently increased respective signaling cascade intermediates. We found no change in the quantity of EVs ([~]200nm diameter), but there were distinct EV miRNA signatures following stimulation of GPCRs. Network analyses of differentially expressed miRNA and their predicted targets validated the linkage between specific receptors and cell function and pathological states. The data can be used to reverse engineer mechanisms involving EV miRNAs for various physiological and pathological processes. GPCRs are major pharmacological targets, so understanding the mechanisms that stimulate or inhibit GPCR-mediated changes in extracellular miRNA signatures could improve long- and short-term therapeutic and unwanted drug effects.

SummaryStimulation of specific GPCRs elicited unique EV miRNA signatures that up- and down-regulated predicted biological and pathological cellular processes.

4. Significance StatementG protein-coupled receptors (GPCR) are therapeutic targets for various human diseases. However, the mechanisms of GPCR signaling through extracellular vesicle (EV) miRNA activity are unknown. Here, we identified unique EV miRNA signatures resulting from GPCR activation. The miRNA functional analyses indicate that the downstream signaling effects and their functional networks differ by receptor. This study has important implications for developing new drugs and their long-term and "off-target" effects.

5. Visual Abstract

O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=186 SRC="FIGDIR/small/659918v2_ufig1.gif" ALT="Figure 1">
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org.highwire.dtl.DTLVardef@1a9812org.highwire.dtl.DTLVardef@1f5cbf3org.highwire.dtl.DTLVardef@20d1fdorg.highwire.dtl.DTLVardef@ba4fad_HPS_FORMAT_FIGEXP  M_FIG C_FIG
]]></description>
<dc:creator>Shi, X.</dc:creator>
<dc:creator>Palumbo, M. C.</dc:creator>
<dc:creator>Benware, S.</dc:creator>
<dc:creator>Wiedrick, J.</dc:creator>
<dc:creator>Markwardt, S.</dc:creator>
<dc:creator>Janowsky, A. J.</dc:creator>
<dc:date>2025-06-20</dc:date>
<dc:identifier>doi:10.1101/2025.06.16.659918</dc:identifier>
<dc:title><![CDATA[Specific GPCRs Elicit Unique Extracellular Vesicle MiRNA Array Signatures: An Exploratory Study]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-06-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.06.24.660981v1?rss=1">
<title>
<![CDATA[
Host iron deficiency protects against Plasmodium infection and drives parasite molecular reprofiling. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.06.24.660981v1?rss=1</link>
<description><![CDATA[
Iron deficiency, anemia and Plasmodium infection represent significant global health challenges with overlapping geographical distributions, particularly affecting pregnant women in Africa, yet the mechanisms underlying their interactions remain poorly understood. We employed a multilayered approach combining clinical data from Malawian pregnant women (n=711) in the REVAMP trial, a genetic mouse model (Tmprss6-knockout), and in vitro P. falciparum cultures to clarify associations between iron status and malaria susceptibility. Iron deficiency was associated with 50% reduced P. falciparum parasitemia in pregnant women (95% CI [30%-64%], p<0.0001), while iron-deficient mice exhibited significantly improved survival against P. berghei (median 15.5 days vs. 7.0 days for WT mice) and protection from cerebral malaria (83% vs 17% survival). Iron chelation induced substantial transcriptomic and proteomic changes in cultured parasites, affecting host cell invasion and nutrient acquisition processes. Importantly, intravenous iron supplementation did not increase subsequent parasitemia when coupled with malaria prevention. These findings demonstrate that iron deficiency protects against Plasmodium infection and support WHO recommendations for iron supplementation in malaria-endemic regions when combined with adequate malaria prevention strategies.
]]></description>
<dc:creator>Clucas, D.</dc:creator>
<dc:creator>Bennett, C.</dc:creator>
<dc:creator>Harding, R.</dc:creator>
<dc:creator>Pettikiriarachchi, A.</dc:creator>
<dc:creator>Baldi, A.</dc:creator>
<dc:creator>Randall, L. M.</dc:creator>
<dc:creator>Steel, R.</dc:creator>
<dc:creator>Mellin, R.</dc:creator>
<dc:creator>Hobbs, M.</dc:creator>
<dc:creator>Caiazzo, S.</dc:creator>
<dc:creator>Mwangi, M. N.</dc:creator>
<dc:creator>Fielding, K. L.</dc:creator>
<dc:creator>Hickey, P. F.</dc:creator>
<dc:creator>Baldwin, T. M.</dc:creator>
<dc:creator>Amann-Zalcenstein, D.</dc:creator>
<dc:creator>Emery-Corbin, S. J.</dc:creator>
<dc:creator>Mzembe, G.</dc:creator>
<dc:creator>Moya, E.</dc:creator>
<dc:creator>Braat, S.</dc:creator>
<dc:creator>Jex, A.</dc:creator>
<dc:creator>Demir, A. Y.</dc:creator>
<dc:creator>Verhoef, H.</dc:creator>
<dc:creator>Phiri, K. S.</dc:creator>
<dc:creator>Biggs, B.-A.</dc:creator>
<dc:creator>Tham, W.-H.</dc:creator>
<dc:creator>Boddey, J. A.</dc:creator>
<dc:creator>Pasricha, S.-R.</dc:creator>
<dc:creator>Ataide, R.</dc:creator>
<dc:date>2025-06-26</dc:date>
<dc:identifier>doi:10.1101/2025.06.24.660981</dc:identifier>
<dc:title><![CDATA[Host iron deficiency protects against Plasmodium infection and drives parasite molecular reprofiling.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-06-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.06.23.661120v1?rss=1">
<title>
<![CDATA[
Sensitizing Immune-Refractory Ovarian Tumors via p53 Mutation-Tailored Immunotherapy 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.06.23.661120v1?rss=1</link>
<description><![CDATA[
High-grade serous ovarian cancer demonstrates limited responsiveness to immune checkpoint inhibitors, owing in part to immunosuppressive environments shaped by nearly universal p53 aberrations. Utilizing an immunocompetent mouse model and individual p53 mutations, we identified a dependence of the p53-R270H mutation (equivalent of human R273H) on regulatory T cells (Tregs) and the PD-1/PD-L1 axis. Analysis of patient datasets associated R273H with elevated levels of two p53 targets, PD-L1 and amphiregulin (AREG), a Tregs growth factor. In contrast to p53-R172H tumors, where there was limited activity, dual antibody therapy targeting AREG and PD-L1 selectively and effectively inhibited R270H tumors. This involved polarization toward M1 macrophages, infiltration of CD8+ T cells, diminished Ly6G+ neutrophils and downregulation of interleukin-4. In patient-derived R273C organoids, the combination treatment reduced the CD4/CD8 ratio. This study is the first to establish a mutation-tailored therapeutic approach that leverages the capacity of p53 to modulate immunosuppressive mechanisms.
]]></description>
<dc:creator>Chatterjee, R.</dc:creator>
<dc:creator>Simoni-Nieves, A.</dc:creator>
<dc:creator>Truong, A.</dc:creator>
<dc:creator>Lindzen, M.</dc:creator>
<dc:creator>Ozmen, F.</dc:creator>
<dc:creator>Cherry, C.</dc:creator>
<dc:creator>Zwicky, P.</dc:creator>
<dc:creator>Mukherjee, S.</dc:creator>
<dc:creator>Selvadurai, B.-R.</dc:creator>
<dc:creator>Salame, T. M.</dc:creator>
<dc:creator>Gupta, N.</dc:creator>
<dc:creator>Giri, S.</dc:creator>
<dc:creator>Kramarski, L.</dc:creator>
<dc:creator>Avraham, Y.</dc:creator>
<dc:creator>Weizman, E.</dc:creator>
<dc:creator>Ozmen, T.</dc:creator>
<dc:creator>Noronha, A.</dc:creator>
<dc:creator>Chakrabarti, P.</dc:creator>
<dc:creator>Ramesh-Kumar, D.</dc:creator>
<dc:creator>Downward, J.</dc:creator>
<dc:creator>Dahan, R.</dc:creator>
<dc:creator>Amit, I.</dc:creator>
<dc:creator>Velculescu, V.</dc:creator>
<dc:creator>Brenton, J.</dc:creator>
<dc:creator>Mills, G.</dc:creator>
<dc:creator>Oren, M.</dc:creator>
<dc:creator>Yarden, Y.</dc:creator>
<dc:date>2025-06-27</dc:date>
<dc:identifier>doi:10.1101/2025.06.23.661120</dc:identifier>
<dc:title><![CDATA[Sensitizing Immune-Refractory Ovarian Tumors via p53 Mutation-Tailored Immunotherapy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-06-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.06.26.661643v1?rss=1">
<title>
<![CDATA[
Methylation dynamics in the decades preceding acute myeloid leukaemia 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.06.26.661643v1?rss=1</link>
<description><![CDATA[
DNA methylation is emerging as a highly sensitive and specific marker of cancer initiation and progression. How these cancer-specific methylation changes are established in the decades before cancer, however, remains largely unknown. Here, we use a unique collection of longitudinal blood samples collected annually up to 15 years prior to a diagnosis of acute myeloid leukaemia to sensitively track the dynamics of DNA methylation changes at high temporal resolution. We identify thousands of differentially methylated regions (DMRs) that exhibit altered patterns of methylation up to 10 years before acute myeloid leukaemia (AML) diagnosis. Most of these DMRs are strongly associated with expanding clones carrying somatic driver mutations. We identify a subset of  epigenetic driver DMRs characterised by recurrent CpG alterations that are highly shared across pre-AML cases. These are likely to reflect early epigenetic reprogramming associated with AML development. We also reveal large numbers of stochastic  passenger CpGs whose differential methylation results from hitch-hiking with clonal expansions driven by somatically acquired genetic events. These passenger CpGs can be exploited for lineage tracing to discover clonal expansions driven by missing driver mutations. Our findings show widespread changes in methylation patterns during the early stages of cancer development which could be utilised for risk prediction and therapeutic intervention.
]]></description>
<dc:creator>Fonseca, A. V.</dc:creator>
<dc:creator>Boniface, C. T.</dc:creator>
<dc:creator>Watson, C.</dc:creator>
<dc:creator>Hackett, S.</dc:creator>
<dc:creator>Gabbutt, C.</dc:creator>
<dc:creator>Ramos-Yamasaki, A. D.</dc:creator>
<dc:creator>Tan, A. B.</dc:creator>
<dc:creator>Montoya, J. M.</dc:creator>
<dc:creator>Strogantsev, R.</dc:creator>
<dc:creator>Hanssen, L.</dc:creator>
<dc:creator>Zhang, Y.</dc:creator>
<dc:creator>Apostolidou, S.</dc:creator>
<dc:creator>Gentry-Maharaj, A.</dc:creator>
<dc:creator>Esener, S.</dc:creator>
<dc:creator>Spellman, P.</dc:creator>
<dc:creator>Graham, T.</dc:creator>
<dc:creator>Menon, U.</dc:creator>
<dc:creator>Mohammed, H.</dc:creator>
<dc:creator>Blundell, J. R.</dc:creator>
<dc:date>2025-06-27</dc:date>
<dc:identifier>doi:10.1101/2025.06.26.661643</dc:identifier>
<dc:title><![CDATA[Methylation dynamics in the decades preceding acute myeloid leukaemia]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-06-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.06.25.661419v1?rss=1">
<title>
<![CDATA[
Long Noncoding RNAs Enforce Pancreatic Cancer Identity and Block Reprogramming 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.06.25.661419v1?rss=1</link>
<description><![CDATA[
The Yamanaka factors (OCT4, SOX2, KLF4, and MYC; OSKM) can rejuvenate aging phenotypes in somatic cell types by resetting the epigenetic landscape. Curiously, most solid tumor cells remain largely resistant to reprogramming despite their well-documented plasticity, and the underlying mechanisms are unclear. Here, we combined genomic profiling and in vivo assays to investigate OSKM-mediated reprogramming of pancreatic ductal adenocarcinoma (PDAC). In the initial stages, we found that cancer-specific genes were refractory while mesodermal/ECM programs, normally silenced by PRC2, were aberrantly upregulated. A CRISPR interference screen for OSKM reprogramming coupled with functional analyses revealed that suppression of cancer-associated long noncoding RNAs (lncRNAs) erased malignant epithelial programs, restored tumor suppressor activity, and impaired tumorigenicity in vivo. We further identified that ATXN7L3-AS1 lncRNA sustains the PDAC malignant identity through its association with active epithelial oncogenic programs and poised PRC2-targeted developmental genes, thereby supporting both plasticity and memory. Thus, by exploring why cancer cells are resistant to reprogramming, we identify lncRNAs as gatekeepers of malignant identity, suggesting that targeting lncRNAs could be a generalizable therapeutic strategy in treating solid tumors.
]]></description>
<dc:creator>Grygoryev, D.</dc:creator>
<dc:creator>Lee, S.-W.</dc:creator>
<dc:creator>Frankston, C. M.</dc:creator>
<dc:creator>Rakshe, S.</dc:creator>
<dc:creator>Berry, M.</dc:creator>
<dc:creator>Hirano, A.</dc:creator>
<dc:creator>Ekstrom, T.</dc:creator>
<dc:creator>Manalo, E.</dc:creator>
<dc:creator>Tessier, J.</dc:creator>
<dc:creator>Chow-Castro, M.</dc:creator>
<dc:creator>Link, J.</dc:creator>
<dc:creator>Keith, D.</dc:creator>
<dc:creator>Sheppard, B. C.</dc:creator>
<dc:creator>Fei, S.</dc:creator>
<dc:creator>Morgan, T.</dc:creator>
<dc:creator>Remotti, H. E.</dc:creator>
<dc:creator>Yang, W.</dc:creator>
<dc:creator>Furth, E.</dc:creator>
<dc:creator>Thakurela, S.</dc:creator>
<dc:creator>Sears, R. C.</dc:creator>
<dc:creator>Kim, J.</dc:creator>
<dc:date>2025-06-28</dc:date>
<dc:identifier>doi:10.1101/2025.06.25.661419</dc:identifier>
<dc:title><![CDATA[Long Noncoding RNAs Enforce Pancreatic Cancer Identity and Block Reprogramming]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-06-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.06.30.662352v1?rss=1">
<title>
<![CDATA[
N-Alkyl Sulfamates as a New Class of nsP2 Cysteine Protease Inhibitors with Broad Spectrum Antialphaviral Activity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.06.30.662352v1?rss=1</link>
<description><![CDATA[
The emergence of mosquito-borne alphaviruses that cause chronic arthritis or encephalitis underscores the urgent need for broad-spectrum antiviral therapeutics. The viral nsP2 cysteine protease, which is essential for alphavirus replication, is a promising antiviral target. Vinyl sulfone-based inhibitors, such as RA-2034, potently inhibit nsP2 protease but suffer from glutathione reactivity and species-dependent systemic clearance catalyzed by glutathione S-transferase. To address these liabilities, we explored alternative electrophilic warheads and identified reverse amide inhibitors bearing N-alkyl sulfamate warheads with improved biochemical and antiviral profiles. N-methyl sulfamate acetamide 5 emerged as a lead compound with potency against both New and Old World alphaviruses, low GSH reactivity, and high proteome-wide selectivity. Despite its promising antialphaviral activity, 5 exhibited rapid clearance due to hepatic glucuronidation. Structure-activity studies revealed modifications that improve metabolic stability while retaining antiviral activity. These findings introduce sulfamate acetamides as a new class of covalent nsP2 protease inhibitors and advance the discovery of direct acting pan-alphavirus drugs.
]]></description>
<dc:creator>Ghosal, A.</dc:creator>
<dc:creator>Sears, J. D.</dc:creator>
<dc:creator>Hossain, M. A.</dc:creator>
<dc:creator>Tse, E.</dc:creator>
<dc:creator>Howell, S.</dc:creator>
<dc:creator>Burdick, J. E.</dc:creator>
<dc:creator>Morales, N. L.</dc:creator>
<dc:creator>Martinez, S. A.</dc:creator>
<dc:creator>Law, I.</dc:creator>
<dc:creator>Streblow, Z. J.</dc:creator>
<dc:creator>Streblow, D. N.</dc:creator>
<dc:creator>Counago, R. M.</dc:creator>
<dc:creator>Moorman, N. J.</dc:creator>
<dc:creator>Heise, M. T.</dc:creator>
<dc:creator>Willson, T. M.</dc:creator>
<dc:date>2025-07-04</dc:date>
<dc:identifier>doi:10.1101/2025.06.30.662352</dc:identifier>
<dc:title><![CDATA[N-Alkyl Sulfamates as a New Class of nsP2 Cysteine Protease Inhibitors with Broad Spectrum Antialphaviral Activity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-07-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.06.30.662483v1?rss=1">
<title>
<![CDATA[
The IFN I response in tumor cells is shaped by PARP7-p300/CBP interactions through distinct loss- and gain-of-function mechanisms 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.06.30.662483v1?rss=1</link>
<description><![CDATA[
PARP7, a mono-ADP-ribosyl (MAR) transferase, is a key suppressor of the type I interferon (IFN-I) IFN{beta} in various tumor cells and a validated drug target. This negative regulation is reversed by small-molecule inhibitors of PARP7 catalytic activity, resulting in increased IFN-{beta} expression. Yet, the mechanism of action of PARP7 inhibitors remains unclear because the relevant substrates of PARP7-mediated MARylation are unknown. Using an optimized analog- sensitive chemical genetic (ASCG) approach, we identified the co-activators, p300 and CBP, as nuclear PARP7 substrates. We identified an -helical domain in PARP7 essential for p300/CBP interaction, MARylation, and proteasome degradation. Disrupting PARP7-p300/CBP interaction prevents PARP7s suppression of IFN{beta} in colorectal cancer cells. p300/CBP reciprocally regulate PARP7s activity and nuclear localization. Intriguingly, treatment with PARP7 inhibitors increased IFN{beta} expression more than PARP7 knockout in a p300/CBP-dependent manner. Our findings suggest that in some contexts, IFN{beta} induction by PARP7 inhibitors occurs via two mechanisms: inhibiting MARylation of p300/CBP (loss-of-function) and stabilizing the PARP7- p300/CBP complex (gain-of-function).

TeaserChemical genetics discovery of p300 and CBP as substrates of PARP7 that are essential for PARP7-mediated regulation of IFN{beta} via a dual mechanism.
]]></description>
<dc:creator>Siordia, I. R.</dc:creator>
<dc:creator>Rieth, S.</dc:creator>
<dc:creator>Jeltema, D.</dc:creator>
<dc:creator>Morgan, R.</dc:creator>
<dc:creator>Tullis, J.</dc:creator>
<dc:creator>Orth, J.</dc:creator>
<dc:creator>Yan, N.</dc:creator>
<dc:creator>Marx, A.</dc:creator>
<dc:creator>Cohen, M. S.</dc:creator>
<dc:date>2025-07-04</dc:date>
<dc:identifier>doi:10.1101/2025.06.30.662483</dc:identifier>
<dc:title><![CDATA[The IFN I response in tumor cells is shaped by PARP7-p300/CBP interactions through distinct loss- and gain-of-function mechanisms]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-07-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.07.01.662163v1?rss=1">
<title>
<![CDATA[
Human induced pluripotent stem cell-derived microglia with a CX3CR1-V249I genetic variant exhibit dysfunctional phenotypes and modulate neuronal growth and function 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.07.01.662163v1?rss=1</link>
<description><![CDATA[
The involvement of microglia in neurodegenerative diseases has drawn increasing attention, as many genetic risk factors are preferentially expressed in microglia. Microglial fractalkine receptor (CX3CR1) signaling regulates many key microglial functions, and the CX3CR1-V249I single nucleotide polymorphism (SNP) has been associated with increased risk for multiple neurodegenerative conditions, including Alzheimers disease, yet its functional consequences in human microglia remain unexplored. In this study, we generated iPSC-derived human microglia-like cells (hMGLs) and found that the CX3CR1-V249I variant increased susceptibility to starvation-induced cell death, reduced amyloid-beta uptake, altered microglial morphology, and impaired migration, with more pronounced effects in homozygous cells. Co-culture with neurons demonstrated that hMGLs with the CX3CR1-V249I variant misregulated neuronal properties, including abnormal neuronal growth as well as an induction of neuronal hyperexcitability. These findings highlight the critical role of CX3CR1 in regulating microglial function and implicate the V249I variant in driving pathogenic microglial states relevant to neurodegeneration.
]]></description>
<dc:creator>Tutrow, K. D.</dc:creator>
<dc:creator>Harkin, J.</dc:creator>
<dc:creator>Varghese, L. D.</dc:creator>
<dc:creator>Hernandez, M.</dc:creator>
<dc:creator>Huang, K.-C.</dc:creator>
<dc:creator>Fang, Y.</dc:creator>
<dc:creator>Wilcox, P.</dc:creator>
<dc:creator>Bedford, L. M.</dc:creator>
<dc:creator>Lin, T.-Y.</dc:creator>
<dc:creator>Zhang, C.</dc:creator>
<dc:creator>Gomes, C.</dc:creator>
<dc:creator>Puntambekar, S. S.</dc:creator>
<dc:creator>Bissel, S. J.</dc:creator>
<dc:creator>Lamb, B. T.</dc:creator>
<dc:creator>Meyer, J. S.</dc:creator>
<dc:date>2025-07-04</dc:date>
<dc:identifier>doi:10.1101/2025.07.01.662163</dc:identifier>
<dc:title><![CDATA[Human induced pluripotent stem cell-derived microglia with a CX3CR1-V249I genetic variant exhibit dysfunctional phenotypes and modulate neuronal growth and function]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-07-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.07.05.663296v1?rss=1">
<title>
<![CDATA[
Progressive Remodeling of Global Protein Interaction Networks in a Mouse Model of Tauopathy 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.07.05.663296v1?rss=1</link>
<description><![CDATA[
Neurodegenerative disease is marked not just by loss of proteins or cells, but by dynamic rewiring of macromolecular interaction networks that precede and drive pathology. Here, we present the first temporally resolved, systems-scale map of multi-protein complex remodeling in a tauopathy model, integrating co-fractionation mass spectrometry, quantitative phosphoproteomics, and machine learning to decode phosphorylation-dependent shifts in protein interactomes across disease progression. This interactomic atlas identifies functionally validated assemblies--including MAPT-Dpysl2 and Cyfip1-actin complexes--that modulate early disease phenotypes in vivo. By revealing how phosphorylation tunes macromolecular complex architecture and function, this work reframes tauopathy as a disease of dynamic network instability, and establishes a generalizable framework for early detection and mechanistic dissection of neurodegeneration.
]]></description>
<dc:creator>Lin, W.</dc:creator>
<dc:creator>Phanse, S.</dc:creator>
<dc:creator>van der Spek, S.</dc:creator>
<dc:creator>Lampl, N.</dc:creator>
<dc:creator>Stephens, M. C.</dc:creator>
<dc:creator>Rondon Ortiz, A.</dc:creator>
<dc:creator>Taylor, A.</dc:creator>
<dc:creator>Hekman, R.</dc:creator>
<dc:creator>Jiang, L.</dc:creator>
<dc:creator>Havugimana, P.</dc:creator>
<dc:creator>Botas, J.</dc:creator>
<dc:creator>Emili, A.</dc:creator>
<dc:creator>Wolozin, B.</dc:creator>
<dc:date>2025-07-06</dc:date>
<dc:identifier>doi:10.1101/2025.07.05.663296</dc:identifier>
<dc:title><![CDATA[Progressive Remodeling of Global Protein Interaction Networks in a Mouse Model of Tauopathy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-07-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.07.07.663405v1?rss=1">
<title>
<![CDATA[
JAK2 signalling to HNRNPA1 represses retrotransposon activity in haematopoietic stem cells 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.07.07.663405v1?rss=1</link>
<description><![CDATA[
Retrotransposon expression must be tightly controlled, particularly in long-lived multipotent stem cells, to prevent deleterious consequences including insertional mutations. Several mechanisms are known to repress retrotransposon transcription during development which are generally thought to persist thereafter. However, integration of retrotransposable elements into host genomes has also provided a major source of genetic variation across evolution. This has generated many sequences that have acquired advantageous host functions, but little is known about retrotransposon expression and mobility in adult stem cells or about how these processes might be dynamically regulated. Here we describe the landscape of somatic retrotransposition in haematopoietic stem cells and identify a previously unrecognised pathway which links cytokine signalling, RNA-modulating HNRNP complexes and repression of retrotransposon activity. Activation of JAK2, by thrombopoietin or gain-of-function JAK2 mutations, triggers tyrosine phosphorylation of HNRNPA1 that represses expression of ERVs, LINEs and SINEs and reduces insertional mutagenesis. This pathway coordinates retrotransposon activity with cellular context and state and provides a mechanism for protecting the haematopoietic stem cell genome.
]]></description>
<dc:creator>Deuel, J. W.</dc:creator>
<dc:creator>Mararovs, A.</dc:creator>
<dc:creator>Goosheh, S.</dc:creator>
<dc:creator>Winder, G.</dc:creator>
<dc:creator>Mohammed, H.</dc:creator>
<dc:creator>Carroll, J.</dc:creator>
<dc:creator>Chapman, M. S.</dc:creator>
<dc:creator>Kapadia, C. D.</dc:creator>
<dc:creator>Campbell, P.</dc:creator>
<dc:creator>Nangalia, J.</dc:creator>
<dc:creator>Schneider, C.</dc:creator>
<dc:creator>Oellerich, T.</dc:creator>
<dc:creator>Green, A. R.</dc:creator>
<dc:creator>Loughran, S. J.</dc:creator>
<dc:date>2025-07-09</dc:date>
<dc:identifier>doi:10.1101/2025.07.07.663405</dc:identifier>
<dc:title><![CDATA[JAK2 signalling to HNRNPA1 represses retrotransposon activity in haematopoietic stem cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-07-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.07.08.663801v1?rss=1">
<title>
<![CDATA[
A Novel FNDC1-NAMPT-NAD axis is Implicated in Small and Large-vessel Arterial Disease and Drives Vascular Calcification 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.07.08.663801v1?rss=1</link>
<description><![CDATA[
Vascular calcification represents a convergent pathological feature of diverse cardiovascular diseases, yet the upstream molecular programs orchestrating this process remain poorly defined. Here, we uncover fibronectin type III domain-containing 1 (FNDC1) as a previously unrecognized regulator of vascular calcification across both microvascular and macrovascular beds. Integrative transcriptomic profiling of human calciphylaxis lesions and atherosclerotic coronaries identified FNDC1 as one of the most significantly upregulated genes. In primary human vascular smooth muscle cells, FNDC1 drove osteogenic phenotype switch and vascular calcification through activation of PI3K/AKT signaling and metabolic reprogramming. Mechanistically, FNDC1 directly binds to nicotinamide phosphoribosyltransferase (NAMPT) resulting in elevated intracellular NAD levels, thus coupling vascular signaling to control of NAD biosynthesis. In murine models, genetic deletion of Fndc1 or pharmacologic inhibition of NAMPT suppressed arterial calcification and prolonged survival. Clinically, circulating FNDC1 levels were elevated in patients with both calciphylaxis and coronary artery disease and independently predicted cardiovascular risk in 42,687 UK Biobank participants. Together, these findings establish FNDC1 as a central mediator of vascular pathology and highlight the FNDC1- NAMPT-NAD+ axis as a promising target for therapeutic intervention.
]]></description>
<dc:creator>Lee, S.</dc:creator>
<dc:creator>Guo, Y.</dc:creator>
<dc:creator>Kajuluri, L. P.</dc:creator>
<dc:creator>Roh, K.</dc:creator>
<dc:creator>Singh, K.</dc:creator>
<dc:creator>Jiang, W.</dc:creator>
<dc:creator>Moore, E.</dc:creator>
<dc:creator>Tattersfield, H.</dc:creator>
<dc:creator>Ostrom, K.</dc:creator>
<dc:creator>Birchenough, C.</dc:creator>
<dc:creator>Johnson, A. L.</dc:creator>
<dc:creator>Krinsky, S.</dc:creator>
<dc:creator>Bouchouari, H.</dc:creator>
<dc:creator>Mahamdeh, M.</dc:creator>
<dc:creator>Lee, L. Y.</dc:creator>
<dc:creator>Wyant, G.</dc:creator>
<dc:creator>Rich, S. S. S.</dc:creator>
<dc:creator>Rotter, J. I. I.</dc:creator>
<dc:creator>Taylor, K. D.</dc:creator>
<dc:creator>Malik, S.</dc:creator>
<dc:creator>Gerszten, R. E.</dc:creator>
<dc:creator>Cruz, D. E.</dc:creator>
<dc:creator>Tracy, R. P.</dc:creator>
<dc:creator>Wu, H.</dc:creator>
<dc:creator>Zhang, H.</dc:creator>
<dc:creator>Lu, H.</dc:creator>
<dc:creator>Kavousi, M.</dc:creator>
<dc:creator>Shanahan, C. M.</dc:creator>
<dc:creator>Duer, M. J.</dc:creator>
<dc:creator>Kramann, R.</dc:creator>
<dc:creator>Nazarian, R. M.</dc:creator>
<dc:creator>Eagleton, M. J.</dc:creator>
<dc:creator>Chen, Y.</dc:creator>
<dc:creator>Miller, C. L.</dc:creator>
<dc:creator>Nigwekar, S. U.</dc:creator>
<dc:creator>Malhotra, R.</dc:creator>
<dc:date>2025-07-10</dc:date>
<dc:identifier>doi:10.1101/2025.07.08.663801</dc:identifier>
<dc:title><![CDATA[A Novel FNDC1-NAMPT-NAD axis is Implicated in Small and Large-vessel Arterial Disease and Drives Vascular Calcification]]></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/10.1101/2025.07.09.662476v1?rss=1">
<title>
<![CDATA[
Evaluation of natural killer cell tumor homing and effector function in response to CDK4/6 and AURKA inhibition in a melanoma tumor-on-a-chip platform 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.07.09.662476v1?rss=1</link>
<description><![CDATA[
Natural killer (NK) cells have emerged as an important clinical tool cellular immunotherapy. Whereas immune checkpoint blockade (ICB) or chimeric antigen receptor (CAR) T-cell therapy (CAR-T) therapy have been adopted as a first line treatments in different malignancies, such as melanoma, these approaches do not work for all patients. T cells require proper antigen presentation on tumor cells for recognition and to carry out their corresponding cytotoxic functions. Deficiency of tumor antigens, or high variability in those present, make T cell-based CAR-T and ICB ineffective. By contrast, NK cells are not limited by antigen presentation deficiencies, offering a potential alternative approach, yet their efficacy can suffer from immunosuppressive signals. Herein, we sought to develop in vitro and on-chip platforms to identify strategies for enhance, rather than suppress, NK cell homing to tumor cells. We explored the use of inhibition of kinases such as CK4/6 and AURKA to induce tumor cell production of chemokines that NK cells migrate towards in aggressive melanoma models. We evaluated chemokine-aided NK cell migration-homing capabilities and their therapeutic efficacy and found that treatment of both melanoma cell line and patient-tumor constructs (PTCs) with CDK4/6 and AURKA generally resulted in improved NK cell homing to tumor cells and accompanying tumor cell killing. Interestingly, this chemokine-guided NK cell migration did not generate as effective outcomes in models using a mildly aggressive melanoma cell line. For our studies, we used 3D tumor constructs in both static Transwell models and then in a bioengineered NK cell-functionalized tumor-on-a-chip (NK-TOC) platform.

Graphical abstract

O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=83 SRC="FIGDIR/small/662476v1_ufig1.gif" ALT="Figure 1">
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]]></description>
<dc:creator>Chakraborty, S.</dc:creator>
<dc:creator>Durham, C.</dc:creator>
<dc:creator>Bharti, V.</dc:creator>
<dc:creator>Capece, M.</dc:creator>
<dc:creator>Davies, A.</dc:creator>
<dc:creator>Vilgelm, A.</dc:creator>
<dc:creator>Skardal, A.</dc:creator>
<dc:date>2025-07-11</dc:date>
<dc:identifier>doi:10.1101/2025.07.09.662476</dc:identifier>
<dc:title><![CDATA[Evaluation of natural killer cell tumor homing and effector function in response to CDK4/6 and AURKA inhibition in a melanoma tumor-on-a-chip platform]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-07-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.07.11.664166v1?rss=1">
<title>
<![CDATA[
Dynamic extracellular interactions with AMPA receptors 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.07.11.664166v1?rss=1</link>
<description><![CDATA[
Synaptic plasticity in the central nervous system enables the encoding, storing, and integrating new information. AMPA-type glutamate receptors (AMPARs) are ligand-gated ion channels that mediate most fast excitatory synaptic transmission in the brain, and plasticity of AMPARs signaling underlies the long-lasting changes in synaptic efficacy and strength important for learning and memory.1,2 Recent work has indicated that the enigmatic N-terminal domain (NTD) of AMPARs may be a critical regulator of synaptic targeting and plasticity of AMPARs. However, few synaptic proteins have been identified that regulate AMPAR plasticity through interactions with AMPAR NTDs. Moreover, the scope of AMPAR NTD interactors that are important for synaptic plasticity remains unknown. Here, we present the dynamic, extracellular interactome for AMPARs during synaptic plasticity. Using surface-restricted proximity labeling and BioSITe-based proteomics, we identified 70 proteins that were differentially labeled by APEX2-tagged AMPARs after induction of chemical Long-term potentiation of synapses (cLTP) in cultured neurons. Included in this list, were four members of the IgLON family of GPI-anchored proteins (Ntm, OBCAM/Opcml, Negr1, Lsamp). We show OBCAM and NTM directly interact with the extracellular domains of AMPARs. Moreover, overexpression of NTM significantly attenuates the mobility of surface AMPARs in dendritic spines. These data represent a significant first step at uncovering the unexplored extracellular regulation of AMPARs, with broad implications for synapse function and synaptic plasticity.

Significance StatementOver the past 30 years, significant effort has been focused on understanding the mechanisms that induce long-lasting changes in synapse strength (synaptic plasticity) that drive learning and memory. While many studies have investigated intracellular mechanisms that enable plasticity, especially those acting on AMPA-type glutamate receptors (AMPARs), significantly less is known regarding extracellular mechanisms that shape changes in synapse function. Here, we identified 70 proteins that differentially associate with the extracellular region of AMPARs during chemically-induced synaptic plasticity. We show that OBCAM and NTM directly interact with the NTD of AMPARs and regulate their mobility on the surface of neurons. These data advance our understanding of extracellular AMPAR regulation, with broad implications for synapse function and synaptic plasticity.
]]></description>
<dc:creator>Merrion, H. G.</dc:creator>
<dc:creator>Barber, C. N.</dc:creator>
<dc:creator>Renuse, S. S.</dc:creator>
<dc:creator>Cutler, J.</dc:creator>
<dc:creator>Kreimer, S.</dc:creator>
<dc:creator>Bygrave, A. M.</dc:creator>
<dc:creator>Meyers, D. J.</dc:creator>
<dc:creator>Hale, W. D.</dc:creator>
<dc:creator>Pandey, A.</dc:creator>
<dc:creator>Huganir, R. L.</dc:creator>
<dc:date>2025-07-14</dc:date>
<dc:identifier>doi:10.1101/2025.07.11.664166</dc:identifier>
<dc:title><![CDATA[Dynamic extracellular interactions with AMPA receptors]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-07-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.07.11.664189v1?rss=1">
<title>
<![CDATA[
Tumor-initiating genetics and therapy drive divergent molecular evolution in IDH-mutant gliomas 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.07.11.664189v1?rss=1</link>
<description><![CDATA[
Astrocytomas and oligodendrogliomas are slow-growing and treatment-sensitive IDH-mutant gliomas diagnosed at ages 30-50. Local tumor regrowth and treatment resistance is inevitable resulting in 3-10 year astrocytoma and up to >20 years oligodendroglioma survival. We sought to identify genetic changes associated with tumor evolution in response to therapy through multi-timepoint whole-genome/whole-exome sequencing of 206 IDH-mutant glioma patient samples collected through the Glioma Longitudinal Analysis (GLASS) Consortium. We validated known genomic markers of tumor progression, including hypermutation and CDKN2A homozygous deletion, and discovered novel genetic alterations that distinguish the response to treatment in astrocytomas compared to oligodendrogliomas. Point mutations in PIK3CA, PIK3R1, and NOTCH1 were newly acquired in recurrent oligodendrogliomas and associated with increased mutation rates. Focal oncogene amplifications, together with CDKN2A homozygous deletions, were associated with an increase in recurrence-specific chromosomal imbalances in astrocytomas. Mutational signature analysis revealed additional differences and detected enrichment for the SBS11, and SBS119 mutational signatures after temozolomide treatment in both IDH-glioma subtypes, whereas astrocytomas showed increased ID8 signatures after radiotherapy. These signatures suggest that the genomes of oligodendroglioma and astrocytoma adapt to the selective pressures of tumor progression and treatment in different ways. However, in both IDH-mutant glioma subtypes we observed a convergence of acquired driver gene alterations with genome-wide changes and worse patient outcomes, signaling selection of treatment-refractory clones. By identifying new prognostic markers and delineating the genomic divergence of oligodendrogliomas and astrocytomas after diagnosis, our results suggest that different DNA damage response mechanisms are engaged following chemo- and radiation therapy.
]]></description>
<dc:creator>Chowdhury, T.</dc:creator>
<dc:creator>Tesileanu, C. M. S.</dc:creator>
<dc:creator>Kocakavuk, E.</dc:creator>
<dc:creator>Johnson, K. C.</dc:creator>
<dc:creator>Lee, J.</dc:creator>
<dc:creator>Erson-Omay, Z.</dc:creator>
<dc:creator>Heo, C.</dc:creator>
<dc:creator>Aldape, K.</dc:creator>
<dc:creator>Amin, S. B.</dc:creator>
<dc:creator>Anderson, K. J.</dc:creator>
<dc:creator>Ashley, D. M.</dc:creator>
<dc:creator>Barnholtz-Sloan, J. S.</dc:creator>
<dc:creator>Brat, D. J.</dc:creator>
<dc:creator>Brodbelt, A. R.</dc:creator>
<dc:creator>Castro, A. V.</dc:creator>
<dc:creator>Claus, E. B.</dc:creator>
<dc:creator>Connelly, J. M.</dc:creator>
<dc:creator>Costello, J.</dc:creator>
<dc:creator>Datta, I.</dc:creator>
<dc:creator>Elliott, C.</dc:creator>
<dc:creator>Finocchiaro, G.</dc:creator>
<dc:creator>French, P. J.</dc:creator>
<dc:creator>Gan, H. K.</dc:creator>
<dc:creator>Garofano, L.</dc:creator>
<dc:creator>Ghospurkar, P. L.</dc:creator>
<dc:creator>Golebiewska, A.</dc:creator>
<dc:creator>Gundla, P. S.</dc:creator>
<dc:creator>Hermes, B.</dc:creator>
<dc:creator>Hong, C.</dc:creator>
<dc:creator>Hoogstrate, Y.</dc:creator>
<dc:creator>Horbinski, C.</dc:creator>
<dc:creator>Huse, J. T.</dc:creator>
<dc:creator>Iavarone, A.</dc:creator>
<dc:creator>Karadag, C.</dc:creator>
<dc:creator>Khasraw, M.</dc:creator>
<dc:creator>Kouwenhoven, M. C.</dc:creator>
<dc:creator>LaViolette, P. S.</dc:creator>
<dc:creator>Li, K.</dc:creator>
<dc:creator>Lowman, A.</dc:creator>
<dc:creator>McCortney, K.</dc:creator>
<dc:creator>Moon, H.-E.</dc:creator>
<dc:creator>Na</dc:creator>
<dc:date>2025-07-15</dc:date>
<dc:identifier>doi:10.1101/2025.07.11.664189</dc:identifier>
<dc:title><![CDATA[Tumor-initiating genetics and therapy drive divergent molecular evolution in IDH-mutant gliomas]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-07-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.07.11.664228v1?rss=1">
<title>
<![CDATA[
MiroSCOPE: An AI-driven digital pathology platform for annotating functional tissue units 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.07.11.664228v1?rss=1</link>
<description><![CDATA[
Cancer tissue analysis in digital pathology is typically conducted across different spatial scales, ranging from high-resolution cell-level modeling to lower-resolution tile-based assessments. However, these perspectives often overlook the structural organization of functional tissue units (FTUs), the small, repeating structures which are crucial to tissue function and key factors during pathological assessment. The incorporation of FTU information is hindered by the need for detailed manual annotations, which are costly and time-consuming to obtain. While artificial intelligence (AI)-based solutions hold great promise to accelerate this process, there is currently no comprehensive workflow for building the large, annotated cohorts required. To remove these roadblocks and advance the development of more interpretable approaches, we developed MiroSCOPE, an end-to-end AI-assisted platform for annotating FTUs at scale, built on QuPath. MiroSCOPE integrates a fine-tunable multiclass segmentation model and curation-specific usability features to enable a human-in-the-loop system that accelerates AI annotation by a pathologist. The system is used to efficiently annotate over 71,900 FTUs on 184 prostate cancer hematoxylin and eosin (H&E)-stained tissue samples and demonstrates ready translation to breast cancer. Furthermore, we publicly release a dataset named Miro-120, consisting of 120 prostate cancer H&E with 30,568 annotations, which can be used by the community as a high-quality resource for FTU-level machine learning aims. In summary, MiroSCOPE provides an adaptable AI-driven platform for annotating functional tissue units, facilitating the use of structural information in digital pathology analyses.
]]></description>
<dc:creator>Fenner, M. R.</dc:creator>
<dc:creator>Sevim, S.</dc:creator>
<dc:creator>Wu, G.</dc:creator>
<dc:creator>Beavers, D.</dc:creator>
<dc:creator>Guo, P.</dc:creator>
<dc:creator>Tang, Y.</dc:creator>
<dc:creator>Eddy, C. Z.</dc:creator>
<dc:creator>Ait-Ahmad, K.</dc:creator>
<dc:creator>Rice-Stitt, T.</dc:creator>
<dc:creator>Thomas, G.</dc:creator>
<dc:creator>Kuykendall, M. J.</dc:creator>
<dc:creator>Stavrinides, V.</dc:creator>
<dc:creator>Emberton, M.</dc:creator>
<dc:creator>Xu, D.</dc:creator>
<dc:creator>Song, X.</dc:creator>
<dc:creator>Eksi, S. E.</dc:creator>
<dc:creator>Demir, E.</dc:creator>
<dc:date>2025-07-17</dc:date>
<dc:identifier>doi:10.1101/2025.07.11.664228</dc:identifier>
<dc:title><![CDATA[MiroSCOPE: An AI-driven digital pathology platform for annotating functional tissue units]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-07-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.07.13.664445v1?rss=1">
<title>
<![CDATA[
Age and early life adversity shape heterogeneity of the epigenome across tissues in macaques 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.07.13.664445v1?rss=1</link>
<description><![CDATA[
Age and early life adversity (ELA) are both key determinants of health, but whether they target similar physiological mechanisms across the body is unknown due to limited multi-tissue datasets from well-characterized cohorts. We generated DNA methylation (DNAm) profiles across 14 tissues in 237 semi-free ranging rhesus macaques, with records of naturally occurring ELA. We show that age-associated DNAm variation is predominantly tissue-dependent, yet tissue-specific epigenetic clocks reveal that the pace of epigenetic aging is relatively consistent within individuals. ELA effects on loci are adversity-dependent, but a given ELA has a coordinated impact across tissues. Finally, ELA targeted many of the same loci as age, but the direction of these effects varied, indicating that ELA does not uniformly contribute to accelerated age in the epigenome. ELA thus imprints a coordinated, tissue-spanning epigenetic signature that is both distinct from and intertwined with age-related change, advancing our understanding of how early environments sculpt the molecular foundations of aging and disease.
]]></description>
<dc:creator>Sadoughi, B.</dc:creator>
<dc:creator>Petersen, R.</dc:creator>
<dc:creator>Patterson, S. K.</dc:creator>
<dc:creator>Slikas, E.</dc:creator>
<dc:creator>Adjandba, C.</dc:creator>
<dc:creator>Ryan, N.</dc:creator>
<dc:creator>Costa, C. E.</dc:creator>
<dc:creator>Newman, L. E.</dc:creator>
<dc:creator>Watowich, M. M.</dc:creator>
<dc:creator>Kelsey, C. R.</dc:creator>
<dc:creator>Greenier, A.</dc:creator>
<dc:creator>Goldman, E. A.</dc:creator>
<dc:creator>Negron-Del Valle, J. E.</dc:creator>
<dc:creator>Phillips, D.</dc:creator>
<dc:creator>Thompson, I.</dc:creator>
<dc:creator>Surratt, S. E. B.</dc:creator>
<dc:creator>Gonzalez, O.</dc:creator>
<dc:creator>Compo, N. R.</dc:creator>
<dc:creator>Burgos-Rodriguez, A. G.</dc:creator>
<dc:creator>Cayo Biobank Research Unit,</dc:creator>
<dc:creator>DeCasien, A. R.</dc:creator>
<dc:creator>Chiou, K. L.</dc:creator>
<dc:creator>Walker, C. S.</dc:creator>
<dc:creator>Lambides, A. V. R.</dc:creator>
<dc:creator>Martinez, M. I.</dc:creator>
<dc:creator>Sterner, K. N.</dc:creator>
<dc:creator>Brent, L. J. N.</dc:creator>
<dc:creator>Higham, J. P.</dc:creator>
<dc:creator>Montague, M. J.</dc:creator>
<dc:creator>Platt, M. L.</dc:creator>
<dc:creator>Snyder-Mackler, N.</dc:creator>
<dc:creator>Lea, A. J.</dc:creator>
<dc:date>2025-07-18</dc:date>
<dc:identifier>doi:10.1101/2025.07.13.664445</dc:identifier>
<dc:title><![CDATA[Age and early life adversity shape heterogeneity of the epigenome across tissues in macaques]]></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/10.1101/2025.07.14.664506v1?rss=1">
<title>
<![CDATA[
Negative Cooperativity in a Polybivalent Complex Allows the Prevalence of a Partial Bound State. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.07.14.664506v1?rss=1</link>
<description><![CDATA[
The word "multivalency" carries with it implications of stabilization and increased affinities, however sometimes evolution has opted for multivalent strategies which imperfectly or even negatively impact the affinity of binding interactions. The dynein intermediate chain (IC) binding to TcTex1 and LC8 provides a system for investigating how biological systems may induce such imperfection and utilize it for biological function. LC8 and Tctex1 are homologous proteins, both fold as rotationally symmetric intertwined homodimers which bind to two copies of a client protein, enabling dimerization of the client. LC8 is known to bind over 100 different proteins and has been shown to bind it clients cooperatively, with the second client binding with higher affinity than the first. It is not known whether Tctex1 binding is cooperative, but it does bind to several different partners. Along IC, the LC8 and Tctex1 binding sites are close to one another resulting in additional cooperativity effects. Herein we successfully model the 7-state model of IC/LC8/Tctex1 binding, thereby extracting more information about biologically relevant transitions than have been previously accessed. Our analysis uses Bayesian inference to combine the results from 39 different isotherms to obtain narrow distributions for thermodynamic values. We report the first measurement of cooperativity across subsequent bindings to TcTex1, along with the weakest known cooperativity across LC8 binding. Significantly, we also show anti-cooperativity between LC8 and TcTex1 when binding together. Using the derived free energies, concentrations of the different species illustrates the relatively high concentration of half-bound IC implying a functional role for this state and motivations for biology to employ two weakly dimerizing protein partners rather than one strongly dimerizing partner.
]]></description>
<dc:creator>Walker, D. R.</dc:creator>
<dc:creator>Otten, L.</dc:creator>
<dc:creator>Zuckerman, D. M.</dc:creator>
<dc:creator>Barbar, E. J.</dc:creator>
<dc:date>2025-07-18</dc:date>
<dc:identifier>doi:10.1101/2025.07.14.664506</dc:identifier>
<dc:title><![CDATA[Negative Cooperativity in a Polybivalent Complex Allows the Prevalence of a Partial Bound State.]]></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/10.1101/2025.07.14.664767v1?rss=1">
<title>
<![CDATA[
Myc and Kras cooperate in adult acinar cells to drive phenotypic heterogeneity, metastasis, and therapeutic resistance in a novel pancreatic cancer mouse model 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.07.14.664767v1?rss=1</link>
<description><![CDATA[
Pancreatic ductal adenocarcinoma (PDAc) is a deadly malignancy, most commonly diagnosed in advanced stages when no curative treatments are available. The development of new models that aid ongoing investigation into the mechanisms by which it initiates, disseminates, and evades treatment is of the utmost importance. In vivo models that accurately recapitulate the features and spectrum of human pancreatic cancer are paramount to make a dent in this disease as two decades of the standard-of-care have failed to substantially improve survival. Here, we take advantage of our finding that post-translational stabiliziation of MYC downstream of the canonical PDAc driver, mutant KRAS, is an early event in PDAc progression to design a novel mouse model of PDAc progression based on deregulated, constituitive expression of Myc and mutant Kras in adult pancreatic acinar cells. Tumors from this KMC model histologically and molecularly recapitulate heterogeneity seen in human PDAc, with a high rate of metastasis to the liver. Cell lines derived from KMC autochthonous PDAc provide new models for orthotopic primary tumors that reliably metastasize to the liver and lung, providing important new tools to efficiently study the metastatic cascade and aid in the develoment of new therapeutics addressing metastatic disease. Cell lines represent distinct molecular subtypes with corresponding differential drug sensitivity. Toghether, this model provides a new and additional tool in the study of pancreatic cancer and the means by which it so deftly evades our best efforts at treatment.
]]></description>
<dc:creator>English, I. A.</dc:creator>
<dc:creator>Worth, P. J.</dc:creator>
<dc:creator>MacPherson-Hawthorne, K. A.</dc:creator>
<dc:creator>Vergara, M.</dc:creator>
<dc:creator>Pelz, K.</dc:creator>
<dc:creator>Kiemen, A. L.</dc:creator>
<dc:creator>Shah, V. M.</dc:creator>
<dc:creator>Blise, K. E.</dc:creator>
<dc:creator>Pelz, C.</dc:creator>
<dc:creator>Tsuda, M.</dc:creator>
<dc:creator>Heskett, M. B.</dc:creator>
<dc:creator>Farrell, A. S.</dc:creator>
<dc:creator>Allen-Petersen, B. L.</dc:creator>
<dc:creator>Jimenez, P.</dc:creator>
<dc:creator>Joly, M. M.</dc:creator>
<dc:creator>Thoma, M. C.</dc:creator>
<dc:creator>Eng, J. R.</dc:creator>
<dc:creator>Daniel, C. J.</dc:creator>
<dc:creator>Wang, X.</dc:creator>
<dc:creator>Cunningham, M.</dc:creator>
<dc:creator>Salgado-Garza, G.</dc:creator>
<dc:creator>Phipps, J. L.</dc:creator>
<dc:creator>Betts, C.</dc:creator>
<dc:creator>Sivagnanam, S.</dc:creator>
<dc:creator>Morgan, T. K.</dc:creator>
<dc:creator>Wood, L. D.</dc:creator>
<dc:creator>Coussens, L. M.</dc:creator>
<dc:creator>Brody, J. R.</dc:creator>
<dc:creator>Langer, E. M.</dc:creator>
<dc:creator>Sears, R. C.</dc:creator>
<dc:date>2025-07-18</dc:date>
<dc:identifier>doi:10.1101/2025.07.14.664767</dc:identifier>
<dc:title><![CDATA[Myc and Kras cooperate in adult acinar cells to drive phenotypic heterogeneity, metastasis, and therapeutic resistance in a novel pancreatic cancer mouse model]]></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/10.1101/2025.07.14.664814v1?rss=1">
<title>
<![CDATA[
Characterization of a Novel Transmembrane Activating STING Agonist using Genetically Humanized Mice 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.07.14.664814v1?rss=1</link>
<description><![CDATA[
STING is a pattern recognition receptor that activates type I interferon and proinflammatory responses in addition to unrelated molecular processes following exposure of DNA to the cytosol. Its pharmacologic stimulation enhances vaccine potency and generates effective antitumor responses but clinical trials evaluating STING agonists have not led to approval for human use. STING activation can occur through ligand engagement of either cytosolic or transmembrane protein domains, processes to which distinct cellular phenotypes are attributed. However, the only transmembrane STING agonist identified is human selective and in vivo testing in conventional models is not feasible. Here we describe synthesis of novel STING agonists efficacious against allelic variants of the protein. We also describe genetically humanized STING mice and demonstrate their suitability as a model to evaluate in vivo responses following exogenous administration of human-selective agonists. Experiments demonstrate that the lead molecule (termed INI3069) functions through binding to the STING transmembrane region and its comparison with conventional agonists reveals significant differences in molecular and immune effects. INI3069 can also enhance antibody responses to co-administered antigens and antitumor responses. This work both represents the first in vivo examination of the effects of transmembrane STING agonism and demonstrates efficacy of a potential novel vaccine adjuvant and oncological therapeutic.
]]></description>
<dc:creator>Mizuno, N.</dc:creator>
<dc:creator>Abraham, J.</dc:creator>
<dc:creator>Jimenez-Perez, K.</dc:creator>
<dc:creator>Rose, I.</dc:creator>
<dc:creator>Springgay, L.</dc:creator>
<dc:creator>Boehm, D.</dc:creator>
<dc:creator>Ando, T.</dc:creator>
<dc:creator>Streblow, D. N.</dc:creator>
<dc:creator>Haddad, E.</dc:creator>
<dc:creator>Ward, J.</dc:creator>
<dc:creator>Miller, S.</dc:creator>
<dc:creator>Pandey, U.</dc:creator>
<dc:creator>Junaid, A.</dc:creator>
<dc:creator>Joyner, D.</dc:creator>
<dc:creator>Muir, R.</dc:creator>
<dc:creator>Burkhart, D.</dc:creator>
<dc:creator>Rasheed, O.</dc:creator>
<dc:creator>DeFilippis, V. R.</dc:creator>
<dc:date>2025-07-18</dc:date>
<dc:identifier>doi:10.1101/2025.07.14.664814</dc:identifier>
<dc:title><![CDATA[Characterization of a Novel Transmembrane Activating STING Agonist using Genetically Humanized Mice]]></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/10.1101/2025.07.15.664474v1?rss=1">
<title>
<![CDATA[
S-Nitrosylated COX-2 is a TME-regulated breast cancer biomarker of mesenchymal phenotypes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.07.15.664474v1?rss=1</link>
<description><![CDATA[
COX-2 is an inducible enzyme key to the production of inflammatory prostaglandins. COX-2 also has tumor intrinsic oncogenic activity in mouse models of breast cancer. Previously, we reported increased expression of Cys-526-nitrosylated COX-2 (SNO-COX-2), but not non-nitrosylated COX-2, with progression of early-stage human breast cancer to invasive ductal carcinoma. Here, we used a 3D culture model of early-stage human breast cancer (MCF10DCIS cells) to investigate the relationship between SNO-COX-2 expression and mesenchymal/invasive tumor cell morphology. We find that SNO-COX-2, but not non-nitrosylated COX-2, closely associated with mesenchymal phenotypes induced by fibrillar type I collagen. Interestingly, invasive phenotypes did not associate with induction of the classic epithelial-to-mesenchymal transition (EMT) markers SNAIL, CDH2 (N-cadherin), and VIM (vimentin). By contrast TGF{beta}-1 strongly induced EMT-related transcripts, but not SNO-COX-2 protein expression or mesenchymal phenotypes. These observations suggest that in MCF10DCIS cells, SNO-COX-2 associates with mesenchymal phenotypes more strongly than non-nitrosylated COX-2 protein, or expression of classic EMT transcripts. In a mouse model with breast tumor heterogeneity, mesenchymal tumor regions also have increased SNO-COX-2 expression. Testing 300 distinct tumor microenvironment conditions, we find SNO-COX-2 protein expression is driven by inflammation, wound resolution, and cancer-associated factors, especially TNC, SPP1, decorin, fibrillar type I and III collagens, INF-{gamma}, and IL-4/13, with evidence for specific extracellular matrix-ligand interactions driving both high and low SNO-COX-2 expression. In sum, in MCF10DCIS cells, expression of SNO-COX-2 is highly microenvironment-dependent and strongly associated with invasive/mesenchymal growth, indicating potential for SNO-COX-2 as a biomarker to assess risk of early-stage breast cancer progression.
]]></description>
<dc:creator>Hoffmann, R. J.</dc:creator>
<dc:creator>Bensen, A.</dc:creator>
<dc:creator>Dane, M.</dc:creator>
<dc:creator>Arterberry, J.</dc:creator>
<dc:creator>Smith, R.</dc:creator>
<dc:creator>Korkola, J.</dc:creator>
<dc:creator>Schedin, P. J.</dc:creator>
<dc:date>2025-07-18</dc:date>
<dc:identifier>doi:10.1101/2025.07.15.664474</dc:identifier>
<dc:title><![CDATA[S-Nitrosylated COX-2 is a TME-regulated breast cancer biomarker of mesenchymal phenotypes]]></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/10.1101/2025.07.18.665554v1?rss=1">
<title>
<![CDATA[
Longitudinal Investigation of Structural and Resting-State Effective Connectivity Alterations in a Non-Human Primate Model of Huntington's Disease 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.07.18.665554v1?rss=1</link>
<description><![CDATA[
Huntingtons disease (HD) is a genetic neurodegenerative disorder caused by expanded CAG repeats in the huntingtin gene which produce a mutant huntingtin (mHTT) protein that contributes to progressive striatal, cortical, and white-matter atrophy, resulting in motor dysfunction and cognitive decline. Recently, a non-human primate (NHP) model of HD was developed via stereotaxic delivery of an adeno-associated viral vector expressing 85 CAG repeats (85Q) into the striatum. This model recapitulates several neuropathological changes and symptoms observed in people with HD (PwHD) including chorea and mild cognitive impairment. A previous longitudinal, multimodal MRI investigation in this model revealed volumetric and resting-state functional connectivity (rs-FC) changes compared to controls, in key regions involved in HD, over the course of 30 months.

