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	<title>bioRxiv Channel: Babraham Institute </title>
	<link>https://biorxiv.org</link>
	<description>
	This feed contains articles for bioRxiv Channel "Babraham Institute "
	</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/cgi/content/short/003608v1?rss=1">
<title>
<![CDATA[
A Comparison of Peak Callers Used for DNase-seq Data 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/003608v1?rss=1"
</link>
<description><![CDATA[
Genome-wide profiling of open chromatin regions using DNase I and high-throughput sequencing (DNase-seq) is an increasingly popular approach for finding and studying regulatory elements. A variety of algorithms have been developed to identify regions of open chromatin from raw sequence-tag data, which has motivated us to assess and compare their performance.nnIn this study, four published, publicly available peak calling algorithms used for DNase-seq data analysis (F-seq, Hotspot, MACS and ZINBA) are assessed at a range of signal thresholds on two published DNase-seq datasets for three cell types. The results were benchmarked against an independent dataset of regulatory regions derived from ENCODE in vivo transcription factor binding data for each particular cell type. The level of overlap between peak regions reported by each algorithm and this ENCODE-derived reference set was used to assess sensitivity and specificity of the algorithms.nnOur study suggests that F-seq has a slightly higher sensitivity than the next best algorithms. Hotspot and the ChIP-seq oriented method, MACS, both perform competitively when used with their default parameters. However the generic peak finder ZINBA appears to be less sensitive than the other three.nnWe also assess accuracy of each algorithm over a range of signal thresholds. In particular, we show that the accuracy of F-Seq can be considerably improved by using a threshold setting that is different from the default value.
]]></description>
<dc:creator>Hashem Koohy</dc:creator>
<dc:creator>Thomas Down</dc:creator>
<dc:creator>Mikhail Spivakov</dc:creator>
<dc:creator>Tim Hubbard</dc:creator>
<dc:creator></dc:creator>
<dc:date>2014-03-27</dc:date>
<dc:identifier>doi:10.1101/003608</dc:identifier>
<dc:title><![CDATA[A Comparison of Peak Callers Used for DNase-seq Data]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2014-03-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/028068v1?rss=1">
<title>
<![CDATA[
CHiCAGO: Robust Detection of DNA Looping Interactions in Capture Hi-C data 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/028068v1?rss=1"
</link>
<description><![CDATA[
Capture Hi-C (CHi-C) is a state-of-the art method for profiling chromosomal interactions involving targeted regions of interest (such as gene promoters) globally and at high resolution. Signal detection in CHi-C data involves a number of statistical challenges that are not observed when using other Hi-C-like techniques. We present a background model, and algorithms for normalisation and multiple testing that are specifically adapted to CHi-C experiments, in which many spatially dispersed regions are captured, such as in Promoter CHi-C. We implement these procedures in CHiCAGO (http://regulatorygenomicsgroup.org/chicago), an open-source package for robust interaction detection in CHi-C. We validate CHiCAGO by showing that promoter-interacting regions detected with this method are enriched for regulatory features and disease-associated SNPs.