We aimed to study longitudinal changes in structural connectivity (SC), obtained from diffusion MRI scans from the same animals, comparing the 85Q animals to the control (Buffer) group. Additionally, going beyond the correlative rs-FC analyses, we investigated changes in causal, inter-regional functional interactions by estimating effective connectivity (EC) from rs functional MRI scans, constrained to strong structural connections. We found that the SC between basal ganglia regions and the cortex was reduced in the 85Q primates compared to the Buffer group at 14-months post virus injection, aligning with the pathological process observed in PwHD at later stages of the disease. EC from the caudate and putamen to the motor cortex was significantly reduced in the 85Q animals as early as 3-months post-injection providing novel insights into early alterations in causal functional interactions.
]]></description>
<dc:creator>Petraityte, G.</dc:creator>
<dc:creator>van Rijswijk, J.</dc:creator>
<dc:creator>Liguore, W.</dc:creator>
<dc:creator>McBride, J. L.</dc:creator>
<dc:creator>Verhoye, M.</dc:creator>
<dc:creator>Bertoglio, D.</dc:creator>
<dc:creator>Weiss, A. R.</dc:creator>
<dc:creator>Adhikari, M. H.</dc:creator>
<dc:date>2025-07-23</dc:date>
<dc:identifier>doi:10.1101/2025.07.18.665554</dc:identifier>
<dc:title><![CDATA[Longitudinal Investigation of Structural and Resting-State Effective Connectivity Alterations in a Non-Human Primate Model of Huntington's Disease]]></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/10.1101/2025.07.18.665576v1?rss=1">
<title>
<![CDATA[
Mild subcortical stroke induces widespread astrogliosis independent of microglia and age 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.07.18.665576v1?rss=1</link>
<description><![CDATA[
Ischemic stroke induces a plethora of pathophysiological changes, including neuroinflammation and chronic cerebrovascular dysfunction. In humans, even small, silent strokes can trigger these pathologies, which can spread to brain regions far beyond the infarct and persist chronically, ultimately worsening prognosis and increasing the risk for vascular dementia and Alzheimers disease. The cause of this extensive pathology is unknown, but reactive astrocytes and microglia are likely contributors. Here, we describe an optimized short-duration middle cerebral artery occlusion model that produces a clinically relevant small stroke confined to subcortical regions, similar to most silent strokes in humans. We termed this model the mild subcortical infarct (MSCI). We then mapped the spatiotemporal extent of reactive astrocytes and microglia during the sub-acute period (1, 3, and 7 days) following MSCI. We observed that reactive astrogliosis develops more rapidly and spreads more extensively, permeating the entire middle cerebral artery territory, compared to the reactive microglial response following this small infarct. Microglial depletion resulted in larger infarct sizes but did not prevent the reactive astrocytes, suggesting that ischemia-driven astrogliosis is largely microglia-independent. Lastly, we show that aging mice exposed to MSCI exhibit a comparably strong response of reactive astrocytes and microglia as young mice. We propose that MSCI is a novel and valuable model for examining the subtle yet highly important chronic effects of stroke. It may be especially useful for investigating the influence of reactive astrogliosis on pathologies like neuroinflammation and cerebrovascular dysfunction in regions distal from the primary injury site.
]]></description>
<dc:creator>Stackhouse, T. L.</dc:creator>
<dc:creator>Marxmiller, B. D.</dc:creator>
<dc:creator>Sullivan, S. J.</dc:creator>
<dc:creator>McConnell, H. L.</dc:creator>
<dc:creator>Knittell, L. M.</dc:creator>
<dc:creator>De La Torre, R.</dc:creator>
<dc:creator>Houser, A. M.</dc:creator>
<dc:creator>Zhang, W.</dc:creator>
<dc:creator>Woltjer, R.</dc:creator>
<dc:creator>Mishra, A.</dc:creator>
<dc:date>2025-07-23</dc:date>
<dc:identifier>doi:10.1101/2025.07.18.665576</dc:identifier>
<dc:title><![CDATA[Mild subcortical stroke induces widespread astrogliosis independent of microglia and age]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-07-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.07.21.665984v1?rss=1">
<title>
<![CDATA[
Distinct antibody-based signatures and functionality distinguish latent and active pediatric tuberculosis 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.07.21.665984v1?rss=1</link>
<description><![CDATA[
BackgroundTuberculosis (TB), caused by Mycobacterium tuberculosis (Mtb), is among the leading causes of death from an infectious agent among children worldwide. Children represent a particularly vulnerable population due to the greater challenges in diagnosis and the higher risk of progression to severe forms of the disease. However, whether different pediatric outcomes relate to distinct immunologic responses remains incompletely understood. Emerging data suggest that Mtb-specific humoral immune responses represent a correlate of protection against Mtb both following natural infection and vaccination.

MethodsTo determine if immune profiles can distinguish children across the spectrum from Mtb infection to TB disease, as well as children with TB from non-TB lower respiratory tract infection, we mapped the humoral immune response across a panel of 4 dozen Mtb antigens across children presenting with symptoms of active TB (ATB), children with evidence of latent TB infection (LTBI) and children exhibiting non-TB lower respiratory tract infection (non-TB LRTI). Using a custom Luminex assay, Mtb-specific antibody subclass/isotype, Fc receptor (FcR) binding profiles, and functions were profiled across the pediatric groups.

FindingsA robust humoral immune response was observed in children with active TB compared to non-TB LRTI, marked by a strong IgA response, that exhibited high FcR binding. Conversely, children with LTBI uniquely elicited Mtb-specific antibodies with enhanced opsinophagocytic Fc{gamma}R2A binding, as well as a higher capacity to activate NK cells and neutrophils.

InterpretationThere are significant differences in humoral immune profiles across the landscape of pediatric TB, potentially contributing to differential mycobacterial control, and highlighting biomarkers that could guide both diagnostic and therapeutic approaches.

FundingUS National Institutes of Health.
]]></description>
<dc:creator>Nziza, N.</dc:creator>
<dc:creator>Jung, W.</dc:creator>
<dc:creator>Chen, T.</dc:creator>
<dc:creator>Deng, Y.</dc:creator>
<dc:creator>Franken, K. L.</dc:creator>
<dc:creator>Ottenhoff, T. H. M.</dc:creator>
<dc:creator>Kiguli, S.</dc:creator>
<dc:creator>Lewinsohn, D.</dc:creator>
<dc:creator>Boom, W. H.</dc:creator>
<dc:creator>Mayanja-Kizza, H.</dc:creator>
<dc:creator>Nsereko, M.</dc:creator>
<dc:creator>Fortune, S. M.</dc:creator>
<dc:creator>Stein, C.</dc:creator>
<dc:creator>McNamara, R.</dc:creator>
<dc:creator>Alter, G.</dc:creator>
<dc:creator>Lancioni, C. L.</dc:creator>
<dc:date>2025-07-24</dc:date>
<dc:identifier>doi:10.1101/2025.07.21.665984</dc:identifier>
<dc:title><![CDATA[Distinct antibody-based signatures and functionality distinguish latent and active pediatric tuberculosis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-07-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.07.21.665563v1?rss=1">
<title>
<![CDATA[
Nerve injury promotes glial immune responses through a Draper/Ninjurin A pathway 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.07.21.665563v1?rss=1</link>
<description><![CDATA[
Degenerating neurons elicit striking immune reactions from glial cells, including directed invasion of injury sites and engulfment of neuronal debris. While these conserved glial immune responses are neuroprotective, our mechanistic understanding of glial immunity in the damaged and diseased brain is still incomplete. Here, using an in vivo nerve injury assay in the adult Drosophila olfactory system, we characterize a novel role for the transmembrane adhesion molecule Ninjurin A (NijA). We show that NijA is transcriptionally upregulated in neuropil ensheathing glia, but not local astrocytes, within hours after olfactory nerve transection. In NijA mutants, glia fail to properly infiltrate areas that contain severed olfactory nerves, and degenerating axonal debris is not cleared from the CNS. One well-defined signaling cascade critical for ensheathing glial clearance of damaged olfactory axons is the conserved MEGF10/Draper pathway, which includes the engulfment receptor Draper, downstream transcriptional complex AP-1, and known gene target MMP-1. We show that injury-induced transcription of NijA in responding glia requires the Draper receptor but is independent of MMP-1, suggesting a parallel signaling cascade is activated downstream of Draper in responding glia. Our findings reveal an essential role for the glial adhesion factor NijA in morphological and phagocytic responses to CNS damage, highlighting this conserved molecule as a new potential glial therapeutic target for neurodegenerative conditions.
]]></description>
<dc:creator>Brashaw, C.</dc:creator>
<dc:creator>Griffin, A.</dc:creator>
<dc:creator>Speese, S. D.</dc:creator>
<dc:creator>Logan, M. A.</dc:creator>
<dc:date>2025-07-25</dc:date>
<dc:identifier>doi:10.1101/2025.07.21.665563</dc:identifier>
<dc:title><![CDATA[Nerve injury promotes glial immune responses through a Draper/Ninjurin A pathway]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-07-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.07.25.666627v1?rss=1">
<title>
<![CDATA[
Developmental time course of social touch, parvalbumin interneurons, perineuronal nets and Mef2c expression reveals a sensitive period of somatosensory cortex development in prairie voles 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.07.25.666627v1?rss=1</link>
<description><![CDATA[
Social touch facilitates our attachment to others, especially early in life, which may be linked to the maturation of parvalbumin interneurons (PVI) in the somatosensory cortex (S1). These neurons respond to social touch, mature in a sensory experience-dependent manner, and influence both somatosensory processing and social behavior in models of Autism Spectrum Disorder. Prairie voles (Microtus ochrogaster) are an ideal rodent model for studying these concepts since they engage in a species-typical social touch called "huddling". This study first showed that over development from juvenile to adult, same-sex siblings huddled less and explored more. Next, we tracked two markers of plasticity indicative of PVI maturation, extracellular perineuronal nets (PNNs) and nuclear transcription factor Myocyte enhancing factor 2C (Mef2c) - across seven developmental timepoints. We found that, while PV expression in S1 was stable by P21, PNNs and Mef2c continued to shift afterwards, indicating a protracted development. Four unique clusters of PVIs converge during development between P14-P21, suggesting a sensitive period of PVI development. Finally, to determine environmental factors affecting these processes, environmental enrichment between P21-P28 led to accelerated PVI maturation. This developmental mapping provides a particularly salient model to investigate the molecular underpinnings of cortical and social development.
]]></description>
<dc:creator>Milman, N. E. P.</dc:creator>
<dc:creator>McGuire, N. M.</dc:creator>
<dc:creator>Loeung, J. M.</dc:creator>
<dc:creator>Bueno-Junior, L. S.</dc:creator>
<dc:creator>Tinsley, C. E.</dc:creator>
<dc:creator>Bronstein, H.</dc:creator>
<dc:creator>Kelly, F. D.</dc:creator>
<dc:creator>Wickham, P. T.</dc:creator>
<dc:creator>Ghimire, A.</dc:creator>
<dc:creator>Johnson, Z.</dc:creator>
<dc:creator>Pantazopoulos, H.</dc:creator>
<dc:creator>Watson, B. O.</dc:creator>
<dc:creator>Sorg, B. A.</dc:creator>
<dc:creator>Lim, M. M.</dc:creator>
<dc:date>2025-07-26</dc:date>
<dc:identifier>doi:10.1101/2025.07.25.666627</dc:identifier>
<dc:title><![CDATA[Developmental time course of social touch, parvalbumin interneurons, perineuronal nets and Mef2c expression reveals a sensitive period of somatosensory cortex development in prairie voles]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-07-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.07.23.666474v1?rss=1">
<title>
<![CDATA[
APOE4 genotype negates the benefits of 17β-estradiol on cerebrovascular endothelial and mitochondrial function 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.07.23.666474v1?rss=1</link>
<description><![CDATA[
BackgroundPostmenopausal females who carry an APOE{varepsilon}4 allele are at higher risk of late-onset Alzheimers Disease compared to age-matched APOE{varepsilon}4 males. Estrogen deficiency predisposes females to an increased risk of vascular, cognitive, and metabolic impairments. While estrogen and APOE genotype are known to impact metabolic and mitochondrial function in the brain, their cerebrovascular effects are less understood. Thus, the purpose of this study was to determine the interaction between APOE genotype and estrogen on cerebrovascular endothelial and mitochondrial function.

MethodsYoung female homozygous APOE{varepsilon}3 and APOE{varepsilon}4 mice (n=19-20/group; ~6 months old) fed a high-fat diet were ovariectomized (OVX), OVX and supplemented with 17{beta}-estradiol, or left intact.

ResultsIn APOE{varepsilon}3 mice, OVX was associated with impaired posterior cerebral artery endothelium-dependent dilation, which was rescued by 17{beta}-estradiol. However, in APOE{varepsilon}4 mice, there was no effect of OVX or 17{beta}-estradiol on cerebral artery endothelial function. Carotid artery passive stiffness was greater with OVX and lower with 17{beta}-estradiol treatment in APOE{varepsilon}3 mice, but there was no impact of OVX or 17{beta}-estradiol in the APOE{varepsilon}4 mice. In cerebral arteries and arterioles, mitochondrial complexes I and I+II respiration were lower in APOE{varepsilon}4 mice compared with APOE{varepsilon}3 mice. 17{beta}-estradiol led to higher mitochondrial complex I respiration in APOE{varepsilon}3 but not APOE{varepsilon}4 mice. These functional differences were concomitant with group differences in mitochondrial DNA copy number, antioxidant enzymes, and pro-inflammatory factors. In contrast to other outcomes, we found that 17{beta}-estradiol treatment was associated with lower cerebral artery stiffness in APOE{varepsilon}4 but not APOE{varepsilon}3 mice.

ConclusionsOverall, these results indicate that the APOE genotype modulates the impact of estrogen on the cerebral vasculature. We found that 17{beta}-estradiol enhances cerebrovascular endothelial and mitochondrial function in APOE{varepsilon}3 mice but not in APOE{varepsilon}4 mice. The results suggest that 17{beta}-estradiol supplementation has more cerebrovascular benefit for APOE{varepsilon}4 non-carriers.

Novelty & SignificanceO_ST_ABSWhat is known?C_ST_ABSO_LIFemales have twice the risk of Alzheimers disease compared with males, and the APOE4 genetic variant is associated with a greater risk for Alzheimers disease compared with the APOE3 variant.
C_LIO_LIThe risk for Alzheimers disease increases after menopause in females, suggesting that the loss of female sex hormones may play a role.
C_LIO_LIThere are highly inconsistent results among past studies examining the interaction of APOE genotype and estrogens on cognitive function and other brain outcomes.
C_LI

What new information does this article contribute?Vascular outcomes were not measured in previous studies examining the interaction between APOE genotype and estrogens. As such, we aimed to determine the impact of APOE4 genotype on the cerebrovascular response to estradiol. We found that estradiol improved cerebral artery endothelial function and mitochondrial respiration in APOE3 mice following ovariectomy. In contrast, APOE4 mice were refractory to the beneficial effects of estradiol on cerebrovascular endothelial and mitochondrial function. The broader implication of this research is that APOE genotype may be a consideration when prescribing hormone replacement therapy to menopausal females due to the impact on vascular outcomes.
]]></description>
<dc:creator>Kehmeier, M. N.</dc:creator>
<dc:creator>Famiano, A.</dc:creator>
<dc:creator>Cullen, A. E.</dc:creator>
<dc:creator>Leonhardt, T.</dc:creator>
<dc:creator>Ferguson, S.</dc:creator>
<dc:creator>Snyder, M.</dc:creator>
<dc:creator>McCurdy, C. E.</dc:creator>
<dc:creator>Tyrrell, D. J.</dc:creator>
<dc:creator>Alkayed, N. J.</dc:creator>
<dc:creator>Walker, A. E.</dc:creator>
<dc:date>2025-07-29</dc:date>
<dc:identifier>doi:10.1101/2025.07.23.666474</dc:identifier>
<dc:title><![CDATA[APOE4 genotype negates the benefits of 17β-estradiol on cerebrovascular endothelial and mitochondrial function]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-07-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.07.26.666969v1?rss=1">
<title>
<![CDATA[
Targeting Tmem63b and Piezo2 in C-fiber low threshold mechanoreceptor: limitation of Vglut3-IRES-Cre 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.07.26.666969v1?rss=1</link>
<description><![CDATA[
Peripheral somatosensory neurons in the dorsal root ganglia (DRG) transduce mechanical force in the skin and other organs into electrical signals using specialized mechanically activated (MA) ion channels that initiate neuronal activation in response to force. Increasing evidence highlights PIEZO2 as the primary transducer of low-threshold mechanical force in DRG neurons. However, in the absence of Piezo2, mice and humans still respond to noxious painful stimuli like pinch, suggesting that additional MA channel(s) likely exist in DRG neurons. Strategies to identify Cre lines and DRG subpopulations that select for non-PIEZO2 expressing neurons is therefore an ongoing effort in the field to discover unknown mechanosensors. Here, we investigated a Vglut3 labeled mouse line as a candidate to identify non-PIEZO2 MA channels in a subtype of DRG neurons called C-fiber low threshold mechanoreceptors (C-LTMRs). Our study carefully demonstrates that the Vglut3-IRES-Cre mouse line specifically and efficiently labels C-LTMR neurons of the DRG. Electrophysiological recordings using two different in vitro mechanical stimulation assays show that the genetically labelled Vglut3 neurons have robust indentation- and stretch-activated MA currents that are exclusively slowly or ultra-slowly adapting. To determine whether the Vglut3-IRES-Cre mouse line can be used to delete genes of interest and identify the underlying MA ion channels in C-LTMRs we attempted to generate a Tmem63b conditional knockout using this Cre-line but detected incomplete loss of Tmem63b transcript and lack of TMEM63B- dependent effect on C-LTMR MA currents. Together, our results emphasize that although the Vglut3-IRES-Cre line is robust in driving expression of a conditional reporter gene, it is inefficient in deleting genes like Tmem63b as well as Piezo2.

Statement of SignificanceIn this study, we identify a previously uncharacterized Vglut3-IRES-Cre as a tool to uncover unknown mechanosensors in C-LTMRs of the DRG. We methodically demonstrate that the Vglut3-IRES-Cre line labels C-LTMRs and express two mechanosensors, Tmem63b and Piezo2. However, we were unable to discern which gene contributes to the MA currents due to poor knockout efficiency induced by the Vglut3-IRES-Cre. Our studies highlight the nuances and challenges associated with Cre lines and alert future investigations to exercise caution while using Cre lines to discover the contribution of specific genes to mechanosensation.
]]></description>
<dc:creator>Orlin, D. J.</dc:creator>
<dc:creator>Munoz, A.</dc:creator>
<dc:creator>Berryman, S.</dc:creator>
<dc:creator>Semidey, D.</dc:creator>
<dc:creator>Murthy, S. E.</dc:creator>
<dc:date>2025-07-31</dc:date>
<dc:identifier>doi:10.1101/2025.07.26.666969</dc:identifier>
<dc:title><![CDATA[Targeting Tmem63b and Piezo2 in C-fiber low threshold mechanoreceptor: limitation of Vglut3-IRES-Cre]]></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/10.1101/2025.08.10.669547v1?rss=1">
<title>
<![CDATA[
Glucagon and GLP-1 Accelerate Pseudo-Islet Assembly and Unmask Sex-Specific Islet Fragmentation Dynamics 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.08.10.669547v1?rss=1</link>
<description><![CDATA[
Pancreatic hormones are best known for their role in regulating blood sugar levels as well as islet cell function and proliferation. However, their impact on maintaining and inducing cell aggregation in culture remains under-explored. In this study, we investigated the effects of glucagon (GCG) and glucagon-like-peptide 1(GLP-1) on the formation and integrity of human islet clusters. Native human islets were dissociated and sorted into pure -, {beta}-, and {delta}-cell populations using antibody-based fluorescence-activated cell sorting (FACS). The sorted cells were then co-cultured with mouse endothelial MS1 cells in suspension to generate pseudo-islets of varying cell composition. Hormonal supplementation with GCG or GLP-1 versus blank was administered during the tissue culture phase. Hormone-treated pseudo-islets formed faster, dependent on the cellular composition and the sex of the donor. In parallel, we also exposed native islets, maintained in suspension without prior dissociation or sorting, to hormone supplementation. These islets exhibited accelerated fragmentation under hormone treatment compared to controls, again dependent on donor sex with islets from female donors fragmenting faster than from male donors. These findings suggest that GCG and GLP-1 enhance pseudo-islet formation and affect the structural integrity of native islets in a sex-specific manner, offering insights into islet biology and implications for diabetes research and therapy.

Article HighlightsWe established a manipulatable, expandable human pseudo-islet platform to investigate islet morphogenesis, architecture, and intercellular signaling. We examined the contribution of individual -, {beta}-, and {delta}-cell populations and assessed how glucagon (GCG) and glucagon-like peptide-1 (GLP-1) modulate islet integrity in culture. In native islets, hormonal supplementation attenuated fragmentation in male donors but accelerated it in females. In pseudo-islets, cellular composition was the predominant determinant of maturation versus fragmentation, with donor sex exerting a secondary influence. We present methodological guidelines for generating and maintaining human pseudo-islets, thereby providing a framework to optimize donor selection, culture conditions, and experimental design in diabetes research.
]]></description>
<dc:creator>Keutler, K.</dc:creator>
<dc:creator>Prady, S. B.</dc:creator>
<dc:creator>Canaday, P.</dc:creator>
<dc:creator>Dorrell, C.</dc:creator>
<dc:creator>Schultz, C.</dc:creator>
<dc:date>2025-08-12</dc:date>
<dc:identifier>doi:10.1101/2025.08.10.669547</dc:identifier>
<dc:title><![CDATA[Glucagon and GLP-1 Accelerate Pseudo-Islet Assembly and Unmask Sex-Specific Islet Fragmentation Dynamics]]></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/10.1101/2025.08.12.669930v1?rss=1">
<title>
<![CDATA[
Altered molecular signaling pathways in the hippocampus of rhesus monkeys following chronic alcohol use 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.08.12.669930v1?rss=1</link>
<description><![CDATA[
Context-induced relapse is a significant factor limiting recovery from alcohol use disorder (AUD). However, the molecular processes in the hippocampus, a critical region for contextual memory impacted by chronic alcohol use, remain poorly understood. We used a non-human primate model to test the hypothesis that chronic alcohol use impacts hippocampal molecular pathways that may serve as therapeutic targets for context-induced relapse and memory processing issues associated with chronic alcohol use. We conducted RNAseq profiling on hippocampal samples from adult male rhesus monkeys with chronic alcohol use (n=7) and controls (n=5) from the Monkey Alcohol Tissue Research Resource (MATRR). We identified 2,575 differentially expressed genes (DEGs) in subjects with chronic alcohol use, including genes implicated in genome-wide association studies (GWAS) of alcohol dependence, such as GLP2R and GABBR2. Downregulated pathways included chemical synaptic transmission, trans-synaptic signaling, and neuron development, and upregulated pathways involved mitochondrial function. Targeted pathway analysis highlighted significant downregulation of synaptic signaling (e.g., axonal fasciculation) and upregulation of mitochondrial processes (e.g., electron transport). Leading-edge gene analysis revealed several downregulated genes involved in synaptic signaling including GRIN2B, CACNA1C, and NLGN1 as well as upregulated genes such as NDUFS3 and MT-ND1 involved in mitochondrial processes. Drug repurposing analysis identified several targets including epidermal growth factor receptor (EGFR) inhibitors, and L-type calcium channel blockers as potential therapeutic targets. Our results provide critical insights into molecular pathways underlying hippocampal pathology in chronic alcohol use, emphasizing the roles of mitochondrial function, synaptic regulation and calcium channels, and offering potential novel therapeutic targets.
]]></description>
<dc:creator>Pareek, T.</dc:creator>
<dc:creator>Vergis, J. M.</dc:creator>
<dc:creator>Zhang, X.</dc:creator>
<dc:creator>Platt, D. M.</dc:creator>
<dc:creator>Grant, K. A.</dc:creator>
<dc:creator>McCullumsmith, R.</dc:creator>
<dc:creator>Gisabella, B.</dc:creator>
<dc:creator>O'Donovan, S. M.</dc:creator>
<dc:creator>Pantazopoulos, H.</dc:creator>
<dc:date>2025-08-15</dc:date>
<dc:identifier>doi:10.1101/2025.08.12.669930</dc:identifier>
<dc:title><![CDATA[Altered molecular signaling pathways in the hippocampus of rhesus monkeys following chronic alcohol use]]></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/10.1101/2025.08.12.669998v1?rss=1">
<title>
<![CDATA[
VO: The Vaccine Ontology 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.08.12.669998v1?rss=1</link>
<description><![CDATA[
With the widespread use of vaccines in research and clinical settings, there is an urgent need to standardize vaccine representation, integrate information across diverse vaccine types, and support computer-assisted reasoning. Accordingly, we have since 2007 developed the community-based Vaccine Ontology (VO), which aligns with the Basic Formal Ontology and adheres to OBO Foundry principles. VO models ontologically vaccines, vaccine components, vaccine immune responses, vaccine investigation studies and other vaccine-related topics. VO represents more than 10,000 vaccines targeting 289 infectious pathogens and cancers in humans and over 30 nonhuman animal species. VO provides mappings to external resources such as RxNorm, CVX, FDA, and USDA. Various VO use cases exist. VO facilitates vaccine standardization in resources such as the VIOLIN vaccine database, ImmPort, and the Vaccine Adjuvant Compendium (VAC). Semantic queries can be made to query VO. VO has been shown to enhance experimental and clinical vaccine data analysis and vaccine literature mining. Overall, VO standardizes vaccine modeling and representation and greatly supports vaccine AI research in the Semantic Web era.
]]></description>
<dc:creator>Zheng, J.</dc:creator>
<dc:creator>Lin, A. Y.</dc:creator>
<dc:creator>Huffman, A.</dc:creator>
<dc:creator>Masci, A. M.</dc:creator>
<dc:creator>Racz, R.</dc:creator>
<dc:creator>Wu, G.</dc:creator>
<dc:creator>Roan, K.</dc:creator>
<dc:creator>Ong, E.</dc:creator>
<dc:creator>Sarntivijai, S.</dc:creator>
<dc:creator>Hu, J.</dc:creator>
<dc:creator>Asfaw, E.</dc:creator>
<dc:creator>Kahn, H.</dc:creator>
<dc:creator>Li, X.</dc:creator>
<dc:creator>Zhang, X.</dc:creator>
<dc:creator>Kosar, N.</dc:creator>
<dc:creator>Li, J.</dc:creator>
<dc:creator>Manuel, W.</dc:creator>
<dc:creator>Abeysinghe, R.</dc:creator>
<dc:creator>Rehana, H.</dc:creator>
<dc:creator>Bansal, B.</dc:creator>
<dc:creator>Pan, Y.</dc:creator>
<dc:creator>Guo, J.</dc:creator>
<dc:creator>He, V.</dc:creator>
<dc:creator>Song, J.</dc:creator>
<dc:creator>Seleznev, A. I.</dc:creator>
<dc:creator>Hur, K.</dc:creator>
<dc:creator>He, A.</dc:creator>
<dc:creator>Davydov, A.</dc:creator>
<dc:creator>Yang, Q.</dc:creator>
<dc:creator>Vita, R.</dc:creator>
<dc:creator>Peters, B.</dc:creator>
<dc:creator>Ruttenberg, A.</dc:creator>
<dc:creator>Diehl, A. D.</dc:creator>
<dc:creator>Hoyt, C. T.</dc:creator>
<dc:creator>Roncaglia, P.</dc:creator>
<dc:creator>Huntley, R. P.</dc:creator>
<dc:creator>Scheuermann, R. H.</dc:creator>
<dc:creator>Courtot, M.</dc:creator>
<dc:creator>Todd, T.</dc:creator>
<dc:creator>Sayers, S.</dc:creator>
<dc:creator>Chen, F.</dc:creator>
<dc:creator>Li, X.</dc:creator>
<dc:creator>Yeh, F.-Y.</dc:creator>
<dc:creator>Xiang, Z.</dc:creator>
<dc:creator>Ozgur, A.</dc:creator>
<dc:creator>Whetzel, P. L.</dc:creator>
<dc:creator>Musen, M. A.</dc:creator>
<dc:creator>Mungall, C. J.</dc:creator>
<dc:creator>Leitner, W. W.</dc:creator>
<dc:creator>Cui, L.</dc:creator>
<dc:creator>Colby, L.</dc:creator>
<dc:date>2025-08-15</dc:date>
<dc:identifier>doi:10.1101/2025.08.12.669998</dc:identifier>
<dc:title><![CDATA[VO: The Vaccine Ontology]]></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/10.1101/2025.08.15.670554v1?rss=1">
<title>
<![CDATA[
Epigenetic Reactivation of CNS Endothelial Developmental Programs Triggers Adult Brain Angiogenesis, Promotes Post-Stroke Revascularization and Neuronal Regeneration. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.08.15.670554v1?rss=1</link>
<description><![CDATA[
Therapeutic angiogenesis is essential for regenerating brain tissue damaged by stroke, yet it remains an unmet clinical challenge. During brain development, pro-angiogenic genes drive the formation of vascular networks, with their expression tightly regulated in later stages. We found that in adult CNS endothelial cells (ECs), angiogenesis-related genes are epigenetically silenced through histone deacetylase 2 (HDAC2) and the polycomb repressive complex 2 (PRC2). Conditional deletion of Hdac2 in ECs reactivated pro-angiogenic signaling, including Wnt/{beta}-catenin target genes, leading to functional neovascularization with preserved blood-brain barrier (BBB) integrity in the adult brain. In contrast, Ezh2 (PRC2 subunit) deletion reduced vessel density and compromised BBB function. Deletion of Hdac2 and Ezh2 immediately after transient ischemic stroke conferred vascular protection by modulating stroke-induced transcriptional programs in CNS ECs. In contrast, delayed deletion, initiated seven days post-stroke, after significant neuronal loss in the infarct region, induced robust revascularization and promoted post-stroke neurogenesis, with differentiation into both excitatory and inhibitory neurons. These findings highlight CNS EC HDAC2 as a promising therapeutic target for inducing adult brain angiogenesis, facilitating revascularization, and supporting neuronal regeneration following stroke.
]]></description>
<dc:creator>Thomas, S.</dc:creator>
<dc:creator>Chandran, C.</dc:creator>
<dc:creator>Ahirwar, L. K.</dc:creator>
<dc:creator>Harmanci, A. O.</dc:creator>
<dc:creator>Chathathayil, S. M.</dc:creator>
<dc:creator>Alkayed, N. J.</dc:creator>
<dc:creator>Dienel, A. C.</dc:creator>
<dc:creator>Blackburn, S. L.</dc:creator>
<dc:creator>McBride, D. W.</dc:creator>
<dc:creator>T, P. K.</dc:creator>
<dc:date>2025-08-21</dc:date>
<dc:identifier>doi:10.1101/2025.08.15.670554</dc:identifier>
<dc:title><![CDATA[Epigenetic Reactivation of CNS Endothelial Developmental Programs Triggers Adult Brain Angiogenesis, Promotes Post-Stroke Revascularization and Neuronal Regeneration.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-08-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.08.18.670895v1?rss=1">
<title>
<![CDATA[
Cerebral blood flow in elastin haploinsufficient and 3xTg-AD mice 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.08.18.670895v1?rss=1</link>
<description><![CDATA[
Artery structural properties and Alzheimers disease (AD) pathology are individually associated with impaired cerebrovascular function; however, the interaction of these factors is unclear. Furthermore, while elastin haploinsufficient (Eln+/-) mice are known to have impaired cerebrovascular function, sex differences for this effect have not been previously studied. To answer these questions, we crossed middle-aged and old Eln+/-mice with 3xTg-AD mice. We measured cerebral blood flow (CBF) using arterial spin labeling MRI at rest and during hypercapnia to calculate cerebrovascular reactivity (CVR). We also assessed neuroinflammation by microglia and astrocyte cell counts. We found that Eln+/- mice had lower resting blood flow rate in the cerebral cortex compared with Eln+/+ mice, but Eln+/- mice had an intact hypercapnic response, resulting in better CVR compared with Eln+/+ in hippocampus. Sex did not impact resting blood flow or CVR. 3xTg-AD mice had a lower resting CBF than non-AD mice, and there was an interaction between Eln genotype and AD mutations on CVR, such that Eln+/-x 3xTg-AD mice had the poorest hippocampal CVR of all groups. Glia cell counts were highly dependent on brain region, with Eln+/- having more microglia but fewer astrocytes, while 3xTg-AD having higher both microglia and astrocytes. While sex also impacted glial cell counts, we found no interactions between sex and Eln genotype. Our results demonstrate that elastin haploinsufficiency and AD mutations individually result in lower resting CBF, and the combination of these leads to impaired CVR.

NEW & NOTEWORTHYThe findings of this study demonstrate that elastin haploinsufficiency leads to lower resting cerebral blood flow, but also greater cerebrovascular reactivity. However, elastin haploinsufficiency interacts with Alzheimers disease mutations to impair cerebrovascular reactivity. These results suggest that multiple insults, such as changes to the extracellular matrix combined with genetic risk factors, are needed to impact cerebrovascular reactivity.
]]></description>
<dc:creator>Cullen, A. E.</dc:creator>
<dc:creator>Reeve, E. H.</dc:creator>
<dc:creator>Winder, N. R.</dc:creator>
<dc:creator>Henson, G. D.</dc:creator>
<dc:creator>Arora, N.</dc:creator>
<dc:creator>Leonhardt, T.</dc:creator>
<dc:creator>Hogan, A. P.</dc:creator>
<dc:creator>Kumaran, S. K.</dc:creator>
<dc:creator>Setthavonsack, N.</dc:creator>
<dc:creator>Krajbich, V.</dc:creator>
<dc:creator>Alkayed, N. J.</dc:creator>
<dc:creator>Pike, M. M.</dc:creator>
<dc:creator>Woltjer, R. L.</dc:creator>
<dc:creator>Walker, A. E.</dc:creator>
<dc:date>2025-08-22</dc:date>
<dc:identifier>doi:10.1101/2025.08.18.670895</dc:identifier>
<dc:title><![CDATA[Cerebral blood flow in elastin haploinsufficient and 3xTg-AD mice]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-08-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.08.25.672167v1?rss=1">
<title>
<![CDATA[
Plasmacytoid dendritic cells in the intestine preferentially produce interferon lambda at homeostasis contributing to tonic localized innate immune responses 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.08.25.672167v1?rss=1</link>
<description><![CDATA[
The healthy intestine maintains homeostasis in part via immune responses to microbiota, which includes basal production of interferon cytokines. Previous work showed that Type III Interferon (IFN-{lambda}) stimulates localized pockets of interferon-stimulated genes (ISGs) in the adult mouse intestinal epithelium at homeostasis that provide preemptive protection from viral pathogens. Here, we demonstrate that a major source of homeostatic IFN-{lambda} production in the intestine is a population of epithelium-associated plasmacytoid dendritic cells (pDC). Depletion of bacterial microbiota in the intestine also reduces pDC abundance, and pDC depletion or bone marrow reconstitution with IFN-{lambda}-deficient pDC results in reduced expression of homeostatic ISGs in the intestinal epithelium. Notably, intestinal pDC preferentially produce IFN-{lambda} over Type I IFNs whereas splenic pDC produce more Type I IFNs. Comparison of intestinal and splenic pDC reveal tissue-specific changes in gene expression and genomic accessibility, including evidence of responses to transforming growth factor beta (TGF-{beta}) in the intestine. Isolated gut pDC produce more IFN-{lambda} than splenic pDC upon stimulation, and pre-treatment of a human pDC cell line with TGF-{beta} results in enhanced production of IFN-{lambda} upon stimulation. This study demonstrates that pDC are an important source of homeostatic IFN-{lambda} in the intestine and defines the role of barrier cytokine TGF-{beta} in regulating IFN types produced by pDC upon stimulation. Reprogramming of recruited pDC by tissue cytokines may have important implications for balancing effective antimicrobial responses with damaging inflammation at barrier tissues.

One sentence summaryThis study demonstrates that pDC are an important source of homeostatic IFN-{lambda} in the intestine and defines the role of barrier cytokine TGF-{beta} in regulating IFN types produced by pDC upon stimulation.
]]></description>
<dc:creator>Constant, D. A.</dc:creator>
<dc:creator>Van Winkle, J. A.</dc:creator>
<dc:creator>Norwood, P. A.</dc:creator>
<dc:creator>Mishra, G.</dc:creator>
<dc:creator>Meyer, K. A.</dc:creator>
<dc:creator>Laney, M. E.</dc:creator>
<dc:creator>Madden, S. R.</dc:creator>
<dc:creator>Griffith, A.</dc:creator>
<dc:creator>Rodrigues, P. F.</dc:creator>
<dc:creator>Savan, R.</dc:creator>
<dc:creator>Nice, T. J.</dc:creator>
<dc:date>2025-08-29</dc:date>
<dc:identifier>doi:10.1101/2025.08.25.672167</dc:identifier>
<dc:title><![CDATA[Plasmacytoid dendritic cells in the intestine preferentially produce interferon lambda at homeostasis contributing to tonic localized innate immune responses]]></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/10.1101/2025.08.30.673277v1?rss=1">
<title>
<![CDATA[
Origin and Correlates of Viral Rebound in SIV-Infected Rhesus Macaques Following ART Discontinuation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.08.30.673277v1?rss=1</link>
<description><![CDATA[
The vast majority of persons living with HIV-1 who discontinue antiretroviral therapy (ART) demonstrate viral rebound, but the tissue-level events that lead to rebound viremia are poorly understood. Here we report the origin, dynamics, and correlates of viral rebound in 16 rhesus macaques (RMs) infected with molecularly barcoded SIVmac239M, treated with ART for 70 weeks, and necropsied on day 12 after ART discontinuation. Barcode analysis of plasma following ART discontinuation identified 1 to 38 rebounding barcode-defined viral lineages per animal, with 1 to 4 rebounding lineages contributing to first measurable rebound viremia. Analysis of barcode viral RNA (vRNA) expression in necropsy tissues revealed presumptive anatomic origin sites for 56 of 175 total rebounding viral lineages, with significant enrichment in the gastrointestinal (GI) tract and GI-associated lymph nodes. Daily transcriptomic and proteomic profiling in peripheral blood following ART discontinuation showed upregulation of pathways related to T cell signaling, cytokine responses, and cellular metabolism prior to detectable rebound viremia. These data suggest that viral rebound following ART discontinuation is initiated by local tissue replication of a limited number of clonal lineages, followed by systemic expansion of the initial rebounding lineages and serial initiation of replication of multiple additional clonal lineages. These findings provide mechanistic insights into the processes that result in viral rebound following ART discontinuation and will contribute to next generation HIV-1 cure strategies.
]]></description>
<dc:creator>King, I.</dc:creator>
<dc:creator>Aid, M.</dc:creator>
<dc:creator>Kose, E.</dc:creator>
<dc:creator>Immonen, T.</dc:creator>
<dc:creator>Goodman, C.</dc:creator>
<dc:creator>Fennessey, C. M.</dc:creator>
<dc:creator>Colarusso, A.</dc:creator>
<dc:creator>Walker-Sperling, V.</dc:creator>
<dc:creator>Borducchi, E.</dc:creator>
<dc:creator>Geleziunas, R.</dc:creator>
<dc:creator>Rinaldi, W.</dc:creator>
<dc:creator>Ferguson, M.</dc:creator>
<dc:creator>Picker, L. J.</dc:creator>
<dc:creator>Lifson, J. D.</dc:creator>
<dc:creator>Keele, B. F.</dc:creator>
<dc:creator>Barouch, D. H.</dc:creator>
<dc:date>2025-09-01</dc:date>
<dc:identifier>doi:10.1101/2025.08.30.673277</dc:identifier>
<dc:title><![CDATA[Origin and Correlates of Viral Rebound in SIV-Infected Rhesus Macaques Following ART Discontinuation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-09-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.09.01.673541v1?rss=1">
<title>
<![CDATA[
SimHumanity: Using SLiM 5.0 to run whole-genome simulations of human evolution 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.09.01.673541v1?rss=1</link>
<description><![CDATA[
The reconstruction of human evolutionary history has undergone repeated advances, each made possible by methodological innovations. In recent decades, genetic and genomic data played a central role in the reconstruction of major evolutionary events such as the out-of-Africa migration, and genetic simulations of human evolutionary history have come to play a major role in testing more specific hypotheses including proposed patterns of migration and admixture with archaic hominins. Increasing computational power has allowed human evolutionary history to be modeled at ever-larger scales, but simulations that encompass the complete human genome, including sex chromosomes and mitochondrial DNA, have been difficult due to the lack of support for whole-genome models in commonly used evolutionary simulation frameworks. With the recent introduction of SLiM 5 such simulations are now straightforward to construct, allowing the easy simulation of humans at whole-genome scale under different demographic models and evolutionary dynamics. We here present three versions of a reusable, customizable, open-source SLiM 5 model for simulating the molecular evolution of the full human genome. We also show some simple analyses of results from the model, to illustrate its utility. We hope this model, which we have nicknamed "SimHumanity" in jest, will facilitate further progress in the field of human evolutionary simulations.
]]></description>
<dc:creator>Haller, B. C.</dc:creator>
<dc:creator>Nelson, C. W.</dc:creator>
<dc:creator>Rodrigues, M. F.</dc:creator>
<dc:creator>Messer, P. W.</dc:creator>
<dc:date>2025-09-02</dc:date>
<dc:identifier>doi:10.1101/2025.09.01.673541</dc:identifier>
<dc:title><![CDATA[SimHumanity: Using SLiM 5.0 to run whole-genome simulations of human evolution]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-09-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.09.01.672585v1?rss=1">
<title>
<![CDATA[
Biological landscape of acute illness in children in sub-Saharan Africa and South Asia 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.09.01.672585v1?rss=1</link>
<description><![CDATA[
Childhood illnesses including pneumonia, diarrhoea and malaria are leading causes of hospitalisation and mortality in resource-limited settings. However, we lack understanding of whether systemic responses to such diverse clinical syndromes are shared or specific, how they are impacted by malnutrition and how they differ from well children. We performed multi-omic profiling of plasma proteins, and serum metabolites and lipids in acutely ill hospitalised and well children in sub-Saharan Africa and South Asia. Using network-based clustering and mixed-effects modelling, we identified common and syndrome-specific omics responses to acute illness. We found that malnutrition often modifies host responses to disease. Although the internal structure of individual omics modules was largely preserved between ill and well children, the interactions between these preserved modules were markedly reorganised during acute illness. Compared to well children, biological systems in hospitalised children were more interconnected, exhibiting denser cross-omics interactions. These findings reveal widespread multisystem mobilisation during paediatric acute illness, offer deeper mechanistic insights and highlight candidate pathways for therapeutic intervention in high-burden settings.
]]></description>
<dc:creator>Mudibo, E. O.</dc:creator>
<dc:creator>Sande, C.</dc:creator>
<dc:creator>Ngari, M. M.</dc:creator>
<dc:creator>Jenkins, B.</dc:creator>
<dc:creator>Singa, B. O.</dc:creator>
<dc:creator>Lancioni, C. L.</dc:creator>
<dc:creator>Diallo, A. H.</dc:creator>
<dc:creator>Mbale, E.</dc:creator>
<dc:creator>Mupere, E.</dc:creator>
<dc:creator>John, M.</dc:creator>
<dc:creator>Bamouni, R. M.</dc:creator>
<dc:creator>Prendergast, A. J.</dc:creator>
<dc:creator>McGrath, C. J.</dc:creator>
<dc:creator>Koulman, A.</dc:creator>
<dc:creator>Bandsma, R. H. J.</dc:creator>
<dc:creator>Tickell, K. D.</dc:creator>
<dc:creator>Walson, J. L.</dc:creator>
<dc:creator>Berkley, J. A.</dc:creator>
<dc:creator>Gonzales, G. B.</dc:creator>
<dc:creator>Njunge, J. M.</dc:creator>
<dc:date>2025-09-03</dc:date>
<dc:identifier>doi:10.1101/2025.09.01.672585</dc:identifier>
<dc:title><![CDATA[Biological landscape of acute illness in children in sub-Saharan Africa and South Asia]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-09-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.09.01.673500v1?rss=1">
<title>
<![CDATA[
Pancreatic Cancer Induces Population-Specific Switching of Myosin Isoforms and Discrete Activation of Cachexia Genes in Skeletal Muscle Myocytes 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.09.01.673500v1?rss=1</link>
<description><![CDATA[
Skeletal muscle loss in pancreatic cancer is a significant cause of morbidity and mortality for patients. In order to understand myocytes changes we examined myonuclei- and myofiber-specific dynamics during pancreatic cancer cachexia progression. Single-nucleus RNA-seq was used to interrogate myonuclear gene expression, and RNAscope and immunofluorescence characterized myofiber-specific changes. Bulk RNA-seq of skeletal muscle provided a whole-muscle transcriptomic profile. Cachexia induces a progressive loss of muscle differentiation factor Maf and its target Myh4, accompanied by increased expression of Myh1 and Myh2. This myofiber dedifferentiation occurs without evidence for fiber type shifting, regeneration, or proliferation. Single-nuclei analysis reveals global shifts in myofiber gene expression identity including the identification of a cachexia only myonuclear subpopulation. Cachexia gene expression was not restricted solely to this PDAC-specific myonuclear subpopulation and did not overlap with Myh1 and Myh2 expressing myonuclei early in cachexia. Altogether, PDAC cachexia elicits distinct transcriptional responses across different myonuclear populations. These results reveal population-specific heterogeneity in cachexia gene activation, rather than a uniform upregulation of cachexia mediators across muscle tissue. Our data suggest that myonuclei fate occurs prior to overt muscle wasting when cachexia gene expression only modestly overlaps with differentiation factors, with a strong association after irreversible muscle wasting. These findings explain the challenge of effectively targeting skeletal muscle wasting in cancer cachexia requires addressing the changing cell population induced through non overlapping mechanisms.
]]></description>
<dc:creator>Counts, B. R.</dc:creator>
<dc:creator>Jean, S.</dc:creator>
<dc:creator>Gaafer, O.</dc:creator>
<dc:creator>Ota, S. K.</dc:creator>
<dc:creator>Inabathini, I.</dc:creator>
<dc:creator>Guo, T.</dc:creator>
<dc:creator>Guttridge, D. C.</dc:creator>
<dc:creator>Ostrowski, M. C.</dc:creator>
<dc:creator>Koniaris, L. G.</dc:creator>
<dc:creator>Cao, S.</dc:creator>
<dc:creator>Roh, H. C.</dc:creator>
<dc:creator>Zimmers, T. A.</dc:creator>
<dc:date>2025-09-05</dc:date>
<dc:identifier>doi:10.1101/2025.09.01.673500</dc:identifier>
<dc:title><![CDATA[Pancreatic Cancer Induces Population-Specific Switching of Myosin Isoforms and Discrete Activation of Cachexia Genes in Skeletal Muscle Myocytes]]></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/10.1101/2025.09.02.673845v1?rss=1">
<title>
<![CDATA[
Clinical and molecular features of primary gliosarcoma with digital spatial whole-transcriptome analysis of glial and mesenchymal components 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.09.02.673845v1?rss=1</link>
<description><![CDATA[
Gliosarcoma is a rare subtype of IDH-wildtype glioblastoma defined by mixed malignant glial and high-grade sarcomatous histological elements. Gliosarcoma is clinically managed similarly to glioblastoma and has a poor clinical outcome. The sarcoma-like regions of gliosarcoma are thought to represent extreme mesenchymal metaplasia of neoplastic glial cells. Factors contributing to this phenomenon are not completely understood. Here we report a single-institution series of 37 gliosarcomas including next-generation sequencing data on 25 cases and digital spatial whole-transcriptome analysis on 4 cases to characterize differential gene expression between glial and mesenchymal components. Gliosarcoma demographic and genetic features were compared to a cohort of 75 primary adult hemispheric IDH-wildtype non-sarcomatous glioblastomas. Patient age, tumor location, sex, and overall survival in gliosarcoma were similar to glioblastoma. Gliosarcomas showed a significantly lower rate of EGFR amplification and a higher rate of NF1 mutation compared to glioblastomas in next-generation sequencing analysis. Digital spatial whole-transcriptome analysis showed a distinct transcriptomic profile in sarcomatous regions with over-expression of genes involved in extracellular matrix development and remodeling. Selected differentially expressed transcripts were examined further by immunohistochemistry. The glial elements of gliosarcomas showed higher immunoreactivity for Chitinase-3-like protein 1 (CHI3L1) than glioblastomas, but low to absent expression within the sarcomatous elements. Lymphoid Enhancer-Binding Factor 1 (LEF1) immunoreactivity was identified within sarcomatous regions of gliosarcoma without detectable nuclear {beta}-catenin, suggesting a role for {beta}-catenin independent wingless (WNT) effector signaling in sarcomatous transformation. This study adds to the growing literature demonstrating differences in the genetic underpinning of gliosarcoma and glioblastoma, establishes feasibility of spatial transcriptomic approaches in gliosarcoma, and validates digital spatial profiling-based results as a discovery platform to identify pathways and immunohistochemical markers for further study.
]]></description>
<dc:creator>Wood, M. D.</dc:creator>
<dc:creator>Zangirolani, G.</dc:creator>
<dc:creator>Lee, J.</dc:creator>
<dc:creator>Neff, T.</dc:creator>
<dc:creator>Zhang, K.</dc:creator>
<dc:creator>Corless, C. L.</dc:creator>
<dc:date>2025-09-07</dc:date>
<dc:identifier>doi:10.1101/2025.09.02.673845</dc:identifier>
<dc:title><![CDATA[Clinical and molecular features of primary gliosarcoma with digital spatial whole-transcriptome analysis of glial and mesenchymal components]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-09-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.09.06.674629v1?rss=1">
<title>
<![CDATA[
S3R: Spatially Smooth and Sparse Regression Reveals High-Dimensional Regulatory Networks in Spatial Transcriptomics 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.09.06.674629v1?rss=1</link>
<description><![CDATA[
Spatial transcriptomics (ST) data demands models that recover how associations among molecular and cellular features change across tissue while contending with noise, collinearity, cell mixing, and thousands of predictors. We present Spatially Smooth Sparse Regression (S3R), a general statistical framework that estimates location-specific coefficients linking a response feature to high-dimensional spatial predictors. S3R unites structured sparsity with a minimum-spanning-tree-guided smoothness penalty, yielding coefficient fields that are coherent within neighborhoods yet permit sharp boundaries. In synthetic data, S3R accurately recovers spatially varying effects, selects relevant predictors, and preserves known boundaries. Applied to Visium-based ST data, S3R recapitulates layer-specific target-TF associations in human dorsolateral prefrontal cortex with concordant layer-wise correlations in matched single-cell data. In acute Haemophilus ducreyi skin infection, S3R converts spot-level gene expression mixtures into cell type-attributed expression fields, revealing per-cell type spatial gradients, and improving concordance of spatially variable gene calls when tests are applied to these demixed fields. In pancreatic ductal adenocarcinoma, S3R builds cross-cell-type, cross-gene co-variation tensors that quantify cell-cell interaction strength at gene-pair resolution and nominate interacting genes whose pathway enrichments align with established stromal-epithelial and immune crosstalk. An efficient implementation scales to large assays, and on a Xenium-based breast cancer dataset, S3R delineates the contributions at gene-gene, local neighborhood, and global context-level to target gene expression. Because responses and predictors in S3R are user-defined, it could flexibly address diverse biological questions within a single, scalable, and interpretable regression framework.
]]></description>
<dc:creator>zhou, x.</dc:creator>
<dc:creator>dang, p.</dc:creator>
<dc:creator>Tang, H.</dc:creator>
<dc:creator>Peng, L. X.</dc:creator>
<dc:creator>Yeh, J. J.</dc:creator>
<dc:creator>Sears, R. C.</dc:creator>
<dc:creator>Neelon, B.</dc:creator>
<dc:creator>Zimmers, T.</dc:creator>
<dc:creator>Zhang, C.</dc:creator>
<dc:creator>Cao, S.</dc:creator>
<dc:date>2025-09-11</dc:date>
<dc:identifier>doi:10.1101/2025.09.06.674629</dc:identifier>
<dc:title><![CDATA[S3R: Spatially Smooth and Sparse Regression Reveals High-Dimensional Regulatory Networks in Spatial Transcriptomics]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-09-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.09.10.674685v1?rss=1">
<title>
<![CDATA[
Splicing neoepitope prediction is sensitive to methodological differences 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.09.10.674685v1?rss=1</link>
<description><![CDATA[
MotivationCancer-specific neoepitopes may arise from abnormal splicing in the transcriptomic landscape (alternative splicing neoepitopes, ASNs), leading to divergent proteins with high potential immunogenicity. Predicting candidate ASNs requires a series of algorithmic and parameter choices, but no previous study has investigated the consistency and interpretability of these choices.

ResultsWe apply two ASN prediction methods with 35 matched parameter sets to generate candidate ASNs for five breast and ovarian cancer samples from The Cancer Genome Atlas (TCGA). We find that: 1) junction 9-mer peptides generated by two similarly designed pipelines differed by, on average, 68.9% and 76.6% in the BRCA and OV cohorts, respectively, and putatively cancer-specific junction 9-mers from the two pipelines diverged further, by an average of 81.5 % in OV and 84.6 % in BRCA; 2) the most lenient filters in the BRCA cohort show the highest divergence, at 97 %; 3) the rate of mass spectrometry validation of ASNs protein presence in cells is dominated by the size of the input space; and 4) putatively cancer-specific ASNs found by the intersection of both pipelines can be validated when accounting for false discovery with an average of 1.74 and 284.4 candidates in the BRCA and OV cohorts, respectively. Taken together, these results highlight that ASN identification is fragile and difficult to reproduce across analysis platforms, with limited cross-pipeline overlap and strong dependence on parameter choices.

Availability and ImplementationPython research code and scripts are available for download at https://github.com/ratschlab/projects2020_ohsu.

Contactthompsre@ohsu.edu, raetsch@inf.ethz.ch
]]></description>
<dc:creator>Prelot, L.</dc:creator>
<dc:creator>David, J. K.</dc:creator>
<dc:creator>Lin, A.</dc:creator>
<dc:creator>Kahles, A.</dc:creator>
<dc:creator>Yurchikova, M.</dc:creator>
<dc:creator>Thompson, R. F.</dc:creator>
<dc:creator>Rätsch, G.</dc:creator>
<dc:date>2025-09-16</dc:date>
<dc:identifier>doi:10.1101/2025.09.10.674685</dc:identifier>
<dc:title><![CDATA[Splicing neoepitope prediction is sensitive to methodological differences]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-09-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.09.12.675849v1?rss=1">
<title>
<![CDATA[
Alcohol Dependence-Induced Astrocyte Immune Activation in the Nucleus Accumbens 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.09.12.675849v1?rss=1</link>
<description><![CDATA[
Astrocytes play many physiological roles in the brain including maintenance of brain homeostasis, modulation of synapse formation and function, and regulation of the blood brain barrier permeability. Upon brain exposure to noxious stimuli, astrocytes can become reactive and activate neuroimmune responses. The nucleus accumbens (NAc) is a critical region involved in reward processing and is integrally involved in the establishment and maintenance of alcohol (ethanol, EtOH) dependence. Here we used the chronic intermittent ethanol - two-bottle choice (CIE-2BC) drinking model to induce EtOH dependence in Aldh1l1-eGFP/Rpl10a mice, which allow the pull-down of astrocyte specific RNA using translating ribosome affinity purification (TRAP) procedure. NAc astrocyte translating RNA and bulk-tissue RNA was analyzed by RNA-Seq to identify genes altered by EtOH dependence or EtOH drinking in astrocytes and the bulk NAc. The number of differentially regulated genes was greater in the astrocyte-specific analysis compared to the bulk-tissue suggesting the cell-type specific approach enables greater resolution of the effects of EtOH. In the astrocyte-specific translatome of EtOH dependent animals, genes related to neuroimmune activation were highly enriched with overall activation of pathways related to interferon and interleukin signaling. In addition, pathways relating to oxidative stress and glutathione related responses were enriched in NAc astrocytes from dependent animals. In contrast, the astrocyte response to EtOH drinking identified pathways related to homeostatic changes. These findings highlight the profound immune activation in NAc astrocytes during EtOH dependence which are distinct from the response to lower levels of EtOH exposure. This study identifies astrocyte pathways and genes involved in the transition from alcohol drinking to alcohol dependence and identify potential novel cell type-specific targets that underlie vulnerability to alcohol use.
]]></description>
<dc:creator>Hashimoto, J. G.</dc:creator>
<dc:creator>Mangieri, R. A.</dc:creator>
<dc:creator>Roberts, A. J.</dc:creator>
<dc:creator>Lime, T.</dc:creator>
<dc:creator>Davis, B. A.</dc:creator>
<dc:creator>Carbone, L.</dc:creator>
<dc:creator>Roberto, M.</dc:creator>
<dc:creator>Guizzetti, M.</dc:creator>
<dc:date>2025-09-17</dc:date>
<dc:identifier>doi:10.1101/2025.09.12.675849</dc:identifier>
<dc:title><![CDATA[Alcohol Dependence-Induced Astrocyte Immune Activation in the Nucleus Accumbens]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-09-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.09.16.676549v1?rss=1">
<title>
<![CDATA[
Engineering NIR probes to enhance affinity and clinical workflow compatibility for prostate cancer imaging 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.09.16.676549v1?rss=1</link>
<description><![CDATA[
Positive surgical margins following radical prostatectomy increase the risk of biochemical recurrence and subsequent disease progression. Fluorescence guided surgery (FGS) using targeted contrast agents has shown clinical benefits for several cancer types. However, current prostate cancer targeted imaging probes exhibit long pharmacokinetic (PK) profiles, necessitating extended waiting periods or repeated hospital visits, limiting their integration into standard clinical workflow. To overcome this critical clinical compatibility challenge, we developed an innovative tri-compartment, chemistry-driven probe design strategy. Specifically, we developed a congeneric library of near infrared (NIR) water soluble fluorescent probes incorporating: (1) a glutamic acid-urea-lysine (EuK) ligand targeting prostate specific membrane antigen (PSMA); (2) a NIR heptamethine cyanine fluorophore optimized for enhanced PSMA binding via secondary binding sites interactions; and (3) distinct PK modulators residing outside the PSMA binding pocket to promote rapid off-target tissue clearance. While molecular docking scores, photophysical properties and live-cell staining results showed similar overall performance, probes bearing PK modulators produced stronger tumor-specific fluorescence in vivo than the control lacking a PK modulator. This effort enabled identification of a lead probe with robust tumor targeting and accelerated off-target clearance, providing optimal tumor-specific signal and contrast in a timeframe, fully compatible with robotic-assisted radical prostatectomy (RARP) timelines.
]]></description>
<dc:creator>Malankar, G. S.</dc:creator>
<dc:creator>Szafran, D. A.</dc:creator>
<dc:creator>Kumar, G.</dc:creator>
<dc:creator>Pacce, J.</dc:creator>
<dc:creator>Devereux, M.</dc:creator>
<dc:creator>Tao, K.</dc:creator>
<dc:creator>Gomes, M.</dc:creator>
<dc:creator>Greer, W. S.</dc:creator>
<dc:creator>Rounds, C. C.</dc:creator>
<dc:creator>Masillati, A. M.</dc:creator>
<dc:creator>Gergis, S.</dc:creator>
<dc:creator>Ledvina, H.</dc:creator>
<dc:creator>Wong, M. H.</dc:creator>
<dc:creator>Niedre, M. J.</dc:creator>
<dc:creator>Wang, L. G.</dc:creator>
<dc:creator>Gibbs, S. L.</dc:creator>
<dc:date>2025-09-18</dc:date>
<dc:identifier>doi:10.1101/2025.09.16.676549</dc:identifier>
<dc:title><![CDATA[Engineering NIR probes to enhance affinity and clinical workflow compatibility for prostate cancer imaging]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-09-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.09.18.676928v1?rss=1">
<title>
<![CDATA[
Temporal Requirement for Stearoyl-CoA Desaturase 1 in Oligodendrocyte Development but Not Myelin Maintenance 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.09.18.676928v1?rss=1</link>
<description><![CDATA[
Stearoyl-CoA desaturase 1 is a rate-limiting enzyme in monounsaturated fatty acid synthesis, which is crucial for membrane biosynthesis. Here we show an early requirement for Scd1 in oligodendroglial cells during developmental myelination. Using oligodendrocyte progenitor cell (OPC) specific conditional knockout model of Scd1, we observed a myelination delay during CNS development. Genetic ablation of OPC-specific Scd1 resulted in oligodendrocyte maturation delay and hypomyelination within forebrain white matter tracts and optic nerve. Interestingly, although expressed at high levels within the mature oligodendrocytes, Scd1 was dispensable in maintenance of oligodendrocytes and axonal myelination, as loss of mature oligodendrocyte specific Scd1 showed no effect on myelin maintenance or oligodendrocyte survival. Together, our results suggest that Scd1 function is temporally restricted to the developmental period when oligodendrocytes undergo differentiation and active myelination but becomes dispensable for maintaining established myelin.
]]></description>
<dc:creator>Farnum, I.</dc:creator>
<dc:creator>Kunkel, T. J.</dc:creator>
<dc:creator>Chavali, M.</dc:creator>
<dc:date>2025-09-19</dc:date>
<dc:identifier>doi:10.1101/2025.09.18.676928</dc:identifier>
<dc:title><![CDATA[Temporal Requirement for Stearoyl-CoA Desaturase 1 in Oligodendrocyte Development but Not Myelin Maintenance]]></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/10.1101/2025.09.19.677394v1?rss=1">
<title>
<![CDATA[
Kidney kallikrein-1 contributes to cleavage of gamma-ENaC in vivo 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.09.19.677394v1?rss=1</link>
<description><![CDATA[
The epithelial sodium channel (ENaC) is essential for sodium reabsorption and potassium homeostasis in the distal nephron, where its activity is controlled by mineralocorticoid signaling and downstream proteolytic processing of channel subunits. While cleavage of the {gamma}-ENaC subunit has been implicated in aldosterone-mediated sodium transport, the identity of mineralocorticoid receptor (MR)-regulated proteases responsible for this process remains uncertain. Here, we investigated the role of kallikrein-1 (encoded by Klk1), a serine protease expressed in the connecting tubule and cortical collecting duct (CNT/CCD), as a mediator of ENaC activation. Using CRISPR/Cas9, we generated a conditional Klk1-floxed allele and established mice with CNT/CCD-specific deletion of Klk1 by crossing with Calb1-Cre (CNT-Klk1-/-). On a low sodium, high potassium diet, CNT-Klk1-/- mice exhibited [~]85% less renal kallikrein-1 expression, yet maintained normal serum electrolytes, urinary potassium excretion, and aldosterone responses. Western blot analysis revealed significantly less cleavage of {gamma}-ENaC and -ENaC in CNT-Klk1-/- kidneys, accompanied by more total NCC abundance. Despite impaired ENaC proteolysis, amiloride-sensitive sodium excretion was preserved, indicating intact ENaC function. These findings identify renal kallikrein-1 as a protease that contributes to ENaC subunit processing in vivo. However, the absence of overt sodium or potassium handling defects in CNT-Klk1-/- mice suggests that kallikrein-1 deficiency is not sufficient to disrupt overall ENaC function, likely due to compensatory mechanisms from redundant proteolytic or non-proteolytic pathways. Together, our results refine the role of kallikrein-1 as a modulator, rather than a sole determinant, of ENaC activation and highlight the complexity of aldosterone-dependent sodium transport in the distal nephron.

New & NoteworthyUsing a novel connecting tubule / cortical collecting duct specific kallikrein-1 knockout model, we show that {gamma}- and -ENaC cleavage is impaired by loss of renal kallikrein-1 without major disturbances in sodium or potassium handling. These findings highlight redundancy among ENaC regulatory pathways and suggest that proteolytic cleavage, while biochemically evident, may not be an accurate marker of ENaC-mediated sodium transport under physiological stress.
]]></description>
<dc:creator>Curry, J. N.</dc:creator>
<dc:creator>Su, X.-T.</dc:creator>
<dc:creator>Wu, Q.</dc:creator>
<dc:creator>Maeoka, Y.</dc:creator>
<dc:creator>Yang, C.-L.</dc:creator>
<dc:creator>Delpire, E.</dc:creator>
<dc:creator>Fenton, R. F.</dc:creator>
<dc:creator>Welling, P. A.</dc:creator>
<dc:creator>Ellison, D. H.</dc:creator>
<dc:date>2025-09-21</dc:date>
<dc:identifier>doi:10.1101/2025.09.19.677394</dc:identifier>
<dc:title><![CDATA[Kidney kallikrein-1 contributes to cleavage of gamma-ENaC in vivo]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-09-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.09.24.678442v1?rss=1">
<title>
<![CDATA[
Neurotype matching in monogamous rodents is modulated by early-life sleep experience 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.09.24.678442v1?rss=1</link>
<description><![CDATA[
Studies of human sociability indicate stronger social affinity in matched-neurotype dyads (e.g., two individuals with autism or two without) compared to mixed-neurotype dyads (e.g., one individual with autism paired with one without). Is this neurotype matching phenomenon also quantifiable in non-human animals? Using deep learning tools, we analyzed dyadic male-female interactions in prairie voles, a highly social rodent species. To simulate "neurotypes", voles were exposed to either control conditions or early-life sleep disruption (ELSD) during a critical neurodevelopmental period (postnatal days 14-21), recapitulating two features of human autism: developmental sleep disruption and later-life atypical sociability. Analogous to human studies, voles showed signs of reduced social affinity in mixed dyads compared to matched dyads, including sex-specific changes in aggression and body orientation toward the conspecific. These findings advance our understanding of social affinity, providing a framework for new studies in both animal models and humans.
]]></description>
<dc:creator>Bueno-Junior, L. S.</dc:creator>
<dc:creator>Milman, N. E. P.</dc:creator>
<dc:creator>Jones-Tinsley, C. E.</dc:creator>
<dc:creator>Ghimire, A.</dc:creator>
<dc:creator>Wickham, P. T.</dc:creator>
<dc:creator>Hu, Y.</dc:creator>
<dc:creator>Ye, B.</dc:creator>
<dc:creator>Lim, M. M.</dc:creator>
<dc:creator>Watson, B. O.</dc:creator>
<dc:date>2025-09-25</dc:date>
<dc:identifier>doi:10.1101/2025.09.24.678442</dc:identifier>
<dc:title><![CDATA[Neurotype matching in monogamous rodents is modulated by early-life sleep experience]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-09-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.09.25.678663v1?rss=1">
<title>
<![CDATA[
A dNmnat-sensitized in vivo platform for unbiased discovery of regulators of neurodegeneration 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.09.25.678663v1?rss=1</link>
<description><![CDATA[
Axon degeneration drives nervous system dysfunction in diverse neurological diseases and injuries. While key regulators of injury-induced Wallerian degeneration have been identified, approaches have largely relied on axotomy, a particularly extreme injury for a neuron. Here, we developed a forward genetic screening platform in the adult Drosophila wing that sensitizes neurons to degeneration through depletion of nicotinamide mononucleotide adenylyltransferase (dNmnat), the essential NAD+ biosynthetic enzyme. We screened 9,393 mutagenized chromosomes and recovered 59 mutations that suppress neurodegeneration induced by dNmant depletion, including the known pro-degenerative molecule dSarm, thereby validating our approach. We show loss of CG4098, the Drosophila homolog of the mammalian enzyme NUDT9, robustly preserved axons and induced widespread remodeling of NAD+-related metabolites, which provides a new link between cADP-ribose hydrolase activity and neuronal survival. We further show mutations in the transcription factor Abrupt resulted in elevated dNmnat protein levels and blocked axon degeneration, suggesting Abrupt normally tunes dNmnat levels and neuronal NAD+ homeostasis. dNmnat depletion is thus a versatile approach for unbiased discovery of axon death pathway components, each of which may provide new entry points for therapeutic strategies to preserve axons in neurodegenerative diseases.
]]></description>
<dc:creator>Avetisyan, A.</dc:creator>
<dc:creator>Manzo, E.</dc:creator>
<dc:creator>Maturana, R. B.</dc:creator>
<dc:creator>Kocia, M.</dc:creator>
<dc:creator>Hsu, J.-M.</dc:creator>
<dc:creator>Larson, A.</dc:creator>
<dc:creator>Weirnick, R.</dc:creator>
<dc:creator>Balasubramanian, K.</dc:creator>
<dc:creator>Blackwell, N.</dc:creator>
<dc:creator>Freeman, M.</dc:creator>
<dc:creator>Bis, D. M.</dc:creator>
<dc:creator>Fazal, S.</dc:creator>
<dc:creator>Schuchner, S.</dc:creator>
<dc:creator>Neukomm, L.</dc:creator>
<dc:creator>Freeman, M.</dc:creator>
<dc:date>2025-09-25</dc:date>
<dc:identifier>doi:10.1101/2025.09.25.678663</dc:identifier>
<dc:title><![CDATA[A dNmnat-sensitized in vivo platform for unbiased discovery of regulators of neurodegeneration]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-09-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.09.26.678830v1?rss=1">
<title>
<![CDATA[
Oncostatin M orchestrates collective epithelial migration via HIF1A activation 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.09.26.678830v1?rss=1</link>
<description><![CDATA[
Extracellular signals strongly influence cell behavior, yet the mechanisms by which specific ligands mediate changes in phenotype remain unclear. The cytokine Oncostatin M (OSM) regulates homeostasis, wound healing, inflammation, and cancer progression. We previously found that OSM induces collective cell migration (CCM), a process where cells move as cohesive units while retaining cell-cell contacts, in MCF10A mammary epithelial cells. Here, we investigated how OSM drives CCM by comparing its effects with those elicited by epidermal growth factor (EGF) and interferon gamma (IFNG), defining ligand-specific phenotypes and molecular networks. Integrative transcriptomic and proteomic analyses identified hypoxia-inducible factor-1 (HIF1A) and signal transducer and activator of transcription 3 (STAT3) as central regulators of OSM responses. Functional validation revealed that HIF1A drives transcriptional programs associated with hypoxia, metabolic reprogramming, and immune pathways. Complement signaling emerged as a downstream effector of HIF1A, and its inhibition disrupted OSM-induced clustering and CCM. These findings establish a mechanistic link between OSM signaling, HIF1A activation, and CCM, demonstrating how cytokine-driven transcriptional reprogramming coordinates epithelial migration. Analysis of public breast cancer data suggests this pathway is active in human tumors and may contribute to tissue remodeling, repair, and metastasis.
]]></description>
<dc:creator>McLean, I. C.</dc:creator>
<dc:creator>Gross, S. M.</dc:creator>
<dc:creator>Liby, T. A.</dc:creator>
<dc:creator>Dane, M. A.</dc:creator>
<dc:creator>Derrick, D. S.</dc:creator>
<dc:creator>Heiser, L. M.</dc:creator>
<dc:date>2025-09-27</dc:date>
<dc:identifier>doi:10.1101/2025.09.26.678830</dc:identifier>
<dc:title><![CDATA[Oncostatin M orchestrates collective epithelial migration via HIF1A activation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-09-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.09.29.678242v1?rss=1">
<title>
<![CDATA[
Increased sensitivity to myopia and altered retinal ON/OFF balance in a mouse model lacking Dusp4 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.09.29.678242v1?rss=1</link>
<description><![CDATA[
Myopia, influenced by environmental and genetic factors, occurs when the emmetropization process fails to stop, causing excessive eyeball growth. Highly myopic animal models lacking a functional ON-pathway identified Dusp4 as a potential gene implicated in myopia. Here, we use a mouse model lacking DUSP4 to gain a better understanding of its retinal role and the mechanisms implicated in myopia development. Dusp4-/- mice have a reduced basal level of retinal dopamine and a higher susceptibility to lens-induced myopia. Dusp4 is expressed in ON-bipolar cells and a subset of OFF-bipolar cells in a light dependent manner. The absence of DUSP4 causes a hyperactivation of the MAPK/ERK pathway. Dusp4-/- mice show a reduced optomotor response, increased ON-bipolar cell responses, reduced oscillatory potentials together with altered OFF and ON-OFF RGC response to light flashes. These data provide new insights into retina-driven mechanisms of myopization, nuancing the impact of ON and OFF pathways upon emmetropization.
]]></description>
<dc:creator>Wilmet, B.</dc:creator>
<dc:creator>Michiels, C.</dc:creator>
<dc:creator>Zhang, J.</dc:creator>
<dc:creator>Louboutin, A.</dc:creator>
<dc:creator>Boranijasevic, S.</dc:creator>
<dc:creator>Frederiksen, H.</dc:creator>
<dc:creator>Callebert, J.</dc:creator>
<dc:creator>Varin, J.</dc:creator>
<dc:creator>Gimenez, M. L.</dc:creator>
<dc:creator>Morgans, C. W.</dc:creator>
<dc:creator>Duvoisin, R.</dc:creator>
<dc:creator>Horan, L.</dc:creator>
<dc:creator>Plevin, R.</dc:creator>
<dc:creator>Picaud, S.</dc:creator>
<dc:creator>Marre, O.</dc:creator>
<dc:creator>Audo, I.</dc:creator>
<dc:creator>Zeitz, C.</dc:creator>
<dc:date>2025-10-01</dc:date>
<dc:identifier>doi:10.1101/2025.09.29.678242</dc:identifier>
<dc:title><![CDATA[Increased sensitivity to myopia and altered retinal ON/OFF balance in a mouse model lacking Dusp4]]></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/10.1101/2025.09.29.678590v1?rss=1">
<title>
<![CDATA[
Endocannabinoid signaling is a critical link between circadian desynchronization and metabolic dysfunction 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.09.29.678590v1?rss=1</link>
<description><![CDATA[
It is well documented that disruption of circadian rhythms can cause metabolic dysregulation, but the specific mechanisms involved remain unclear. Our findings demonstrate that the negative metabolic effects of environmental circadian desynchronization (ECD) are dependent upon the cannabinoid receptor 1 (CB1r). The endocannabinoid system has not previously been implicated in mediating the effects of circadian disruption. We showed that ECD induced a positive correlation between the levels of the endocannabinoids AEA and 2-AG in both plasma and liver. While global CB1r knockout protects against the metabolic effects of ECD, behavioral and physiological response to ECD was strikingly similar between WT and CB1r KO mice and could not account for their distinct metabolic outcomes. Using liver-specific CB1r KO mice, we further specified that the ECD-induced metabolic hormone disruption, but not weight gain, is mediated through liver CB1r signaling. Finally, we showed that ECD upregulated transcription of genes involved in oxidative phosphorylation in the liver of WT, but not liver-specific CB1r KO mice. In summary, ECD led to modular metabolic dysfunction through CB1r signaling in multiple tissues, with the liver playing a critical role.
]]></description>
<dc:creator>Baca, B. A.</dc:creator>
<dc:creator>Denaroso, G. E.</dc:creator>
<dc:creator>Akli, S.</dc:creator>
<dc:creator>Pearson, G. L.</dc:creator>
<dc:creator>Wang, J.</dc:creator>
<dc:creator>Bowles, N. P.</dc:creator>
<dc:creator>Phillips, D.</dc:creator>
<dc:creator>Sorensen, W.</dc:creator>
<dc:creator>Hume, C.</dc:creator>
<dc:creator>Hill, M.</dc:creator>
<dc:creator>Karatsoreos, I. N.</dc:creator>
<dc:date>2025-10-01</dc:date>
<dc:identifier>doi:10.1101/2025.09.29.678590</dc:identifier>
<dc:title><![CDATA[Endocannabinoid signaling is a critical link between circadian desynchronization and metabolic dysfunction]]></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/10.1101/2025.09.30.679676v1?rss=1">
<title>
<![CDATA[
Bridging forward-in-time and coalescent simulations using pyslim 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.09.30.679676v1?rss=1</link>
<description><![CDATA[
Ancestral Recombination Graphs (ARGs) provide an expressive and compact way to represent genetic variation data generated by simulations embedded within its genealogical history, and it can dramatically speed up simulations. The fact that the ARG records genealogical as well as genomic information opens up the possibility for a number of new analysis and simulation techniques. Here, we aim to introduce the reader to this deep source of information as produced by the forwards simulator SLiM. SLiM records the ARG using the tree sequence format, which can be manipulated using the tskit and pyslim python packages. We first describe the information that SLiM records in the tree sequence, then provide several examples that use the tree sequence as a format to losslessly pass population states between simulators: recapitation of a forwards simulation with coalescent simulation; initiation of a forwards simulation using the results of a coalescent simulation; and parallelization of simulations, first across branches of a phylogenetic tree, and then across the populations of parasites infecting different hosts.
]]></description>
<dc:creator>Gopalan, S.</dc:creator>
<dc:creator>Rodrigues, M. F.</dc:creator>
<dc:creator>Ralph, P. L.</dc:creator>
<dc:creator>Haller, B. C.</dc:creator>
<dc:date>2025-10-01</dc:date>
<dc:identifier>doi:10.1101/2025.09.30.679676</dc:identifier>
<dc:title><![CDATA[Bridging forward-in-time and coalescent simulations using pyslim]]></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/10.1101/2025.09.30.679577v1?rss=1">
<title>
<![CDATA[
Chronic Ethanol Drinking Alters Medial Prefrontal Cortex and Nucleus Accumbens Astrocyte Translatome and Extracellular Matrix Glycosaminoglycans 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.09.30.679577v1?rss=1</link>
<description><![CDATA[
Alcohol Use Disorder is a leading preventable cause of morbidity and mortality, yet knowledge of mechanisms driving ethanol-related neuroplasticity remains incomplete. While research has traditionally focused on neuronal signaling, emerging evidence implicates astrocytes in addiction-related adaptations. Here, we investigated the astrocyte-specific molecular consequences of chronic ethanol consumption in the prefrontal cortex and nucleus accumbens, two brain regions critical for executive control and reward processing. Using Translating Ribosome Affinity Purification RNA-seq and bulk RNA-seq in Aldh1l1-EGFP/Rpl10a mice, expressing an EGFP tag on astrocyte ribosomes, we identified hundreds of differentially translated astrocytic genes following chronic continuous two-bottle choice ethanol drinking. Sex-specific analyses revealed greater astrocytic changes in the female PFC and male NAc. Pathway enrichment highlighted extracellular matrix remodeling, synaptic signaling, mitochondrial function, and immune-related pathways. Analyses of individual drinking levels further demonstrated distinct correlations between ethanol intake and astrocytic translation. The major components of the brain extracellular matrix are chondroitin sulfate proteoglycans, produced primarily by astrocytes and covalently bound to chondroitin sulfate glycosaminoglycan chains. Complementary mass spectrometry/liquid chromatography analyses of chondroitin sulfate, heparan sulfate, and hyaluronic acid glycosaminoglycan disaccharides revealed ethanol-induced alterations in chondroitin sulfate glycosaminoglycan sulfation patterns, with additional baseline differences identified between selectively bred high- and low-ethanol preference lines. Together, these findings indicate that astrocytes undergo profound sex- and region-specific adaptations to chronic ethanol, implicating extracellular matrix and glycosaminoglycan remodeling as key risk-factors for and mediators of chronic ethanol-related neuroplasticity.
]]></description>
<dc:creator>Hashimoto, J. G.</dc:creator>
<dc:creator>Ozburn, A. R.</dc:creator>
<dc:creator>Reed, C.</dc:creator>
<dc:creator>Erk, J.</dc:creator>
<dc:creator>Song, Y.</dc:creator>
<dc:creator>Yang, J.</dc:creator>
<dc:creator>Xia, K.</dc:creator>
<dc:creator>Zhang, F.</dc:creator>
<dc:creator>Yu, Y.</dc:creator>
<dc:creator>Fei, S. S.</dc:creator>
<dc:creator>Gao, L.</dc:creator>
<dc:creator>Linhardt, R. J.</dc:creator>
<dc:creator>Phillips, T. J.</dc:creator>
<dc:creator>Guizzetti, M.</dc:creator>
<dc:date>2025-10-02</dc:date>
<dc:identifier>doi:10.1101/2025.09.30.679577</dc:identifier>
<dc:title><![CDATA[Chronic Ethanol Drinking Alters Medial Prefrontal Cortex and Nucleus Accumbens Astrocyte Translatome and Extracellular Matrix Glycosaminoglycans]]></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/10.1101/2025.09.30.679624v1?rss=1">
<title>
<![CDATA[
Interferon Restores Antigen Presentation and Sensitizes Medulloblastoma to T Cell Killing 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.09.30.679624v1?rss=1</link>
<description><![CDATA[
Medulloblastomas are commonly considered immunologically cold and refractory to immunotherapy. One contributing factor to their low immunogenicity is impaired antigen presentation, which allows tumor cells to escape from cytotoxic T cells. Here we use a syngeneic mouse model of medulloblastoma to study the role of CD8+ T cells in medulloblastoma growth. We demonstrate that despite low expression of MHC Class I on tumor cells, depletion of CD8+ T cells accelerates tumor growth, whereas adoptive transfer of tumor-reactive CD8+ T cells prolongs survival. These anti-tumor effects rely on T cells secreting interferon gamma (IFN{gamma}), which induces MHC class I on tumor cells and facilitates tumor cell killing by T cells. Notably, this response is essential for CD8+ T cell-mediated tumor attack, as blocking IFN{gamma} signaling in vivo abrogates MHC class I induction and eliminates the beneficial effect of T cells. Importantly, delivering IFN{gamma} directly into tumors via convection-enhanced delivery (CED) enhances CD8+ T cell-mediated killing of tumor cells and significantly prolongs survival in tumor-bearing mice. These studies highlight the importance of T cells in controlling brain tumor growth and the value of IFN{gamma} as an adjuvant for T cell-based immunotherapy.
]]></description>
<dc:creator>Eisemann, T.</dc:creator>
<dc:creator>Masihi, M. B.</dc:creator>
<dc:creator>Tzaridis, T.</dc:creator>
<dc:creator>Pister, V.</dc:creator>
<dc:creator>Youm, I.</dc:creator>
<dc:creator>Chambers, K. R.</dc:creator>
<dc:creator>Dutta, A.</dc:creator>
<dc:creator>Wenzel, A. T.</dc:creator>
<dc:creator>Chin, K.</dc:creator>
<dc:creator>Pomeroy, S. L.</dc:creator>
<dc:creator>Mesirov, J. P.</dc:creator>
<dc:creator>Fraenkel, E.</dc:creator>
<dc:creator>Bagchi, A.</dc:creator>
<dc:creator>Chavez, L.</dc:creator>
<dc:creator>Wechsler-Reya, R. J.</dc:creator>
<dc:date>2025-10-02</dc:date>
<dc:identifier>doi:10.1101/2025.09.30.679624</dc:identifier>
<dc:title><![CDATA[Interferon Restores Antigen Presentation and Sensitizes Medulloblastoma to T Cell Killing]]></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/10.1101/2025.10.07.680309v1?rss=1">
<title>
<![CDATA[
Deveolopment of patient-derived neuroprogenitor cells (hNPCs), neurons, and astrocytes to explore the etiology of Guam Parkinsonism-dementia complex (PDC) 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.10.07.680309v1?rss=1</link>
<description><![CDATA[
Parkinsonism-Dementia Complex (PDC) is one phenotype of a disappearing neurodegenerative disease (Guam ALS-PDC) that shows clinical and neuropathological relationships with amyotrophic lateral sclerosis (ALS), atypical parkinsonism and Alzheimers disease. ALS-PDC has been linked with exposure to environmental factors (notably cycad plant neurotoxins), but evidence from human and animal studies is inconclusive. Patient-derived induced pluripotent stem cells (iPSCs) provide a powerful in vitro system to explore the underlying cause of PDC. iPSC lines were derived from lymphocytes of a PDC-affected Guamanian Chamorro female patient and an age- and gender-matched healthy Chamorro resident of PDC-unaffected Saipan using non-integrating episomal plasmids. iPSCs derived from both patients expressed pluripotency markers (Oct4, SSEA-4, TRA-1-60, Sox2) prior to the generation of neuroprogenitor cells (hNPCs), neurons and astrocytes. An embryoid body protocol was used to derive hNPCs from both iPSC lines while a differentiation media was used to generate neurons from hNPCs. hNPCs derived from both iPSC patients lines displayed established neuroprogenitor markers (nestin, Sox2), while the differentiated hNPCs exhibited both neuronal (beta-tubulin III, Map2, doublecortin) and synaptic (synaptophysin, PSD-95) markers. Expression of these protein markers in hNPCs and neurons by dot blotting was also observed for both lines. Astrocyte progenitor cells and mature astrocytes with appropriate markers were also developed from the hNPCs of both lines using commercial kits. Development of these patient-derived iPSCs provides a human model for evaluating the role of environmental (e.g., cycad toxins) and genetic factors in ALS-PDC and possibly other related neurodegenerative diseases.
]]></description>
<dc:creator>Chlebowski-Giulietti, A.</dc:creator>
<dc:creator>Yang, Y.</dc:creator>
<dc:creator>Siddique, N. A.</dc:creator>
<dc:creator>Siddique, T.</dc:creator>
<dc:creator>Spencer, P. S.</dc:creator>
<dc:creator>Steele, J. C.</dc:creator>
<dc:creator>KISBY, G. E.</dc:creator>
<dc:date>2025-10-07</dc:date>
<dc:identifier>doi:10.1101/2025.10.07.680309</dc:identifier>
<dc:title><![CDATA[Deveolopment of patient-derived neuroprogenitor cells (hNPCs), neurons, and astrocytes to explore the etiology of Guam Parkinsonism-dementia complex (PDC)]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-10-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.10.08.681215v1?rss=1">
<title>
<![CDATA[
A Rapid Gene Expression Profiler Classifies AML Tumor Responsiveness to Standard Therapies 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.10.08.681215v1?rss=1</link>
<description><![CDATA[
The emergence of transcriptional signatures that define cell types and pathways has made it possible to guide cancer therapy selection through gene expression profiling. We developed a rapid qPCR-based platform to profile cell state, stemness, and BCL2 family gene expression as a companion diagnostic test for acute myeloid leukemia (AML). We validated the stability and utility of the signatures across multiple measurement platforms and using patient samples from two centers. Integrating these signatures with clinical features enables an expedient means to predict the likelihood of patient responses to two standard-of-care therapies: intensive chemotherapy and hypomethylating agent plus venetoclax (HMA+Ven). For patients treated with HMA+Ven, expression levels of the promonocyte-like signature and BCL2 add predictive value for response and overall survival in multivariable models that include genetic features. The incorporation of the rapid profiler into the prospective evaluation of newly diagnosed AML patients may enhance treatment stratification and improve outcomes.
]]></description>
<dc:creator>Kurtz, S. E.</dc:creator>
<dc:creator>Eide, C. A.</dc:creator>
<dc:creator>Kaempf, A.</dc:creator>
<dc:creator>Long, N.</dc:creator>
<dc:creator>Miller, A.</dc:creator>
<dc:creator>Bottomly, D.</dc:creator>
<dc:creator>McWeeney, S.</dc:creator>
<dc:creator>Anand, S.</dc:creator>
<dc:creator>Ng, S. W.</dc:creator>
<dc:creator>Wang, J. C.</dc:creator>
<dc:creator>Dick, J. E.</dc:creator>
<dc:creator>Tyner, J. W.</dc:creator>
<dc:date>2025-10-09</dc:date>
<dc:identifier>doi:10.1101/2025.10.08.681215</dc:identifier>
<dc:title><![CDATA[A Rapid Gene Expression Profiler Classifies AML Tumor Responsiveness to Standard Therapies]]></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/10.1101/2025.10.09.681336v1?rss=1">
<title>
<![CDATA[
Spatial organization of the tumor-immune microenvironment in ER-positive breast cancer: remodeling during treatment and associations with clinical response 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.10.09.681336v1?rss=1</link>
<description><![CDATA[
BackgroundThe tumor microenvironment influences treatment response in ER-positive breast cancer, but what distinguishes responders from non-responders and how it changes during treatment is poorly understood.