]]></description>
<dc:creator>Jonathan Cairns</dc:creator>
<dc:creator>Paula Freire-Pritchett</dc:creator>
<dc:creator>Steven W. Wingett</dc:creator>
<dc:creator>Csilla Várnai</dc:creator>
<dc:creator>Andrew Dimond</dc:creator>
<dc:creator>Vincent Plagnol</dc:creator>
<dc:creator>Daniel Zerbino</dc:creator>
<dc:creator>Stefan Schoenfelder</dc:creator>
<dc:creator>Biola-Maria Javierre</dc:creator>
<dc:creator>Cameron Osborne</dc:creator>
<dc:creator>Peter Fraser</dc:creator>
<dc:creator>Mikhail Spivakov</dc:creator>
<dc:creator></dc:creator>
<dc:date>2015-10-01</dc:date>
<dc:identifier>doi:10.1101/028068</dc:identifier>
<dc:title><![CDATA[CHiCAGO: Robust Detection of DNA Looping Interactions in Capture Hi-C data]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2015-10-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/094466v1?rss=1">
<title>
<![CDATA[
Cell cycle dynamics of chromosomal organisation at single-cell resolution 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/094466v1?rss=1"
</link>
<description><![CDATA[
Chromosomes in proliferating metazoan cells undergo dramatic structural metamorphoses every cell cycle, alternating between a highly condensed mitotic structure facilitating chromosome segregation, and a decondensed interphase structure accommodating transcription, gene silencing and DNA replication. These cyclical structural transformations have been evident under the microscope for over a century, but their molecular-level analysis is still lacking. Here we use single-cell Hi-C to study chromosome conformations in thousands of individual cells, and discover a continuum of cis-interaction profiles that finely position individual cells along the cell cycle. We show that chromosomal compartments, topological domains (TADs), contact insulation and long-range loops, all defined by ensemble Hi-C maps, are governed by distinct cell cycle dynamics. In particular, DNA replication correlates with build-up of compartments and reduction in TAD insulation, while loops are generally stable from G1 through S and G2. Analysing whole genome 3D structural models using haploid cell data, we discover a radial architecture of chromosomal compartments with distinct epigenomic signatures. Our single-cell data creates an essential new paradigm for the re-interpretation of chromosome conformation maps through the prism of the cell cycle.
]]></description>
<dc:creator>Nagano, T.</dc:creator>
<dc:creator>Lubling, Y.</dc:creator>
<dc:creator>Varnai, C.</dc:creator>
<dc:creator>Dudley, C.</dc:creator>
<dc:creator>Leung, W.</dc:creator>
<dc:creator>Baran, Y.</dc:creator>
<dc:creator>Mandelson Cohen, N.</dc:creator>
<dc:creator>Wingett, S.</dc:creator>
<dc:creator>Fraser, P.</dc:creator>
<dc:creator>Tanay, A.</dc:creator>
<dc:date>2016-12-15</dc:date>
<dc:identifier>doi:10.1101/094466</dc:identifier>
<dc:title><![CDATA[Cell cycle dynamics of chromosomal organisation at single-cell resolution]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2016-12-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/107250v1?rss=1">
<title>
<![CDATA[
SBpipe: a collection of pipelines for automating repetitive simulation and analysis tasks 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/107250v1?rss=1"
</link>
<description><![CDATA[
Background: The rapid growth of the number of mathematical models in Systems Biology fostered the development of many tools to simulate and analyse them. The reliability and precision of these tasks often depend on multiple repetitions and they can be optimised if executed as pipelines. In addition, new formal analyses can be performed on these repeat sequences, revealing important insights about the accuracy of model predictions.nnResults: Here we introduce SBpipe, an open source software tool for automating repetitive tasks in model building and simulation. Using basic configuration files, SBpipe builds a sequence of repeated model simulations or parameter estimations, performs analyses from this generated sequence, and finally generates a LaTeX/PDF report. The parameter estimation pipeline offers analyses of parameter profile likelihood and parameter correlation using samples from the computed estimates. Specific pipelines for scanning of one or two model parameters at the same time are also provided. Pipelines can run on multicore computers, Sun Grid Engine (SGE), or Load Sharing Facility (LSF) clusters, speeding up the processes of model building and simulation. SBpipe can execute models implemented in Copasi, Python or coded in any other programming language using Python as a wrapper module. Future support for other software simulators can be dynamically added without affecting the current implementation.nnConclusions: SBpipe allows users to automatically repeat the tasks of model simulation and parameter estimation, and extract robustness information from these repeat sequences in a solid and consistent manner, facilitating model development and analysis. The source code and documentation of this project are freely available at the web site: https://pdp10.github.io/sbpipe/.