MethodsER-positive breast tumors treated with neoadjuvant chemotherapy with or without bevacizumab were profiled with bulk proteomics pre-(n = 95), on-(n = 84) and post-treatment (n = 100). A subset of tumors was profiled with spatial single-cell proteomics pre-(n = 13) and on-treatment (n = 11). Cell phenotypes, spatial location and activation states were determined, and cellular colocalization assessed with spatial metrics. Bulk and spatial features were evaluated against treatment response defined by residual cancer burden.

ResultsTreatment with bevacizumab amplified chemotherapy effects on proteomic signaling. The immune contexture shifted from suppressive to supportive during treatment through decreased macrophage, regulatory and anergic T-cell density and increased colocalization between epithelial cells and CD8+, CD4+ T-cells and dendritic cells. At baseline, responders had high density of effector memory T-cells, while non-responders had more naive T-cells. In addition, responders had increased colocalization of epithelial cells with macrophages, and effector memory T-cells with M1-like macrophages compared to non-responders.

ConclusionsSpatially distinct tumor-immune microenvironments influence response to neoadjuvant treatment, offering valuable insights for guiding treatment decisions.
]]></description>
<dc:creator>Dahle, M. A.</dc:creator>
<dc:creator>Foerde, J.-L.</dc:creator>
<dc:creator>Egeland, E. V.</dc:creator>
<dc:creator>Creason, A. L.</dc:creator>
<dc:creator>Watson, C.</dc:creator>
<dc:creator>Garred, O.</dc:creator>
<dc:creator>Prasmickaite, L.</dc:creator>
<dc:creator>Maelandsmo, G. M.</dc:creator>
<dc:creator>Mills, G. B.</dc:creator>
<dc:creator>Engebraaten, O.</dc:creator>
<dc:creator>Haugen, M. H.</dc:creator>
<dc:date>2025-10-10</dc:date>
<dc:identifier>doi:10.1101/2025.10.09.681336</dc:identifier>
<dc:title><![CDATA[Spatial organization of the tumor-immune microenvironment in ER-positive breast cancer: remodeling during treatment and associations with clinical response]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-10-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.10.09.681353v1?rss=1">
<title>
<![CDATA[
BCG Vaccination at Birth Shapes the TCR Usage and Functional Profile of MR1T Cells at 9 Weeks of Age 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.10.09.681353v1?rss=1</link>
<description><![CDATA[
Tuberculosis (TB) is the leading infectious disease killer worldwide and children suffer disproportionately. The ongoing burden of disease, despite widespread vaccination with BCG, highlights the need for novel vaccines. MR1-restricted T (MR1T) cells recognize small molecules, including microbial-derived molecules, presented by the monomorphic MHC class 1- related molecule (MR1). They have both "innate" effector capacity, allowing them to quickly respond to pathogens including Mycobacterium tuberculosis (Mtb), while also having adaptive features (effector memory cell surface phenotype and selective TCR usage). Feasibility of an MR1T cell-based vaccination remains unexplored and critical to this is whether or not MR1T cells possess the capacity for immunological memory. To begin to address this question, peripheral blood mononuclear cells (PBMC) were collected at 9-weeks of age from healthy term- infants in South Africa, who had either received BCG vaccination at birth (n=10) or who had BCG vaccination delayed (n=10). MR1/5-OP-RU tetramer positive cells were sorted using flow cytometry and single-cell RNA and TCR-seq was performed. Ex-vivo MR1T cells from vaccinated infants demonstrated increased expression of type I interferon response genes consistent with a cytokine mediated response to BCG vaccination. Using the TCR clustering algorithm TCRdist3, similar TCRs were grouped together, revealing a cluster significantly enriched in BCG-vaccinated infants. This cluster exhibited elevated expression of pro- inflammatory and cytotoxic genes, consistent with a recall response to prior vaccination and evidence of possible recognition of Mtb. This work provides the first step in addressing if MR1T cells demonstrate immunological memory, however, further work is needed to understand if these clonal expansions persist and possess the capacity for antigenic recall.
]]></description>
<dc:creator>Kain, D.</dc:creator>
<dc:creator>McElfresh, G.</dc:creator>
<dc:creator>Swarbrick, G.</dc:creator>
<dc:creator>Rott, K.</dc:creator>
<dc:creator>Boggy, G.</dc:creator>
<dc:creator>Walzl, G.</dc:creator>
<dc:creator>Du Plessis, N.</dc:creator>
<dc:creator>Hanekom, W.</dc:creator>
<dc:creator>Nemes, E.</dc:creator>
<dc:creator>Shey, M.</dc:creator>
<dc:creator>Scriba, T.</dc:creator>
<dc:creator>Bimber, B. N.</dc:creator>
<dc:creator>Lewinsohn, D.</dc:creator>
<dc:creator>Lewinsohn, D.</dc:creator>
<dc:date>2025-10-10</dc:date>
<dc:identifier>doi:10.1101/2025.10.09.681353</dc:identifier>
<dc:title><![CDATA[BCG Vaccination at Birth Shapes the TCR Usage and Functional Profile of MR1T Cells at 9 Weeks of Age]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-10-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.10.14.682328v1?rss=1">
<title>
<![CDATA[
Prognostic and Therapeutic Implications of BRAF Mutations in Acute Myeloid Leukemia 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.10.14.682328v1?rss=1</link>
<description><![CDATA[
Mutations in the RAS/MAPK signaling pathway are recurrent in acute myeloid leukemia (AML), primarily involving NRAS and KRAS. In contrast, mutations in the gene encoding an effector protein, BRAF, occur at relatively lower frequencies in AML and are associated with poor outcomes. To date, no comprehensive analysis has assessed the clinical and molecular characteristics of BRAF-mutated AML. In this study, we report the identification of canonical and non-canonical BRAF mutations in [~]1% of 5,779 consecutive clinically and molecularly fully-annotated AML patients treated at two major United States Cancer Centers (50/5779 AML patients: 21 newly diagnosed AML; 9 relapsed/refractory; 20 newly diagnosed secondary AML). We performed single-cell multiomic analysis on a subset of AML samples. BRAF mutations were enriched in myelodysplasia-related AML (AML-MR), and most mutations were located outside the V600 hotspot. Single-cell multiomic profiling delineated BRAF mutation class-specific patterns of co-mutations, clonality, and immunophenotypes. Notably, BRAF mutations and other signaling co-mutation(s) could be found in the same cell, a finding that significantly diverges from prior studies of RAS-mutant AML. In this cohort, BRAF-mutant AML patients had poor overall survival with currently available treatments, including venetoclax-based regimens. Drug sensitivity data suggest possible avenues for targeted treatment of BRAF-mutated AML.

Statement of SignificanceCanonical and non-canonical BRAF mutations are enriched in AML-MR and associate with poor survival outcomes. Single-cell multiomic profiling revealed unique co-mutation patterns and immunophenotypes that highlight RAS pathway addiction and nominate BRAF-mutated disease as a distinct subtype within RAS pathway-aberrated leukemias. Drug sensitivity screens suggest broad CDK or HSP90 inhibition in addition to BRAF/RAS-directed inhibition may be effective targeted therapies in this prognostically poor AML subtype.
]]></description>
<dc:creator>Lee, D.</dc:creator>
<dc:creator>Abu-Shihab, Y.</dc:creator>
<dc:creator>Plas, K.</dc:creator>
<dc:creator>Nicolet, D.</dc:creator>
<dc:creator>Mrozek, K.</dc:creator>
<dc:creator>Routbort, M. J.</dc:creator>
<dc:creator>Patel, K. P.</dc:creator>
<dc:creator>Walker, C. J.</dc:creator>
<dc:creator>Buss, J.</dc:creator>
<dc:creator>Stiff, A. R.</dc:creator>
<dc:creator>Laganson, A.</dc:creator>
<dc:creator>DiNardo, C. D.</dc:creator>
<dc:creator>Daver, N. G.</dc:creator>
<dc:creator>Kadia, T. M.</dc:creator>
<dc:creator>Ravandi, F.</dc:creator>
<dc:creator>Carroll, A. J.</dc:creator>
<dc:creator>Kolitz, J. E.</dc:creator>
<dc:creator>Powell, B. L.</dc:creator>
<dc:creator>Blum, W. G.</dc:creator>
<dc:creator>Baer, M. R.</dc:creator>
<dc:creator>Marcucci, G.</dc:creator>
<dc:creator>Uy, G. L.</dc:creator>
<dc:creator>Stock, W.</dc:creator>
<dc:creator>Stone, R. M.</dc:creator>
<dc:creator>Medeiros, L. J.</dc:creator>
<dc:creator>Byrd, J. C.</dc:creator>
<dc:creator>Blachly, J. S.</dc:creator>
<dc:creator>Bowman, R. L.</dc:creator>
<dc:creator>Tyner, J.</dc:creator>
<dc:creator>Loghavi, S.</dc:creator>
<dc:creator>Eisfeld, A.-K.</dc:creator>
<dc:creator>Miles, L. A.</dc:creator>
<dc:date>2025-10-15</dc:date>
<dc:identifier>doi:10.1101/2025.10.14.682328</dc:identifier>
<dc:title><![CDATA[Prognostic and Therapeutic Implications of BRAF Mutations in Acute Myeloid Leukemia]]></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/10.1101/2025.10.16.682761v1?rss=1">
<title>
<![CDATA[
A Zika Virus-Like Particle Vaccine Mitigates Early Pregnancy Loss In Rhesus Macaques 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.10.16.682761v1?rss=1</link>
<description><![CDATA[
Zika virus (ZIKV) is an arthropod-borne Orthoflavivirus that caused a major outbreak in the Americas in 2015-16. In Brazil, up to 46% of ZIKV positive pregnancies resulted in congenital Zika syndrome (CZS). CZS is characterized by a wide range of neurologic birth defects and miscarriage in up to 7.6% of affected pregnancies. With no current licensed ZIKV vaccines, we sought to evaluate a Zika virus-like particle (VLP) vaccine candidate in a rhesus macaque (RM) pregnancy model. VLPs were produced in mammalian cells expressing the pre-membrane-envelope region of the Asian-lineage ZIKV strain PRVABC59, which belongs to the Asian ZIKV lineage that is associated with outbreaks of congenital disease. To evaluate vaccine protection against adverse pregnancy complications, two cohorts of female RM were vaccinated with ZIKV-VLP with adjuvant Alhydrogel (alum) or adjuvant alone prior to mating. At gestational day (GD) 30 (early first trimester), pregnant animals were challenged with ZIKV-DAK 41524, an African-lineage strain shown to induce 1st-trimester fetal demise in 78% (n=11/14 animals) of RM, making it an ideal and stringent model for evaluating ZIKV vaccines. Within the vaccinated cohort, 2 of 3 animals reached the study endpoint of GD 90 with no observed adverse pregnancy outcomes. The third animal experienced pregnancy loss at GD 49 (18 d post infection), although no infectious virus was detected in placental or fetal tissues. In the unvaccinated cohort, two animals had severe adverse events. One animal experienced preterm labor, and another developed early-onset hydrops fetalis with widespread ZIKV-RNA detected via RNAscope and extensive placental damage. These results confirm a significant risk for early pregnancy loss in RM infected with ZIKV-DAK 41524. This model can be further used to understand the complexities of placental immunological features underlying stillbirth and miscarriage following infection. Our findings indicate that this ZIKV-VLP vaccine candidate protected pregnant macaques against fetal demise associated with highly pathogenic ZIKV challenge.

Author SummaryZika virus (ZIKV) infection during pregnancy is associated with pregnancy loss, severe birth defects, including microcephaly and developmental delays, and other subtle neurologic changes. Although vaccine development efforts have been ongoing since the 2015/2016 ZIKV outbreak, no approved vaccine is currently available. Many existing studies have tested vaccines in animal models using strains such as ZIKV-PR that cause mild or moderate pregnancy complications at similar rates to human cases. Using challenge strains that only cause mild, or moderate pregnancy complications makes it difficult to rigorously evaluate vaccine efficacy. In this study, we tested a virus-like particle (VLP) vaccine, a safe and effective method for use during pregnancy as it is replication-deficient and only contains viral structural antigens. We found that the VLP vaccine, when paired with an adjuvant (alum), induced strong antibody responses in mice and controlled viral dissemination following challenge in nonpregnant macaques. To evaluate the protective efficacy of the ZIKV-VLP vaccine against in utero infection and disease, we used a stringent model of ZIKV infection during early pregnancy in rhesus macaques that is associated with high rates of fetal demise. In pregnant macaques, the vaccine reduced maternal viremia, limited viral spread to fetal and placental tissues, and conferred protection against virus-mediated fetal demise and placental damage. This is the first study to evaluate a VLP-based vaccine in a consistent pregnancy loss model of ZIKV infection. These findings support the continued development of VLP vaccines as a safe and effective strategy for protecting pregnant individuals and their developing fetuses from ZIKV.
]]></description>
<dc:creator>Jaeger, H. K.</dc:creator>
<dc:creator>Smith, J. L.</dc:creator>
<dc:creator>Labriola, C. S.</dc:creator>
<dc:creator>Pung, L. J.</dc:creator>
<dc:creator>Hagen, O. L.</dc:creator>
<dc:creator>Denton, M.</dc:creator>
<dc:creator>D'Mello, R. J.</dc:creator>
<dc:creator>Parkins, C. J.</dc:creator>
<dc:creator>Weber, W. C.</dc:creator>
<dc:creator>Medica, S.</dc:creator>
<dc:creator>Kreklywich, C. N.</dc:creator>
<dc:creator>DeFilippis, V. R.</dc:creator>
<dc:creator>Bondoc, S.</dc:creator>
<dc:creator>Busman-Sahay, K.</dc:creator>
<dc:creator>Castro, J. N.</dc:creator>
<dc:creator>Zilverberg, G.</dc:creator>
<dc:creator>White, R.</dc:creator>
<dc:creator>Terry, M.</dc:creator>
<dc:creator>Barber-Axthelm, A. M.</dc:creator>
<dc:creator>Axthelm, M. K.</dc:creator>
<dc:creator>Smedley, J.</dc:creator>
<dc:creator>Aliota, M. T.</dc:creator>
<dc:creator>Weiler, A. M.</dc:creator>
<dc:creator>Friedrich, T. C.</dc:creator>
<dc:creator>Estes, J.</dc:creator>
<dc:creator>Morgan, T. K.</dc:creator>
<dc:creator>Lo, J. O.</dc:creator>
<dc:creator>Roberts, V. H. J.</dc:creator>
<dc:creator>Streblow, D. N.</dc:creator>
<dc:creator>Hirsch, A. J.</dc:creator>
<dc:date>2025-10-16</dc:date>
<dc:identifier>doi:10.1101/2025.10.16.682761</dc:identifier>
<dc:title><![CDATA[A Zika Virus-Like Particle Vaccine Mitigates Early Pregnancy Loss In Rhesus Macaques]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-10-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.10.21.683745v1?rss=1">
<title>
<![CDATA[
Tumor-intrinsic MHC-II activation in pancreatic ductal adenocarcinoma enhances immune response and treatment efficacy 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.10.21.683745v1?rss=1</link>
<description><![CDATA[
Pancreatic ductal adenocarcinoma (PDAC) is characterized by an immunosuppressive tumor microenvironment (TME) and poor prognosis. While major histocompatibility complex class II (MHC-II) expression is traditionally associated with professional antigen-presenting cells, its role in PDAC malignant cells remains underexplored. Herein, we utilized single-cell RNA sequencing (scRNA-seq), spatial transcriptomics, bulk RNA sequencing, multiplex immunohistochemistry (mIHC) and ex vivo studies in culture with both human and murine models to investigate the prognostic relevance of MHC-II expression in malignant PDAC cells. Elevated MHC-II expression in malignant cells was strongly associated with increased infiltration of CD4+ T and CD8+ T cells in human PDAC, and pronounced co-localization with plasma cells, indicative of an antigen-activated immune microenvironment. In the KPC mouse model of PDAC, pharmacologic induction of MHC-II expression by cobimetinib treatment in malignant epithelial cells significantly enhanced the therapeutic response to immune checkpoint blockade (ICB). These findings highlight the role of malignant cell- intrinsic MHC-II expression in promoting antigen presentation and fostering an anti-tumor immune microenvironment. Our results position MHC-II as a promising prognostic biomarker and therapeutic target in PDAC, paving the way for novel immunomodulatory strategies.

Summary of highlightsSingle-cell and spatial transcriptomic analyses reveal that elevated MHC-II expression in malignant PDAC cells correlates with increased infiltration of CD4 and CD8 T cells.

Stimulating MHC-II expression in tumors effectively enhances immunotherapeutic responses to ICB in the PDAC KPC mouse model, including PDAC tumors previously resistant to therapeutic interventions.

MHC-II serves as a prognostic biomarker and a promising target for immunotherapy in PDAC.
]]></description>
<dc:creator>Chen, C.</dc:creator>
<dc:creator>Gribbin, K.</dc:creator>
<dc:creator>Li, X.</dc:creator>
<dc:creator>Ozmen, T.</dc:creator>
<dc:creator>Ozmen, F.</dc:creator>
<dc:creator>Sivagnanam, S.</dc:creator>
<dc:creator>Kim, J.</dc:creator>
<dc:creator>Blise, K.</dc:creator>
<dc:creator>Yang, X.</dc:creator>
<dc:creator>Zhang, Y.</dc:creator>
<dc:creator>Dai, R.</dc:creator>
<dc:creator>Keith, D.</dc:creator>
<dc:creator>Sherman, M.</dc:creator>
<dc:creator>Dai, M.-s.</dc:creator>
<dc:creator>Coussens, L.</dc:creator>
<dc:creator>Lopez, C.</dc:creator>
<dc:creator>Sears, R.</dc:creator>
<dc:creator>Mills, G.</dc:creator>
<dc:creator>Byrne, K.</dc:creator>
<dc:creator>Xia, Z.</dc:creator>
<dc:date>2025-10-22</dc:date>
<dc:identifier>doi:10.1101/2025.10.21.683745</dc:identifier>
<dc:title><![CDATA[Tumor-intrinsic MHC-II activation in pancreatic ductal adenocarcinoma enhances immune response and treatment efficacy]]></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/10.1101/2025.10.23.683697v1?rss=1">
<title>
<![CDATA[
Neuronal architecture of the mouse insular cortex underlying its diverse functions 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.10.23.683697v1?rss=1</link>
<description><![CDATA[
The insular cortex integrates interoceptive and exteroceptive information to mediate bodily homeostasis, emotion, learning, and potentially consciousness.1-4 However, the cellular and circuit substrates governing the insula and other associative cortices are poorly understood compared to primary cortices. Here, we quantify the dendritic morphology together with electrical properties, local inputs, and/or projections of 1,093 insular pyramidal neurons. These neurons are mapped onto a quantitative anatomical model of the insula based on a Nissl-staining coordinate framework. Using improved algorithms, we define 21 morphological, 12 electrical, and 9 input neuronal types, and identify several morphological and input types that are unique to the insula. Further, we find that morphological properties constrain and often predict inputs, electrical properties, or projection targets. Several morphological types are differentially distributed between the functionally distinct anterior and posterior insula, providing the substrates for a quantitative demarcation between the anterior and posterior insular subregions. Surprisingly, certain neuronal types receive intra-insular inputs originating far beyond canonical cortical columns. Functionally, these connections bridge a long-range thalamus-to-amygdalar circuit that potentially links sensory information to valence. Our work establishes a structure-and-function foundation for investigating the insular cortex.
]]></description>
<dc:creator>Jongbloets, B. C.</dc:creator>
<dc:creator>Chen, Y.</dc:creator>
<dc:creator>Gingerich, I. K.</dc:creator>
<dc:creator>Muniak, M. A.</dc:creator>
<dc:creator>Mao, T.</dc:creator>
<dc:date>2025-10-24</dc:date>
<dc:identifier>doi:10.1101/2025.10.23.683697</dc:identifier>
<dc:title><![CDATA[Neuronal architecture of the mouse insular cortex underlying its diverse functions]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-10-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.10.24.684446v1?rss=1">
<title>
<![CDATA[
GRASS-NB: Group-structured variable selection for spatial negative binomial data with applications to cancer registry and spatial omics 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.10.24.684446v1?rss=1</link>
<description><![CDATA[
Spatially structured, overdispersed count data with high-dimensional predictors are increasingly observed across studies from population-level epidemiology to cellular-level spatial omics. Feature selection is critical to identify influential predictors, such as key risk factors or biomarkers. Few Bayesian studies have assessed negative binomial regression (NBR) models with standard variable selection priors, like the mixture spike-and-slab (SS) or continuous horseshoe (HS), but mostly under aspatial settings. Features often form groups; for instance, in population surveys, caloric intake and physical activity may fall under "Diet & Exercise", while cigarette use and smoking laws belong to "Smoking". We propose a flexible NBR model that accommodates spatial autocorrelation and introduces a novel group-structured prior by hybridizing SS and HS shrinkage. The models performance with different priors is evaluated in terms of specificity, precision, and computational cost under challenging scenarios, including "large p, small n" cases. We further apply the model to CDC state-level cancer data, comprising demographic, screening, and behavioral covariates, to identify key drivers and population-level risk factors, and to a melanoma spatial omics dataset for predictive modeling expression of gene. An efficient R package is provided on GitHub.
]]></description>
<dc:creator>Mattila, C.</dc:creator>
<dc:creator>Neelon, B.</dc:creator>
<dc:creator>Sonawane, K.</dc:creator>
<dc:creator>Cao, S.</dc:creator>
<dc:creator>Angel, P.</dc:creator>
<dc:creator>Hill, E.</dc:creator>
<dc:creator>Seal, S.</dc:creator>
<dc:date>2025-10-25</dc:date>
<dc:identifier>doi:10.1101/2025.10.24.684446</dc:identifier>
<dc:title><![CDATA[GRASS-NB: Group-structured variable selection for spatial negative binomial data with applications to cancer registry and spatial omics]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-10-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.10.23.684176v1?rss=1">
<title>
<![CDATA[
Loss of cell cycle gatekeeping by 1 CNOT3 impairs hematopoietic stem and progenitor cell division and repopulating activity 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.10.23.684176v1?rss=1</link>
<description><![CDATA[
Adult mammalian hematopoietic stem cells (HSCs) constitute a heterogeneous population responsible for generating various cell types in the blood throughout adulthood. Gene expression programs underlying regulation of self-renewal and differentiation of HSCs are tightly regulated. However, how post-transcriptional regulation of gene expression influences HSCs and hematopoiesis remains largely unexplored. Here, we report the critical role of CNOT3, a subunit of the CCR4-NOT complex, in regulating hematopoietic stem cells (HSCs) function in adult hematopoiesis. We observed that Cnot3 mRNA is highly expressed in HSCs and CNOT3 ablation in the murine Cnot3 conditional knockout mouse model resulted in anemia, reduced bone marrow cellularity and enhanced extramedullary hematopoiesis in spleen. Deletion of Cnot3 resulted in the early expansion of immunophenotypic HSCs which were then progressively lost over time. Cnot3 knockout hematopoietic stem/progenitor cells (HSPCs) failed to reconstitute hematopoietic systems of recipient animals in transplantation assays. Single-cell RNA sequencing (scRNA-seq) analysis of HSPCs revealed disruptions in lineage development and loss of HSCs. Transcriptomic profiling and cell cycle analysis demonstrated that Cnot3 deletion led to increased cycling activity in HSCs. Our results indicate that CNOT3 is critical for maintenance of homeostasis in HSCs and the hematopoietic system.
]]></description>
<dc:creator>Vu, L. P.</dc:creator>
<dc:creator>Yuen, K. A.</dc:creator>
<dc:creator>Taibi, T.</dc:creator>
<dc:creator>Hoang, Q. A.</dc:creator>
<dc:creator>Yue, M.</dc:creator>
<dc:creator>Liu, Z.</dc:creator>
<dc:creator>Habibi, E.</dc:creator>
<dc:creator>Spiliotopoulos, E.</dc:creator>
<dc:creator>Ramkumar, N.</dc:creator>
<dc:creator>Nation, N.</dc:creator>
<dc:creator>Edin, G.</dc:creator>
<dc:creator>Ngo, T. T.</dc:creator>
<dc:date>2025-10-26</dc:date>
<dc:identifier>doi:10.1101/2025.10.23.684176</dc:identifier>
<dc:title><![CDATA[Loss of cell cycle gatekeeping by 1 CNOT3 impairs hematopoietic stem and progenitor cell division and repopulating activity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-10-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.10.28.685173v1?rss=1">
<title>
<![CDATA[
Brain insulin signaling restores deficits in striatal dopamine release in overweight male mice with preexisting low D2-receptor expression 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.10.28.685173v1?rss=1</link>
<description><![CDATA[
Obesity is characterized by insulin resistance, motivational impairments, and, in some cases, reduced availability of dopamine D2 receptors in the brain. However, whether the low D2 receptor levels represent a predisposing factor or a consequence of obesity, and how these processes are mechanistically linked, remains unclear. Here, we directly tested this causal relationship by selectively reducing D2 receptor density in striatal neurons. Male, but not female, mice with a low density of striatal D2 receptors consumed more food, gained more weight, and developed metabolic features of peripheral insulin resistance despite being maintained on standard chow. Motivational deficits preceded weight gain, manifesting as delayed circadian locomotor onset, reduced physical activity, and diminished effort to obtain food. In the brain, male mice with low D2 receptor density showed reduced dopamine release capacity and age-dependent alterations in brain insulin sensitivity. Prior to weight gain, brain insulin responses were blunted compared to those of controls, in which insulin potentiates dopamine release and enhances striatal acetylcholine signaling. Once overweight, however, these mice exhibited brain insulin hypersensitivity, with insulin strongly restoring dopamine release capacity. Together, these findings demonstrate that low striatal D2 receptor density predisposes male mice to an obesity-like phenotype through early dopaminergic dysfunction that precedes weight gain and is later compensated by insulin hypersensitivity in the brain.
]]></description>
<dc:creator>Bocarsly, M. E.</dc:creator>
<dc:creator>Mehr, J. B.</dc:creator>
<dc:creator>Swanson, E. S.</dc:creator>
<dc:creator>Sriramoji-Virdi, S.</dc:creator>
<dc:creator>Authement, M. E.</dc:creator>
<dc:creator>Shashikiran, S.</dc:creator>
<dc:creator>Goldbach, H.</dc:creator>
<dc:creator>Matsui, A.</dc:creator>
<dc:creator>Rimondini, R.</dc:creator>
<dc:creator>Bock, R.</dc:creator>
<dc:creator>Alvarez, V. A.</dc:creator>
<dc:date>2025-10-30</dc:date>
<dc:identifier>doi:10.1101/2025.10.28.685173</dc:identifier>
<dc:title><![CDATA[Brain insulin signaling restores deficits in striatal dopamine release in overweight male mice with preexisting low D2-receptor expression]]></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/10.1101/2025.10.29.685365v1?rss=1">
<title>
<![CDATA[
Vaginal epithelial estrogen receptor α coordinates glycogen deposition, microbial stability, and pH regulation in mice 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.10.29.685365v1?rss=1</link>
<description><![CDATA[
Estrogen plays a central role in regulating the vaginal environment, but the specific contribution of epithelial estrogen receptor  (ESR1) to microbial and biochemical homeostasis has not been fully defined. In our previous work, we showed that epithelial ESR1 is indispensable for estrogen-induced epithelial proliferation, cornification, and MUC1 expression. Here, using mice with conditional deletion of Esr1 in vaginal epithelial cells, called epithelial Esr1d/d, we extend these findings to demonstrate that epithelial ESR1 also regulates glycogen deposition, luminal pH, and microbial stability. Compared to control littermates, epithelial Esr1d/d mice reduced glycogen abundance, elevated vaginal pH, and a compositional shift in the vaginal microbiome, marked by enrichment of Comamonadaceae and loss of Lactobacillus species, without significant differences in alpha diversity. These changes parallel features of postmenopausal dysbiosis in women. Together, our findings identify epithelial ESR1 as a master regulator of multiple pathways that sustain vaginal homeostasis, integrating epithelial metabolism, barrier function, and host-microbe interactions. This work provides a mechanistic framework to understand postmenopausal vaginal dysbiosis and suggests epithelial estrogen signaling as a potential therapeutic target for genitourinary syndrome of menopause.

Significance StatementThe vaginal environment is essential for reproductive and gynecologic health, yet the mechanisms by which estrogen shapes this niche remain incompletely understood. We show that epithelial estrogen receptor  (ESR1) regulates glycogen deposition, luminal pH, and microbial composition in the murine vagina. Loss of epithelial ESR1 reduced glycogen and increased luminal pH without altering overall microbial diversity, but shifted community structure toward enrichment of Comamonadaceae, a family associated with neutral to mildly alkaline environments. These findings identify epithelial ESR1 as a key regulator of the metabolic and physicochemical conditions that maintain vaginal microbial balance and provide a mechanistic framework for understanding postmenopausal dysbiosis.
]]></description>
<dc:creator>Mahalingam, S.</dc:creator>
<dc:creator>Carulli, E. M.</dc:creator>
<dc:creator>Mao, J.</dc:creator>
<dc:creator>Stephens, K. K.</dc:creator>
<dc:creator>Erickson, J. A.</dc:creator>
<dc:creator>Winuthayanon, W.</dc:creator>
<dc:date>2025-10-30</dc:date>
<dc:identifier>doi:10.1101/2025.10.29.685365</dc:identifier>
<dc:title><![CDATA[Vaginal epithelial estrogen receptor α coordinates glycogen deposition, microbial stability, and pH regulation in mice]]></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/10.1101/2025.10.31.685426v1?rss=1">
<title>
<![CDATA[
The ID93 + GLA-3M-052-LS vaccine candidate elicits mucosal and systemic immunogenicity and protective efficacy against Mycobacterium tuberculosis challenge in BCG-primed Collaborative Cross inbred mice 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.10.31.685426v1?rss=1</link>
<description><![CDATA[
New vaccine approaches are needed against tuberculosis (TB). We sought to optimize mucosal immunogenicity and protective efficacy by modulating the adjuvant component and route of immunization of a next-generation TB vaccine using the recombinant TB vaccine antigen (Ag) ID93. ID93-specific mucosal and systemic immunogenicity and protective efficacy were assessed in the Collaborative Cross 004 mouse strain, a mouse strain susceptible to Mycobacterium tuberculosis (Mtb) infection, as a suitable model of Mtb susceptible populations. Immunogenicity data from various vaccine candidates were used to select lead vaccine candidates with the most preferred immunostimulatory profiles using a pre-determined desirability index. A liposomal adjuvant system containing synthetic TLR4 and TLR7/8 ligands (GLA-3M-052-LS), administered by a heterologous intramuscular-intranasal regimen, induced an optimal comprehensive immune response profile including high levels of mucosal antibody and Th1 CD4+ T cells in the lungs. In BCG-primed mice, immunization with intramuscular followed by intranasal ID93 + GLA-3M-052-LS boosts significantly reduced Mtb burden in the lungs after challenge vs. BCG vaccinated mice alone. Thus, ID93 + GLA-3M-052-LS represents a promising next-generation TB vaccine candidate suitable for testing in additional preclinical models.
]]></description>
<dc:creator>Voigt, E. A.</dc:creator>
<dc:creator>Alsharaydeh, A.</dc:creator>
<dc:creator>Kasal, D. N.</dc:creator>
<dc:creator>Jennewein, M. F.</dc:creator>
<dc:creator>Brandt, D. S.</dc:creator>
<dc:creator>Lin, S.</dc:creator>
<dc:creator>Singh, J.</dc:creator>
<dc:creator>Bakken, J.</dc:creator>
<dc:creator>Mohamath, R.</dc:creator>
<dc:creator>Fusco, P.</dc:creator>
<dc:creator>Torrelles, J. B.</dc:creator>
<dc:creator>Beamer, G.</dc:creator>
<dc:creator>Fox, C. B.</dc:creator>
<dc:date>2025-11-01</dc:date>
<dc:identifier>doi:10.1101/2025.10.31.685426</dc:identifier>
<dc:title><![CDATA[The ID93 + GLA-3M-052-LS vaccine candidate elicits mucosal and systemic immunogenicity and protective efficacy against Mycobacterium tuberculosis challenge in BCG-primed Collaborative Cross inbred mice]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-11-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.10.31.685861v1?rss=1">
<title>
<![CDATA[
Contextual control of CD8+ T cell priming by dendritic cell subsets in tumor and inflammatory microenvironments 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.10.31.685861v1?rss=1</link>
<description><![CDATA[
Conventional dendritic cells orchestrate adaptive immunity by trafficking peripheral antigens to draining lymph nodes and presenting peptide-MHC complexes to prime naive T cells. Migratory and lymph node resident conventional dendritic cell subsets occupy distinct anatomical niches and have been shown to shape T cell activation in a variety of immunologic contexts including infection, vaccination and cancer. How peripheral tissue context and dendritic cell subset specific transcriptional programs collaborate to determine CD8+ T cell priming remains incompletely defined. Using fluorescent antigen, we tracked antigen distribution, dendritic cell transcriptional programming, and functional cross-presentation across tumor, inflammatory, and steady state tissue contexts. We find that skin tumor antigen is more widely distributed amongst draining lymph node conventional dendritic cells than skin antigen derived from either steady state or inflamed skin tissue. Comparing across dendritic cell subsets, migratory type 1 dendritic cells display higher expression of MHCI antigen presentation machinery and genes associated with cross-presentation compared with lymph node resident type 1 dendritic cells in tumor and inflamed tissue contexts. Similarly, they exhibited superior per-cell cross-presentation and stronger induction of naive CD8+ T cell responses. We find that both antigen access in the lymph node and cell-intrinsic cross-presentation efficiency together predict the magnitude and quality of CD8+ T cell priming regardless of tissue context. These results identify migratory dendritic cells, particularly type 1, as central mediators of antitumor CD8+ T cell responses and support therapeutic strategies that either restrict antigen dispersal from migratory dendritic cells or augment the efficiency of resident dendritic cell cross-presentation.

SYNOPSISAntigen transfer from migratory to resident conventional dendritic cell subsets is increased in melanoma compared to steady state or inflamed skin draining lymph nodes. Superior T cell proliferation and expression of cross-presentation machinery by migratory dendritic cells following antigen uptake of tumor suggests retaining antigen within these dendritic cell subsets may improve antitumor CD8+ T cell responses.
]]></description>
<dc:creator>Schuster, V. P.</dc:creator>
<dc:creator>Brown, K.</dc:creator>
<dc:creator>Unsworth, J. R.</dc:creator>
<dc:creator>Berkowitz, N.</dc:creator>
<dc:creator>Ruhland, M. K.</dc:creator>
<dc:date>2025-11-03</dc:date>
<dc:identifier>doi:10.1101/2025.10.31.685861</dc:identifier>
<dc:title><![CDATA[Contextual control of CD8+ T cell priming by dendritic cell subsets in tumor and inflammatory microenvironments]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-11-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.11.03.685725v1?rss=1">
<title>
<![CDATA[
CD38hiCD19dim cells in lymph nodes predict favorable prognosis in patients with stage III melanoma receiving adjuvant PD-1-blockade 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.11.03.685725v1?rss=1</link>
<description><![CDATA[
BackgroundAdjuvant immunotherapy has significantly improved survival for patients with stage III cutaneous melanoma, yet a fraction of patients will not benefit from immune checkpoint inhibitors (ICI). The tumor microenvironment plays a pivotal role in generating durable responses to ICI. By analyzing the cellular composition of tumor-associated subsets, key immune components essential for promoting an anti-tumor environment can be pinpointed. This will allow for both patient stratification and identification of biomarkers associated with improved patient outcome.

MethodRegional lymph nodes were obtained from patients with stage III melanoma at surgery (n=29). Patients eligible for anti-PD-1 therapy (PD-1; pembrolizumab or nivolumab) received adjuvant treatment for up to one year. CyTOF was used to determine cellular composition in pre-treatment surgical specimens. Bulk gene expression data generated by NanoString from patients receiving surgery without adjuvant therapy (n=125) was implemented for evaluating trends observed in the CyTOF dataset.

ResultsAlthough no significant differences were observed across major hierarchical immune cell types between patients who developed distant metastasis after surgery and those that did not, an increased proportion of CD103+PD-1+CD8+ (TRM) T cells and plasmablast-like CD38hiCD19dim cells were associated with improved prognosis in the CyTOF cohort. In the untreated cohort, a subset of patients defined as "Ultra-cold" (< 2.5 % tumor-infiltrating lymphocyte (TIL) scored by a pathologist) had significantly worse outcome than those with higher TIL infiltration. This Ultra-cold TIL group was associated with reduced B cell score, but not CD8+ T cell score, as well as reduced expression of activation genes like CD38.

ConclusionIn this study, CD103+PD-1+CD8+ (TRM) T cells and plasmablast-like CD38hiCD19dim cell populations were found to be strongly associated with prolonged distant metastasis-free survival in regional lymph nodes from patients with stage III melanoma treated with PD-1. This suggests an association between progression and infiltration of these cell types at baseline and highlights the potential of using immune cell subsets as prognostic biomarkers.

What is already known on this topic - Patients being diagnosed with stage III melanoma will receive immune checkpoint inhibitors but will often be cured by surgery alone. Selection of which patients might benefit from treatment is still unresolved. Accurate biomarkers would aid in treatment stratification, to avoid overtreatment, and unnecessary toxicities.

What this study adds - This study highlights how baseline CD103+PD-1+CD8+ (TRM) T cells and plasmablast-like CD38hiCD19dim cell populations in the regional lymph node, is strongly associated with improved outcome in patients receiving anti-PD-1 therapy.

How this study might affect research, practice or policy - The strong association between baseline plasmablast-like cell infiltration in RLN and prolonged distant metastasis free- survival, highlights this cell type as a potential treatment stratification criterion to identify patients with good prognosis.
]]></description>
<dc:creator>Ailte, I.</dc:creator>
<dc:creator>Chauhan, S. K.</dc:creator>
<dc:creator>Prasmickaite, L.</dc:creator>
<dc:creator>Bassarova, A. V.</dc:creator>
<dc:creator>Poissonnier, A.</dc:creator>
<dc:creator>Feenstra, M.</dc:creator>
<dc:creator>Nyakas, M.</dc:creator>
<dc:creator>Johannessen, B.</dc:creator>
<dc:creator>Ryder, T.</dc:creator>
<dc:creator>Hermann, R.</dc:creator>
<dc:creator>Frich, L.</dc:creator>
<dc:creator>Holth, A.</dc:creator>
<dc:creator>Inderberg, E. M.</dc:creator>
<dc:creator>Jespersen, H.</dc:creator>
<dc:creator>Florenes, V. A.</dc:creator>
<dc:creator>Kyte, J. A.</dc:creator>
<dc:creator>Maelandsmo, G. M.</dc:creator>
<dc:creator>Egeland, E. V.</dc:creator>
<dc:date>2025-11-04</dc:date>
<dc:identifier>doi:10.1101/2025.11.03.685725</dc:identifier>
<dc:title><![CDATA[CD38hiCD19dim cells in lymph nodes predict favorable prognosis in patients with stage III melanoma receiving adjuvant PD-1-blockade]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-11-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.11.04.686352v1?rss=1">
<title>
<![CDATA[
GNL3 SUMOylation is essential for DNA double-strand break repair by homologous recombination 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.11.04.686352v1?rss=1</link>
<description><![CDATA[
DNA double-strand break (DSB) repair via homologous recombination (HR) is critical for maintaining genomic integrity and requires proper DNA end resection to generate single-stranded DNA (ssDNA) overhangs. However, the mechanisms governing this critical step in cells remains poorly understood. Here, we report that GNL3, a nucleolar GTP-binding protein, plays a key role in HR repair via regulating DNA end resection dependently on its SUMOylation. Ectopic expression of wild-type, but not the SUMO-defective K196R mutant, GNL3 completely abolished DNA damage response induced by knockdown of endogenous GNL3. GNL3 interacts with the BLM-DNA2 helicase-nuclease complex and is critical for DNA end resection and subsequent loading of RPA and RAD51. This interaction requires SUMOylation and SUMO-interacting motifs (SIMs) in both proteins. We further demonstrate that USP36, a nucleolar deubiquitinating enzyme, functions as a novel SUMO ligase for GNL3, while the SUMO protease SENP3 deSUMOylates GNL3. Notably, several breast cancer-derived GNL3 variants that disrupt its SUMOylation or SIM fail to interact with the BLM-DNA2 complex. Knockdown of GNL3 sensitizes HR-proficient breast cancer cells to etoposide or Olaparib treatment. Together, our results reveal that GNL3 SUMOylation is crucial for HR repair and suggest that targeting GNL3 SUMOylation may induce HR deficiency, thereby sensitizing breast cancers to DNA damage-inducing agents.
]]></description>
<dc:creator>Yang, Y.</dc:creator>
<dc:creator>Li, Y.</dc:creator>
<dc:creator>Dai, R. S.</dc:creator>
<dc:creator>Chen, C.</dc:creator>
<dc:creator>Zientek, K.</dc:creator>
<dc:creator>Reddy, A.</dc:creator>
<dc:creator>Xia, Z.</dc:creator>
<dc:creator>Sears, R. C.</dc:creator>
<dc:creator>Sun, X.-X.</dc:creator>
<dc:creator>Dai, M.-S.</dc:creator>
<dc:date>2025-11-05</dc:date>
<dc:identifier>doi:10.1101/2025.11.04.686352</dc:identifier>
<dc:title><![CDATA[GNL3 SUMOylation is essential for DNA double-strand break repair by homologous recombination]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-11-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.11.05.686857v1?rss=1">
<title>
<![CDATA[
The Christchurch point mutation in mouse APOE reduces Aβ-induced tau and α-synuclein pathologies 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.11.05.686857v1?rss=1</link>
<description><![CDATA[
Apolipoprotein E (APOE) genotype is well known to influence both amyloid-{beta} (A{beta}) and tau pathologies and risk for Alzheimers disease (AD), but it also affects -synuclein (-syn) levels, Lewy pathology and risk of dementia in Parkinsons disease (PD) and dementia with Lewy bodies (DLB). The APOE-R136S (Christchurch, CC) point mutation has been shown to protect against AD pathology and dementia, however, the molecular mechanisms underlying this protection and its effects on -syn pathology are not well understood. Using CRISPR/Cas9 technology, we created a CC arginine-to-serine point mutation at the conserved location in mouse APOE (R128S) to understand its effects on A{beta}, tau and -syn pathologies. We crossed these APOE CC mice to 5xFAD, PS19 and A53T-Syn-GFP (A53T) mice. Using these various double mutant mice, we tested the effect of mouse APOE CC on different proteinopathies, including A{beta}, tau, A{beta}-induced tau after paired helical filament (PHF)-tau intracortical injections, and -syn after preformed fibril (PFF) intracortical and intramuscular injections. We used immunohistochemical, biochemical and behavioral measures to test for protective effects of APOE CC on these different proteinopathies. Heterozygous (Het) and homozygous (Hom) APOE CC mice showed increased plasma cholesterol and triglyceride levels, as seen in humans, but no differences in body or brain weight, or life expectancy. APOE CC decreased A{beta}-induced tau pathologies in PHF-tau injected 5xFAD;Hom mice but did not change A{beta}-plaque pathology in 5xFAD mice or tau pathology in PS19 mice. Although A{beta} levels, tau levels and mouse sex correlated strongly with the behavioral performance, we only detected subtle effects of APOE CC on anxiety-like behaviors in crosses with 5xFAD, PS19 and PHF-tau injected 5xFAD mice. Interestingly, Het and Hom APOE CC mice both showed reduced formation and spread of Lewy pathology in brain after intracortical -syn PFF injection and reduced formation in spinal cord after -syn PFF injection into the hindlimb gastrocnemius muscle in A53T mice. Our study emphasizes the protective effects of the APOE CC variant against different proteinopathies important for dementia and movement disorders, including A{beta} plaque, tau and -syn, and suggests that targeting APOE CC could provide new therapeutic strategies for AD, DLB and PD.