]]></description>
<dc:creator>Dalle Pezze, P.</dc:creator>
<dc:creator>Le Novere, N.</dc:creator>
<dc:date>2017-02-09</dc:date>
<dc:identifier>doi:10.1101/107250</dc:identifier>
<dc:title><![CDATA[SBpipe: a collection of pipelines for automating repetitive simulation and analysis tasks]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-02-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/119206v1?rss=1">
<title>
<![CDATA[
Multi-Tissue DNA Methylation Age Predictor In Mouse 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/119206v1?rss=1"
</link>
<description><![CDATA[
BackgroundDNA-methylation changes at a discrete set of sites in the human genome are predictive of chronological and biological age. However, it is not known whether these changes are causative or a consequence of an underlying ageing process. It has also not been shown whether this  epigenetic clock is unique to humans or conserved in the more experimentally tractable mouse.nnResultsWe have generated a comprehensive set of genome-scale base-resolution methylation maps from multiple mouse tissues spanning a wide range of ages. Many CpG sites show significant tissue-independent correlations with age and allowed us to develop a multi-tissue predictor of age in the mouse. Our model, which estimates age based on DNA methylation at 329 unique CpG sites, has a median absolute error of 3.33 weeks, and has similar properties to the recently described human epigenetic clock. Using publicly available datasets, we find that the mouse clock is accurate enough to measure effects on biological age, including in the context of interventions. While females and males show no significant differences in predicted DNA methylation age, ovariectomy results in significant age acceleration in females. Furthermore, we identify significant differences in age-acceleration dependent on the lipid content of the offspring diet.nnConclusionsHere we identify and characterize an epigenetic predictor of age in mice, the mouse epigenetic clock. This clock will be instrumental for understanding the biology of ageing and will allow modulation of its ticking rate and resetting the clock in vivo to study the impact on biological age.
]]></description>
<dc:creator>Stubbs, T. M.</dc:creator>
<dc:creator>Bonder, M. J.</dc:creator>
<dc:creator>Stark, A.-K.</dc:creator>
<dc:creator>Krueger, F.</dc:creator>
<dc:creator>BI Ageing Clock Team,</dc:creator>
<dc:creator>von Meyenn, F.</dc:creator>
<dc:creator>Stegle, O.</dc:creator>
<dc:creator>Reik, W.</dc:creator>
<dc:date>2017-03-22</dc:date>
<dc:identifier>doi:10.1101/119206</dc:identifier>
<dc:title><![CDATA[Multi-Tissue DNA Methylation Age Predictor In Mouse]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-03-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/177626v1?rss=1">
<title>
<![CDATA[
A non-catalytic role of TET3 promotes open chromatin and enhances global transcription 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/177626v1?rss=1"
</link>
<description><![CDATA[
The methylcytosine dioxygenase Tet3 is highly expressed as a specific isoform in oocytes and zygotes but essentially absent from later stages of mouse preimplantation development. Here, we show that Tet3 expression promotes transdifferentiation of embryonic stem cells to trophoblast-like stem cells. By genome-wide analyses we demonstrate that TET3 associates with and co-occupies chromatin with RNA Polymerase II. Tet3 expression induces a global increase of transcription and total RNA levels, and its presence further enhances chromatin accessibility in regions open for transcription. This novel function of TET3 is not specific to the oocyte isoform, independent of its catalytic activity, the CXXC domain, or its interaction with OGT, and is localised in its highly conserved exon 4. We propose a more general role for TET3 promoting open chromatin and enhancing global transcription during changes of cellular identity, separate from its catalytic function.
]]></description>
<dc:creator>Krueger, C.</dc:creator>
<dc:creator>Peat, J. R.</dc:creator>
<dc:creator>Eckersley-Maslin, M. A.</dc:creator>
<dc:creator>Hore, T. A.</dc:creator>
<dc:creator>Mohammed, H.</dc:creator>
<dc:creator>Andrews, S. R.</dc:creator>
<dc:creator>Dean, W.</dc:creator>
<dc:creator>Reik, W.</dc:creator>
<dc:date>2017-08-17</dc:date>
<dc:identifier>doi:10.1101/177626</dc:identifier>
<dc:title><![CDATA[A non-catalytic role of TET3 promotes open chromatin and enhances global transcription]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-08-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/196147v1?rss=1">
<title>
<![CDATA[
Spatial RNA proximities reveal a bipartite nuclear transcriptome and territories of differential density and transcription elongation rates 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/196147v1?rss=1"
</link>
<description><![CDATA[
Spatial transcriptomics aims to understand how the ensemble of RNA molecules in tissues and cells is organized in 3D space. Here we introduce Proximity RNA-seq, which enriches for nascent transcripts, and identifies contact preferences for individual RNAs in cell nuclei. Proximity RNA-seq is based on massive-throughput RNA-barcoding of sub-nuclear particles in water-in-oil emulsion droplets, followed by sequencing. We show a bipartite organization of the nuclear transcriptome in which compartments of different RNA density correlate with transcript families, tissue specificity and extent of alternative splicing. Integration of proximity measurements at the DNA and NA level identify transcriptionally active genomic regions with increased nucleic acid density and faster RNA polymerase II elongation located close to compact chromatin.
]]></description>
<dc:creator>Morf, J.</dc:creator>
<dc:creator>Wingett, S. W.</dc:creator>
<dc:creator>Farabella, I.</dc:creator>
<dc:creator>Cairns, J.</dc:creator>
<dc:creator>Furlan-Magaril, M.</dc:creator>
<dc:creator>Liu, X.</dc:creator>
<dc:creator>Craig, F. F.</dc:creator>
<dc:creator>Andrews, S.</dc:creator>
<dc:creator>Marti-Renom, M. A.</dc:creator>
<dc:creator>Fraser, P.</dc:creator>
<dc:date>2017-09-29</dc:date>
<dc:identifier>doi:10.1101/196147</dc:identifier>
<dc:title><![CDATA[Spatial RNA proximities reveal a bipartite nuclear transcriptome and territories of differential density and transcription elongation rates]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-09-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/238048v1?rss=1">
<title>
<![CDATA[
Tri-methylation of Histone H3 Lysine 4 Facilitates Gene Expression in Ageing Cells 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/238048v1?rss=1"
</link>
<description><![CDATA[
Transcription of protein coding genes is accompanied by recruitment of COMPASS to promoter-proximal chromatin, which deposits di- and tri-methylation on histone H3 lysine 4 (H3K4) to form H3K4me2 and H3K4me3. Here we determine the importance of COMPASS in maintaining gene expression across lifespan in budding yeast. We find that COMPASS mutations dramatically reduce replicative lifespan and cause widespread gene expression defects. Known repressive functions of H3K4me2 are progressively lost with age, while hundreds of genes become dependent on H3K4me3 for full expression. Induction of these H3K4me3 dependent genes is also impacted in young cells lacking COMPASS components including the H3K4me3-specific factor Spp1. Remarkably, the genome-wide occurrence of H3K4me3 is progressively reduced with age despite widespread transcriptional induction, minimising the normal positive correlation between promoter H3K4me3 and gene expression. Our results provide clear evidence that H3K4me3 is required to attain normal expression levels of many genes across organismal lifespan.
]]></description>
<dc:creator>Cruz, C.</dc:creator>
<dc:creator>Della Rosa, M.</dc:creator>
<dc:creator>Krueger, C.</dc:creator>
<dc:creator>Gao, Q.</dc:creator>
<dc:creator>Field, L.</dc:creator>
<dc:creator>Houseley, J.</dc:creator>
<dc:date>2017-12-22</dc:date>
<dc:identifier>doi:10.1101/238048</dc:identifier>
<dc:title><![CDATA[Tri-methylation of Histone H3 Lysine 4 Facilitates Gene Expression in Ageing Cells]]></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/cgi/content/short/138685v1?rss=1">
<title>
<![CDATA[
Joint Profiling Of Chromatin Accessibility, DNA Methylation And Transcription In Single Cells 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/138685v1?rss=1"
</link>
<description><![CDATA[
Parallel single-cell sequencing protocols represent powerful methods for investigating regulatory relationships, including epigenome-transcriptome interactions. Here, we report a novel single-cell method for parallel chromatin accessibility, DNA methylation and transcriptome profiling. scNMT-seq (single-cell nucleosome, methylation and transcription sequencing) uses a GpC methyltransferase to label open chromatin followed by bisulfite and RNA sequencing. We validate scNMT-seq by applying it to differentiating mouse embryonic stem cells, finding links between all three molecular layers and revealing dynamic coupling between epigenomic layers during differentiation.