Graphical Abstract

O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=123 SRC="FIGDIR/small/686857v1_ufig1.gif" ALT="Figure 1">
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]]></description>
<dc:creator>Soto-Faguas, C. M.</dc:creator>
<dc:creator>O'Niel, A.</dc:creator>
<dc:creator>Mueller, P. A.</dc:creator>
<dc:creator>Sanchez-Molina, P.</dc:creator>
<dc:creator>Woltjer, R. L.</dc:creator>
<dc:creator>Raber, J.</dc:creator>
<dc:creator>Unni, V. K.</dc:creator>
<dc:date>2025-11-07</dc:date>
<dc:identifier>doi:10.1101/2025.11.05.686857</dc:identifier>
<dc:title><![CDATA[The Christchurch point mutation in mouse APOE reduces Aβ-induced tau and α-synuclein pathologies]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-11-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.11.07.687263v1?rss=1">
<title>
<![CDATA[
Combining the alpha-2 adrenergic agonist clonidine with naloxone rescues fentanyl-induced physiologic dysfunction and increases survival 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.11.07.687263v1?rss=1</link>
<description><![CDATA[
Fentanyl leads to tens of thousands of overdose deaths every year despite widespread availability of naloxone. Like other opioids, fentanyl causes respiratory depression. Unlike morphine, high dose fentanyl rapidly produces airway obstruction, muscle rigidity, and cardiovascular failure. Using a rat model of opioid overdose, we compared the physiological effects of fentanyl and morphine and studied the efficacy of a novel rescue strategy. In contrast to morphine, we report that fentanyl more frequently causes respiratory failure secondary to vocal cord closure and leads to more severe cardiovascular disruption, including the blockade of baroreflex-like rebound in blood pressure. We also show that administration of intramuscular naloxone immediately after intravenous infusion of fentanyl did not improve survival. However, combining intramuscular naloxone with the alpha-2 adrenergic agonist clonidine rescued vocal cord function and stabilized cardiovascular and respiratory physiology from fentanyl-induced effects. Our findings demonstrate that fentanyl is associated with a unique and more severe toxidrome compared to morphine. Also, supplementing naloxone with drugs targeting the adrenergic system improves survival primarily by reopening the upper airway, implicating airway obstruction as a significant component of fentanyl-induced respiratory depression. Therefore, reversal of vocal cord closure appears to be the necessary precursor to the restoration of not only respiration, but also vascular autoregulation, a significant determinant of survival from fentanyl overdose.
]]></description>
<dc:creator>Rivera, A.</dc:creator>
<dc:creator>Schutzer, W. E.</dc:creator>
<dc:creator>Sonneborn, A.</dc:creator>
<dc:creator>Shotwell, S. L.</dc:creator>
<dc:creator>Janowsky, A.</dc:creator>
<dc:creator>Abbas, A.</dc:creator>
<dc:creator>Torralva, R.</dc:creator>
<dc:date>2025-11-10</dc:date>
<dc:identifier>doi:10.1101/2025.11.07.687263</dc:identifier>
<dc:title><![CDATA[Combining the alpha-2 adrenergic agonist clonidine with naloxone rescues fentanyl-induced physiologic dysfunction and increases survival]]></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/10.1101/2025.11.11.687916v1?rss=1">
<title>
<![CDATA[
A Population-Specific PARP1 Gene Variation Modulates PARP Trapping 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.11.11.687916v1?rss=1</link>
<description><![CDATA[
Poly-(ADP-ribose) polymerase inhibitors (PARPi) block NAD+-binding pocket of PARP1, inhibiting PAR synthesis. However, they differ in their ability to retain PARP1 on damaged DNA to induce synthetic lethality in homologous recombination-deficient (HRD) cancer. Allosteric enzymatic activation requires destabilization of the helical domain (HD) of PARP1 and is indispensable for activation and chromatin retention induced by distinct PARPi. Here we report that the effect of the clinical PARPi talazoparib is robustly impacted by a common human polymorphism within the HD. PARP1V762 greatly enhanced talazoparib-driven allosteric retention on chromatin, prolonged XRCC1 recruitment, and enhanced cell killing. Talazoparib switches from Type-II PARPi behavior in PARP1A762 to allosteric, pro-retention Type-I behavior for PARP1V762. Thus, both PARPi efficacy and dose-limiting tolerability depends on PARP1 allele, motivating variant-guided cancer therapies.

TEASEROne of the four FDA-approved PARPi drugs, talazoparib, is modulated by a PARP1 SNP that is widespread in the population.
]]></description>
<dc:creator>Cai, J.</dc:creator>
<dc:creator>Billur, R.</dc:creator>
<dc:creator>Cohen, M. S.</dc:creator>
<dc:creator>Ladurner, A. G.</dc:creator>
<dc:creator>Black, B. E.</dc:creator>
<dc:date>2025-11-13</dc:date>
<dc:identifier>doi:10.1101/2025.11.11.687916</dc:identifier>
<dc:title><![CDATA[A Population-Specific PARP1 Gene Variation Modulates PARP Trapping]]></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/10.1101/2025.11.11.687936v1?rss=1">
<title>
<![CDATA[
CREG1 promotes autophagy and protects the heart against nutritional stress-induced injury and age-associated hypertrophy, fibrosis and diastolic dysfunction 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.11.11.687936v1?rss=1</link>
<description><![CDATA[
BackgroundCellular repressor of E1A-stimulated genes 1 (CREG1) is an evolutionarily conserved endolysosomal glycoprotein that enhances lysosomal biogenesis and autophagy, suppresses proliferation, and promotes differentiation. A prior gene targeting strategy that produced truncated N-terminal fragments resulted in embryonic lethality, limiting the ability to assess the physiological role of complete CREG1 loss. We hypothesized that CREG1 regulates cardiac autophagy, thereby maintaining cardiac structure and function under both physiological and stress conditions.

MethodsWe generated true Creg1 knockout (KO) mice by deleting the entire open reading frame and established a gain-of-function model by inserting human CREG1 into the Rosa26 locus. Cardiac structure and function were assessed in global and cardiomyocyte-specific Creg1 knockout (cmCreg1KO) and knock-in (cmCREG1KI) mice. Autophagy was evaluated using biochemical assays, immunofluorescence, electron microscopy, and the CAG-EGFP-RFP-LC3 reporter analysis.

ResultsGlobal Creg1 knockout mice developed progressive cardiac hypertrophy, fibrosis, and diastolic dysfunction at [~]80 weeks of age. At younger ages, CREG1 deficiency increased susceptibility to nutritional stress, resulting in mitochondrial damage and myofiber disruption in cardiomyocytes. cmCreg1KO mice exhibited dilated cardiomyopathy, left atrial thrombosis, and lethality around 50 weeks of age; however, interpretation of disease severity is confounded by Myh6-Cre-associated cardiotoxicity, which may mask additional pathogenic effects attributable to CREG1 loss. In contrast, cmCREG1KI mice demonstrated enhanced exercise capacity under nutritional stress. Mechanistically, CREG1 was localized to endolysosomal and autophagosomal compartments. Loss of CREG1 impaired autophagy flux and mitophagy, likely due to defective autophagosome membrane expansion and degradation. In contrast, CREG1 overexpression enhanced autophagy in cardiomyocytes.

ConclusionsCREG1 is a key regulator of cardiac autophagy, protecting the heart against nutritional stress-induced injury and age-associated cardiac hypertrophy, fibrosis, and diastolic dysfunction.

Graphic Abstract

O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=171 SRC="FIGDIR/small/687936v1_ufig1.gif" ALT="Figure 1000">
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]]></description>
<dc:creator>Qi, Y.</dc:creator>
<dc:creator>Pepe, R. J.</dc:creator>
<dc:creator>Tejeda, G.</dc:creator>
<dc:creator>Joo, Y.</dc:creator>
<dc:creator>Yang, N. K.</dc:creator>
<dc:creator>Hayes, K. B.</dc:creator>
<dc:creator>rahimi, s.</dc:creator>
<dc:creator>Zhou, Z.</dc:creator>
<dc:creator>Li, S.</dc:creator>
<dc:creator>Lee, L.</dc:creator>
<dc:date>2025-11-13</dc:date>
<dc:identifier>doi:10.1101/2025.11.11.687936</dc:identifier>
<dc:title><![CDATA[CREG1 promotes autophagy and protects the heart against nutritional stress-induced injury and age-associated hypertrophy, fibrosis and diastolic dysfunction]]></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/10.1101/2025.11.13.687902v1?rss=1">
<title>
<![CDATA[
The retinal light response is modulated by an mGluR5-mediated retrograde signal from ON-bipolar cells to photoreceptors 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.11.13.687902v1?rss=1</link>
<description><![CDATA[
The ON visual pathway is initiated by the deactivation of mGluR6, coupled to the opening of TRPM1 channels in retinal ON-bipolar cell dendrites. Here, we show that a second metabotropic glutamate receptor, mGluR5, is localized with TRPM1 and mGluR6 in the dendrites of ON-bipolar cells. To examine the function of mGluR5, we performed electroretinogram (ERG) recordings and found that the amplitude of the b-wave, which is primarily a measure of ON-bipolar cell light-driven activity, is reduced in mGluR5 knock-out mice compared to wild type. In the mGluR5-/- retina, we observed weaker mGluR6 immunofluorescence in the dendritic tips of ON-bipolar cells that could explain the smaller ERG b-wave. To observe the effect of mGluR5 without perturbing mGluR6 expression, wild type mice were injected with MTEP, an allosteric antagonist of mGluR5. MTEP increased the amplitude of the b-wave in response to dim stimuli and caused an inflection in the intensity-response plot for flashes in the mesopic range. In the brain, postsynaptic mGluR5 regulates presynaptic glutamate release via endocanabinoid-mediated retrograde signaling. Therefore, we tested the effect of the CB1 receptor antagonist, SR1417A, on the ERG and found that the b-wave was affected as by MTEP, including an inflection in the intensity-response. We further showed that the CB1 receptor agonist, ACEA, reversed the effects of MTEP. Together, our results indicate that mGluR5 plays a role in gain-control at the photoreceptor to ON-bipolar cell synapses, likely via an endocannabinoid-mediated retrograde feedback.

Significance StatementLight-ON stimuli are transmitted from photoreceptors to ON-bipolar cells via three pathways, which are used under different light intensities, from dim to bright light. For each pathway, glutamate is the neurotransmitter released by rods and cones, and the postsynaptic role of the mGluR6-TRPM1 signaling cascade in ON-bipolar cells is well studied, with defects known to cause congenital stationary night blindness. We show here that mGluR5 is also present in ON-bipolar cell dendrites and regulates synaptic transmission between photoreceptors and ON-bipolar cells. In particular, we find that mGluR5 facilitates the transition between the different photoreceptor to ON-bipolar cell pathways via mGluR5-controlled retrograde release of endocannabinoids.
]]></description>
<dc:creator>Morgans, C. W.</dc:creator>
<dc:creator>Haley, T. L.</dc:creator>
<dc:creator>Ren, G.</dc:creator>
<dc:creator>Taylor, I. A. M.</dc:creator>
<dc:creator>Randall, K. L.</dc:creator>
<dc:creator>Duvoisin, R. M.</dc:creator>
<dc:date>2025-11-14</dc:date>
<dc:identifier>doi:10.1101/2025.11.13.687902</dc:identifier>
<dc:title><![CDATA[The retinal light response is modulated by an mGluR5-mediated retrograde signal from ON-bipolar cells to photoreceptors]]></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/10.1101/2025.11.17.688858v1?rss=1">
<title>
<![CDATA[
Degraded neural coding of temporal fine structure with age predicts effortful listening in multi-talker environments 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.11.17.688858v1?rss=1</link>
<description><![CDATA[
Middle age represents a critical window for early detection of neurophysiological decline. Hearing loss is increasingly recognized as both an early marker of neural degeneration and a modifiable risk factor for dementia. Yet many adults report difficulty understanding speech in noise despite normal audiograms, highlighting the limitations of current clinical tests that fail to capture the underlying physiology or effort required for real-world listening. Beyond hearing thresholds, speech comprehension in complex environments depends on precise neural encoding of temporal fine structure (TFS) cues that convey pitch and spatial information. Here, we use a noninvasive EEG-based measure of neural phase-locking (frequency modulation following responses or FMFRs) to quantify TFS encoding in young and middle-aged adults with normal hearing thresholds. Middle-aged listeners exhibited reduced FMFR amplitudes and shallower discriminability slopes, reflecting diminished neural synchrony despite preserved hearing thresholds. Using a multi-talker speech task we further found that pupil-indexed listening effort was significantly greater in middle-aged adults despite matched accuracy across groups. Further, increases in listening effort were predicted by decreases in TFS encoding. Together, these results reveal that degraded neural encoding of TFS underlies subclinical listening difficulties and increased cognitive load, establishing the FMFR as a sensitive biomarker of hidden auditory neural decline.

SignificanceUnderstanding speech in noisy environments depends on precise neural encoding of temporal fine structure (TFS) cues. Using a noninvasive EEG metric, the FMFR, we show that neural TFS coding in the peripheral auditory system declines markedly by midlife, even when hearing thresholds and speech performance are normal. These neural deficits predict elevated pupil-indexed listening effort during multi-talker speech perception, revealing that subclinical degradation of temporal coding increases cognitive load in everyday listening. The FMFR thus provides a promising biomarker for early auditory neural decline and its downstream cognitive consequences.
]]></description>
<dc:creator>Zhen, L.</dc:creator>
<dc:creator>Parida, S.</dc:creator>
<dc:creator>McHaney, J.</dc:creator>
<dc:creator>Zink, M.</dc:creator>
<dc:creator>Chandrasekaran, B.</dc:creator>
<dc:creator>Parthasarathy, A.</dc:creator>
<dc:date>2025-11-17</dc:date>
<dc:identifier>doi:10.1101/2025.11.17.688858</dc:identifier>
<dc:title><![CDATA[Degraded neural coding of temporal fine structure with age predicts effortful listening in multi-talker environments]]></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/10.1101/2025.11.18.689169v1?rss=1">
<title>
<![CDATA[
Lipid Tail Length Determines Nano-Bio Interactions of Peptide Amphiphile Nanostructures 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.11.18.689169v1?rss=1</link>
<description><![CDATA[
Understanding the interactions of nanomaterials with biological systems is essential to designing effective nanomedicines. However, most of our understanding originates from studies with solid nanoparticles, and nano-bio interactions of self-assembled nanomaterials have remained largely unexplored. To address this knowledge gap, we develop a series of self-assembling peptide amphiphiles (PAs) with different lipid modifications and investigate their interactions with biological systems. We find that PA nanostructures rapidly disassemble and reassemble with albumin and lipoproteins in blood plasma. While PAs with shorter lipid tails mainly assemble with albumin, increasing lipid length shifts binding to lipoproteins. All PAs show strong tumor accumulation in 4T1 tumor-bearing mice with tumor to liver ratios of [~]3-6. Overall, albumin-binding improves blood circulation and tumor accumulation compared to lipoprotein-binding, but also increases off-target accumulation. This study shows that the biointeractions of self-assembled nanomaterials can be controlled through molecular design, which may lead to the development of effective nanomedicines.
]]></description>
<dc:creator>Bonic, K.</dc:creator>
<dc:creator>Stewart, M. R.</dc:creator>
<dc:creator>Xiang, L.</dc:creator>
<dc:creator>Mooney, K.</dc:creator>
<dc:creator>Armstrong, R.</dc:creator>
<dc:creator>Esener, S.</dc:creator>
<dc:creator>Fischer, J. M.</dc:creator>
<dc:creator>Yildirim, A.</dc:creator>
<dc:date>2025-11-19</dc:date>
<dc:identifier>doi:10.1101/2025.11.18.689169</dc:identifier>
<dc:title><![CDATA[Lipid Tail Length Determines Nano-Bio Interactions of Peptide Amphiphile Nanostructures]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-11-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.11.19.689361v1?rss=1">
<title>
<![CDATA[
RNA switch model for localization and translation of the myelin basic protein mRNA 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.11.19.689361v1?rss=1</link>
<description><![CDATA[
Oligodendrocytes myelinate the central nervous system by extending cellular projections that ensheath axons and elongate to form lipid-rich myelin. Classic studies visualizing RNA dynamics showed that myelin basic protein (MBP), one of the most abundant myelin proteins, is locally synthesized at the myelin sheath through the transport and local translation of Mbp mRNA. Mbp transport requires its 1.5-kb 3 untranslated region (3 UTR) and prior work identified candidate sub-sequences that may act as cis-acting transport stimulating RNA elements, including one with putative secondary structure. Here, a high-throughput reporter assay, dimethyl sulfate (DMS)-based RNA structure probing, and microscopy in primary rat oligodendrocytes identify a structured 127-nt region that we name the Mbp localization signal (MLS) as both necessary and sufficient for RNA enrichment to oligodendrocyte projections. Lysate pulldown experiments further identify hnRNP-F - a known constituent of the Mbp RNA granule that can suppress mRNA translation - as associated with the MLS; paradoxically, binding of this protein should compete with the ordered MLS RNA structure. These results suggest a model in which the MLS switches between two RNA conformations with distinct protein partners during the transition from Mbp mRNA transport to Mbp translation at the myelin sheath. Such regulation of RNA behavior by structure switching may generalize to other eukaryotic mRNAs whose behaviors shift across space and time.

Significance StatementIn the brain, oligodendrocyte cells generate myelin, a type of insulation that wraps around neuronal axons in order to facilitate fast electrical signaling. A critical step in myelination is the local translation of MBP (myelin basic protein) in the myelin sheath. This requires the transport of Mbp mRNA, an incompletely understood phenomenon that we revisit using two recent approaches for mRNA structure and function. We refine a 127-nt region that is necessary and sufficient for mRNA transport to the myelin sheath. A proteomic screen reveals that this myelin localization signal (MLS) associates with a translation-suppressing protein called hnRNP-F, suggesting a model where Mbp mRNA switches between two states, one for transport and one for translation at the myelin sheath.
]]></description>
<dc:creator>Topkar, V. V.</dc:creator>
<dc:creator>Wu, V.</dc:creator>
<dc:creator>Ho, L. T.</dc:creator>
<dc:creator>Ambiel, N.</dc:creator>
<dc:creator>Valenzuela, A.</dc:creator>
<dc:creator>Yoshimura, K.</dc:creator>
<dc:creator>Zuchero, J. B.</dc:creator>
<dc:creator>Fu, M.-m.</dc:creator>
<dc:creator>Das, R.</dc:creator>
<dc:date>2025-11-20</dc:date>
<dc:identifier>doi:10.1101/2025.11.19.689361</dc:identifier>
<dc:title><![CDATA[RNA switch model for localization and translation of the myelin basic protein mRNA]]></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/10.1101/2025.11.21.689510v1?rss=1">
<title>
<![CDATA[
Sphingosine-1-phosphate receptor modulators resensitize FLT3-ITD acute myeloid leukemia cells with NRAS mutations to FLT3 inhibitors 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.11.21.689510v1?rss=1</link>
<description><![CDATA[
FLT3 inhibitor efficacy in AML with FLT3-ITD is short-lived, frequently due to new mutations, most commonly in NRAS. Sphingosine kinase 1 (SPHK1), which phosphorylates sphingosine to generate sphingosine-1-phosphate (S1P), is upregulated and localized to the plasma membrane in RAS-mutated cells. We studied S1P and FLT3 co-targeting to overcome FLT3 inhibitor resistance in NRAS-mutated FLT3-ITD AML cells. NRAS-mutated FLT3-ITD AML cell lines and patient blasts were treated with FLT3 inhibitors and/or S1P receptor (S1PR) modulators. FLT3 inhibitor sensitivity was assessed by immunoblotting, cytotoxicity and apoptosis assays. Co-treatment was also assessed in vivo in an orthotopic mouse model. Downstream RAS and SPHK1 effectors were measured by immunoblotting and qRT-PCR. The S1PR modulators fingolimod (FTY720) and mocravimod (KRP-203) resensitized FLT3-ITD-expressing MOLM-14 and MV4-11 human AML cells with G12D, G12S, Q61K or Q61H, but not G12C, and patient blasts with G13D or G13V NRAS mutations to FLT3 inhibitors. Moreover, FTY720 co-treatment resensitized G12D NRAS-mutated M14(R)701 cells to gilteritinib in vivo. Co-treatment inactivated ERK, transcriptionally downregulated SPHK1, and inactivated downstream AKT, p70S6K and BAD, with inactivation abrogated by constitutive SPHK1 expression. The clinically applicable S1PR modulators fingolimod and mocravimod resensitize NRAS-mutated FLT3-ITD AML cells to FLT3 inhibitors, supporting potential clinical efficacy of these combinations.
]]></description>
<dc:creator>Baer, M. R.</dc:creator>
<dc:creator>Chatterjee, A.</dc:creator>
<dc:creator>Ali, M. K. M.</dc:creator>
<dc:creator>Bailey, C. M.</dc:creator>
<dc:creator>Liu, Y.</dc:creator>
<dc:creator>Small, D.</dc:creator>
<dc:creator>Smith, C. C.</dc:creator>
<dc:creator>Traer, E. C.</dc:creator>
<dc:creator>Wang, Y.</dc:creator>
<dc:creator>Silvestri, G.</dc:creator>
<dc:date>2025-11-24</dc:date>
<dc:identifier>doi:10.1101/2025.11.21.689510</dc:identifier>
<dc:title><![CDATA[Sphingosine-1-phosphate receptor modulators resensitize FLT3-ITD acute myeloid leukemia cells with NRAS mutations to FLT3 inhibitors]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-11-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.11.21.689800v1?rss=1">
<title>
<![CDATA[
In vivo assessment of differential toxicity of cancer treatment drugs in Fanconi Anemia 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.11.21.689800v1?rss=1</link>
<description><![CDATA[
Fanconis Anemia (FA) is a DNA repair disorder with a very elevated risk of cancer, especially squamous cell carcinomas (SCC). Many cancer chemotherapy agents induce DNA damage, are highly toxic in FA, and cannot be safely used in this population. The potential differential toxicity to FA patients of many new drugs being explored for use in SCC is unknown. To evaluate such compounds of unknown toxicity for use in FA cancers, we developed a sensitive in vivo bone marrow repopulation competition assay in mice. We found that afatinib, alisertib and everolimus exhibited no significant differential toxicity in this system. Therefore, these drugs are candidates for chemotherapy of cancers in human FA patients. Our competitive repopulation assay provides a robust method to screen novel chemotherapy agents for their safety in FA.

Summary- We developed a mouse model system for evaluating differential toxicity of cancer chemotherapy drugs on Fanconi Anemia mutant hematopoietic cells in vivo.
- Different drugs resulted in either increases, decreases, or neutral wildtype to Fanconi mutant cell ratios permitting robust assessment of their safety in FA.
]]></description>
<dc:creator>Grompe, M.</dc:creator>
<dc:creator>Dorrell, C.</dc:creator>
<dc:creator>Burtness, B.</dc:creator>
<dc:creator>Cruz-Gomez, S.</dc:creator>
<dc:creator>MacMillan, A.</dc:creator>
<dc:date>2025-11-24</dc:date>
<dc:identifier>doi:10.1101/2025.11.21.689800</dc:identifier>
<dc:title><![CDATA[In vivo assessment of differential toxicity of cancer treatment drugs in Fanconi Anemia]]></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/10.1101/2025.11.20.689439v1?rss=1">
<title>
<![CDATA[
Infection Of Rhesus Macaques With Onyong-nyong Virus UVIR-O804 Recapitulates Key Aspects of Human Clinical Disease 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.11.20.689439v1?rss=1</link>
<description><![CDATA[
1.Onyong-nyong virus (ONNV) is a mosquito-borne alphavirus first isolated in Uganda in 1959. Since its discovery, ONNV has caused several outbreaks in Africa, manifesting clinically as fever, rash, and joint/muscle pain lasting months. Currently, we have a limited understanding of ONNV infection and disease in relevant animal models, which restricts the evaluation of vaccines and therapeutics. In 1967, Binn et al. reported that infection of rhesus macaques (RMs) with ONNV failed to induce viremia in two animals. This may be attributed to the potential attenuation of the virus through extensive passaging. To mitigate this issue, we constructed an infectious clone from the sequence of ONNV-UVRI0804 (ONNV0804), a 2017 clinical isolate from a febrile patient in Uganda. This strain demonstrated high pathogenicity in immunocompetent mice, resulting in an earlier and more severe onset of disease and significantly higher viremia compared to a highly passaged control strain ONNVUgMP30. In the current study, three male and three female rhesus macaques were subcutaneously inoculated with ONNV0804. All animals became viremic at 2 days post inoculation (dpi). Both classical and nonclassical monocytes were activated (CD169+), peaking at 3 dpi, which corresponded with the peak of viremia. Additionally, CD4+ and CD8+ effector memory T cells and memory B cells began proliferating in peripheral blood by day 7, peaking at day 10, which also corresponded to the timing of neutralizing antibody development, indicating a robust adaptive immune response to ONNV0804. Finally, key clinical disease manifestations were recapitulated, including lymphadenopathy and histological features of early-stage arthritis. Taken together, rhesus macaque infection with ONNV0804 clinical isolate is a promising model for investigating immune responses to alphaviruses and evaluating vaccines to protect against future epidemics.

2. Author summaryOnyong-nyong virus (ONNV) is a mosquito-transmitted alphavirus that causes fever, rash, and prolonged joint and muscle pain, similar to chikungunya virus and other arthritogenic alphaviruses. Despite its capacity to cause outbreaks in Africa, ONNV remains understudied, and there are currently no approved vaccines or therapeutics to prevent or treat infection and disease. A major barrier to advancing ONNV research has been the lack of suitable animal models to study the virus and investigate host immune responses. We engineered an ONNV infectious clone of a recent clinical isolate sequenced from a patient in Uganda (ONNV0804) that causes robust infection and disease in immunocompetent mice. In the current study, we provide data demonstrating that this contemporary ONNV strain infects rhesus macaques. Notably, rhesus macaques developed detectable viremia, rash, lymphadenopathy, joint and muscle inflammation, and strong innate and adaptive immune responses following subcutaneous ONNV0804 infection. These findings suggest that ONNV0804 infection in macaques is a promising model for studying ONNV pathogenesis and immunity. This model will be instrumental for evaluating future vaccine and therapeutic candidates aimed at preventing ONNV infection and related viral disease.
]]></description>
<dc:creator>Jaeger, H. K.</dc:creator>
<dc:creator>Denton, M.</dc:creator>
<dc:creator>Andoh, T. F.</dc:creator>
<dc:creator>Kreklywich, C. N.</dc:creator>
<dc:creator>Gao, L.</dc:creator>
<dc:creator>Pung, L. J.</dc:creator>
<dc:creator>Streblow, Z. J.</dc:creator>
<dc:creator>McMonigal, A.</dc:creator>
<dc:creator>Ray, K.</dc:creator>
<dc:creator>Graves, B.</dc:creator>
<dc:creator>Streblow, M. M.</dc:creator>
<dc:creator>Barber-Axthelm, A. M.</dc:creator>
<dc:creator>Zilverberg, G.</dc:creator>
<dc:creator>Terry, M.</dc:creator>
<dc:creator>Fei, S. S.</dc:creator>
<dc:creator>Hogan, G.</dc:creator>
<dc:creator>Schultz, D. C.</dc:creator>
<dc:creator>Cherry, S.</dc:creator>
<dc:creator>Axthelm, M. K.</dc:creator>
<dc:creator>Labriola, C. S.</dc:creator>
<dc:creator>Heise, M. T.</dc:creator>
<dc:creator>Streblow, D. N.</dc:creator>
<dc:date>2025-11-26</dc:date>
<dc:identifier>doi:10.1101/2025.11.20.689439</dc:identifier>
<dc:title><![CDATA[Infection Of Rhesus Macaques With Onyong-nyong Virus UVIR-O804 Recapitulates Key Aspects of Human Clinical Disease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-11-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.11.24.690244v1?rss=1">
<title>
<![CDATA[
Impaired synaptic inhibition and enhanced aversion encoding by lateral habenula neurons during Δ9-tetrahydrocannabinol withdrawal. 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.11.24.690244v1?rss=1</link>
<description><![CDATA[
The lateral habenula (LHb) integrates cortical and basal forebrain (BF) input to control monoaminergic outflow and is implicated in depression, anxiety, impulsivity, and aversion. Although cannabis withdrawal is associated with negative affect and heightened anxiety, LHb involvement is not established. Here, effects of withdrawal from the psychoactive cannabis constituent {Delta}9-tetrahydrocannabinol ({Delta}9-THC) on LHb neurons were assessed with photometric calcium measurement during fear conditioning and in vitro electrophysiology. LHb calcium signals were larger during footshock, and presentation of a tone paired with footshock in {Delta}9-THC withdrawn rats. Fear-induced freezing to the tone was also larger during {Delta}9-THC withdrawal. Electrophysiology revealed larger LHb excitatory-inhibitory (E-I) ratios during {Delta}9-THC withdrawal, resulting from impaired synaptic GABA release. Moreover, GABA release via optogenetic activation of BF-LHb inputs was impaired during {Delta}9-THC withdrawal, whereas no changes occurred at ventral tegmental area-LHb inputs. Recovery of BF-LHb GABA release and cannabinoid CB1 receptor desensitization were incomplete at 30 days of {Delta}9-THC withdrawal. The data show that LHb responsivity to primary aversive and conditioned stimuli are increased during {Delta}9-THC withdrawal, and that this is likely mediated by altered E-I balance. We propose negative affect experienced during cannabis withdrawal is related to LHb hypersensitivity to aversive stimuli and this facilitates encoding of associated environmental cues.
]]></description>
<dc:creator>Hwang, E.-K.</dc:creator>
<dc:creator>Daphne, D.</dc:creator>
<dc:creator>Maddox, C. C.</dc:creator>
<dc:creator>Zapata, A.</dc:creator>
<dc:creator>Hoffman, A. F.</dc:creator>
<dc:creator>Lupica, C. R.</dc:creator>
<dc:date>2025-11-28</dc:date>
<dc:identifier>doi:10.1101/2025.11.24.690244</dc:identifier>
<dc:title><![CDATA[Impaired synaptic inhibition and enhanced aversion encoding by lateral habenula neurons during Δ9-tetrahydrocannabinol withdrawal.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-11-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.1101/2025.11.28.691231v1?rss=1">
<title>
<![CDATA[
Gene flow weakens genomic clines while selection maintains adaptive loci: two decades of evolution in Drosophila melanogaster natural populations 
]]>
</title>
<link>https://biorxiv.org/content/10.1101/2025.11.28.691231v1?rss=1</link>
<description><![CDATA[
Using spatiotemporal sampling to understand evolutionary changes over time is highly effective, particularly to gain insights into changes driven by environmental shifts. The latitudinal clines in Drosophila melanogaster in North America provide an excellent system for studying temporal changes. Due to D. melanogasters short generational times, it is possible to observe changes spanning up to 100 generations within a single decade. Using temporal samples across the latitudinal transect could shed light on the processes responsible for the establishment and maintenance of those clines, and help to disentangle demographic and selective forces. To do this, we used pool-seq to obtain genomewide data of 16 new D. melanogaster natural populations collected along the North American east coast from 1997 to 2023. Our data suggest that there has been a homogenization over time. Consistent with this, we observed a reduction in the number of clinal single nucleotide polymorphisms. Polymorphisms that remained clinal had smaller slopes on chromosome 3R, aligned with the reduction in slope of inversions In(3R)Payne and In(3R)Mo. Clinal SNPs tended to be lost more frequently in genomic regions with higher recombination rates, and clinal SNPs identified in 2009/2010 were enriched in functional classes with greater phenotypic impact relative to those from 1997. We also investigated signs of selective sweeps that were shared among locations using a window FST approach, which corroborated previously found signals of selection in the insecticide resistance-linked region of the P450 gene family. Overall, the results indicate that gene flow gradually reduces clinality over time, while the remaining clinal loci are likely shaped by spatially varying selection. Together, these findings highlight how gene flow and selection jointly shape spatial genomic patterns over time in natural populations.
]]></description>
<dc:creator>Horvath Miranda, V.</dc:creator>
<dc:creator>da Silva Ribeiro, T.</dc:creator>
<dc:creator>Alves, J. M.</dc:creator>
<dc:creator>Eanes, W. F.</dc:creator>
<dc:creator>Brud, E.</dc:creator>
<dc:creator>Veeramah, K. R.</dc:creator>
<dc:creator>Pool, J. E.</dc:creator>
<dc:creator>Rodrigues, M. F.</dc:creator>
<dc:creator>Cogni, R.</dc:creator>
<dc:date>2025-11-29</dc:date>
<dc:identifier>doi:10.1101/2025.11.28.691231</dc:identifier>
<dc:title><![CDATA[Gene flow weakens genomic clines while selection maintains adaptive loci: two decades of evolution in Drosophila melanogaster natural populations]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-11-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2025.12.03.691670v1?rss=1">
<title>
<![CDATA[
Sorting endosomes play key roles in presentation of Mycobacterium tuberculosis-derived ligands to MAIT cells 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2025.12.03.691670v1?rss=1</link>
<description><![CDATA[
The immune system has developed specialized mechanisms to recognize intracellular pathogens such as Mycobacterium tuberculosis (Mtb). Major Histocompatibility Complex Class I-Related molecule (MR1) is a conserved nonclassical antigen presenting molecule that presents ligands derived from microbial riboflavin synthesis to Mucosal Associated Invariant T (MAIT) cells. While endosomal trafficking facilitates MR1 antigen presentation during Mtb infection, the exact mechanisms by which MR1 loading of Mtb-derived ligands occurs are not known. We found that trafficking through sorting endosomes mediates MR1 antigen presentation during Mtb infection. Sorting endosomes utilize trafficking proteins such as Syntaxin 6, Syntaxin 12, Syntaxin 16 and VAMP4. Prior work demonstrates the importance of VAMP4 for MR1 presentation during Mtb infection; we have found that Stx12 and Stx16 are also important. Interference with Stx12 or Stx16 via siRNA-mediated knockdown reduces MR1 antigen presentation of Mtb. Using RFP-tagged constructs, we found Stx16 co-localized more with MR1 vesicles compared to Stx12 in MR1-GFP expressing airway epithelial cells. Stx12 and Stx16 blockade increase MR1 surface stabilization and total expression, indicating that impaired endosomal trafficking hinders MR1 internalization. Together, these findings support a role for sorting endosomes in the selective sampling of the intracellular environment and MR1-mediated recognition of Mtb-infected cells.
]]></description>
<dc:creator>Tammen, A. E.</dc:creator>
<dc:creator>Peterson, J. C.</dc:creator>
<dc:creator>Worley, A.</dc:creator>
<dc:creator>Lewinsohn, D. M.</dc:creator>
<dc:creator>Karamooz, E.</dc:creator>
<dc:date>2025-12-04</dc:date>
<dc:identifier>doi:10.64898/2025.12.03.691670</dc:identifier>
<dc:title><![CDATA[Sorting endosomes play key roles in presentation of Mycobacterium tuberculosis-derived ligands to MAIT cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-12-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2025.12.04.692159v1?rss=1">
<title>
<![CDATA[
Differential targeting of human pyroptotic Caspase-5 and Caspase-4 by Shigella OspC2 and OspC3 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2025.12.04.692159v1?rss=1</link>
<description><![CDATA[
Pyroptosis is an inflammatory cell death pathway that is a key defense mechanism of intestinal epithelial cells. To successfully establish an infection, intracytosolic Gram-negative pathogens must block this host response. Indeed, Shigella OspC3 suppresses epithelial pyroptosis by targeting and inactivating Caspase-4 (CASP4). Here, we demonstrate that OspC2, which shares 96% identity with OspC3, targets Caspase-5 (CASP5), a close paralog of CASP4. Through a combination of yeast two-hybrid, transfection, and bacterial infection assays, we show that the distinct pyroptotic caspase specificities of OspC2 and OspC3 are determined by a short -helical region, designated the Pyroptotic Caspase Specificity (PCS) domain. This domain is located upstream from the Ankyrin Rich Repeat (ARR) region previously established to promote OspC3 binding to CASP4. Swapping PCS domains between OspC2 and OspC3 is sufficient to redirect their caspase targeting. Evidence for CASP5-driven pyroptosis in response to infection has not yet been established. However, CASP5 displays signatures of positive selection at residues predicted to interact with the PCS domain of OspC2. Notably, the introduction of orangutan-specific residues into human CASP5 disrupt its interaction with and modification by OspC2, demonstrating that CASP5 natural variation can cause key functional differences in this host-microbe molecular interaction. These findings highlight the evolutionary interplay between bacterial effectors and host proteins, support a likely role for CASP5 in responding to Gram-negative bacteria, and identify promising therapeutic targets for enhancing epithelial defense against bacterial pathogens.

ImportanceShigella species are human-specific Gram-negative pathogens that establish a replicative niche in intestinal epithelial cells by blocking pyroptosis, a key inflammatory cell death pathway. We reveal that two closely related Shigella type III secreted effectors, OspC2 and OspC3, specifically inactivate the human caspase paralogs CASP5 and CASP4, respectively. This specificity is determined by their newly identified Pyroptotic Caspase Specificity (PCS) domains. In addition, we find that positively selected residues in CASP5 alter the OspC2/CASP5 interaction, underscoring the impacts of ongoing evolutionary arms races between bacterial effectors and host immune proteins. By elucidating the molecular basis of caspase targeting and adaptation, this work provides new insight into the diversification of host defense mechanisms and identifies potential therapeutic targets for enhancing epithelial resistance to bacterial infection.
]]></description>
<dc:creator>Kim, K.</dc:creator>
<dc:creator>Valdespino Diaz, M. A.</dc:creator>
<dc:creator>Karr, R. M.</dc:creator>
<dc:creator>muscolo, m. E.</dc:creator>
<dc:creator>Goers, L.</dc:creator>
<dc:creator>Yu, T.</dc:creator>
<dc:creator>Levin, T.</dc:creator>
<dc:creator>Pruneda, J. N.</dc:creator>
<dc:creator>Lesser, C.</dc:creator>
<dc:date>2025-12-04</dc:date>
<dc:identifier>doi:10.64898/2025.12.04.692159</dc:identifier>
<dc:title><![CDATA[Differential targeting of human pyroptotic Caspase-5 and Caspase-4 by Shigella OspC2 and OspC3]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-12-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2025.12.04.692306v1?rss=1">
<title>
<![CDATA[
The human cytomegalovirus chemokine binding protein UL22A is necessary for efficient reactivation from latency in CD34+ hematopoietic progenitor cells. 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2025.12.04.692306v1?rss=1</link>
<description><![CDATA[
Herpesviruses and Poxviruses encode secreted chemokine binding proteins that prevent the interaction between chemokines and their cognate G protein coupled receptors to alter chemotactic gradients and intracellular signaling pathways. Human cytomegalovirus (HCMV) encodes the secreted protein UL22A (formerly UL21.5), which is described as a CCL5 (RANTES) binding protein and requires sulfation at two tyrosine residues (Y65 and Y69) for efficient RANTES interaction. In this report, we show that the UL22A protein, and the UL22A Y65 and Y69 residues are necessary for efficient HCMV reactivation from latency in CD34+ hematopoietic progenitor cells and that UL22A expression is essential for reactivation in a humanized mouse model of latency. However, RANTES neutralization is not sufficient to complement the in vitro reactivation defect of UL22A mutant viruses. These data suggest that UL22A plays an important role in latency, possibly through interactions with additional chemokines or other types of ligands via its tyrosine residues, in order to mediate efficient HCMV reactivation.

IMPORTANCEHCMV is a ubiquitous herpesvirus that infects 60-90% of the population worldwide. In immunocompetent individuals, primary infection is asymptomatic and results in lifelong latent infection in CD34+ hematopoietic progenitor cells (HPCs). Viral reactivation remains a major complication for immunosuppressed individuals, but current therapeutics targeting HCMV replication show significant toxicity. Thus, a better understanding of the mechanisms controlling latency and reactivation is necessary to develop new therapeutics targeting these stages of the HCMV lifecycle. We show that virus lacking the HCMV chemokine binding protein UL22A is incapable of efficient reactivation in CD34+ HPCs and in vivo. UL22A tyrosine residues important for interaction with the chemokine RANTES are necessary for reactivation. However, neutralizing RANTES does not complement the reactivation defect of UL22A mutant viruses, demonstrating that UL22A has functions other than RANTES binding. Together, our results reveal a novel role for UL22A in HPCs and a new understanding of UL22A-chemokine interactions.
]]></description>
<dc:creator>Turner, R.</dc:creator>
<dc:creator>Diggins, N.</dc:creator>
<dc:creator>Slind, L. E.</dc:creator>
<dc:creator>Mitchell, J.</dc:creator>
<dc:creator>Pham, A.</dc:creator>
<dc:creator>Parkins, C.</dc:creator>
<dc:creator>Perez, W.</dc:creator>
<dc:creator>Medica, S.</dc:creator>
<dc:creator>Denton, M.</dc:creator>
<dc:creator>Andoh, T. F.</dc:creator>
<dc:creator>Webb, G. M.</dc:creator>
<dc:creator>Andrade-Vera, D.</dc:creator>
<dc:creator>Streblow, D. N.</dc:creator>
<dc:creator>Caposio, P.</dc:creator>
<dc:creator>Hancock, M. H.</dc:creator>
<dc:date>2025-12-04</dc:date>
<dc:identifier>doi:10.64898/2025.12.04.692306</dc:identifier>
<dc:title><![CDATA[The human cytomegalovirus chemokine binding protein UL22A is necessary for efficient reactivation from latency in CD34+ hematopoietic progenitor cells.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-12-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2025.12.02.691930v1?rss=1">
<title>
<![CDATA[
Prodromal pathogenesis of CLN7 Batten Disease revealed by multimodal biomarkers in macaques 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2025.12.02.691930v1?rss=1</link>
<description><![CDATA[
Neuronal ceroid lipofuscinosis type 7 (CLN7) is a devastating paediatric neurodegenerative disorder with no cure and limited natural history data to guide therapeutic development. Here, we present the first multimodal characterization of prodromal and early-stage CLN7 disease in Japanese macaques carrying a spontaneous CLN7-/- mutation.

Using structural T2-weighted MRI for volumetry, [18F]FDG PET for glucose metabolism, and [11C]PBR28 PET for neuroinflammation, we observed region-dependent patterns in volumetric, molecular, and metabolic alterations.

MRI confirmed the presence of disease-associated atrophy in many cortical and subcortical brain regions, consistent with human pathology, [18F]FDG PET revealed early cortical and subcortical widespread hypometabolism, and [11C]PBR28 PET imaging detected progressive neuroinflammation in the same brain areas. CSF analyses further showed age-dependent increases in neurofilament light (NfL), providing convergent evidence for neurodegeneration.

Together, these results define a prodromal trajectory in CLN7 disease, establish sensitive imaging and fluid biomarkers, and validate the macaque model as a powerful platform for testing interventions.
]]></description>
<dc:creator>Liguore, W. A.</dc:creator>
<dc:creator>Hilven, W.</dc:creator>
<dc:creator>Lurye, L.</dc:creator>
<dc:creator>Akkermans, J.</dc:creator>
<dc:creator>Decrop, M.</dc:creator>
<dc:creator>Zweig, R.</dc:creator>
<dc:creator>Sherman, L. S.</dc:creator>
<dc:creator>McBride, J. L.</dc:creator>
<dc:creator>Bertoglio, D.</dc:creator>
<dc:creator>Weiss, A. R.</dc:creator>
<dc:date>2025-12-05</dc:date>
<dc:identifier>doi:10.64898/2025.12.02.691930</dc:identifier>
<dc:title><![CDATA[Prodromal pathogenesis of CLN7 Batten Disease revealed by multimodal biomarkers in macaques]]></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/10.64898/2025.12.08.693047v1?rss=1">
<title>
<![CDATA[
Endogenous Osteocyte-Osteoclast Signaling Enables Growth Factor-Free Bone Remodeling, Drug Response, and Cancer Invasion in a Nanoscale Calcified Bone-on-a-Chip Model 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2025.12.08.693047v1?rss=1</link>
<description><![CDATA[
Bone homeostasis depends on spatially orchestrated interactions among osteoclasts, osteoblasts, and osteocytes that are embedded within a unique extracellular matrix that is mineralized on the nanoscale. Reconstructing these interactions to enable autonomous cell differentiation and tissue remodeling has remained a significant challenge towards mimicking adequate bone physiology and disease in-vitro. Here, we present an engineered model that spatially defines the paracrine communication of heterogeneous cell populations within bone tissue that supports the rapid maturation of primary osteoblasts into osteocytes, the differentiation of immune cells into osteoclasts, and calcified tissue resorption within a mineralized cell-laden bone-like tissue. We demonstrate that nanoscale mineralization of cell-laden collagen hydrogels on-a-chip enhances osteoblast to osteocyte differentiation, whereas osteocytes in the matrix accelerate osteoclastogenesis and remodeling in a spatially defined manner without the need for exogenous growth factors. Osteocyte-dependent osteoclastogenesis on-a-chip outperformed conventional stimulation with RANKL and M-CSF, reproduced the clinical response of anti-resorptive drugs, and mimicked established tumor-bone interactions observed in invasive oral cancer. By replicating essential aspects of bone composition and function, this system provides a robust, self-regulated microphysiologic model to investigate bone remodeling, cancer-bone crosstalk, and therapeutic interventions.
]]></description>
<dc:creator>Sousa, M.</dc:creator>
<dc:creator>Athirasala, A.</dc:creator>
<dc:creator>Roth, D.</dc:creator>
<dc:creator>Fraga, M. A.</dc:creator>
<dc:creator>Vignolo, S.</dc:creator>
<dc:creator>Doe, A.</dc:creator>
<dc:creator>Lee, J.</dc:creator>
<dc:creator>Romanowicz, G.</dc:creator>
<dc:creator>Nguyen, J.</dc:creator>
<dc:creator>Lin, A.</dc:creator>
<dc:creator>Franca, C. M.</dc:creator>
<dc:creator>Guldberg, R.</dc:creator>
<dc:creator>Bertassoni, L. E.</dc:creator>
<dc:date>2025-12-11</dc:date>
<dc:identifier>doi:10.64898/2025.12.08.693047</dc:identifier>
<dc:title><![CDATA[Endogenous Osteocyte-Osteoclast Signaling Enables Growth Factor-Free Bone Remodeling, Drug Response, and Cancer Invasion in a Nanoscale Calcified Bone-on-a-Chip Model]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-12-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2025.12.10.693544v1?rss=1">
<title>
<![CDATA[
Activation of cannabinoid receptor CB1 leads to aberrant myelination in development 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2025.12.10.693544v1?rss=1</link>
<description><![CDATA[
The endocannabinoid system (ECS) has a widespread role in the development and function of the central nervous system (CNS). Cannabinoid receptors like CB1 and CB2 can be activated with exogenous cannabinoids most popularly known as tetrahydrocannabinol (THC) or cannabis and cannabidiol (CBD). The components of the ECS are expressed early in fetal development, and prenatal exposure to cannabis can lead to structural changes in white matter. White matter is composed of neuronal axons ensheathed in myelin, a lipid-rich insulation that facilitates saltatory conduction and maintains axon integrity. In the CNS, myelin is made by specialized glial cells called oligodendrocytes (OLs), which in addition to neurons also express components of the ECS. However, while several studies have focused on how the ECS regulates neuronal development, there is a limited understanding of its impact on OL development or myelin formation.

Therefore, our current study set out to understand how pharmacological activation of the ECS alters OL differentiation and myelin formation in vivo. We administered WIN 55,212-2 (WIN 55), a CB1 and CB2 agonist, to larval zebrafish and longitudinally analyzed OL development and myelination in vivo. Interestingly, we observed an increase in non-axonal ensheathments in the spinal cord, which appeared to be surrounding neuronal cell bodies. These non-axonal ensheathments were dependent on CB1, as the addition of WIN 55 in a global CB1 mutant prevented this phenotype. Furthermore, this ectopic cell body ensheathment occurred independently from normal myelination processes, as individual OLs did not exhibit changes in the number of myelin sheaths, sheath length, or total myelin output. This study shows that activation of CB receptors in vivo leads to increased non-axonal ensheathment without significantly changing OL differentiation or normal myelin formation. Future studies can further investigate the pathways that drive this phenotype to better understand how exogenous cannabinoid activation can regulate the precision of oligodendrocyte ensheathment.
]]></description>
<dc:creator>Miramontes, T. G.</dc:creator>
<dc:creator>Hamling, K. R.</dc:creator>
<dc:creator>Doan, R. A.</dc:creator>
<dc:creator>Singh, S.</dc:creator>
<dc:creator>Collins, H. Y.</dc:creator>
<dc:creator>Emery, B.</dc:creator>
<dc:creator>Call, C. L.</dc:creator>
<dc:creator>Monk, K.</dc:creator>
<dc:date>2025-12-11</dc:date>
<dc:identifier>doi:10.64898/2025.12.10.693544</dc:identifier>
<dc:title><![CDATA[Activation of cannabinoid receptor CB1 leads to aberrant myelination in development]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-12-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2025.12.11.693769v1?rss=1">
<title>
<![CDATA[
Decoding the interactome for Cyclic-di-AMP-producing enzyme diadenylate cyclase (DacA) 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2025.12.11.693769v1?rss=1</link>
<description><![CDATA[
Diadenylate cyclase (DacA) synthesizes the second messenger cyclic di-AMP (c-di-AMP), which regulates essential cellular processes across many Gram-positive and select Gram-negative bacteria/archaeal lineages. Although DacA is known to interact with regulators such as GlmM and CdaR, the breadth and functional relevance of its interactome remains poorly defined. Our study seeks to identify novel protein-protein interactions to further elucidate their unknown regulatory mechanisms and cellular roles. Using Streptococcus mutans as a model, we engineered a Flag-tagged strain (DacA-FLAG) and then performed co-immunoprecipitation under non-crosslinked and crosslinked conditions followed by mass spectrometry. We identified 22 candidates interacting proteins in non-crosslinked samples, 18 in crosslinked samples, and 6 shared between conditions. Selected partners were validated in vivo using split luciferase complementation. Notably, SMU_723 emerged as a key binding partner. AlphaFold-guided modeling predicated a direct DacA and SMU_723 interaction interface involving threonine 147, glutamine 148, and threonine 149 in DacA . Site-directed mutagenesis of these residues impaired binding, confirming their critical role. An SMU_723 deletion phenocopied a dacA deletion strain, sharing prolonged lag phase, aberrant cell morphology, reduced acid production and acid tolerance, impaired sorbitol metabolism, decreased colonization in a Drosophila model, and delayed growth upon calcium stimulation. These shared phenotypes suggest a functional and possibly regulatory link between DacA and SMU_723. Given SMU_723 sequence homology consistent with a calcium transporter, these data suggest that the DacA-723 interaction contributes to calcium homeostasis and/or calcium-responsive signaling. Collectively, our findings expand the DacA interaction network and implicate calcium transport in c-di-AMP-mediated regulation in S. mutans.

ImportanceMapping the DacA interactome reveals how environmental and intracellular cues tune c-di-AMP signaling to control stress adaptation, ion balance, and virulence traits in S. mutans. By identifying new DacA-associated proteins and validating SMU_723 as a previously unrecognized interactor with genetic and phenotypic linkage to DacA, this study provides new insights into the mechanistic framework for c-di-AMP regulation in S. mutans. The connection between DacA and a putative calcium transporter highlights a plausible axis that couples second-messenger signaling to calcium homeostasis, with implications for biofilm physiology and pathogenesis. These insights open new avenues to therapeutically modulate c-di-AMP pathways.
]]></description>
<dc:creator>Mu, R.</dc:creator>
<dc:creator>Xie, B.</dc:creator>
<dc:creator>Momeni, S.</dc:creator>
<dc:creator>Wu, H.</dc:creator>
<dc:date>2025-12-12</dc:date>
<dc:identifier>doi:10.64898/2025.12.11.693769</dc:identifier>
<dc:title><![CDATA[Decoding the interactome for Cyclic-di-AMP-producing enzyme diadenylate cyclase (DacA)]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-12-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2025.12.10.693492v1?rss=1">
<title>
<![CDATA[
The cyclic nucleotide binding sites of Swiss-Cheese, the Drosophila orthologue of human PNPLA6, are required for its catalytic function 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2025.12.10.693492v1?rss=1</link>
<description><![CDATA[
Mutations in Swiss-cheese (SWS) or its vertebrate ortholog PNLPA6, also called Neuropathy Target Esterase (NTE), cause progressive neuronal degeneration in Drosophila and mice and several complex syndromes in humans. These include mental retardation, spastic paraplegia, ataxia and blindness and several other symptoms. SWS and PNPLA6 are widely expressed in neurons and in several glial cell types in Drosophila and mice and both cell types require SWS/PNPLA6 function autonomously. SWS and PNPLA6 are structurally and functionally conserved because expression of human or mouse PNPLA6 can replace SWS in flies. These orthologues share several domains, including the highly conserved phospholipase domain that mediates its function in deacetylating phosphatidylcholine (PC) to lysophosphatidylcholine and glycerophosphocholine. In addition, they share three cyclic nucleotide binding sites and although about 10% of the known disease-causing mutations occur in these sites, their function is still unknown. We therefore generated mutations in these sites in SWS to address what consequences this has for the function of the protein. Mutating only one site (SWSG558E) results in a partially functional protein that rescues the sws knockdown and that decreases PC when overexpressed. However, mutating all three sites (SWS{Delta}CNB) renders SWS non-functional and results in an increase of PC when overexpressed, suggesting that cyclic nucleotide binding can regulate the phospholipase function.
]]></description>
<dc:creator>Law, A.</dc:creator>
<dc:creator>Wentzell, J.</dc:creator>
<dc:creator>da Cruz, A.</dc:creator>
<dc:creator>Marney, L.</dc:creator>
<dc:creator>Kretzschmar, D.</dc:creator>
<dc:date>2025-12-13</dc:date>
<dc:identifier>doi:10.64898/2025.12.10.693492</dc:identifier>
<dc:title><![CDATA[The cyclic nucleotide binding sites of Swiss-Cheese, the Drosophila orthologue of human PNPLA6, are required for its catalytic function]]></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/10.64898/2025.12.12.694003v1?rss=1">
<title>
<![CDATA[
P/Q-type voltage-gated calcium channels regulate calcium signaling and developmental myelination in oligodendrocyte lineage cells 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2025.12.12.694003v1?rss=1</link>
<description><![CDATA[
Oligodendrocyte lineage cells (OLCs) are glia that arise as oligodendrocyte progenitor cells (OPCs) in the central nervous system (CNS) and may persist as progenitors or differentiate into myelin-producing oligodendrocytes (OLs). OLCs are sensitive to neuronal activity, which can influence myelin formation via activity-dependent myelination. Calcium influx in OLCs regulates many developmental processes, including stabilizing newly formed sheaths. OLCs possess P/Q-type voltage-gated calcium channels (VGCC) that contribute to calcium influx and mutations in these channels are implicated in a spectrum of neurological disorders, yet the functional significance of P/Q-type channels in OLCs is not well understood. In this study, we employ zebrafish to investigate the role of P/Q-type channels in OLCs in vivo during development. We use global and cell-type specific CRISPR/Cas9-mediated genome editing approaches in conjunction with live imaging and physiology to characterize the morphology and signaling properties of OLCs with mutated P/Q-type channel genes. P/Q-type channels are required for normal myelination in the developing CNS and mutants present with decreased amplitude sheath calcium transients, reduced myelin production, shorter myelin sheaths and blebbing membrane structures during development. These findings provide new insight into the role of P/Q-type calcium channels in regulating OLC development and myelination.
]]></description>
<dc:creator>Piller, M.</dc:creator>
<dc:creator>Doan, R.</dc:creator>
<dc:creator>Call, C. L.</dc:creator>
<dc:creator>Smith, S.</dc:creator>
<dc:creator>Monk, K.</dc:creator>
<dc:date>2025-12-13</dc:date>
<dc:identifier>doi:10.64898/2025.12.12.694003</dc:identifier>
<dc:title><![CDATA[P/Q-type voltage-gated calcium channels regulate calcium signaling and developmental myelination in oligodendrocyte lineage cells]]></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/10.64898/2025.12.11.693731v1?rss=1">
<title>
<![CDATA[
Selective infection and loss of PRDM1+ LN Tfh cells in uncontrolled HIV infection precludes formation of Tfh reservoirs under ART 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2025.12.11.693731v1?rss=1</link>
<description><![CDATA[
The multifaceted and long-lived nature of HIV-1 reservoirs has proven to be a formidable obstacle in the development of a therapeutic cure of HIV-1. One of the major dimensions of the HIV-1 reservoir is its prevalence and persistence in tissue environments, including in lymph nodes (LN). Within LN, T follicular helper (Tfh) cells are widely considered as the dominant sub-reservoir in viremic people with HIV (PWH). However, whether Tfh cells survive to establish a reservoir in PWH undergoing suppressive antiretroviral therapy (ART) has remained controversial. To address these issues, we deeply phenotyped over 500,000 cells and identified over 2,000 HIV-1 infected cells by employing single-cell multiomics on LN from PWH during viremia and suppressed on ART. While we detected HIV-1 infected Tfh cells, the majority of infected cells during viremia and ART had non-follicular phenotypes and were instead heterogeneously distributed between various CD4+ T-cell subsets. Within-subset comparisons of HIV+ and HIV- cells revealed heightened activation signatures and altered cell cycle states during viremia, but largely similar features during ART. Furthermore, the comparison between viremia and ART in PWH highlighted a noncanonical Tfh subset - defined by high locus accessibility and transcription of PRDM1 - that is selectively depleted during viremia, recovers after ART, and is highly susceptible to infection in vitro. Our work suggests a revised direction for the HIV-1 cure field where a mandate of any comprehensive strategy will be to address the high proportional burden of non-follicular cells within the HIV reservoir.
]]></description>
<dc:creator>Wu, V. H.</dc:creator>
<dc:creator>Nordin, J. M.</dc:creator>
<dc:creator>Pampena, M. B.</dc:creator>
<dc:creator>Rachimi, S.</dc:creator>
<dc:creator>Burgess, W. L.</dc:creator>
<dc:creator>Clark, M.</dc:creator>
<dc:creator>Robinson, J. A.</dc:creator>
<dc:creator>Estrada, P. M. d. R.</dc:creator>
<dc:creator>Schleimann, M. H.</dc:creator>
<dc:creator>Torres-Ruiz, F.</dc:creator>
<dc:creator>Gonzalez-Navarro, M.</dc:creator>
<dc:creator>Salgado, G.</dc:creator>
<dc:creator>Luna-Villalobos, Y. A.</dc:creator>
<dc:creator>Reyes-Teran, G.</dc:creator>
<dc:creator>Bar, K. J.</dc:creator>
<dc:creator>Estes, J. D.</dc:creator>
<dc:creator>Sogaard, O. S.</dc:creator>
<dc:creator>Romberg, N.</dc:creator>
<dc:creator>Avila-Rios, S.</dc:creator>
<dc:creator>Betts, M. R.</dc:creator>
<dc:date>2025-12-15</dc:date>
<dc:identifier>doi:10.64898/2025.12.11.693731</dc:identifier>
<dc:title><![CDATA[Selective infection and loss of PRDM1+ LN Tfh cells in uncontrolled HIV infection precludes formation of Tfh reservoirs under ART]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-12-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2025.12.12.693851v1?rss=1">
<title>
<![CDATA[
A window trial in metastatic pancreatic ductal adenocarcinoma reveals resistance mechanisms to targeting the KRAS-MEK pathway 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2025.12.12.693851v1?rss=1</link>
<description><![CDATA[
Copy number alterations of KRAS, mutated in over 90% of pancreatic ductal adenocarcinomas (PDAC), and MYC occur in 30-40% of PDAC. Here we demonstrate that KRAS and MYC are frequently co-gained and accompanied with worse prognosis in PDAC. In a Window-of-Opportunity clinical trial for metastatic PDAC, serial biopsies and deep multi-omics analyses were utilized to explore resistance mechanisms to MEK inhibition, as a surrogate for KRAS inhibition. Tumors from four of 14 patients showed Ki-67/CA19-9-based biomarker response (BR). Non-BR tumors were enriched for KRAS/MYC co-gain and KRASG12D variant. A transcriptomic signature of BR tumors was inversely correlated with KRASG12D/MYC co-gain in a large PDAC dataset and predictive for KRAS inhibitor response in multiple models. Finally, co-targeting KRAS and MYC was synergistic in KRASG12D/MYC co-gain PDAC. Together, this study provides insight into KRAS inhibitor resistance and supports MYC as an important target to improve patient outcomes in this deadly disease.
]]></description>
<dc:creator>Tsuda, M.</dc:creator>
<dc:creator>Keith, D.</dc:creator>
<dc:creator>Daniel, C. J.</dc:creator>
<dc:creator>Pelz, C.</dc:creator>
<dc:creator>Blise, K. E.</dc:creator>
<dc:creator>Ozmen, T. Y.</dc:creator>
<dc:creator>Ozmen, F.</dc:creator>
<dc:creator>Hawthone, K.</dc:creator>
<dc:creator>Eng, J. R.</dc:creator>
<dc:creator>Li, X.</dc:creator>
<dc:creator>Wang, X.</dc:creator>
<dc:creator>Zimny, H.</dc:creator>
<dc:creator>Sivagnanam, S.</dc:creator>
<dc:creator>Betre, K.</dc:creator>
<dc:creator>Kirchberger, N.</dc:creator>
<dc:creator>Chong, B.</dc:creator>
<dc:creator>Lee, J.</dc:creator>
<dc:creator>Matter, _.</dc:creator>
<dc:creator>Smith, A.</dc:creator>
<dc:creator>Agritelley, E. S.</dc:creator>
<dc:creator>Waugh, T.</dc:creator>
<dc:creator>Sannecy, A.</dc:creator>
<dc:creator>Alarcon, K.</dc:creator>
<dc:creator>Youm, I.</dc:creator>
<dc:creator>Protzek, S.</dc:creator>
<dc:creator>Egger, J.</dc:creator>
<dc:creator>English, I. A.</dc:creator>
<dc:creator>Yang, M.</dc:creator>
<dc:creator>Shah, V. M.</dc:creator>
<dc:creator>Link, J. M.</dc:creator>
<dc:creator>Creason, A. L.</dc:creator>
<dc:creator>Worth, P. J.</dc:creator>
<dc:creator>Goodyear, S. M.</dc:creator>
<dc:creator>Chin, K.</dc:creator>
<dc:creator>Muschler, J. L.</dc:creator>
<dc:creator>Suciu, C. G.</dc:creator>
<dc:creator>Corless, C. L.</dc:creator>
<dc:creator>Cao, S.</dc:creator>
<dc:creator>Soucek, L.</dc:creator>
<dc:creator>Kardosh, A.</dc:creator>
<dc:creator>Coussens, L. M.</dc:creator>
<dc:creator>Brody, J. R.</dc:creator>
<dc:creator>Lopez, C. D.</dc:creator>
<dc:creator>Mills, G. B.</dc:creator>
<dc:creator>Sears, R. C.</dc:creator>
<dc:date>2025-12-15</dc:date>
<dc:identifier>doi:10.64898/2025.12.12.693851</dc:identifier>
<dc:title><![CDATA[A window trial in metastatic pancreatic ductal adenocarcinoma reveals resistance mechanisms to targeting the KRAS-MEK pathway]]></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/10.64898/2025.12.15.694292v1?rss=1">
<title>
<![CDATA[
Chemoproteomic elucidation of β-lactam drug targets in Mycobacterium abscessus 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2025.12.15.694292v1?rss=1</link>
<description><![CDATA[
The pathogen Mycobacterium abscessus (Mab) can cause severe and difficult to treat chronic lung infections. Despite the rising incidence and clinical concern of Mab infections, treatment options are limited and often ineffective. Treatment is complicated by Mabs ability to persist in a non-replicative, drug-resistant state. Several {beta}-lactam antibiotics are potently bactericidal against Mab but are underutilized because their molecular mechanisms of action against Mab are incompletely understood. In the current study, we used {beta}-lactam-derived activity-based probes and chemoproteomics to report the first comprehensive list of enzymes in Mab targeted by {beta}-lactams. We compared {beta}-lactam targets across two Mab subspecies in actively replicating and non-replicative cultures, using a new carbon starvation model of persistence. We identified 17 targets that were active in every condition tested, seven of which were previously unknown to bind {beta}-lactams. Lastly, we characterized the {beta}-lactamase activity and {beta}-lactam inhibition profiles of nine Mab enzymes, demonstrating that imipenem inhibits these targets more effectively than cefoxitin. These findings provide clarity on the mechanisms of action of clinically relevant {beta}-lactams in Mab, a crucial step toward fully realizing their potential for treating infections caused by this opportunistic pathogen.
]]></description>
<dc:creator>Devlin, K. L.</dc:creator>
<dc:creator>Hutchinson, E.</dc:creator>
<dc:creator>Leach, D. T.</dc:creator>
<dc:creator>Nelson, W. C.</dc:creator>
<dc:creator>Gorham, L. J.</dc:creator>
<dc:creator>Lamichhane, G.</dc:creator>
<dc:creator>Lin, V. S.</dc:creator>
<dc:creator>Beatty, K. E.</dc:creator>
<dc:date>2025-12-15</dc:date>
<dc:identifier>doi:10.64898/2025.12.15.694292</dc:identifier>
<dc:title><![CDATA[Chemoproteomic elucidation of β-lactam drug targets in Mycobacterium abscessus]]></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/10.64898/2025.12.15.693905v1?rss=1">
<title>
<![CDATA[
Multi-timescale Rhythmic Dynamics in Rostral Ventromedial Medulla Neurons 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2025.12.15.693905v1?rss=1</link>
<description><![CDATA[
Neural circuits involved in sensory control must integrate fast, reflexive responses with slower, state-dependent modulation. This coordination is especially critical in the pain system, where rapid withdrawal must be balanced against slower adjustments in nociceptive sensitivity that reflect arousal, physiology, and internal state. We hypoth-esized that neurons of the rostral ventromedial medulla (RVM)--the principal brainstem node for descending pain control--operate across multiple temporal scales, combining fast stimulus-evoked responses with slower intrinsic dynamics. Using single-unit recordings from identified RVM ON-, OFF-, and NEUTRAL-cells and probabilistic modeling, we find that ON- and OFF-cells exhibit multi-phase population responses characterized by rapid activation and prolonged recovery dynamics lasting tens of seconds. In the absence of stimulation, the same neurons display coherent, quasi-periodic oscillations on the order of minutes, captured by Gaussian process models as predictable, low-frequency fluctuations. These rhythmic dynamics were specific to ON- and OFF-cells and occasionally coherent with autonomic parameters, suggesting coupling between nociceptive and homeostatic control loops. Together, these results identify a multi-timescale organizational principle within descending brainstem circuits, in which fast, reflex-related and slow, rhythmic processes jointly structure sensory modulation and internal state control.
]]></description>
<dc:creator>Ashworth, C.</dc:creator>
<dc:creator>Martenson, M.</dc:creator>
<dc:creator>Shi, Z.</dc:creator>
<dc:creator>De Preter, C. C.</dc:creator>
<dc:creator>Heinricher, M. M.</dc:creator>
<dc:creator>Mancini, F.</dc:creator>
<dc:date>2025-12-17</dc:date>
<dc:identifier>doi:10.64898/2025.12.15.693905</dc:identifier>
<dc:title><![CDATA[Multi-timescale Rhythmic Dynamics in Rostral Ventromedial Medulla Neurons]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-12-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2025.12.18.695265v1?rss=1">
<title>
<![CDATA[
In vitro evidence for bisphenol A as a human liver carcinogen: Environmentally relevant doses inhibit cancer protective ESR1 signaling in a human liver cell line 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2025.12.18.695265v1?rss=1</link>
<description><![CDATA[
I.Several environmentally ubiquitous endocrine disrupting chemicals (EDCs) are suspected carcinogens, but their mechanism(s) of action are unknown. In this study, we test the potential for a model EDC, bisphenol A (BPA), to both initiate (via oxidative mutagenesis) and promote (via endocrine disruption) liver carcinogenesis. This study is motivated by our prior finding that developmental BPA exposure caused hepatocellular carcinoma (HCC) in mice. Here, we provide in vitro evidence supporting a mechanism for BPA as a non-genotoxic carcinogen. Using a highly sensitive, error-corrected sequencing method, we demonstrate that human population-relevant doses of BPA cause mutations that are consistent with oxidative DNA damage; however, overall mutation frequency does not differ substantially from controls. In contrast, we show that BPA inhibits cancer-protective, estrogen-induced transcription of estrogen receptor 1 (ESR1) target genes in the presence of pre-pubertal, but not post-pubertal, levels of estradiol. These results constitute strong initial evidence supporting BPA as a liver cancer promoting agent. This mechanism may be generalizable to a wide range of environmental EDCs that are weak agonists for ESR1. This finding is critically important to prevention of HCC, which is prevalent, lethal, and poorly responsive to therapy.
]]></description>
<dc:creator>Weeks, E.</dc:creator>
<dc:creator>Kennedy, S. R.</dc:creator>
<dc:creator>Searles, R.</dc:creator>
<dc:creator>Carrothers, S.</dc:creator>
<dc:creator>Davis, B.</dc:creator>
<dc:creator>Carbone, L.</dc:creator>
<dc:creator>Turker, M.</dc:creator>
<dc:creator>Lloyd, R. S.</dc:creator>
<dc:creator>Weinhouse, C.</dc:creator>
<dc:date>2025-12-22</dc:date>
<dc:identifier>doi:10.64898/2025.12.18.695265</dc:identifier>
<dc:title><![CDATA[In vitro evidence for bisphenol A as a human liver carcinogen: Environmentally relevant doses inhibit cancer protective ESR1 signaling in a human liver cell line]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-12-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2025.12.18.695277v1?rss=1">
<title>
<![CDATA[
Epigenetic plasticity is a driver of heritable pollution tolerance in Atlantic killifish 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2025.12.18.695277v1?rss=1</link>
<description><![CDATA[
Heritable epigenetic adaptation to environmental stressors is a compelling but highly contested possibility. Previously, we showed evidence of a generationally heritable epigenetic memory at the cytochrome P450 1a (cyp1a) gene in wild Atlantic killifish (Fundulus heteroclitus) with acquired tolerance to polycyclic aromatic hydrocarbons (PAHs). This memory leads to blunted induction of cyp1a by PAHs; this blunted response protects against PAH-induced cancer. Here, using Oxford Nanopore long-read sequencing in PAH-tolerant and -sensitive F. heteroclitus embryos, we show that PAH-tolerant embryos displayed reduced plasticity in DNA methylation response to PAH, as compared to PAH-sensitive embryos, that was not due to mutational loss of CpG sites. Notably, we observed population differences in DNA methylation of genes in pathways linked to the PAH tolerance phenotype, including aryl hydrocarbon receptor (ahr) and voltage-gated potassium channel signaling, as well as developmental processes and energy metabolism. Specifically, we observed PAH-induced loss of cyp1a gene body methylation in PAH-sensitive but not-tolerant embryos. We observed similar patterns at cyp1b1, cyp1c1, and the aryl hydrocarbon receptor repressor, ahrr, which show similarly blunted expression in response to PAH challenge. The reduced loss in genic methylation in tolerant embryos was correlated with greater induction of natural anti-sense RNA transcripts in cis (cis-NATs), which may regulate transcription of these genes. Our data support the existence of stable epigenetic responses to chronic environmental stressors in a natural experimental setting, with broad implications for natural or directed adaptation strategies for other populations.
]]></description>
<dc:creator>Colwell, M.</dc:creator>
<dc:creator>Drown, M.</dc:creator>
<dc:creator>Flack, N.</dc:creator>
<dc:creator>Walls, C.</dc:creator>
<dc:creator>Faulk, C.</dc:creator>
<dc:creator>Carrothers, S.</dc:creator>
<dc:creator>Weeks, E.</dc:creator>
<dc:creator>Weinhouse, C.</dc:creator>
<dc:date>2025-12-22</dc:date>
<dc:identifier>doi:10.64898/2025.12.18.695277</dc:identifier>
<dc:title><![CDATA[Epigenetic plasticity is a driver of heritable pollution tolerance in Atlantic killifish]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-12-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2025.12.19.695559v1?rss=1">
<title>
<![CDATA[
Dysfunctional memory B cell responses to the protective repeat region of the Plasmodium circumsporozoite protein are associated with waning humoral immunity 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2025.12.19.695559v1?rss=1</link>
<description><![CDATA[
Malaria vaccines provide waning protection from disease that is correlated with the production of antibodies to the repeat region of the liver-stage circumsporozoite protein (CSP). CSP-based vaccines display limited durability in malaria-naive individuals yet are even less effective in malaria-experienced individuals, suggesting the generation of non-optimal humoral immunity in response to both infection and vaccination. To address this hypothesis, we performed a cross-species, comprehensive analysis of B cell responses to CSP after Plasmodium infection or immunization, focusing our analysis on the repeat and C-terminus domains included in malaria subunit vaccines. Herein we demonstrate that while multiple factors may impinge on strong, lasting protective CSP-specific immunity, two forces predominantly impact proper memory B cell differentiation; a) inhibition of germinal center formation by infection and b) skewed B cell differentiation due to the repetitive nature of the protective region of the CSP protein.
]]></description>
<dc:creator>McDougal, C. E.</dc:creator>
<dc:creator>Mkindi, C. G.</dc:creator>
<dc:creator>Rodda, L. B.</dc:creator>
<dc:creator>Lucarelli, C. V.</dc:creator>
<dc:creator>Langowski, M. D.</dc:creator>
<dc:creator>King, N. P.</dc:creator>
<dc:creator>Jongo, S.</dc:creator>
<dc:creator>Daubenberger, C.</dc:creator>
<dc:creator>Pepper, M.</dc:creator>
<dc:date>2025-12-22</dc:date>
<dc:identifier>doi:10.64898/2025.12.19.695559</dc:identifier>
<dc:title><![CDATA[Dysfunctional memory B cell responses to the protective repeat region of the Plasmodium circumsporozoite protein are associated with waning humoral immunity]]></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/10.64898/2025.12.20.695723v1?rss=1">
<title>
<![CDATA[
Application of Large Language Models for Annotating Genes into Reactome Pathways 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2025.12.20.695723v1?rss=1</link>
<description><![CDATA[
Reactome is the most comprehensive, open source, open access biological pathway knowledgebase, widely used in the research community. To ensure the highest quality of its content, human pathway data in Reactome is manually curated. However, manual curation is labor-intensive, time-consuming, and increasingly difficult to keep up with the ever-growing biomedical literature. Large language model (LLM)-driven artificial intelligence (AI) technologies are transforming many fields, including bioinformatics resource development. Applying LLM/AI technologies in Reactome may offer a powerful way to scale curation and consolidate pathway-related data into a single resource. This manuscript describes the first stage of our attempt to adopt LLM/AI technologies for Reactome manual curation. We developed an LLM workflow that can assist curators in adding new genes to existing pathways and refining the functional annotations of existing ones. The workflow predicts pathways in which genes are likely to function, identifies PubMed-indexed literature that may support these predictions, generates text summaries describing potential molecular mechanisms, and extracts functional relationships among biological entities from full-text PDF papers. To validate the workflow output, we used a computational approach based on semantic similarity between LLM workflow-generated summaries and Reactome manual annotations. The results show significant enrichment of high-similarity matches. Manual evaluation of 19 genes indicated that more than half of the outputs are useful for supporting curation. Based on these results, we developed an enhanced workflow that incorporates protein-protein interaction data, facilitating Reactomes reaction-based annotation. In summary, our initial adoption of LLM/AI technologies produced encouraging results and provides a practical framework for integrating AI-assisted methods into Reactomes curation pipeline. The strategies described here may be broadly applicable to community knowledgebases in general.
]]></description>
<dc:creator>Wu, G.</dc:creator>
<dc:creator>Matthews, L.</dc:creator>
<dc:creator>Boyer, N.</dc:creator>
<dc:creator>Milacic, M.</dc:creator>
<dc:creator>Beavers, D.</dc:creator>
<dc:creator>Li, N. T.</dc:creator>
<dc:creator>May, B.</dc:creator>
<dc:creator>Rothfels, K.</dc:creator>
<dc:creator>Shamovsky, V.</dc:creator>
<dc:creator>Stephan, R.</dc:creator>
<dc:creator>Gillespie, M.</dc:creator>
<dc:creator>Hermjakob, H.</dc:creator>
<dc:creator>DEustachio, P.</dc:creator>
<dc:creator>Stein, L. D.</dc:creator>
<dc:date>2025-12-23</dc:date>
<dc:identifier>doi:10.64898/2025.12.20.695723</dc:identifier>
<dc:title><![CDATA[Application of Large Language Models for Annotating Genes into Reactome Pathways]]></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/10.64898/2025.12.20.695739v1?rss=1">
<title>
<![CDATA[
Identification of senescence-associated drivers of tumour growth and progression using a novel microarray platform 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2025.12.20.695739v1?rss=1</link>
<description><![CDATA[
Senescence and the senescence associated secretory phenotype (SASP) are implicated in promoting early tumorigenesis but due to the complexity of SASP it has been difficult to identify the responsible factors. We used canonical SASP factors on our microenvironment microarray (MEMA) platform to systematically identify SASP-associated drivers of tumorigenesis in breast and lung cancer cells. We found multiple SASP factors enhanced the proliferation and overall cell numbers for both lung and breast cells grown on the MEMA, and that there was significant overlap in SASP-associated growth-promoting factors between the two different cell types. We validated the ability of several factors, including IL-6, TGF-{beta} and EGF, to drive growth in in vitro assays. Interestingly, these factors were effective in driving growth and survival in cells that were altered (either immortalized or fully transformed) but not in normal cells and impacted breast cells differently depending on the age of the patient. RNAseq identified upregulation of wound-healing and stem-cell programs in SASP factor-treated cells. Many of these same SASP factors were present in conditioned media collected from senescent cells, which enhanced the growth of both lung and breast cancer cells, and inhibitors of the specific SASP factors partially reduced growth. Similarly, targeted inhibition of EGF partially reduced lung tumour growth in xenografts when senescent but not normal fibroblasts were co-implanted. Our findings have identified core SASP drivers of tumorigenesis and suggest that effective tumorigenesis driven by SASP is multifactorial and requires alterations in the target cells to achieve maximal response.
]]></description>
<dc:creator>Ou, H.-L.</dc:creator>
<dc:creator>Hoffmann, R. J.</dc:creator>
<dc:creator>Smith, R.</dc:creator>
<dc:creator>Devlin, K. L.</dc:creator>
<dc:creator>Dane, M.</dc:creator>
<dc:creator>Munoz-Espin, D.</dc:creator>
<dc:creator>Korkola, J.</dc:creator>
<dc:date>2025-12-23</dc:date>
<dc:identifier>doi:10.64898/2025.12.20.695739</dc:identifier>
<dc:title><![CDATA[Identification of senescence-associated drivers of tumour growth and progression using a novel microarray platform]]></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/10.64898/2025.12.22.695544v1?rss=1">
<title>
<![CDATA[
FHIR as a Unifying Format for Genomic Research Data Tracking, Aggregation, and Integration 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2025.12.22.695544v1?rss=1</link>
<description><![CDATA[
The increasing complexity of genomic research demands standardized data sharing and integration. The Fast Healthcare Interoperability Resources (FHIR) specification has become a well-established standard for exchanging data among health data systems. While designed primarily for clinical and patient data in health care environments, it also has applicability to represent genomic research data and offers a path for aggregating and integrating extremely rich datasets that have traditionally remained siloed and disparate. To study this potential, we developed FHIR Aggregator, an integration of seven major biomedical repositories, including the Genomic Data Commons, GTEx, HTAN, and DepMap, that covered 142334 patients, 819251 specimens, 1096491 observations and 711166 documents. We explore the various ways the FHIR standard can be applied to structure genomic research data and enable new possibilities. We demonstrate how FHIR can be used, where it succeeds or falls short, and which concepts must be extended to better support large-scale clinical and genomics research projects.
]]></description>
<dc:creator>Sanati, N.</dc:creator>
<dc:creator>Walsh, B.</dc:creator>
<dc:creator>Gray, P.</dc:creator>
<dc:creator>Hagen, L.</dc:creator>
<dc:creator>Carroll, R. J.</dc:creator>
<dc:creator>Heath, A. P.</dc:creator>
<dc:creator>Charbonneau, A.</dc:creator>
<dc:creator>Ellrott, K.</dc:creator>
<dc:date>2025-12-24</dc:date>
<dc:identifier>doi:10.64898/2025.12.22.695544</dc:identifier>
<dc:title><![CDATA[FHIR as a Unifying Format for Genomic Research Data Tracking, Aggregation, and Integration]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-12-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2025.12.23.696181v1?rss=1">
<title>
<![CDATA[
The synaptic vesicle priming protein Munc13 mediates evoked somatodendritic dopamine release 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2025.12.23.696181v1?rss=1</link>
<description><![CDATA[
Midbrain dopamine neurons secrete dopamine from their somata and dendrites in addition to their axonal release. Shared and distinct properties have been proposed for somatodendritic and axonal release, but the mechanisms of somatodendritic release remain unclear. We here used mouse genetics, electrophysiology, and imaging to define roles of the synaptic vesicle priming protein Munc13 in somatodendritic release in comparison to axonal secretion. Munc13 ablation in dopamine neurons decreased evoked but not spontaneous somatodendritic dopamine transmission measured as D2 receptor-mediated currents. Imaging with a fluorescent dopamine sensor confirmed this finding and established comparable importance for Munc13 in somatodendritic and axonal secretion. Pharmacological experiments revealed that release from adrenergic terminals contributes to D2 receptor-mediated currents, and their relative contribution was enhanced after Munc13 knockout. Altogether, these data establish important roles for Munc13 in evoked somatodendritic release. These roles are similar to Munc13 functions in axonal dopamine release and at fast synapses. Spontaneous midbrain dopamine release does not necessitate Munc13 in dopamine neurons and may rely on a release pathway that is independent of the prototypical release machinery employed at synapses.
]]></description>
<dc:creator>Lebowitz, J. J.</dc:creator>
<dc:creator>Banerjee, A.</dc:creator>
<dc:creator>Handy, G.</dc:creator>
<dc:creator>Williams, J. T.</dc:creator>
<dc:creator>Kaeser, P. S.</dc:creator>
<dc:date>2025-12-24</dc:date>
<dc:identifier>doi:10.64898/2025.12.23.696181</dc:identifier>
<dc:title><![CDATA[The synaptic vesicle priming protein Munc13 mediates evoked somatodendritic dopamine release]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-12-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2025.12.26.696629v1?rss=1">
<title>
<![CDATA[
A New Paradigm of Transcriptional Regulation by the SufR-Like Iron-Sulfur Transcription Factors 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2025.12.26.696629v1?rss=1</link>
<description><![CDATA[
SufR is an iron-sulfur ([4Fe-4S]) cluster-containing transcription factor belonging to an uncharacterized domain family (COG2345). It has been shown to negatively regulate the sulfur utilization factor (SUF) Fe-S biogenesis system in several Gram-negative and Gram-positive bacteria including Cyanobacteria, Mycobacteria and Streptomyces. The structural basis for its DNA recognition and transcriptional regulation by the SufR-like proteins remains enigmatic. In this study, we present the cryo-EM structure of Mycobacterium tuberculosis SufR bound to its promoter, revealing a new domain architecture. Our structural, biochemical and molecular analyses show that SufR possesses an unusual [4Fe-4S] cluster coordination environment in the sensory domain and recognizes its promoter via a dual-module mechanism using both the AT-hook and the helix-turn-helix (HTH) DNA-binding motif in the DNA-binding domain. This DNA recognition strategy differs from a canonical winged HTH transcription factor. Moreover, our bioinformatic analysis and structural modeling suggest that SufR and SufR-like proteins in the COG2345 family represent a large, previously uncharacterized family of transcription factors widely distributed across bacteria and archaea. Together, these findings establish SufR-like proteins as a new model for transcriptional regulation by Fe-S transcription factors in prokaryotes and uncover the underappreciated evolutionary versatility of AT-hooks.
]]></description>
<dc:creator>Lu, Z.</dc:creator>
<dc:creator>Ong, C.</dc:creator>
<dc:creator>Zhang, L.</dc:creator>
<dc:creator>Wan, T.</dc:creator>
<dc:creator>Davulcu, O.</dc:creator>
<dc:creator>de Farias, M.</dc:creator>
<dc:creator>Zhang, L.-M.</dc:creator>
<dc:date>2025-12-27</dc:date>
<dc:identifier>doi:10.64898/2025.12.26.696629</dc:identifier>
<dc:title><![CDATA[A New Paradigm of Transcriptional Regulation by the SufR-Like Iron-Sulfur Transcription Factors]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-12-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.01.07.698299v1?rss=1">
<title>
<![CDATA[
Evaluating Acridones as Novel Therapeutics for Human Babesiosis 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.01.07.698299v1?rss=1</link>
<description><![CDATA[
BackgroundHuman babesiosis is an emerging tick-borne disease caused by Babesia parasites, most notably Babesia microti and Babesia duncani in North America and Babesia divergens in Europe. Infections can be severe, persistent, or relapse despite treatment, and current therapeutic options remain limited, underscoring the urgent need for new and effective treatment options.

MethodsAcridone derivatives, originally developed as potent antimalarial agents against multiple life cycle stages of the malaria parasites, were evaluated for their antibabesial activity using continuous in vitro culture systems of B. duncani and B. divergens. Lead candidates were assessed for selectivity against human cell lines to establish preliminary safety profiles, and select compounds were further advanced into preliminary in vivo efficacy studies using murine models of B. duncani and B. microti babesiosis to assess their therapeutic potential.

ResultsNine prioritized acridone derivatives demonstrated potent in vitro activity against both B. duncani and B. divergens, accompanied by favorable selectivity indices relative to human cell lines. However, in vivo evaluation of representative compounds did not achieve parasite clearance in murine models. Structure-activity relationship (SAR) analyses highlighted key structural features that are critical for maintaining antibabesial potency and offer guidance for further lead optimization.

ConclusionsAcridone derivatives show strong in vitro antibabesial activity and represent promising lead chemotypes for therapeutic development. To advance these candidates, future studies focused on optimizing their pharmacokinetic properties, improving in vivo efficacy, and evaluating synergistic combination regimens will be essential for progressing toward effective treatments for human babesiosis.
]]></description>
<dc:creator>Vydyam, P.</dc:creator>
<dc:creator>Zhang, E.</dc:creator>
<dc:creator>Chattopadhyay Pal, A.</dc:creator>
<dc:creator>Dodean, R.</dc:creator>
<dc:creator>Kancharla, P.</dc:creator>
<dc:creator>Kelly, J. X.</dc:creator>
<dc:creator>Ben Mamoun, C.</dc:creator>
<dc:date>2026-01-08</dc:date>
<dc:identifier>doi:10.64898/2026.01.07.698299</dc:identifier>
<dc:title><![CDATA[Evaluating Acridones as Novel Therapeutics for Human Babesiosis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-01-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.01.08.697228v1?rss=1">
<title>
<![CDATA[
MYST acetyltransferases are a targetable therapeutic vulnerability in SETBP1-mutant leukemia 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.01.08.697228v1?rss=1</link>
<description><![CDATA[
Mutations in SET binding protein 1 (SETBP1) are associated with an adverse prognosis in myeloid malignancies. These mutations stabilize SETBP1 protein, driving increased expression of a progenitor-associated gene expression program through incompletely described mechanisms. A proteomic screen revealed interactions between SETBP1 and members of the MYST acetyltransferase complexes, including the catalytic subunits--KAT6A and KAT7. Mutant SETBP1 increases the localization of MYST complexes at known SETBP1 target genes, including the Hoxa cluster, where it drives increased histone acetylation and gene expression. Treatment of SETBP1D868N-expressing myeloid progenitors with MYST inhibitors reduced target gene expression. To establish the efficacy of MYST inhibition in vivo, we treated mice harboring a syngeneic SETBP1-mutant leukemia with the clinical-grade MYST inhibitor--PF-9363. This resulted in complete hematologic control and increased survival. MYST inhibition was also highly effective against a SETBP1-mutant PDX model. These studies identify MYST acetyltransferases as promising therapeutic targets in SETBP1-mutant malignancies.

Statement of SignificanceSETBP1 mutations are markers of high-risk myeloid malignancies, but we lack any targeted therapies to improve outcomes. In this study, we identify MYST acetyltransferases as key drivers of mutant SETBP1-driven transcription. MYST inhibitors are highly effective against SETBP1-mutant leukemia and represent a promising avenue for clinical translation.
]]></description>
<dc:creator>Maxson, J. E.</dc:creator>
<dc:creator>Braun, T. P.</dc:creator>
<dc:creator>Carlson, H.</dc:creator>
<dc:creator>Nguyen, T.</dc:creator>
<dc:creator>Tauchmann, S.</dc:creator>
<dc:creator>Carratt, S. A.</dc:creator>
<dc:creator>Kruer, T. L.</dc:creator>
<dc:creator>Dankyi, N. A.</dc:creator>
<dc:creator>Balasis, M.</dc:creator>
<dc:creator>Kim, H.</dc:creator>
<dc:creator>Tsai, C.-F.</dc:creator>
<dc:creator>Liu, T.</dc:creator>
<dc:creator>Shrestha, S.</dc:creator>
<dc:creator>Fischer, J. M.</dc:creator>
<dc:creator>Padron, E.</dc:creator>
<dc:date>2026-01-08</dc:date>
<dc:identifier>doi:10.64898/2026.01.08.697228</dc:identifier>
<dc:title><![CDATA[MYST acetyltransferases are a targetable therapeutic vulnerability in SETBP1-mutant leukemia]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-01-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.01.13.699315v1?rss=1">
<title>
<![CDATA[
Mutant ASXL1 Drives Transcriptional Activation and Repression in Human Hematopoiesis 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.01.13.699315v1?rss=1</link>
<description><![CDATA[
Mutations in the epigenetic regulator ASXL1 are common in myeloid malignancies and portend a near-universally poor prognosis. While multiple mechanisms for mutant ASXL1-dependent oncogenesis have been proposed, none have been functionally validated in the context of the human hematopoietic stem cell, where these mutations almost certainly arise. Here, we extensively characterized a CRISPR-engineered human hematopoietic stem and progenitor cell model of ASXL1 mutations. In this context, mutant ASXL1 expression decreases differentiation, increases clonogenicity in serial replating experiments, and improves engraftment in immunodeficient mice. We also show that endogenous truncating mutations in ASXL1 drive protein stabilization and confirm that mutant ASXL1 is resistant to proteasomal degradation. At the transcriptional level, these phenotypes are driven by significant repression of the stress-response genes and by increased expression of bromodomain and extra-terminal family protein targets. Using protein-interaction screens, genomic and functional approaches, we link the positive transcriptional changes in ASXL1-mutant cells to BRD4-dependent RNA polymerase II pause release and identify a mechanism for transcriptional repression via a previously uncharacterized interaction with the transcription factor MECOM. Finally, we demonstrate that ASXL1-mutant AML exhibits increased MECOM activity consistent with our gene-editing models. Collectively, these studies highlight a highly reproducible model of mutant ASXL1 in the appropriate cell context. Further, they are the first to functionally describe the mutant ASXL1 interactome in the context of the human HSC, identifying new dependencies with therapeutic potential.
]]></description>
<dc:creator>Hall, M. L.</dc:creator>
<dc:creator>Quintal, A.</dc:creator>
<dc:creator>Worme, S.</dc:creator>
<dc:creator>Nguyen, T.</dc:creator>
<dc:creator>Schonrock, Z.</dc:creator>
<dc:creator>Tsuchiya, M.</dc:creator>
<dc:creator>Acharya, S. N.</dc:creator>
<dc:creator>Carlson, H. L.</dc:creator>
<dc:creator>Pacentine, I. V.</dc:creator>
<dc:creator>Shrestha, S.</dc:creator>
<dc:creator>Macaraeg, J.</dc:creator>
<dc:creator>Armstrong, R.</dc:creator>
<dc:creator>McGann, J.</dc:creator>
<dc:creator>Evans-Dutson, S.</dc:creator>
<dc:creator>Lusardi, T. A.</dc:creator>
<dc:creator>O'Connell, B. L.</dc:creator>
<dc:creator>Adey, A. C.</dc:creator>
<dc:creator>Maxson, J. E.</dc:creator>
<dc:creator>Yardımcı, G. G.</dc:creator>
<dc:creator>Braun, T. P.</dc:creator>
<dc:date>2026-01-14</dc:date>
<dc:identifier>doi:10.64898/2026.01.13.699315</dc:identifier>
<dc:title><![CDATA[Mutant ASXL1 Drives Transcriptional Activation and Repression in Human Hematopoiesis]]></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/10.64898/2026.01.13.699360v1?rss=1">
<title>
<![CDATA[
Defects in tissue-resident macrophages lead to smaller eardrums and abnormal neurovascular networks with increased middle-ear infection 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.01.13.699360v1?rss=1</link>
<description><![CDATA[
The tympanic membrane (TM), or eardrum, is essential for hearing. Macrophages, the primary innate immune cells, are densely distributed in the eardrum after birth, especially near blood vessels and nerve fibers. During postnatal development, neonatal tissue-resident macrophages (TRMs) gradually polarize, and their population declines. These dynamic changes closely parallel the maturation of neurovascular networks. What are the precursors of the early  wave of TRMs? Are they critical for postnatal TM development? Using fate mapping, single-cell RNA sequencing, and macrophage depletion, this study reveals for the first time that postnatal eardrum TRMs are heterogeneous, originating mainly from embryonic myeloid lineages, with increasing input from postnatal monocyte progenitor-derived cells. Single-cell RNA sequencing identifies gene signatures vital for vascular and neuronal development. Depleting TRMs results in smaller eardrums, disrupted vascular-neuronal networks, and increased risk of middle-ear infection. This study offers new insight into how the innate immune system supports TM maturation and protects against middle-ear infection during a critical postnatal window.