]]></description>
<dc:creator>Clark, S. J.</dc:creator>
<dc:creator>Argelaguet, R.</dc:creator>
<dc:creator>Kapourani, C.-A.</dc:creator>
<dc:creator>Stubbs, T. M.</dc:creator>
<dc:creator>Lee, H. J.</dc:creator>
<dc:creator>Krueger, F.</dc:creator>
<dc:creator>Sanguinetti, G.</dc:creator>
<dc:creator>Kelsey, G.</dc:creator>
<dc:creator>Marioni, J. C.</dc:creator>
<dc:creator>Stegle, O.</dc:creator>
<dc:creator>Reik, W.</dc:creator>
<dc:date>2017-05-17</dc:date>
<dc:identifier>doi:10.1101/138685</dc:identifier>
<dc:title><![CDATA[Joint Profiling Of Chromatin Accessibility, DNA Methylation And Transcription In Single Cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-05-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/165449v1?rss=1">
<title>
<![CDATA[
Comparison of whole-genome bisulfite sequencing library preparation strategies identifies sources of biases affecting DNA methylation data 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/165449v1?rss=1"
</link>
<description><![CDATA[
BackgroundWhole-genome bisulfite sequencing (WGBS) is becoming an increasingly accessible technique, used widely for both fundamental and disease-oriented research. Library preparation methods benefit from a variety of available kits, polymerases and bisulfite conversion protocols. Although some steps in the procedure, such as PCR amplification, are known to introduce biases, a systematic evaluation of biases in WGBS strategies is missing.nnResultsWe perform a comparative analysis of several commonly used pre-and post-bisulfite WGBS library preparation protocols for their performance and quality of sequencing outputs. Our results show that bisulfite conversion per se is the main trigger of pronounced sequencing biases, and PCR amplification builds on these underlying artefacts. The majority of standard library preparation methods yield a significantly biased sequence output and overestimate global methylation. Importantly, both absolute and relative methylation levels at specific genomic regions vary substantially between methods, with clear implications for DNA methylation studies.nnConclusionsWe show that amplification-free library preparation is the least biased approach for WGBS. In protocols with amplification, the choice of BS conversion protocol or polymerase can significantly minimize artefacts. To aid with the quality assessment of existing WGBS datasets, we have integrated a bias diagnostic tool in the Bismark package and offer several approaches for consideration during the preparation and analysis of WGBS datasets.
]]></description>
<dc:creator>Olova, N.</dc:creator>
<dc:creator>Krueger, F.</dc:creator>
<dc:creator>Andrews, S.</dc:creator>
<dc:creator>Oxley, D. O.</dc:creator>
<dc:creator>Branco, M. R.</dc:creator>
<dc:creator>Reik, W.</dc:creator>
<dc:date>2017-07-18</dc:date>
<dc:identifier>doi:10.1101/165449</dc:identifier>
<dc:title><![CDATA[Comparison of whole-genome bisulfite sequencing library preparation strategies identifies sources of biases affecting DNA methylation data]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2017-07-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/283747v1?rss=1">
<title>
<![CDATA[
Nucleosome remodelling at origins of Global Genome-Nucleotide Excision Repair occurs at the boundaries of higher-order chromatin structure 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/283747v1?rss=1"
</link>
<description><![CDATA[
Repair of UV-induced DNA damage requires chromatin remodeling. How repair is initiated in chromatin remains largely unknown. We recently demonstrated that Global Genome Nucleotide Excision Repair (GG-NER) in chromatin is organized into domains around open reading frames. Here, we identify these domains, and by examining DNA damage-induced changes in the linear structure of nucleosomes, we demonstrate how chromatin remodeling is initiated during repair. In undamaged cells, we show that the GG-NER complex occupies chromatin at nucleosome free regions of specific gene promoters. This establishes the nucleosome structure at these genomic locations, which we refer to as GG-NER complex binding sites (GCBSs). We demonstrate that these sites are frequently located at genomic boundaries that delineate chromasomally interacting domains (CIDs). These boundaries define domains of higher-order nucleosome-nucleosome interaction. We show that efficient repair of DNA damage in chromatin is initiated following disruption of H2A.Z-containing nucleosomes adjacent to GCBSs by the GG-NER complex.