TeaserTissue-resident macrophages guide eardrum neurovascular maturation and help prevent middle-ear abnormalities.
]]></description>
<dc:creator>Zhang, Y.</dc:creator>
<dc:creator>Wang, P.</dc:creator>
<dc:creator>Neng, L.</dc:creator>
<dc:creator>Burwood, G.</dc:creator>
<dc:creator>Sharma, K.</dc:creator>
<dc:creator>Kachelmeier, A.</dc:creator>
<dc:creator>Shi, X.</dc:creator>
<dc:date>2026-01-14</dc:date>
<dc:identifier>doi:10.64898/2026.01.13.699360</dc:identifier>
<dc:title><![CDATA[Defects in tissue-resident macrophages lead to smaller eardrums and abnormal neurovascular networks with increased middle-ear infection]]></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/10.64898/2026.01.14.699534v1?rss=1">
<title>
<![CDATA[
Aging macaques bridge the translational gap in perivascular space biology 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.01.14.699534v1?rss=1</link>
<description><![CDATA[
The prevalence of MR-visible perivascular spaces (PVS) is age-dependent in rhesus macaques. Automated quantification in 94 male and female animals ranging 5-28 years of age demonstrated a robust association between age and PVS burden, with an anatomical distribution paralleling that of humans. Preliminary ex vivo validation confirmed that the MRI-detected tubular structures correspond to perivascular spaces. These findings establish the rhesus macaque as a tractable model for understanding the role of perivascular dysfunction in age-related brain vulnerability.
]]></description>
<dc:creator>Liguore, W. A.</dc:creator>
<dc:creator>Schwartz, D. L.</dc:creator>
<dc:creator>Piantino, J.</dc:creator>
<dc:creator>Rajendran, A.</dc:creator>
<dc:creator>Jensen, G.</dc:creator>
<dc:creator>Karpf, J. A.</dc:creator>
<dc:creator>Chernov, M. M.</dc:creator>
<dc:creator>Woltjer, R. L.</dc:creator>
<dc:creator>Kohama, S. G.</dc:creator>
<dc:creator>Silbert, L. C.</dc:creator>
<dc:creator>Weiss, A. R.</dc:creator>
<dc:date>2026-01-15</dc:date>
<dc:identifier>doi:10.64898/2026.01.14.699534</dc:identifier>
<dc:title><![CDATA[Aging macaques bridge the translational gap in perivascular space biology]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-01-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.01.14.699484v1?rss=1">
<title>
<![CDATA[
Monounsaturated fatty acid biosynthesis is critical for streptococcal envelope homeostasis and stress tolerance 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.01.14.699484v1?rss=1</link>
<description><![CDATA[
AbstractThe genus Streptococcus contains some of the most important commensals and pathogens of the human microbiome. To obtain the fatty acids required for cell membranes, Streptococcus either produce fatty acids de novo through the fatty acid biosynthesis (fab) pathway or uptake host fatty acids through the fatty acid kinase (fak) pathway. Although both the fab and fak pathways represent potential therapeutic targets to prevent or treat infection, progress is limited because of an incomplete understanding of taxon-to-taxon variability in streptococcal lipid metabolism. Here, we examined the role of de novo monounsaturated fatty acid (MUFA) synthesis in physiology and virulence-associated traits in Streptococcus mutans, Streptococcus pyogenes, and Streptococcus pneumoniae, three major pathogens that cause disease at distinct body sites. In all three species, deletion of fabM abolished MUFA production and caused severe growth defects, decreased stress tolerance, increased antibiotic susceptibility, and defects in cell viability, morphology, and division. In S. mutans, loss of fabM also markedly reduced competence signaling and production of the mutacin IV bacteriocin. Deletion of fabM increased susceptibility to killing by human neutrophils in S. mutans and S. pneumoniae, but not S. pyogenes. Together, these findings illustrate that MUFA synthesis is broadly important for streptococcal physiology and cell membrane homeostasis, while its contribution to pathogenesis is strongly species- and context-dependent, providing leads to guide development of novel therapeutic and/or preventative strategies.
]]></description>
<dc:creator>Baker, J. L.</dc:creator>
<dc:creator>Tang, J.</dc:creator>
<dc:creator>Guo, M.</dc:creator>
<dc:creator>Farias-da-Silva, F. F.</dc:creator>
<dc:creator>Barbisan, M.</dc:creator>
<dc:creator>Burnside, M.</dc:creator>
<dc:creator>Crofton, K.</dc:creator>
<dc:creator>Williams, S.</dc:creator>
<dc:creator>Rao, S.</dc:creator>
<dc:creator>Lee, M.</dc:creator>
<dc:creator>Drucker, S. G.</dc:creator>
<dc:creator>Higashi, D. L.</dc:creator>
<dc:creator>Merrit, J.</dc:creator>
<dc:creator>Hirose, Y.</dc:creator>
<dc:creator>Jan-Willem, V.</dc:creator>
<dc:creator>Nizet, V.</dc:creator>
<dc:date>2026-01-16</dc:date>
<dc:identifier>doi:10.64898/2026.01.14.699484</dc:identifier>
<dc:title><![CDATA[Monounsaturated fatty acid biosynthesis is critical for streptococcal envelope homeostasis and stress tolerance]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-01-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.01.17.700062v1?rss=1">
<title>
<![CDATA[
An Integrated Method for Profiling Lipid-Protein Interactions Using Multifunctional Lipid Probes 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.01.17.700062v1?rss=1</link>
<description><![CDATA[
Cellular lipids shape health and disease through specific protein interactions, yet lipid-protein networks remain poorly defined. Despite rapid advances in functional lipid probes, the field still lacks a practical, dedicated protocol for conducting lipid-protein interaction studies. We describe detailed methods for determining lipid interactomes from cells using multifunctionalized lipid derivatives. We provide protocols that detail 1) how to treat cells with lipid derivatives and perform photochemistry to obtain lipid-protein conjugates; 2) how to perform click chemistry with a fluorophore and observe lipid-protein conjugates by in-gel fluorescence; 3) how to perform click chemistry with azide beads and prepare lipid-protein conjugates for proteomic analysis. We provide context on important parameters for each step and include guidelines for controls, as well as suggestions for troubleshooting based on common problems encountered during the preparation of this protocol. This protocol enables mapping lipid interactomes across diverse biological systems. The entire workflow from cell treatment to complete proteomic sample preparation requires [~]15 hours over four days, depending on the type of experimental readout (in-gel fluorescence or proteomics), and the usage of pause points. Practitioners are expected to be familiar with standard biochemical techniques, such as sterile sample handling and tissue culture and gel electrophoresis. Additional skills are needed for mass spectrometric analysis, and collaboration with a proteomics core facility is recommended. The described procedures uniquely enable the identification of the protein interactors (the interactome) of select lipid species, providing for a major shift in the characterization of the biological roles of lipids in cellular systems.
]]></description>
<dc:creator>Farley, S. E.</dc:creator>
<dc:creator>Guzman, G.</dc:creator>
<dc:creator>Blume, B.</dc:creator>
<dc:creator>Stein, F.</dc:creator>
<dc:creator>Schultz, C.</dc:creator>
<dc:creator>Tafesse, F. G.</dc:creator>
<dc:date>2026-01-17</dc:date>
<dc:identifier>doi:10.64898/2026.01.17.700062</dc:identifier>
<dc:title><![CDATA[An Integrated Method for Profiling Lipid-Protein Interactions Using Multifunctional Lipid Probes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-01-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.01.16.699339v1?rss=1">
<title>
<![CDATA[
Selective Pharmacological Blockade of GPR39 Markedly Reduces No Reflow and Infarct Volumes in a Rat Model of Acute Myocardial Infarction 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.01.16.699339v1?rss=1</link>
<description><![CDATA[
Our aim was to determine whether selective pharmacological blockade of GPR39 by the novel drug, VC108, reduces no reflow (NRV) and infarct (INV) volumes acute myocardial infarction (AMI). Rats underwent 1 h of coronary occlusion and 1 h of reperfusion. Groups 1 and 2 animals received drug/vehicle prior to or during coronary occlusion. Group 3 received drug/vehicle 5 min prior to reperfusion. Readouts also included tissue pO2, hemodynamics, and wall thickening. In Groups 4 and 5 animals, drug was injected for measurement of plasma and tissue levels. Immunohistochemistry and western blot of rat myocardium as well as immuocytochemistry and qPCR of cardiac cells was performed for presence of GPR39. There was marked reduction in NRV and INV in groups 1 and 3 animals where both were measured and in Group 2 where INV was measured. The reduction in NRV in all animals was associated with higher tissue pO2 during VC108 compared to vehicle. Similar results were obtained for INV in Group 2 animal, but not in Groups 1 and 3. Immunohistochemistry and western blot of heart tissue as well as immunocytochemistry and qPCR of cells revealed high expression of GPR39 in cardiomyocytes, a hithertofore unknown finding. We conclude that VC108 is very effective in reducing INV and NRV in a rat AMI model when given before coronary occlusion or just prior to reperfusion. VC108 acts by blocking GPR39, resulting in vasodilation by relaxing pericytes and VSMCs. It also directly protects cardiomyocytes by preventing downstream effects of GPR39 stimulation.
]]></description>
<dc:creator>Methner, C.</dc:creator>
<dc:creator>Le, D. E.</dc:creator>
<dc:creator>Liu, L.</dc:creator>
<dc:creator>Plascencia, M.</dc:creator>
<dc:creator>Kajimoto, M.</dc:creator>
<dc:creator>thompson, a.</dc:creator>
<dc:creator>Cianciulli, A.</dc:creator>
<dc:creator>Pellacani, A.</dc:creator>
<dc:creator>Micheli, F.</dc:creator>
<dc:creator>Kaul, S.</dc:creator>
<dc:date>2026-01-20</dc:date>
<dc:identifier>doi:10.64898/2026.01.16.699339</dc:identifier>
<dc:title><![CDATA[Selective Pharmacological Blockade of GPR39 Markedly Reduces No Reflow and Infarct Volumes in a Rat Model of Acute Myocardial Infarction]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-01-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.01.20.700677v1?rss=1">
<title>
<![CDATA[
PU.1 inhibition sensitizes stem-monocytic AML to BCL2 blockade 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.01.20.700677v1?rss=1</link>
<description><![CDATA[
Acute myeloid leukemia (AML) exhibits substantial transcriptional heterogeneity across differentiation states that influences therapeutic response to BCL2 inhibition with venetoclax. While hematopoietic stem cell (HSC)-like AMLs show high sensitivity to venetoclax and monocytic-like AMLs demonstrate resistance, the therapeutic behavior of leukemias harboring both transcriptional programs remains poorly defined. Analysis of a large AML cohort reveals a distinct patient population exhibiting concurrent HSC- and monocyte-like transcriptional signatures, which we term stem-monocytic AML. Ex vivo drug sensitivity profiling demonstrates that stem-monocytic AMLs exhibit venetoclax resistance comparable to pure monocytic disease, despite expressing HSC-like transcriptional features. Using a leukemia cell line model that recapitulates stem-monocytic AML characteristics, we show through immunophenotyping and single-cell lineage tracing that venetoclax preferentially depletes immature blasts while sparing differentiated monocytic populations. Single-cell transcriptomic and chromatin accessibility analyses identify enrichment of myeloid differentiation transcription factors, particularly PU.1, in resistant populations. A targeted CRISPR knockout screen confirms that PU.1 disruption induces differentiation arrest and enhances venetoclax sensitivity primarily in the immature immunophenotypic compartments. Pharmacologic PU.1 inhibition with the small molecule DB2313 synergizes with venetoclax in both cell line models and primary patient samples. These findings establish stem-monocytic AML as a transcriptionally and functionally distinct subtype and nominate combined PU.1 and BCL2 inhibition as a rational therapeutic strategy for improving venetoclax response in this patient population.
]]></description>
<dc:creator>Yashar, W. M.</dc:creator>
<dc:creator>Pacentine, I. V.</dc:creator>
<dc:creator>Taherinasab, A.</dc:creator>
<dc:creator>Nguyen, T.</dc:creator>
<dc:creator>Worme, S.</dc:creator>
<dc:creator>Tsuchiya, M.</dc:creator>
<dc:creator>Lusardi, T.</dc:creator>
<dc:creator>Hutchinson, C.</dc:creator>
<dc:creator>Fillmore, T. L.</dc:creator>
<dc:creator>Posso, C.</dc:creator>
<dc:creator>Gosline, S. J.</dc:creator>
<dc:creator>Piehowski, P. D.</dc:creator>
<dc:creator>Yardimci, G. G.</dc:creator>
<dc:creator>Adey, A. C.</dc:creator>
<dc:creator>Maxson, J. E.</dc:creator>
<dc:creator>Braun, T. P.</dc:creator>
<dc:date>2026-01-23</dc:date>
<dc:identifier>doi:10.64898/2026.01.20.700677</dc:identifier>
<dc:title><![CDATA[PU.1 inhibition sensitizes stem-monocytic AML to BCL2 blockade]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-01-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.01.21.700911v1?rss=1">
<title>
<![CDATA[
miniMTI: minimal multiplex tissue imaging enhances biomarker expression prediction from histology 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.01.21.700911v1?rss=1</link>
<description><![CDATA[
Virtual multiplexing from routine histology has advanced rapidly, yet morphology alone provides limited access to molecular state, imposing an intrinsic ceiling on H&E-only inference. Here, we introduce miniMTI, a molecularly anchored virtual staining framework that determines the minimal set of experimentally measured markers required, alongside H&E, to accurately reconstruct large multiplex tissue imaging (MTI) panels while preserving biologically and clinically relevant information. miniMTI learns from paired same-section H&E-MTI data using a unified multimodal generative model that can condition on arbitrary combinations of measured marker channels, coupled with an iterative panel selection strategy to rank informative molecular anchors. Across colorectal and prostate cancer cohorts spanning two MTI platforms and over 40 million cells, miniMTI reduces a 40-marker MTI assay to H&E plus as few as three measured molecular markers, while accurately recovering withheld markers, preserving cellular phenotypes and spatial tissue architecture, and disease-associated molecular programs, including Gleason grade-linked signatures. By integrating histology context with sparse molecular grounding, miniMTI overcomes the limitations of morphology-only virtual staining and provides a scalable, cost-effective approach for expanding MTI-level biomarker coverage with retained biological interpretability and clinical relevance.
]]></description>
<dc:creator>Sims, Z.</dc:creator>
<dc:creator>Govindarajan, S.</dc:creator>
<dc:creator>Ait-Ahmad, K.</dc:creator>
<dc:creator>Ak, C.</dc:creator>
<dc:creator>Kuykendall, M.</dc:creator>
<dc:creator>Mills, G. B.</dc:creator>
<dc:creator>Eksi, E.</dc:creator>
<dc:creator>Chang, Y. H.</dc:creator>
<dc:date>2026-01-23</dc:date>
<dc:identifier>doi:10.64898/2026.01.21.700911</dc:identifier>
<dc:title><![CDATA[miniMTI: minimal multiplex tissue imaging enhances biomarker expression prediction from histology]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-01-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.01.22.700703v1?rss=1">
<title>
<![CDATA[
A First-In-Class Broad Spectrum Inhibitor of Copper Exporting P1B-type ATPases 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.01.22.700703v1?rss=1</link>
<description><![CDATA[
Copper (Cu) transporting ATPases represent a highly conserved subclass of P-type ATPases with critical roles in Cu export and metalloenzyme synthesis. Despite their important biological roles and association with a wide range of human diseases, no high-affinity small-molecule inhibitors have been described. Here, we identify MKV3 as a first-in-class inhibitor of Cu-transporting P-type ATPases that targets a conserved Cu+ entry site to the translocation pathway. In silico docking against the Xenopus ATP7B structure revealed a highly conserved pocket suitable for pharmacological inhibition. MKV3 bound human ATP7A and ATP7B with nanomolar affinity, competed with N-terminal metal-binding domains for access to the Cu+ entry site, and selectively inhibited Escherichia coli CopA ATPase activity and Cu+ transport. Mechanistically, MKV3 blocked chaperone-mediated Cu+ delivery to the intramembranous CPC site of CopA that is essential for its transport function. We further identified a single charged P-domain residue that governed MKV3 affinity and potency across species. Functionally, MKV3 phenocopied the genetic loss of Cu+-ATPases in bacteria, fungi, plants, zebrafish, and mammals, impairing copper-dependent enzymes, transporter trafficking, and copper tolerance. These findings establish a conserved, druggable vulnerability in Cu+-ATPases and introduce MKV3 as a broadly active chemical tool to modulate copper homeostasis across biological kingdoms.

Significance StatementCopper-transporting P1B-type ATPases are essential for copper homeostasis in all domains of life, yet have lacked pharmacological inhibitors. This work identifies MKV3 as the first small-molecule inhibitor of Cu+-ATPases in bacteria, fungi, plants and animals, and defines a conserved, druggable Cu+ entry pocket that governs metal delivery to the transmembrane pathway. MKV3s ability to potentiate copper-mediated killing in multidrug-resistant bacterial pathogens highlights its potential as an antimicrobial adjuvant, while its attenuation of mammalian ATP7A/B function offers promise in oncology and copper-related diseases. Collectively, these findings establish a new tool for targeting of Cu+-ATPases with wide-ranging applications across biological systems.
]]></description>
<dc:creator>Shanbhag, V.</dc:creator>
<dc:creator>Dinguyella, S. A.</dc:creator>
<dc:creator>Gudekar, N.</dc:creator>
<dc:creator>Conrad, K.</dc:creator>
<dc:creator>Azubuogu, C.</dc:creator>
<dc:creator>Probst, C.</dc:creator>
<dc:creator>Ralle, M.</dc:creator>
<dc:creator>Mediavilla, M. G.</dc:creator>
<dc:creator>Cricco, J. A.</dc:creator>
<dc:creator>Garza, N. M.</dc:creator>
<dc:creator>Gohil, V. M.</dc:creator>
<dc:creator>Peck, S.</dc:creator>
<dc:creator>Kumar, S.</dc:creator>
<dc:creator>Natarajan, A.</dc:creator>
<dc:creator>Horadigala-Gamage, M.</dc:creator>
<dc:creator>Meloni, G.</dc:creator>
<dc:creator>Singh, K.</dc:creator>
<dc:creator>Petris, M. J.</dc:creator>
<dc:date>2026-01-24</dc:date>
<dc:identifier>doi:10.64898/2026.01.22.700703</dc:identifier>
<dc:title><![CDATA[A First-In-Class Broad Spectrum Inhibitor of Copper Exporting P1B-type ATPases]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-01-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.01.23.701393v1?rss=1">
<title>
<![CDATA[
Differential Assembly of Native ENaC Complexes Across Mouse Epithelial Tissues 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.01.23.701393v1?rss=1</link>
<description><![CDATA[
The epithelial sodium channel (ENaC) governs sodium and fluid absorption in the lung, kidney, and colon, but the organization of native ENaC complexes has remained difficult to define because of their low abundance and biochemical instability. To enable direct analysis of native assemblies, we generated a knock-in mouse in which the endogenous {gamma} subunit is fused at its C terminus to mVenus, a 3C protease cleavage site, and a 3xFLAG epitope (ENaC{gamma}-VF). The tag preserves physiological ENaC function, as ENaC{gamma}-VF mice display normal electrolyte handling, benzamil affinity, and amiloride-sensitive Na:K responses indistinguishable from wild-type animals. Using fluorescence-detection size-exclusion chromatography and single-molecule pull-down, we directly monitor intact native ENaC complexes from lung, kidney, and colon and uncover marked tissue-to-tissue differences in channel abundance and apparent complex size. Dual-color analysis with a fluorescent Fab against ENaC marks fully assembled {beta}{gamma} channels, while {gamma}-based fluorescence reports the broader population of {gamma}-containing assemblies. In combination, the ENaC{gamma}-VF line provides a biochemical anchor for identifying regulatory and trafficking proteins that co-purify with native ENaC complexes. These data show that ENaC architecture in vivo is heterogeneous, and establish ENaC{gamma}-VF mice as a platform for dissecting how epithelial environments shape ENaC assembly, composition, and regulation.
]]></description>
<dc:creator>Bharadwaj, A.</dc:creator>
<dc:creator>Curry, J. N.</dc:creator>
<dc:creator>Su, X.-T.</dc:creator>
<dc:creator>Barria Maturana, R.</dc:creator>
<dc:creator>McCormick, J. A.</dc:creator>
<dc:creator>Ellison, D. H.</dc:creator>
<dc:creator>Baconguis, I.</dc:creator>
<dc:date>2026-01-25</dc:date>
<dc:identifier>doi:10.64898/2026.01.23.701393</dc:identifier>
<dc:title><![CDATA[Differential Assembly of Native ENaC Complexes Across Mouse Epithelial Tissues]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-01-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.01.24.701475v1?rss=1">
<title>
<![CDATA[
Transcriptomic profiling reveals neurophysiological gene candidates underlying vocal evolution in African clawed frogs 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.01.24.701475v1?rss=1</link>
<description><![CDATA[
Neurophysiologists have discovered many mechanisms underlying the production of animal behaviors in specific species; these involve a collection of neuromuscular systems, neuronal membrane and neural network properties, as well as the hormones and neuromodulators known to modify them. However, the mechanistic basis of behavioral evolution is less well-studied, and causal links between differences in gene expression, cellular mechanisms and species-typical behaviors are rare. Vertebrate vocal behaviors are an excellent system for studying the evolution of behaviors because they are ancient, diverse and readily quantifiable. Xenopus frogs are particularly well-suited to the study of vocal evolution due to the temporal diversity of male advertisement calls between closely related species and the well-described vocal pattern generating circuitry. Here we focus on two species, X. laevis and X. petersii, that diverged 8.5 million years ago and produce advertisement calls with distinct timing. To begin bridging the gap between behavioral and mechanistic diversity in Xenopus vocal behaviors, we performed RNA sequencing of the parabrachial nucleus, a vocal premotor hindbrain area known to encode species-typical temporal patterns in X. laevis and X. petersii. We identified hundreds of differentially expressed genes between the two species, including many genes related to hormone signaling, neuromodulation, neuronal and synaptic functions, ion channels and neurotransmitter receptors. We explore several testable hypotheses emerging from these results that may explain mechanisms by which candidate genes and gene families may contribute to vocal pattern differences between X. laevis and X. petersii.
]]></description>
<dc:creator>Barkan, C. L.</dc:creator>
<dc:creator>Binder, L.</dc:creator>
<dc:creator>Davis, B. A.</dc:creator>
<dc:creator>Carbone, L.</dc:creator>
<dc:creator>Zornik, E.</dc:creator>
<dc:date>2026-01-26</dc:date>
<dc:identifier>doi:10.64898/2026.01.24.701475</dc:identifier>
<dc:title><![CDATA[Transcriptomic profiling reveals neurophysiological gene candidates underlying vocal evolution in African clawed frogs]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-01-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.01.26.701810v1?rss=1">
<title>
<![CDATA[
Genome-wide CRISPRi screen identifies basigin loss as protective in cardiac hypoxia 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.01.26.701810v1?rss=1</link>
<description><![CDATA[
Cardiac function depends on continuous oxidative metabolism, rendering cardiomyocytes highly vulnerable to oxygen deprivation. Here, we performed a genome-wide CRISPR interference (CRISPRi) screen in human iPSC-derived cardiomyocytes to identify genes that modulate survival during chronic hypoxia. This screen revealed that knockdown of basigin (BSG), a chaperone for the monocarboxylate transporters MCT1 and MCT4, confers robust protection. Canonically, hypoxic cells suppress pyruvate dehydrogenase (PDH) activity to reduce the oxidation of major fuel sources, thereby limiting TCA cycle flux, lowering oxygen consumption, and minimizing reactive oxygen species generated by an overly reduced electron transport chain (ETC). In contrast, we found that BSG inhibition reverses this response, prioritizing ATP maintenance during hypoxia and enhancing cardiomyocyte survival. Mechanistically, BSG loss restricts lactate efflux, leading to decreased PDH phosphorylation and increased glucose uptake for oxidation. Consistent with this, ETC subunits are more essential under hypoxia, highlighting cardiomyocytes unusual reliance on aerobic ATP production even when oxygen is limited. These findings challenge prevailing models of hypoxic adaptation by revealing cardiomyocyte-specific bioenergetic requirements and motivating future therapeutic efforts.
]]></description>
<dc:creator>Flanigan, W. R.</dc:creator>
<dc:creator>Midha, A. D.</dc:creator>
<dc:creator>Blume, S. Y.</dc:creator>
<dc:creator>Marti-Mateos, Y.</dc:creator>
<dc:creator>Costa, M. W.</dc:creator>
<dc:creator>Huang, Y.</dc:creator>
<dc:creator>Baik, A. H.</dc:creator>
<dc:creator>Huynh, H.</dc:creator>
<dc:creator>Susarla, G.</dc:creator>
<dc:creator>Bennett, N. K.</dc:creator>
<dc:creator>Nowak, R. A.</dc:creator>
<dc:creator>Srivastava, D.</dc:creator>
<dc:creator>Nakamura, K.</dc:creator>
<dc:creator>Jain, I. H.</dc:creator>
<dc:date>2026-01-27</dc:date>
<dc:identifier>doi:10.64898/2026.01.26.701810</dc:identifier>
<dc:title><![CDATA[Genome-wide CRISPRi screen identifies basigin loss as protective in cardiac hypoxia]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-01-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.01.26.701894v1?rss=1">
<title>
<![CDATA[
Mapping SARS-CoV-2 Nucleocapsid Function with Nanobodies 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.01.26.701894v1?rss=1</link>
<description><![CDATA[
The SARS-CoV-2 nucleocapsid (N) protein is essential for viral RNA packaging, replication, and immune modulation. Despite its central role, the mechanistic contributions of its individual domains, the N-terminal domain (NTD), C-terminal domain (CTD), and the intrinsically flexible linker (LINK), remain poorly defined, largely due to the proteins structural complexity. In this study, we developed a panel of twelve alpaca-derived nanobodies (VHHs) targeting the NTD, CTD, and LINK regions of N. Using ELISA and biolayer interferometry, we characterized their binding affinities, and we mapped their epitopes via hydrogen-deuterium exchange-mass spectrometry (HDX-MS) and structural modeling. When expressed intracellularly, these VHHs inhibited SARS-CoV-2 infection. In vitro, they disrupted phase separation of the N protein, a critical step in viral replication. Strikingly, VHHs targeting each domain independently blocked both phase condensation and viral replication, underscoring the functional importance of all three regions. These findings establish domain-specific VHHs as versatile tools for dissecting N biology, with promising therapeutic potential.

ImportanceSARS-CoV-2 and emerging coronaviruses remain a major global health threat, yet critical gaps persist in our understanding of their molecular pathogenesis. The nucleocapsid (N) protein, the most abundantly expressed SARS-CoV-2 antigen, plays essential roles beyond genome packaging, including immune evasion and intracellular organization. Here, we generate and characterize a panel of domain-specific nanobodies (VHHs) that enable precise dissection of Ns functional architecture. Using integrated biochemical, structural, and virological approaches, we uncover distinct mechanisms of viral inhibition, including disruption of phase condensation through a conserved linker region. These findings address long-standing knowledge gaps about a multifunctional viral protein and establish VHHs as powerful, modular tools for probing coronavirus biology, with broad potential for therapeutic, diagnostic, and cell biology applications.
]]></description>
<dc:creator>Weinstein, J. B.</dc:creator>
<dc:creator>Bates, T. A.</dc:creator>
<dc:creator>Anastas, A.</dc:creator>
<dc:creator>Alfahdli, A.</dc:creator>
<dc:creator>Trank-Green, M.</dc:creator>
<dc:creator>McBride, S. K.</dc:creator>
<dc:creator>Garcia, M.</dc:creator>
<dc:creator>Parson, M. A.</dc:creator>
<dc:creator>Jenkins, M. L.</dc:creator>
<dc:creator>Burke, J. E.</dc:creator>
<dc:creator>Barklis, E.</dc:creator>
<dc:creator>Tafesse, F. G.</dc:creator>
<dc:date>2026-01-27</dc:date>
<dc:identifier>doi:10.64898/2026.01.26.701894</dc:identifier>
<dc:title><![CDATA[Mapping SARS-CoV-2 Nucleocapsid Function with Nanobodies]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-01-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.01.26.701898v1?rss=1">
<title>
<![CDATA[
Pre-myelinating oligodendrocyte ADGRG1 is required for axon ensheathment and CNS myelin formation 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.01.26.701898v1?rss=1</link>
<description><![CDATA[
Myelin is essential for axonal health and rapid propagation of action potentials. In the central nervous system (CNS), myelination is initiated with axon ensheathment by pre-myelinating oligodendrocytes (preOLs), followed by iterative membrane wrapping and longitudinal extension along the axon. The molecular mechanisms that govern preOL development remain largely unknown. In this study, we identify the adhesion G protein-coupled receptor ADGRG1 (also called GPR56) as a key, evolutionarily conserved regulator of this process. We show that ADGRG1 is highly expressed in preOLs and that its conditional deletion in mouse preOLs leads to simplified preOL morphology and defective axon ensheathment, resulting in CNS hypomyelination. Live imaging in zebrafish demonstrated conserved function of Adgrg1 in axon ensheathment. Mechanistically, we show that ADGRG1 promotes preOL maturation by RhoA activation. Collectively, these findings reveal an ADGRG1-mediated RhoA signaling pathway that governs axon ensheathment and myelin formation.
]]></description>
<dc:creator>Zhu, B.</dc:creator>
<dc:creator>Li, T.</dc:creator>
<dc:creator>Rodrigues, B. V.</dc:creator>
<dc:creator>Piller, M. V.</dc:creator>
<dc:creator>Call, C. L.</dc:creator>
<dc:creator>Mogha, A.</dc:creator>
<dc:creator>Belloso-Iguerategui, A.</dc:creator>
<dc:creator>Chiou, B.</dc:creator>
<dc:creator>Schmidt, R.</dc:creator>
<dc:creator>Wangzhou, A.</dc:creator>
<dc:creator>Thurber, A.</dc:creator>
<dc:creator>Dee, M.</dc:creator>
<dc:creator>Larson, C.</dc:creator>
<dc:creator>Monk, K.</dc:creator>
<dc:creator>Piao, X.</dc:creator>
<dc:date>2026-01-27</dc:date>
<dc:identifier>doi:10.64898/2026.01.26.701898</dc:identifier>
<dc:title><![CDATA[Pre-myelinating oligodendrocyte ADGRG1 is required for axon ensheathment and CNS myelin formation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-01-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.01.26.701774v1?rss=1">
<title>
<![CDATA[
Identification of clofibric acid as a SYVN1 ligand for PROTAC development 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.01.26.701774v1?rss=1</link>
<description><![CDATA[
Targeted protein degradation (TPD) is an emerging therapeutic modality for numerous diseases. PROteolysis-TArgeting Chimeras (PROTACs) represent a potentially generalizable strategy to achieve TPD. A PROTAC is composed of a ligand for a protein of interest, a linker and a ligand for E3 ligase. As such, PROTACs can bring the E3 ligase into the close proximity of a protein target leading to polyubiquitination followed by target protein degradation. While the human genome encodes over 600 E3 ligases, only a handful of them have been harnessed for developing PROTACs. In order to expand the repertoire of E3 ligases for PROTAC development, we developed clickable photoaffinity probes based on clinically used drugs and metabolites to identify potential E3 ligases as the targets. In this paper, we report the discovery of clofibric acid with a molecular weight of only 214 Daltons as a ligand for synoviolin (SYVN1). We demonstrate its utility by developing clofibric acid-based BRD4 PROTACs. The linker length and architecture play a critical role in target degradation efficiency. The clofibric acid-derived BRD4 PROTACs achieve selective BRD4 degradation in a SYVN1-dependent manner. Our findings establish clofibric acid as a robust addition to the TPD toolbox, offering a novel E3 ligase recruitment strategy for the development of next-generation degraders.

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]]></description>
<dc:creator>Warren, J.</dc:creator>
<dc:creator>Munakala, A.</dc:creator>
<dc:creator>Zientek, K.</dc:creator>
<dc:creator>Kim, K.</dc:creator>
<dc:creator>Wilmarth, P.</dc:creator>
<dc:creator>Reddy, A.</dc:creator>
<dc:creator>Li, B. X.</dc:creator>
<dc:creator>Xiao, X.</dc:creator>
<dc:date>2026-01-28</dc:date>
<dc:identifier>doi:10.64898/2026.01.26.701774</dc:identifier>
<dc:title><![CDATA[Identification of clofibric acid as a SYVN1 ligand for PROTAC development]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-01-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.01.27.702041v1?rss=1">
<title>
<![CDATA[
Antigenic stimulation in conjunction with cytokine is required for mediating IL-17A production in human MAIT cells 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.01.27.702041v1?rss=1</link>
<description><![CDATA[
Mucosal-associated invariant T (MAIT) cells are donor unrestricted T cells capable of both antigen-specific adaptive responses and cytokine driven innate-like functions. Although human MAIT cells uniformly express ROR{gamma}t and IL23R, they generally produce IFN-{gamma}, and only a small fraction produces IL-17. Recent studies show that combined TCR and cytokine stimulation can elicit functional heterogeneity in blood-derived MAIT cells. Here, we investigate the role of IL-23/IL-23R signaling in mediating the function and transcriptional profiles of lung MAIT cell clones. We demonstrate that BAL-derived lung MAIT cell clones exhibit distinct cytokine profiles and variable IL23R expression. Short-term IL-23 stimulation triggers clone-specific transcriptional programs and IL23R-dependent upregulation of type 17-associated genes. Prolonged conditioning of lung MAIT cell clones with TCR (5-OP-RU) and cytokine (IL-23) stimulation induces stable IL-17A production along with unique transcriptional changes. TCR + IL-23 conditioning alone upregulates clone-specific and shared cytoskeletal/structural gene programs, whereas subsequent PMA/Ionomycin stimulation further induces IL-12 family signaling and metabolic genes. Together, these findings demonstrate that IL23R expression and TCR signaling are required for IL-17A production, highlighting that these conditions may be met in tissue environments where MR1-specific antigens and proinflammatory cytokines coexist.
]]></description>
<dc:creator>Kim, S.-J.</dc:creator>
<dc:creator>Kain, D.</dc:creator>
<dc:creator>Lewinsohn, D. A.</dc:creator>
<dc:creator>Swarbrick, G. M.</dc:creator>
<dc:creator>Cansler, M. E.</dc:creator>
<dc:creator>Bimber, B. N.</dc:creator>
<dc:creator>McElfresh, G.</dc:creator>
<dc:creator>Wong, E. B.</dc:creator>
<dc:creator>Khuzwayo, S.</dc:creator>
<dc:creator>Riffelmacher, T.</dc:creator>
<dc:creator>Lewinsohn, D. M.</dc:creator>
<dc:date>2026-01-30</dc:date>
<dc:identifier>doi:10.64898/2026.01.27.702041</dc:identifier>
<dc:title><![CDATA[Antigenic stimulation in conjunction with cytokine is required for mediating IL-17A production in human MAIT cells]]></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/10.64898/2026.01.30.702659v1?rss=1">
<title>
<![CDATA[
Small molecule agonists of 8-oxoguanine DNA glycosylase, OGG1 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.01.30.702659v1?rss=1</link>
<description><![CDATA[
Base excision repair (BER) is the primary pathway that removes oxidatively-induced DNA base damage from the nuclear and mitochondrial genomes, with 8-oxoguanine DNA glycosylase (OGG1) initiating repair at the two most frequently-formed base lesions: 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxoGua) and 2,6-diamino-4-oxo-5-formamidopyrimidine (FapyGua). Humans expressing a catalytically-compromised variant of OGG1 (S326C) are at increased risk for type 2 diabetes, Alzheimers disease, and Parkinsons disease. To potentially enhance the overall catalytic efficiency of this variant, a prior medicinal chemistry screen discovered seven chemically distinct agonists of OGG1 that stimulated activity in vitro and attenuated a paraquat (PQ) challenge in cultured cells. Herein, we developed structure-activity relationships around one specific core structure, F01. Using fluorescence-based DNA cleavage assays, we assessed the abilities of these compounds to stimulate the overall rate of OGG1 catalysis. Multiple compounds were identified that increased OGG1 activity on DNAs containing a site-specific 8-oxoGua by 2-fold or greater, with 9 compounds showing EC50 concentrations lower than F01 and were specific for OGG1. Selected agonists were shown to enhance OGG1-catalyzed release of 8-oxoGua and FapyGua from {gamma}-irradiated high-molecular-weight DNA using gas chromatography tandem mass spectrometry analyses. Since these assays did not reveal which step in the overall reaction was stimulated, we used a separation-of-function OGG1 mutant that possessed glycosylase, but not abasic-site (AP) lyase activity to demonstrate that the glycosylase step was not enhanced. In contrast, all agonists stimulated the AP lyase activity to levels equal to or greater than the magnitude of stimulation observed for overall chemistry, implicating agonist-mediated turnover as a potential contributor to the overall rate stimulation. The biological activities of selected agonists were evaluated in OGG1-deficient Kasumi-1 cells under conditions of paraquat (PQ)-induced oxidative stress, with several compounds mitigating PQ challenge.
]]></description>
<dc:creator>Luzadder, M. M.</dc:creator>
<dc:creator>Minko, I. G.</dc:creator>
<dc:creator>Moellmer-Gomez, S. A.</dc:creator>
<dc:creator>Tozaki, N. N.</dc:creator>
<dc:creator>Jaruga, P.</dc:creator>
<dc:creator>Dizdaroglu, M.</dc:creator>
<dc:creator>Jin, H.</dc:creator>
<dc:creator>Devereaux, J.</dc:creator>
<dc:creator>Nilsen, A.</dc:creator>
<dc:creator>Lloyd, R. S.</dc:creator>
<dc:creator>McCullough, A. K.</dc:creator>
<dc:date>2026-01-31</dc:date>
<dc:identifier>doi:10.64898/2026.01.30.702659</dc:identifier>
<dc:title><![CDATA[Small molecule agonists of 8-oxoguanine DNA glycosylase, OGG1]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-01-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.01.30.702880v1?rss=1">
<title>
<![CDATA[
Transcriptomics-based modeling of methionine metabolism effectively estimates sample-wise DNA methylation activity and epigenetic aging 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.01.30.702880v1?rss=1</link>
<description><![CDATA[
DNA methylation is a central epigenetic modification that regulates gene expression, maintains genomic stability, and guides cellular differentiation. However, direct measurements of DNA methylation, such as whole-genome bisulfite sequencing or DNA methylation arrays, are costly and require substantial DNA input, limiting their scalability for large cohorts and their applicability to emerging modalities such as single-cell and spatially resolved transcriptomics. In this study, motivated by the fact that DNA methylation is fundamentally a metabolic process, we investigate whether sample-wise DNA methylation activity can be inferred directly from transcriptomic profiles of genes involved in methionine and one-carbon metabolism. We show that a compact metabolic model comprising seven core reaction steps and 98 genes accurately predicts total DNA methylation activity across matched transcriptomic and methylation datasets from CCLE, TCGA, GTEx, and an independent single-cell multi-omics data set. Building on this framework, we develop Total DNA Methylation Activity (TDMA), a physics-informed neural network-based score that enables robust estimation of DNA methylation activity from bulk, single-cell, and spatial transcriptomics data. We demonstrate that TDMA captures methylation-dependent transcriptional regulation and identifies genes and pathways under epigenetic control. Applying TDMA to GTEx, we further reveal strong associations between the predicted total methylation activity, chronological aging, and established epigenetic clocks. We also demonstrated that TDMA can serve as a transcriptomics-derived epigenetic clock and highlights age-dependent roles of folate and glutathione metabolism in epigenetic aging. Applying TDMA to single cell and spatial transcriptomics data collected from pancreatic adenocarcinoma (PDAC), we identified that methionine metabolism and DNA methylation regulates T cell cytotoxicity in the tumor microenvironment of PDAC. Together, this work establishes a scalable, modality-agnostic framework for estimating DNA methylation activity from transcriptomics and provides new insights into the metabolic regulation of epigenetic aging.
]]></description>
<dc:creator>Guo, T.</dc:creator>
<dc:creator>Dang, P.</dc:creator>
<dc:creator>Fang, Y.</dc:creator>
<dc:creator>Zhu, H.</dc:creator>
<dc:creator>Wang, X.</dc:creator>
<dc:creator>Wang, J.</dc:creator>
<dc:creator>Ma, A.</dc:creator>
<dc:creator>Ma, Q.</dc:creator>
<dc:creator>Cao, S.</dc:creator>
<dc:creator>Zhang, C.</dc:creator>
<dc:date>2026-02-02</dc:date>
<dc:identifier>doi:10.64898/2026.01.30.702880</dc:identifier>
<dc:title><![CDATA[Transcriptomics-based modeling of methionine metabolism effectively estimates sample-wise DNA methylation activity and epigenetic aging]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-02-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.01.29.702548v1?rss=1">
<title>
<![CDATA[
Maternal Obesity Reprograms Differentiation Trajectories of Fetal Hematopoietic Stem and Progenitor Cells Through Altered Inflammatory Signaling 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.01.29.702548v1?rss=1</link>
<description><![CDATA[
BackgroundMaternal obesity is a global health challenge with profound consequences for offspring health. While its impact on metabolic programming has been widely studied, far less is known about how maternal obesity shapes the fetal immune system. The fetal bone marrow (FBM) is the central site of hematopoietic stem and progenitor cell (HSPC) development, and disruptions in this niche can have lifelong effects on immunity, infection susceptibility, and inflammatory disease risk. In this study, we examined FBM hematopoiesis in a nonhuman primate model of spontaneous maternal obesity.

MethodsUsing spectral flow cytometry, single-cell RNA sequencing, and functional differentiation assays, we mapped progenitor composition, lineage trajectories, and immune function in offspring exposed to maternal obesity compared with lean controls. These complementary approaches allowed us to capture cellular frequencies and transcriptional programs, while trajectory and signaling analyses provided insight into how progenitor maturation and intercellular communication are disrupted by maternal obesity.

ResultsOur findings reveal that maternal obesity decreases CD34+ HSPCs and common lymphoid progenitor populations, while expanding megakaryocyte-erythroid and granulocyte-monocyte progenitors. Pseudotime analysis demonstrated altered maturation, with cells accumulating at early differentiation states. Transcriptional profiling uncovered a strong inflammatory bias, with myeloid progenitors upregulating alarmins, interferon-stimulated genes, and proinflammatory mediators. Functionally, monocytes derived from obese FBM showed impaired migratory and colony-stimulating capacity, coupled with exaggerated TNF responses to LPS stimulation.

ConclusionTogether, these results demonstrate that maternal obesity, even in the absence of obesogenic diet, disrupts fetal bone marrow hematopoiesis by altered HSPC maturation, reprogramming lineage trajectories, and inducing inflammatory bias.
]]></description>
<dc:creator>Doratt, B. M.</dc:creator>
<dc:creator>Hemati, H.</dc:creator>
<dc:creator>Wagner, S. B.</dc:creator>
<dc:creator>Blanton, M. B.</dc:creator>
<dc:creator>Avila, U.</dc:creator>
<dc:creator>Varlamov, O.</dc:creator>
<dc:creator>Messaoudi, I.</dc:creator>
<dc:date>2026-02-03</dc:date>
<dc:identifier>doi:10.64898/2026.01.29.702548</dc:identifier>
<dc:title><![CDATA[Maternal Obesity Reprograms Differentiation Trajectories of Fetal Hematopoietic Stem and Progenitor Cells Through Altered Inflammatory Signaling]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-02-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.02.01.703134v1?rss=1">
<title>
<![CDATA[
ImmunoPheno: A Computational Framework for Data-Driven Design and Analysis of Immunophenotyping Experiments 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.02.01.703134v1?rss=1</link>
<description><![CDATA[
Immunophenotyping is fundamental to characterizing tissue cellular composition, pathogenic processes, and immune infiltration, yet its accuracy and reproducibility remain constrained by heuristic antibody panel design and manual gating. Here, we present ImmunoPheno, an open-source computational platform that repurposes large-scale single-cell proteo-transcriptomic data to guide immunophenotyping experimental design and analysis. ImmunoPheno integrates existing datasets to automate the design of optimal antibody panels, gating strategies, and cell identity annotation. We used ImmunoPheno to construct a harmonized reference (HICAR) comprising 390 monoclonal antibodies and 93 human immune cell populations. Leveraging this resource, we algorithmically designed minimal panels to isolate rare populations, such as MAIT cells and pDCs, which we validated experimentally. We further demonstrate accurate cell identity annotation across publicly available and newly generated cytometry datasets spanning diverse technologies, including spatial platforms like CODEX. ImmunoPheno complements expert curation and supports continual expansion, providing a scalable framework to enhance the accuracy, reproducibility, and resolution of immunophenotyping.
]]></description>
<dc:creator>Wu, L.</dc:creator>
<dc:creator>Nguyen, M. A.</dc:creator>
<dc:creator>Yang, Z.</dc:creator>
<dc:creator>Potluri, S.</dc:creator>
<dc:creator>Sivagnanam, S.</dc:creator>
<dc:creator>Kirchberger, N.</dc:creator>
<dc:creator>Joshi, A.</dc:creator>
<dc:creator>Ahn, K. J.</dc:creator>
<dc:creator>Tumulty, J. S.</dc:creator>
<dc:creator>Cruz Cabrera, E.</dc:creator>
<dc:creator>Romberg, N.</dc:creator>
<dc:creator>Tan, K.</dc:creator>
<dc:creator>Coussens, L. M.</dc:creator>
<dc:creator>Camara, P. G.</dc:creator>
<dc:date>2026-02-03</dc:date>
<dc:identifier>doi:10.64898/2026.02.01.703134</dc:identifier>
<dc:title><![CDATA[ImmunoPheno: A Computational Framework for Data-Driven Design and Analysis of Immunophenotyping Experiments]]></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/10.64898/2026.02.06.704446v1?rss=1">
<title>
<![CDATA[
Unwinding of an RNA duplex by the Orthoflavivirus NS3 helicase requires translocation beyond the displaced strand and is stimulated by the NS5 RdRp 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.02.06.704446v1?rss=1</link>
<description><![CDATA[
The NS3 helicases from the Flaviviridae family of viruses exhibit nucleotide-hydrolysis-dependent, nucleic-acid-unwinding activity. The RNA unwinding activity for NS3 helicases from the Orthoflavivirus genus has not been fully explored and contrasts with NS3 helicase from Hepatitis C virus (HCV) of the Hepacivirus genus, which has thus far served as the prototypical model enzyme from this family of viruses. To begin to understand the functional differences between flavivirus NS3 helicases, we first developed an expression and purification system for full-length untagged NS3 protein from West Nile virus (WNV) and Zika virus (ZIKV). Both enzymes exhibit RNA-stimulated ATPase activity and are dependent on the nucleoside triphosphatase active site of the enzyme. Unlike HCV NS3, orthoflavivirus NS3s do not efficiently pre-assemble on a 3-ssRNA-tailed dsRNA substrate in the absence of ATP-Mg which is a prerequisite for formation of a productive HCV NS3-RNA complex that can exhibit a rapid burst of RNA unwinding. Instead, to observe RNA unwinding by WNV and ZIKV NS3s, low Mg-ATP concentrations are required at a time coincident when NS3 encounters the RNA substrate. In addition, we find that orthoflavivirus NS3s require translocation beyond the displaced strand to completely unwind a dsRNA substrate. Last, we find that orthoflavivirus NS5 stimulates the ability of NS3 to unwind dsRNA. These results suggest that functional differences exist between the flavivirus NS3 helicases and illuminate that orthoflavivirus NS3s require a functional interaction with the NS5 protein for coordination of its activity, as it is believed these two proteins constitute the viral replicase.
]]></description>
<dc:creator>Arnold, J. J.</dc:creator>
<dc:creator>Chib, S.</dc:creator>
<dc:creator>Cameron, C. E.</dc:creator>
<dc:date>2026-02-07</dc:date>
<dc:identifier>doi:10.64898/2026.02.06.704446</dc:identifier>
<dc:title><![CDATA[Unwinding of an RNA duplex by the Orthoflavivirus NS3 helicase requires translocation beyond the displaced strand and is stimulated by the NS5 RdRp]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-02-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.02.05.704050v1?rss=1">
<title>
<![CDATA[
MetaKnogic-Alpha: A Hyper-Relational Knowledge Base for Grounded Metabolic Reasoning 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.02.05.704050v1?rss=1</link>
<description><![CDATA[
The exponential trajectory of biomedical literature has precipitated a fundamental "synthesis gap" in metabolic research, where critical mechanistic insights remain fragmented across hundreds of thousands of disjointed full-text articles, preventing the consolidation of a global mechanistic view. Here, we present MetaKnogic-Alpha, a foundational mechanistic knowledge substrate designed to bridge this gap by transforming unstructured literature into a navigable, logic-based resource. MetaKnogic-Alpha synthesizes over 100K full-text articles into a hyper-relational hypergraph structure, preserving the n-ary relational logic inherent in complex metabolic pathways. To ensure biological rigor, we implemented a hierarchical discovery protocol: an autonomous reasoning agent first enriches query nomenclature for domain-specific precision, followed by a multi-hop topological expansion within the hypergraph to surface functional neighbors, such as enzymatic co-factors and distal regulators, often lost in traditional search paradigms. Crucially, the system subjects all literature-derived insights to a deterministic biochemical grounding against a curated metabolic reaction network, significantly mitigating the risk of probabilistic hallucinations common in standalone generative models. In rigorous benchmarking, MetaKnogic-Alpha achieved a mechanistic accuracy of 0.98 in scenarios where supporting evidence was present, providing a robustly attributable audit trail back to the primary literature via PubMed Central Identifiers. We designate this primary release as "alpha" to establish the foundational architectural logic for a burgeoning million-scale resource. By compressing the synthesis of thousands of papers from a multi-month manual effort into several hours of automated discovery, MetaKnogic-Alpha serves as a high-fidelity research companion that augments the human experts ability to resolve complex metabolic interactions and identify novel therapeutic drivers in precision oncology.
]]></description>
<dc:creator>Dang, P.</dc:creator>
<dc:creator>Swaminathan, P.</dc:creator>
<dc:creator>Guo, T.</dc:creator>
<dc:creator>Wan, C.</dc:creator>
<dc:creator>Cao, S.</dc:creator>
<dc:creator>Zhang, C.</dc:creator>
<dc:date>2026-02-09</dc:date>
<dc:identifier>doi:10.64898/2026.02.05.704050</dc:identifier>
<dc:title><![CDATA[MetaKnogic-Alpha: A Hyper-Relational Knowledge Base for Grounded Metabolic Reasoning]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-02-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.02.06.704439v1?rss=1">
<title>
<![CDATA[
Evaluation and application of chemical decrosslinking in the context of histopathological spatial proteomics 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.02.06.704439v1?rss=1</link>
<description><![CDATA[
Laser capture microdissection (LCM) - based spatial mass spectrometry proteomics is a rapidly emerging technique with strong potential for use in formalin-fixed, paraffin-embedded (FFPE) tissues. Several sample-preparation methods have been developed to decrosslink FFPE proteins for spatial proteomics; however, residual crosslinks often remain, and depth can remain impaired relative to fresh frozen tissue samples. To increase proteome coverage in spatially resolved LCM-FFPE samples, we investigated a panel of chemical compounds with the potential to catalyze the decrosslinking of nucleophilic functional groups on proteins. Systematic screening and optimization of temperature, incubation time, and reagent concentration led to the identification of 3,4-diaminobenzoic acid as an effective agent for improving proteome coverage in FFPE pancreatic tissue. This compound could boost precursor identifications by more than 10% at both reduced (70 {degrees}C) and high (90 {degrees}C) temperatures. Application of this chemical-decrosslinking strategy to a pancreatic ductal adenocarcinoma tissue section enabled the identification of numerous cell-type-enriched proteins with clinical and therapeutic relevance. Taken together, our findings show that chemical decrosslinking can increase proteome coverage in FFPE tissues, thereby advancing our understanding of tissue microenvironments in physiological and pathological contexts.
]]></description>
<dc:creator>Nwosu, A. J.</dc:creator>
<dc:creator>Chen, L.</dc:creator>
<dc:creator>Kumar, R.</dc:creator>
<dc:creator>Kwon, Y.</dc:creator>
<dc:creator>Goodyear, S. M.</dc:creator>
<dc:creator>Kardosh, A.</dc:creator>
<dc:creator>Fulcher, J. M.</dc:creator>
<dc:creator>Pasa-Tolic, L.</dc:creator>
<dc:date>2026-02-09</dc:date>
<dc:identifier>doi:10.64898/2026.02.06.704439</dc:identifier>
<dc:title><![CDATA[Evaluation and application of chemical decrosslinking in the context of histopathological spatial proteomics]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-02-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.02.10.704454v1?rss=1">
<title>
<![CDATA[
A Framework for Comparing Mouse Neoantigen Immunogenicity 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.02.10.704454v1?rss=1</link>
<description><![CDATA[
Cytotoxic CD8+ T cell responses targeting tumor neoantigens are critical for immunotherapy efficacy and are widely studied across different preclinical mouse tumor models. Defined neoantigens are commonly introduced to enable tracking of tumor-specific T cells; however, variation in neoantigen choice may yield immune phenotypes attributable to differences in neoantigen immunogenicity, complicating interpretation of tumor-intrinsic mechanisms. Here, we determined the relative immunogenicity of a set of 25 commonly used mouse tumor-derived and model neoantigens to facilitate comparison of neoantigens across studies. We found that in silico predicted major histocompatibility complex (MHC) binding affinity poorly stratified in vivo immunogenicity. In contrast, experimental measurement of peptide-MHC complex stability (Koff), more so than measured affinity (KD), closely correlated with the relative magnitude of neoantigen-targeted vaccine responses in vivo. Thus, we report the relative stability of a known set of commonly used neoantigens as a reference and provide a simple method to benchmark novel neoantigens against this library. This framework will allow contextualization of the level of immunogenicity of newly identified neoantigens and aid in comparative interpretation of tumor-immune phenotypes across studies.
]]></description>
<dc:creator>Matulich, P. J.</dc:creator>
<dc:creator>Sprague, C. N.</dc:creator>
<dc:creator>Schuster, V. P.</dc:creator>
<dc:creator>Granados, A. M.</dc:creator>
<dc:creator>Chaudhari, R. B.</dc:creator>
<dc:creator>Burger, M. L.</dc:creator>
<dc:date>2026-02-12</dc:date>
<dc:identifier>doi:10.64898/2026.02.10.704454</dc:identifier>
<dc:title><![CDATA[A Framework for Comparing Mouse Neoantigen Immunogenicity]]></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/10.64898/2026.02.12.705407v1?rss=1">
<title>
<![CDATA[
Chronic ethanol self-administration alters dopamine in the caudate nucleus and putamen of rhesus macaques in a sex-dependent manner 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.02.12.705407v1?rss=1</link>
<description><![CDATA[
Alcohol use disorder (AUD) affects over 28 million people in the U.S and is associated with neurobiological alterations, including in the basal ganglia. Within the basal ganglia, the caudate nucleus (caudate) and putamen are implicated in AUD due to their roles in ethanol reinforcement, with the caudate receiving inputs from cortico-associative areas and the putamen receiving inputs from somatosensory areas, supporting goal-directed and habitual behaviors respectively. These distinct behavioral roles are supported by dopamine signaling, including phasic dopamine, involved in assessing action-outcome associations, and tonic dopamine, which reflects ongoing dopaminergic tone that biases action initiation. Intrastriatal dopamine release is modulated by cholinergic interneurons via nicotinic acetylcholine receptors. Dysregulation of these mechanisms can contribute to the transition from occasional to habitual ethanol drinking. Here, we used in-vitro fast-scan cyclic voltammetry to measure dopamine signaling in male (n=6) and female (n=6) rhesus macaques following six months of ethanol self-administration. In putamen, ethanol increased tonic dopamine in both sexes, with females exhibiting greater release and faster dopamine uptake rates than males. In the caudate, ethanol self-administration enhanced dopamine uptake rates only in males. Phasic dopamine release was enhanced in caudate of both sexes but only putamen in males. nAChR blockade revealed that phasic dopamine release in males, but not females, was dependent on cholinergic modulation. These results demonstrate basal and sex-specific dopamine release and uptake are uniquely altered in rhesus macaque caudate and putamen in conjunction with chronic ethanol drinking.
]]></description>
<dc:creator>Levy, C. C.</dc:creator>
<dc:creator>Carlson, V. C.</dc:creator>
<dc:creator>Grant, K. A.</dc:creator>
<dc:creator>Salinas, A. G.</dc:creator>
<dc:date>2026-02-15</dc:date>
<dc:identifier>doi:10.64898/2026.02.12.705407</dc:identifier>
<dc:title><![CDATA[Chronic ethanol self-administration alters dopamine in the caudate nucleus and putamen of rhesus macaques in a sex-dependent manner]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-02-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.02.18.706493v1?rss=1">
<title>
<![CDATA[
Opposing roles for SNAP23 and SNAP25 in mediating MR1 trafficking and antigen presentation 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.02.18.706493v1?rss=1</link>
<description><![CDATA[
MHC class I-related protein 1 (MR1) is a highly conserved antigen presenting molecule that presents small molecule metabolites derived from diverse microbial pathogens to mucosal-associated invariant T (MAIT) cells. We have shown previously that MR1 traffics through endosomal compartments via soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) proteins, including Syntaxin 4 and vesicle-associated membrane protein (VAMP) 4. Here, we investigate the role of synaptosome-associated proteins (SNAPs), which pair with Syntaxins and VAMPs to form functional SNARE complexes, in MR1-mediated antigen presentation. Among SNAP homologs, we identify that SNAP23 contributes to the presentation of Mycobacterium tuberculosis (Mtb)-derived antigens and loss of SNAP23 reduces the number of MR1-containing vesicles during infection. In contrast, SNAP25 suppresses MR1 presentation for both intracellular pathogens Mtb and Mycobacterium avium, as well as extracellular pathogen Candida albicans. This study demonstrates opposing roles for SNAP23 and SNAP25 in MR1 antigen presentation to MAIT cells, and extends our understanding of how SNAP family proteins regulate MR1 trafficking.
]]></description>
<dc:creator>Kim, S.-J.</dc:creator>
<dc:creator>Kulicke, C. A.</dc:creator>
<dc:creator>Lewinsohn, D. M.</dc:creator>
<dc:creator>Karamooz, E.</dc:creator>
<dc:date>2026-02-19</dc:date>
<dc:identifier>doi:10.64898/2026.02.18.706493</dc:identifier>
<dc:title><![CDATA[Opposing roles for SNAP23 and SNAP25 in mediating MR1 trafficking and antigen presentation]]></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/10.64898/2026.02.19.706768v1?rss=1">
<title>
<![CDATA[
The archaic CUP pilus SMF-1 uses an antiparallel bundling mechanism to initiate biofilm formation 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.02.19.706768v1?rss=1</link>
<description><![CDATA[
Bacterial biofilms represent a survival strategy that enables microbial communities to withstand environmental stress. Stenotrophomonas maltophilia is an emerging, antibiotic-resistant, Gram-negative, opportunistic pathogen that frequently colonizes the lungs of individuals with cystic fibrosis. Its chaperone-usher pathway (CUP) pilus, SMF-1, is present in nearly all clinical isolates and is essential for biofilm development; however, its molecular architecture has remained unknown. Here, we present a 4.0 [A] cryo-EM structure revealing that SMF-1 is an archaic, rather than a classic, CUP pilus. SMF-1 forms thin, zigzag filaments that assemble into defined antiparallel "pili-couples," which further aggregate into thick bundles. These bundles act as robust intercellular tethers, facilitating the rapid cell-to-cell aggregation required for biofilm initiation. Despite high sequence and structural similarity to classic CUP systems, SMF-1 lacks the interfaces required to form a rod-like architecture, suggesting it may represent an evolutionary intermediate between the CUP classes. Finally, we demonstrate that SMF-1 producing bacteria initiate biofilm formation within 24 hours and that flagella can further accelerate this process. Together, these findings uncover a conserved bundling mechanism that promotes bacterial colonization and may contribute to pathogenicity.
]]></description>
<dc:creator>Fields, J. L.</dc:creator>
<dc:creator>Sebastian, C. C.</dc:creator>
<dc:creator>Zhang, H.</dc:creator>
<dc:creator>Zia, A.</dc:creator>
<dc:creator>Robertson, A. N.</dc:creator>
<dc:creator>Wu, H.</dc:creator>
<dc:creator>Kiedrowski, M. R.</dc:creator>
<dc:creator>Wang, F.</dc:creator>
<dc:date>2026-02-20</dc:date>
<dc:identifier>doi:10.64898/2026.02.19.706768</dc:identifier>
<dc:title><![CDATA[The archaic CUP pilus SMF-1 uses an antiparallel bundling mechanism to initiate biofilm formation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-02-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.02.20.706901v1?rss=1">
<title>
<![CDATA[
Paired oral clinical specimens reveal the underlying ecology supporting the emergence of inflammophilic microbiome communities 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.02.20.706901v1?rss=1</link>
<description><![CDATA[
BackgroundInflammatory oral diseases are associated with reproducible shifts from commensal-dominated microbiota toward pathobiont-enriched communities, yet the ecological mechanisms underlying the emergence of inflammophiles remain poorly understood. This study aims to investigate if host-derived inflammatory environments act as selective pressures that restructure microbial metabolism and community organization during disease progression.

MethodsWe performed 16S rRNA gene sequencing of patient-matched pediatric dental plaque and odontogenic abscess specimens to capture microbial community transitions across an inflammatory ecological gradient. Community ecology modeling and inferred metagenomic analyses were used to identify taxa and functional programs associated with commensal and inflammophilic states.

ResultsPatient-matched comparisons revealed a reproducible ecological selection gradient linking inflammatory environments to expansion of metabolically specialized inflammophiles and depletion of carbohydrate-utilizing commensals. Commensal-dominated plaque communities exhibited anabolic, carbohydrate-centered metabolic capacity, whereas abscess microbiota were enriched for catabolic metabolism, amino acid fermentation, and antimicrobial resistance, consistent with adaptation to inflammation-driven nutrient landscapes and immune pressure.

ConclusionsThese findings support a model in which host inflammation drives ecological restructuring of the oral microbiome toward metabolically adapted inflammophilic communities. Defining the metabolic requirements and selective pressures governing these transitions provides a framework for microbiome-directed therapeutic strategies aimed at restoring ecological stability during inflammatory dysbiosis.
]]></description>
<dc:creator>Krieger, M.</dc:creator>
<dc:creator>Kerns, K. A.</dc:creator>
<dc:creator>Palmer, E. A.</dc:creator>
<dc:creator>McLean, J. S.</dc:creator>
<dc:creator>Kreth, J.</dc:creator>
<dc:creator>Yardimci, G. G.</dc:creator>
<dc:creator>Merritt, J.</dc:creator>
<dc:date>2026-02-21</dc:date>
<dc:identifier>doi:10.64898/2026.02.20.706901</dc:identifier>
<dc:title><![CDATA[Paired oral clinical specimens reveal the underlying ecology supporting the emergence of inflammophilic microbiome communities]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-02-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.02.22.707119v1?rss=1">
<title>
<![CDATA[
Biased synaptic activation of dentate granule cells by exercise reflects inputs from the lateral entorhinal cortex. 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.02.22.707119v1?rss=1</link>
<description><![CDATA[
Hippocampal dentate granule cells receive multisensory information from the entorhinal cortex in a laminated and functionally segregated manner. We previously reported that brief periods of voluntary exercise in mice increased EPSCs and dendritic spines for inputs from the lateral, but not the medial, entorhinal cortex. Here we asked whether laminar specificity was due to molecular changes specific to distal granule cell dendrites or rather was dependent on upstream drive from the entorhinal cortex. Selective chemogenetic stimulation of either lateral entorhinal cortex (LEC) or the medial entorhinal cortex (MEC) increased granule cell dendritic spine density in the selected pathway. However, the preponderance of exercise-activated cells originated from LEC based on expression of an activity-dependent retrograde virus in Fos-TRAP mice. Our results indicate that the preferential activation by exercise reflects the drive of locomotor-related inputs from the lateral entorhinal cortex rather than selective molecular mechanisms in distal dendrites of dentate granule cells. How this activation pattern affects other salient stimuli involving contextual or spatial cues may underlie the benefits of exercise on learning and memory.
]]></description>
<dc:creator>Chatzi, C.</dc:creator>
<dc:creator>Simmonds, A.</dc:creator>
<dc:creator>Veshagh, A.</dc:creator>
<dc:creator>Ellingson, A.</dc:creator>
<dc:creator>Krush, M.</dc:creator>
<dc:creator>McLean, T.</dc:creator>
<dc:creator>Schnell, E.</dc:creator>
<dc:creator>Westbrook, G.</dc:creator>
<dc:date>2026-02-23</dc:date>
<dc:identifier>doi:10.64898/2026.02.22.707119</dc:identifier>
<dc:title><![CDATA[Biased synaptic activation of dentate granule cells by exercise reflects inputs from the lateral entorhinal cortex.]]></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/10.64898/2026.02.23.704410v1?rss=1">
<title>
<![CDATA[
Bradykinin Contributes to Vasogenic Edema in Murine Experimental Cerebral Malaria 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.02.23.704410v1?rss=1</link>
<description><![CDATA[
Cerebral malaria (CM) due to Plasmodium falciparum (Pf) infection is a major cause of death in African children. Bradykinin (BK) is a mediator of vasogenic edema. It could contribute to the pathogenesis of central nervous system malaria in Kenyan children and P. berghei ANKA (PbA) infected C57BL/6J mice with experimental cerebral malaria. Cleaved plasma high molecular weight kininogen (cHK) is a marker for prior BK release. 40% of children with central nervous system malaria had plasma cHK versus 18% of children with uncomplicated malaria. Wild-type PbA-infected mice had circulating plasma cHK, elevated BK levels, and reduced HK and prekallikrein levels. HK null (Kng1-/-), combined BK B1 and B2 receptor null (Bdkrb1-/- / Bdkrb2-/-), BK B2 (Bdkrb2-/-) or BK B1 (Bdkrb1-/-) receptor null mice were protected from neurologic deterioration and brain edema compared to wild-type mice. F12-/-mice were not protected from neurological deterioration.

Prekallikrein null (Klkb1-/-), prolylcarboxypeptidase hypomorphs (Prcpgt/gt), and brain endothelial cell conditional knockout of PRCP (Prcpfl/fl Cre) mice had reduced neurologic deterioration and brain edema. Adjuvant plasma kallikrein inhibition combined with artesunate treatment of PbA-infected mice reversed neurologic deterioration and brain edema and prolonged survival relative to artesunate alone. BK-induced vasogenic edema contributes to human and murine CM.
]]></description>
<dc:creator>Pinheiro, A. d. S.</dc:creator>
<dc:creator>Teixeira, D. E.</dc:creator>
<dc:creator>Silva-Aguiar, R. P.</dc:creator>
<dc:creator>Shim, Y. J.</dc:creator>
<dc:creator>Merkulova, A.</dc:creator>
<dc:creator>Silbak, S.</dc:creator>
<dc:creator>Skomorovska-Prokvolit, Y.</dc:creator>
<dc:creator>Midem, D.</dc:creator>
<dc:creator>Ogolla, S.</dc:creator>
<dc:creator>Burckhardt, B. B.</dc:creator>
<dc:creator>Gangnus, T.</dc:creator>
<dc:creator>Scharfstein, J.</dc:creator>
<dc:creator>Caruso-Neves, C.</dc:creator>
<dc:creator>McCarty, O. J.</dc:creator>
<dc:creator>Gailani, D.</dc:creator>
<dc:creator>Bader, M.</dc:creator>
<dc:creator>Rosenthal, P.</dc:creator>
<dc:creator>Dent, A. E.</dc:creator>
<dc:creator>Janse, C. J.</dc:creator>
<dc:creator>McCrae, K.</dc:creator>
<dc:creator>Pinheiro, A. A. d. S.</dc:creator>
<dc:creator>Kazura, J. W.</dc:creator>
<dc:creator>Schmaier, A. H.</dc:creator>
<dc:date>2026-02-26</dc:date>
<dc:identifier>doi:10.64898/2026.02.23.704410</dc:identifier>
<dc:title><![CDATA[Bradykinin Contributes to Vasogenic Edema in Murine Experimental Cerebral Malaria]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-02-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.03.02.709058v1?rss=1">
<title>
<![CDATA[
Polycyclic aromatic hydrocarbons, gut microbiome composition, impulsivity, and attention covary in a human cohort 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.03.02.709058v1?rss=1</link>
<description><![CDATA[
Polycyclic aromatic hydrocarbons (PAHs) are pervasive environmental pollutants linked to adverse neurobehavioral outcomes, yet the biological pathways coupling exposure to behavior are poorly defined. The gut microbiome is both sensitive to PAH exposure and a modulator of central nervous system function, suggesting it may mediate how PAH exposure influences neurobehavior. We tested whether PAH exposure, gut microbiome composition, and neurobehavioral function covary in a statewide sample of 34 adults stratified into high-impulsivity/poor-attention (HH) and low-impulsivity/fast-attention (LL) groups. Participants provided fecal samples for 16S rRNA profiling and wore silicone wristbands for 30 days to passively sample PAH exposure. Higher PAH exposure associated with HH group membership in a sex-dependent manner, with the largest elevations among HH males. At the community level, PAH exposure profiles correlated with microbiome dissimilarity, and HH membership associated with increased alpha-diversity and altered community composition relative to LL members. At the taxon level, 21 genera were significantly associated with 14 PAH compounds (FDR < 0.1). No individual genera were significantly associated with neurobehavioral group after multiple testing correction. Nevertheless, cross-referencing PAH-responsive genera (FDR < 0.1) against those with nominal neurobehavioral associations (p < 0.05) identified two candidate genera, Hydrogenoanaerobacterium and Methanobrevibacter, whose abundance covaries with both PAH exposure and neurobehavioral phenotype. Both have been independently linked to cognitive or neurological outcomes in prior work. These findings support a three-way relationship among environmental chemical exposure, gut microbiome composition, and neurobehavioral function, establishing an empirical foundation for testing microbiome-mediated links between PAH exposure and neurobehavioral outcomes.

IMPORTANCEPAH exposure is widespread and associates with impulsivity and attention problems, but how exposure translates into neurobehavioral risk is unclear. The gut microbiome is a plausible intermediary: gut microbes biotransform environmental chemicals and produce metabolites that influence brain function. In a statewide adult cohort, we show that higher PAH exposure tracks with greater impulsivity and poorer attention in a sex-dependent manner, and that both PAH exposure and neurobehavioral phenotype associate with distinct gut microbiome features at the community and taxon levels. We identify candidate genera at the intersection of PAH exposure and neurobehavioral group whose biology independently implicates them in cognitive and neurological function. By demonstrating that all three domains covary within a single human cohort, this work moves beyond pairwise associations to identify candidate microbial intermediaries for mechanistic investigation. Defining the microbiome constituents that respond to PAH exposure and co-associate with neurobehavioral phenotypes creates opportunities to test microbiome-targeted or exposure-reduction strategies for mitigating PAH-related neurobehavioral impacts.
]]></description>
<dc:creator>Hammer, A. J.</dc:creator>
<dc:creator>Kasschau, K. D.</dc:creator>
<dc:creator>Alexiev, A.</dc:creator>
<dc:creator>Davis, E. W.</dc:creator>
<dc:creator>Hoffman, P.</dc:creator>
<dc:creator>Johnson-Camacho, K.</dc:creator>
<dc:creator>Anderson, K.</dc:creator>
<dc:creator>Marriott, L. K.</dc:creator>
<dc:creator>Shannon, J.</dc:creator>
<dc:creator>Sharpton, T. J.</dc:creator>
<dc:date>2026-03-02</dc:date>
<dc:identifier>doi:10.64898/2026.03.02.709058</dc:identifier>
<dc:title><![CDATA[Polycyclic aromatic hydrocarbons, gut microbiome composition, impulsivity, and attention covary in a human cohort]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-03-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.02.27.702183v1?rss=1">
<title>
<![CDATA[
Multi-Omic, Multi-Tissue Responses to Acute Exercise in Sedentary Adults: Findings from the Molecular Transducers of Physical Activity Consortium 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.02.27.702183v1?rss=1</link>
<description><![CDATA[
Regular physical activity represents one of the greatest mechanisms for maintaining human health, yet the underlying molecular transducers of these benefits remain incompletely understood. Multi-omic assays now provide new opportunities to study the coordinated molecular responses of body tissues to different exercise modalities. The Molecular Transducers of Physical Activity Consortium (MoTrPAC) was established to address this need by creating a molecular map of the response to physical activity. Described here is the first human cohort of MoTrPAC: sedentary adults enrolled prior to study suspension during the COVID-19 pandemic (N=175) randomized to either endurance or resistance exercise, or non-exercise control. From these participants, we detail their global acute molecular response in skeletal muscle, adipose tissue, and blood, integrated at multiple levels: tissue, exercise modality, timepoint, and omic category. These analyses characterize key molecular pathways, identify central regulators, and implicate novel candidate exerkines in mediating multi-organ exercise effects.
]]></description>
<dc:creator>MoTrPAC Study Group,</dc:creator>
<dc:creator>Katz, D. H.</dc:creator>
<dc:creator>Jin, C. A.</dc:creator>
<dc:creator>Many, G. M.</dc:creator>
<dc:creator>Smith, G. R.</dc:creator>
<dc:creator>Keshishian, H.</dc:creator>
<dc:creator>Clark, N. M.</dc:creator>
<dc:creator>Iyer, G.</dc:creator>
<dc:creator>Ahn, C.</dc:creator>
<dc:creator>Lindholm, M. E.</dc:creator>
<dc:creator>Sagendorf, T. J.</dc:creator>
<dc:creator>Amar, D.</dc:creator>
<dc:creator>Barber, J. L.</dc:creator>
<dc:creator>Brandt, A. R.</dc:creator>
<dc:creator>Coen, P. M.</dc:creator>
<dc:creator>Ge, Y.</dc:creator>
<dc:creator>Hart, P.</dc:creator>
<dc:creator>Hsu, F.-C.</dc:creator>
<dc:creator>Jaeger, B. C.</dc:creator>
<dc:creator>Jimenez-Morales, D.</dc:creator>
<dc:creator>Leach, D. T.</dc:creator>
<dc:creator>Mani, D. R.</dc:creator>
<dc:creator>Montalvo, S.</dc:creator>
<dc:creator>Pincas, H.</dc:creator>
<dc:creator>Rao, P.</dc:creator>
<dc:creator>Sanford, J. A.</dc:creator>
<dc:creator>Smith, K. S.</dc:creator>
<dc:creator>Vetr, N. G.</dc:creator>
<dc:creator>Adkins, J. N.</dc:creator>
<dc:creator>Ashley, E. A.</dc:creator>
<dc:creator>Carr, S. A.</dc:creator>
<dc:creator>Miller, M. E.</dc:creator>
<dc:creator>Montgomery, S. B.</dc:creator>
<dc:creator>Nair, V. D.</dc:creator>
<dc:creator>Robbins, J. M.</dc:creator>
<dc:creator>Snyder, M. P.</dc:creator>
<dc:creator>Sparks, L. M.</dc:creator>
<dc:creator>Tracy, R.</dc:creator>
<dc:creator>Walsh, M. J.</dc:creator>
<dc:creator>Wheeler, M. T.</dc:creator>
<dc:creator>Xia, A. Y.</dc:creator>
<dc:creator>Sealfon, S. C.</dc:creator>
<dc:creator>Gerszten, R</dc:creator>
<dc:date>2026-03-03</dc:date>
<dc:identifier>doi:10.64898/2026.02.27.702183</dc:identifier>
<dc:title><![CDATA[Multi-Omic, Multi-Tissue Responses to Acute Exercise in Sedentary Adults: Findings from the Molecular Transducers of Physical Activity Consortium]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-03-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.03.10.710924v1?rss=1">
<title>
<![CDATA[
Combined Menin and XPO1 inhibition drive synergistic antileukemic activity in KMT2Ar and NPM1-m AML 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.03.10.710924v1?rss=1</link>
<description><![CDATA[
Menin scaffolds the oncogenic histone-lysine-N-methyltransferase (KMT2A)-fusion protein (FP) complex in KMT2A-r and wild-type KMT2A complex in NPM1-m acute myeloid leukemia (AML). Menin inhibitors (MIs) are effective in KMT2A-r AML and NPM1-m AML. However, not all patients respond to MIs as monotherapy. In this preclinical study, we demonstrate that the MI ziftomenib, in combination with the XPO1 inhibitor selinexor, synergistically inhibited the growth of multiple KMT2A-r and NPM1-m AML cell lines (CI<1). The combination suppressed colony formation in primary CD34+ KMT2A-r progenitor cells without affecting normal stem cells. Robust apoptosis and decreased G2/M populations were also evident. The combination downregulated HOXA9 and MEIS1 while upregulating monocytic differentiation marker CD11b in both the AML molecular signatures. RNA sequencing and proteomic analysis in KMT2A-r revealed suppression of multiple bona fide menin-KMT2A target genes. Our mechanistic studies also identified a novel role of XPO1 in stabilizing menins binding to chromatin and its interactions with KMT2A and KMT2A/MLLT3. XPO1 inhibitor-mediated disruption of these interactions, particularly in combination with ziftomenib, synergistically impairs oncogenic transcriptional programs. In vivo, combination therapy improved survival in both MV4;11 and OCI-AML3 cell line and primary patient-derived KMT2A-r and NPM1-m AML xenograft models in NSG mice, effective even at reduced drug doses. These preclinical findings demonstrate that simultaneous inhibition of the menin-KMT2A interaction and XPO1 can be a more effective translational strategy for treating KMT2A-r and NPM1-m AML than MI monotherapy to deepen responses and delay/prevent relapses.
]]></description>
<dc:creator>Uddin, M. H.</dc:creator>
<dc:creator>Dhiman, S.</dc:creator>
<dc:creator>Han, Y.</dc:creator>
<dc:creator>Aboukameel, A.</dc:creator>
<dc:creator>Dhillon, V.</dc:creator>
<dc:creator>Aguillar, J.</dc:creator>
<dc:creator>Buck, S.</dc:creator>
<dc:creator>Deol, A.</dc:creator>
<dc:creator>Boerner, J. L.</dc:creator>
<dc:creator>Polin, L.</dc:creator>
<dc:creator>Kessler, L.</dc:creator>
<dc:creator>Burrows, F.</dc:creator>
<dc:creator>Yang, J.</dc:creator>
<dc:creator>Azmi, A. S.</dc:creator>
<dc:creator>Maciejewski, J.</dc:creator>
<dc:creator>Cutler, J.</dc:creator>
<dc:creator>Du, Y.</dc:creator>
<dc:creator>Balasubramanian, S. K.</dc:creator>
<dc:date>2026-03-13</dc:date>
<dc:identifier>doi:10.64898/2026.03.10.710924</dc:identifier>
<dc:title><![CDATA[Combined Menin and XPO1 inhibition drive synergistic antileukemic activity in KMT2Ar and NPM1-m AML]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-03-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.03.12.711395v1?rss=1">
<title>
<![CDATA[
SynAPSeg: A novel dataset and image analysis framework for deep learning-based synapse detection and quantification 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.03.12.711395v1?rss=1</link>
<description><![CDATA[
Synapses are the fundamental units of neural computation, yet quantifying their organization across circuit-level scales remains a critical bottleneck in neuroscience. While advances in fluorescent labeling and imaging can generate vast datasets, analysis is often the limiting factor. Several deep learning-based tools have been proposed to ameliorate these issues. However, existing applications primarily focus on dendritic spines and lack robust solutions for segmenting synaptic puncta in dense tissue preparations. To address this, we introduce SynAPSeg, which encompasses an open-source framework for deep learning-based analysis and, to the best of our knowledge, the first large-scale, publicly available instance segmentation dataset specifically curated for synaptic puncta. We use this dataset to train deep learning models that reach the performance of human experts across a unique benchmark dataset. SynAPSeg integrates these models into an interactive interface, with support for multi-dimensional data, enabling fully automated segmentation and quantification pipelines alongside an annotation module for refinement and validation. We demonstrate the frameworks scalability by performing the first comprehensive mapping of nearly 4 million excitatory postsynaptic PSD95 puncta within inhibitory interneurons across the dorsal hippocampus, revealing regional differences in synapse properties. Finally, we show SynAPSegs utility for 3D quantification by applying these models to study aging-associated synaptic changes in CA1 parvalbumin (PV)-positive inhibitory neurons. Through this approach, we uncover a reduction in PSD95 density along PV dendrites in the aged CA1, indicating reduced glutamatergic recruitment of PV neurons which could contribute to age-related cognitive decline. Collectively, these results demonstrate that SynAPSeg provides a scalable solution for comprehensively studying synaptic architecture in health and disease.
]]></description>
<dc:creator>Schamber, P.</dc:creator>
<dc:creator>Darbhamulla, S.</dc:creator>
<dc:creator>Boyer, M.</dc:creator>
<dc:creator>Pelletier, M.</dc:creator>
<dc:creator>Hartman, H.</dc:creator>
<dc:creator>Friedman, O.</dc:creator>
<dc:creator>Zhang, S.</dc:creator>
<dc:creator>Blais, A.</dc:creator>
<dc:creator>Oh, S.</dc:creator>
<dc:creator>Zhong, H.</dc:creator>
<dc:creator>Bygrave, A. M.</dc:creator>
<dc:date>2026-03-16</dc:date>
<dc:identifier>doi:10.64898/2026.03.12.711395</dc:identifier>
<dc:title><![CDATA[SynAPSeg: A novel dataset and image analysis framework for deep learning-based synapse detection and quantification]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-03-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.03.13.711472v1?rss=1">
<title>
<![CDATA[
CROCHET: a versatile pipeline for automated analysis and visual atlas creation from single-cell spatialomic data 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.03.13.711472v1?rss=1</link>
<description><![CDATA[
Spatial biology technologies offer a unique opportunity to link tissue composition with function. However, analytical methods for quantifying and interpreting highly complex spatial data remain limited. We present CROCHET (ChaRacterization Of Cellular HEterogeneity in Tissues), an end-to-end analysis pipeline for construction of spatially resolved cell atlases from raw data covering millions of cells across large sample cohorts. Its modular architecture supports the integration of diverse data modalities and novel analytical methods for image processing and segmentation, spatialomics quantification and downstream analyses. With comprehensive, open-source, user-friendly, interactive, and visual analysis modules, CROCHET aims to democratize spatial omics for a broad community of users.
]]></description>
<dc:creator>Bozorgui, B.</dc:creator>
<dc:creator>Thibault, G.</dc:creator>
<dc:creator>Yuan, C.</dc:creator>
<dc:creator>Dereli, Z.</dc:creator>
<dc:creator>Wang, H.</dc:creator>
<dc:creator>Overman, M. J.</dc:creator>
<dc:creator>Weinstein, J. N.</dc:creator>
<dc:creator>Korkut, A.</dc:creator>
<dc:date>2026-03-17</dc:date>
<dc:identifier>doi:10.64898/2026.03.13.711472</dc:identifier>
<dc:title><![CDATA[CROCHET: a versatile pipeline for automated analysis and visual atlas creation from single-cell spatialomic data]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-03-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.03.20.713247v1?rss=1">
<title>
<![CDATA[
A critical signaling role for diacylglycerol in phagocytosis of M. tuberculosis 
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</title>
<link>https://biorxiv.org/content/10.64898/2026.03.20.713247v1?rss=1</link>
<description><![CDATA[
Mycobacterium tuberculosis (Mtb) establishes infection by entering host phagocytes through phagocytosis. While host lipids are known to influence this process, the specific contribution of the signaling lipid diacylglycerol (DAG) remains poorly defined. Here, we identify DAG as a critical regulator of phagocytosis. Disruption of DAG production, through inhibition or genetic deletion of adipose triglyceride lipase (ATGL) and phospholipase C gamma 2 (PLC{gamma}2), two major pathways that generate cellular DAG pools, markedly reduced uptake of both Mtb and zymosan-coated beads. Notably, loss of ATGL or PLC{gamma}2 did not impair receptor trafficking to the cell surface or cargo binding, indicating that DAG is not required for phagocytic recognition or initiation, but instead for a later step in phagosome formation. Mechanistically, cells lacking ATGL or PLC{gamma}2 displayed constitutive phosphoinositide 3-kinase (PI3K) phosphorylation, suggesting that dysregulated intracellular signaling prevents completion of phagocytosis. These findings uncover a previously unappreciated role for DAG biosynthesis in coordinating intracellular signaling required for phagocytosis and provide new insight into host pathways that govern Mtb entry.
]]></description>
<dc:creator>Griffith, A. M.</dc:creator>
<dc:creator>Garcia, M.</dc:creator>
<dc:creator>Guzman, G.</dc:creator>
<dc:creator>Tafesse, F.</dc:creator>
<dc:date>2026-03-23</dc:date>
<dc:identifier>doi:10.64898/2026.03.20.713247</dc:identifier>
<dc:title><![CDATA[A critical signaling role for diacylglycerol in phagocytosis of M. tuberculosis]]></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/10.64898/2026.03.21.713346v1?rss=1">
<title>
<![CDATA[
Metaplastic sleep regulation in Drosophila determined by microscale circadian neural dynamics 
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</title>
<link>https://biorxiv.org/content/10.64898/2026.03.21.713346v1?rss=1</link>
<description><![CDATA[
The biophysical mechanisms by which circadian clock neurons integrate temporal coding signals to regulate sleep remain elusive. Here, using Drosophila, we identify Rabphilin (Rph) in DN1p clock neurons as a key stabilizer of the metaplasticity setpoint governing circadian regulation of sleep. Rph protein levels are elevated at night relative to daytime and modulate stochastic process of DN1p membrane potential dynamics linked to variability in synaptic activity at connections between DN1p neurons and their downstream postsynaptic partners. We find that Rph acts as a bidirectional regulator of synaptic plasticity thresholds. Under dim nocturnal light stimulation, Rph knockdown permits synaptic potentiation, whereas synthesized Rph introduction induces synaptic depression. In contrast, under optogenetic manipulation mimicking daytime spiking in DN1p neurons, these effects are reversed. We further show that spike-timing-dependent plasticity emerges when postsynaptic spiking is engaged, with nocturnal dim light conditions determining the direction of plasticity. Together, these findings establish a mechanistic link between microscale circadian neural dynamics and hierarchical metaplastic regulation, demonstrating how circadian regulation of sleep dynamically balances stability and adaptive flexibility through circadian setpoints and environmental nocturnal light interactions.

Significance StatementWe show that circadian metaplasticity regulates sleep through membrane potential dynamics. Circadian clock neurons implement flexible metaplasticity, whereby the direction can be determined by internal circadian setpoints and interactions with nocturnal environmental light. This mechanism engages spike-timing-dependent plasticity to determine plasticity polarity. Our findings identify membrane potential dynamics as a computational substrate for physiological state control, linking molecular mechanisms to circuit-level circadian regulation of sleep. Together, they reframe sleep regulation as an active metaplastic process that hierarchically integrates microscale circadian neural dynamics to optimize circuit function.
]]></description>
<dc:creator>Hutson, A. N.</dc:creator>
<dc:creator>Zarjetskiy, A. N.</dc:creator>
<dc:creator>Zhang, Y. J.</dc:creator>
<dc:creator>Pokaleva, N.</dc:creator>
<dc:creator>Paul, E. M.</dc:creator>
<dc:creator>Xie, Y.</dc:creator>
<dc:creator>Chong, B.</dc:creator>
<dc:creator>Sanchez Franco, V. M.</dc:creator>
<dc:creator>Zukowski, L. H.</dc:creator>
<dc:creator>Faulk, E. E.</dc:creator>
<dc:creator>Walker, J. A.</dc:creator>
<dc:creator>Brown, A. M.</dc:creator>
<dc:creator>Nguyen, D. L.</dc:creator>
<dc:creator>Ferry, F. S.</dc:creator>
<dc:creator>Snyder, E. M.</dc:creator>
<dc:creator>Tabuchi, M.</dc:creator>
<dc:date>2026-03-24</dc:date>
<dc:identifier>doi:10.64898/2026.03.21.713346</dc:identifier>
<dc:title><![CDATA[Metaplastic sleep regulation in Drosophila determined by microscale circadian neural dynamics]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-03-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.03.27.714739v1?rss=1">
<title>
<![CDATA[
Cascading periods of language-related brain plasticity across early childhood 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.03.27.714739v1?rss=1</link>
<description><![CDATA[
Language is thought to have multiple sensitive periods in early childhood, but the neural basis of these sensitive periods is less understood. We leverage advances in in-vivo neuroimaging of plasticity, measuring the neural inhibition across the brain via Hurst exponent. Using two large datasets with children ages 10 months to 15 years (Baby Connectome Project: 10m-3y6m, 458 observations across n=222 children; Human Connectome Project-Development: 5-15y, n=324), we characterize the development of the Hurst exponent in language-related brain regions. In early childhood, Hurst increases in temporal and frontal language areas, and posterior regions develop earlier than anterior regions. In contrast, thalamic Hurst plateaus earlier, perhaps underlying the earliest language-related sensitive periods. Children with higher language-related skills show slower increases in cortical Hurst in early childhood, suggesting protracted plasticity. Later in childhood, cortical Hurst plateaus around age 9, suggesting a potential neural mechanism for age-related declines syntax learning. These results highlight a potential neural basis for cascading language-related sensitive periods.

Research highlightsO_LILanguage has multiple, cascading sensitive periods, but the neural basis of these sensitive periods is not well-understood.
C_LIO_LIWe leverage advances in in-vivo neuroimaging to quantify plasticity in language-related brain regions across childhood via Hurst, a measure of inhibition.
C_LIO_LIWe find that Hurst increases (plasticity decreases) in a graded fashion, with posterior regions maturing before anterior regions.
C_LIO_LIThalamic hurst plateaus in the first year of life, while cortical Hurst plateaus at age 9, suggesting a neural basis for distinct language-related sensitive periods.
C_LI
]]></description>
<dc:creator>Ellwood-Lowe, M. E.</dc:creator>
<dc:creator>Nishio, M.</dc:creator>
<dc:creator>Dufford, A. E.</dc:creator>
<dc:creator>Arcaro, M.</dc:creator>
<dc:creator>Satterthwaite, T. D.</dc:creator>
<dc:creator>Mackey, A. P.</dc:creator>
<dc:date>2026-03-27</dc:date>
<dc:identifier>doi:10.64898/2026.03.27.714739</dc:identifier>
<dc:title><![CDATA[Cascading periods of language-related brain plasticity across early childhood]]></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/10.64898/2026.03.26.714521v1?rss=1">
<title>
<![CDATA[
Tumor-immune trajectory context connects static tissue architecture to clinical outcomes 
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</title>
<link>https://biorxiv.org/content/10.64898/2026.03.26.714521v1?rss=1</link>
<description><![CDATA[
Multiplexed tissue imaging (MTI) has revealed recurrent tumor microenvironment (TME) architectures with prognostic value, yet these measurements are inherently static, obscuring dynamic changes in the TME that govern therapeutic response. Here, we introduce a trajectory-centric framework that reconstructs continuous TME dynamics by integrating agent-based mathematical modeling and simulation with state space analysis. This approach yields a mechanistically constrained reference landscape built entirely from in silico simulation, and onto which static patient biospecimens can be projected and mapped onto simulated TME trajectories. Systematic simulation of tumor-immune interactions in triple-negative breast cancer identifies six metastable TME states connected by transition pathways spanning immune control to immune escape. Mapping MTI data from two independent patient cohorts, including longitudinal samples from a randomized immunotherapy trial, validates this landscape by positioning individual biospecimens along inferred TME trajectories rather than in static states. We show that treatment-phase TME states, but not pre-treatment configurations, robustly predict immunotherapy response, and identical terminal states can arise from distinct trajectory histories corresponding to immune failure or resolved inflammation. Thus, this framework enables mechanistic simulations to define a reference dynamical landscape that serves as a coordinate system for interpreting static clinical spatial data, providing a principled basis for evaluating consistency, predictiveness, and clinical relevance across independent patient cohorts. Altogether, this study advances spatial tumor profiling from static state classification of human tissues to dynamic trajectory inference, establishing a quantitative framework for trajectory-informed, state-guided, and temporally adaptive immunotherapy strategies.
]]></description>
<dc:creator>Cramer, E. M.</dc:creator>
<dc:creator>Heiland, R.</dc:creator>
<dc:creator>Lima da Rocha, H.</dc:creator>
<dc:creator>Bergman, D. R.</dc:creator>
<dc:creator>Gray, J. W.</dc:creator>
<dc:creator>Mills, G. B.</dc:creator>
<dc:creator>Fertig, E. J.</dc:creator>
<dc:creator>Macklin, P.</dc:creator>
<dc:creator>Heiser, L. M.</dc:creator>
<dc:creator>Chang, Y. H.</dc:creator>
<dc:date>2026-03-30</dc:date>
<dc:identifier>doi:10.64898/2026.03.26.714521</dc:identifier>
<dc:title><![CDATA[Tumor-immune trajectory context connects static tissue architecture to clinical outcomes]]></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/10.64898/2026.03.27.714838v1?rss=1">
<title>
<![CDATA[
rAAV prostaglandin-based gene therapy lowers intraocular pressure and preserves optic nerve health in glaucomatous DBA/2J mice 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.03.27.714838v1?rss=1</link>
<description><![CDATA[
Open-angle glaucoma (OAG) affects approximately 57.5 million individuals worldwide and is characterized by the progressive loss of retinal ganglion cells (RGC) and irreversible optic nerve damage resulting from chronic ocular hypertension. Intraocular pressure (IOP) is the only major modifiable risk factor in OAG and clinical treatments necessarily aim to lower IOP in order to preserve RGCs and prevent vison loss. Pharmacological therapies, such as prostaglandin analog containing eye drops, are known to be effective at reducing IOP, but are critically undermined by poor patient compliance and are unable to control for potentially damaging diurnal fluctuations in IOP, leading to vision loss even in patients diagnosed early. Herein we evaluate the effectiveness of a long-acting, single use, prostaglandin-based recombinant adeno-associated virus (rAAV)-mediated IOP-lowering gene therapy treatment in glaucomatous DBA/2J mice and demonstrate that sustained IOP reduction leads to preservation of both optic nerve anatomy and function in end-stage glaucomatous disease.

One Sentence SummaryIOP-lowering gene therapy provides partial anatomical and functional rescue in glaucomatous mouse model following single dose treatment
]]></description>
<dc:creator>Chern, K. J.</dc:creator>
<dc:creator>Fehrman, R. L.</dc:creator>
<dc:creator>Marcoe, G. J.</dc:creator>
<dc:creator>Lipinski, D. M.</dc:creator>
<dc:date>2026-03-31</dc:date>
<dc:identifier>doi:10.64898/2026.03.27.714838</dc:identifier>
<dc:title><![CDATA[rAAV prostaglandin-based gene therapy lowers intraocular pressure and preserves optic nerve health in glaucomatous DBA/2J mice]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-03-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/content/10.64898/2026.03.27.714880v1?rss=1">
<title>
<![CDATA[
Spontaneous Pregravid Obesity Reshapes Fetal Immune Ontogeny in a Nonhuman Primate Model 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.03.27.714880v1?rss=1</link>
<description><![CDATA[
Pregravid obesity is associated with longterm immune alterations in the offspring; however, the mechanisms remain poorly defined. To address this gap, we investigated the impact of spontaneous pregravid obesity, independent of obesogenic diet, on fetal immune ontogeny in a rhesus macaque model. Using spectral flow cytometry, multiplex cytokine profiling, functional stimulation assays, and singlecell RNA sequencing, we profiled immune composition, function, transcriptional profiles, and intercellular communication in umbilical cord blood as well as fetal spleen and lung. Pregravid obesity was associated with altered fetal organ growth, elevated inflammatory mediators, altered frequencies of immune cell populations, and hyperresponsiveness to stimulation by splenic and lung leukocytes. Singlecell transcriptomic analyses revealed tissuespecific reprogramming of innate immune cells, including heightened inflammatory, migratory, and metabolic signatures with impaired antigen presentation. Moreover, there was evidence of impaired T cell differentiation, premature effector differentiation, and B cell dysfunction. Cell-cell communication analysis identified loss of tolerogenic signaling and enhanced proinflammatory pathways across spleen and lung myeloid cells. These findings demonstrate that spontaneous pregravid obesity fundamentally reshapes fetal circulating and tissueresident immune cells, providing mechanistic insight into the increased susceptibility to infection, respiratory diseases, and immune dysregulation observed in offspring of mothers with obesity.
]]></description>
<dc:creator>Doratt, B. M.</dc:creator>
<dc:creator>Wagner, S. B.</dc:creator>
<dc:creator>Avila, U.</dc:creator>
<dc:creator>Hodge, T.</dc:creator>
<dc:creator>Martin, L. D.</dc:creator>
<dc:creator>Varlamov, O.</dc:creator>
<dc:creator>Messaoudi, I.</dc:creator>
<dc:date>2026-03-31</dc:date>
<dc:identifier>doi:10.64898/2026.03.27.714880</dc:identifier>
<dc:title><![CDATA[Spontaneous Pregravid Obesity Reshapes Fetal Immune Ontogeny in a Nonhuman Primate Model]]></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/10.64898/2026.03.24.714034v1?rss=1">
<title>
<![CDATA[
Rectifying AI-generated protein structure ensembles for equilibrium using physics-based computations 
]]>
</title>
<link>https://biorxiv.org/content/10.64898/2026.03.24.714034v1?rss=1</link>
<description><![CDATA[
Recently, a number of tools have been released that generate ensembles of protein structures based on artificial intelligence (AI) approaches. Although ensembles generated by the tools differ significantly, we demonstrate a computational path to harmonizing the various outputs under a stationary condition using two complementary physics-based approaches. In the first stage, the AI ensemble is used to seed a weighted ensemble (WE) simulation, promoting relaxation toward the steady state. In the second stage, trajectory segments generated by WE are reweighted to steady state using the recently developed RiteWeight (RW) algorithm. We applied this approach to generate an atomically- detailed equilibrium ensemble of unliganded adenylate kinase conformations, starting from ensembles produced by three AI tools: AFSample2, ESMFlow-PDB (trained from PDB structures), and ESMFlow-MD (trained from molecular dynamics simulation data). Dramatic differences in the AI-generated ensembles are largely erased during the WE-RW process, yielding a consistent description of the equilibrium ensemble for a given force field.
]]></description>
<dc:creator>Otten, L.</dc:creator>
<dc:creator>Leung, J. M. G.</dc:creator>
<dc:creator>Chong, L.</dc:creator>
<dc:creator>Zuckerman, D. M.</dc:creator>
<dc:date>2026-04-03</dc:date>
<dc:identifier>doi:10.64898/2026.03.24.714034</dc:identifier>
<dc:title><![CDATA[Rectifying AI-generated protein structure ensembles for equilibrium using physics-based computations]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-04-03</prism:publicationDate>
<prism:section></prism:section>
</item>
</rdf:RDF>