]]></description>
<dc:creator>van Eijk, P.</dc:creator>
<dc:creator>Nandi, S. P.</dc:creator>
<dc:creator>Yu, S.</dc:creator>
<dc:creator>Bennett, M.</dc:creator>
<dc:creator>Leadbitter, M.</dc:creator>
<dc:creator>Teng, Y.</dc:creator>
<dc:creator>Reed, S.</dc:creator>
<dc:date>2018-03-16</dc:date>
<dc:identifier>doi:10.1101/283747</dc:identifier>
<dc:title><![CDATA[Nucleosome remodelling at origins of Global Genome-Nucleotide Excision Repair occurs at the boundaries of higher-order chromatin structure]]></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/cgi/content/short/280651v1?rss=1">
<title>
<![CDATA[
PI3Kδ hyper-activation promotes the development of B cells that exacerbate Streptococcus pneumoniae infection in an antibody-independent manner 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/280651v1?rss=1"
</link>
<description><![CDATA[
Streptococcus pneumoniae is a major cause of pneumonia and a leading cause of death world-wide. Antibody-mediated immune responses can offer protection against repeated exposure to S. pneumoniae, yet vaccines only offer partial protection. Patients with Activated PI3K{delta} Syndrome (APDS) are highly susceptible to S. pneumoniae. We generated a conditional knockin mouse model of this disease and identified a CD19+B220- B cell subset that is induced by PI3K{delta} signaling, is resident in the lungs, and which promotes increased susceptibility to S. pneumoniae during the early phase of infection via an antibody-independent mechanism. We show that an inhaled PI3K{delta} inhibitor improves survival rates following S. pneumoniae infection in wild-type mice and in mice with activated PI3K{delta}. These results suggest that a subset of B cells in the lung can promote the severity of S. pneumoniae infection, representing a novel therapeutic target.
]]></description>
<dc:creator>Stark, A.-K.</dc:creator>
<dc:creator>Chandra, A.</dc:creator>
<dc:creator>Chakraborty, K.</dc:creator>
<dc:creator>Alam, R.</dc:creator>
<dc:creator>Carbonaro, V.</dc:creator>
<dc:creator>Clark, J.</dc:creator>
<dc:creator>Sriskantharajah, S.</dc:creator>
<dc:creator>Bradley, G.</dc:creator>
<dc:creator>Richter, A. G.</dc:creator>
<dc:creator>Banham-Hall, E.</dc:creator>
<dc:creator>Clatworthy, M. R.</dc:creator>
<dc:creator>Nejentsev, S.</dc:creator>
<dc:creator>Hamblin, J. N.</dc:creator>
<dc:creator>Hessel, E. M.</dc:creator>
<dc:creator>Condliffe, A. M.</dc:creator>
<dc:creator>Okkenhaug, K.</dc:creator>
<dc:date>2018-03-18</dc:date>
<dc:identifier>doi:10.1101/280651</dc:identifier>
<dc:title><![CDATA[PI3Kδ hyper-activation promotes the development of B cells that exacerbate Streptococcus pneumoniae infection in an antibody-independent manner]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-03-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/338822v1?rss=1">
<title>
<![CDATA[
Genome-scale oscillations in DNA methylation during exit from pluripotency 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/338822v1?rss=1"
</link>
<description><![CDATA[
Pluripotency is accompanied by the erasure of parental epigenetic memory with naive pluripotent cells exhibiting global DNA hypomethylation both in vitro and in vivo. Exit from pluripotency and priming for differentiation into somatic lineages is associated with genome-wide de novo DNA methylation. We show that during this phase, coexpression of enzymes required for DNA methylation turnover, DNMT3s and TETs, promotes cell-to-cell variability in this epigenetic mark. Using a combination of single-cell sequencing and quantitative biophysical modelling, we show that this variability is associated with coherent, genome-scale, oscillations in DNA methylation with an amplitude dependent on CpG density. Analysis of parallel single-cell transcriptional and epigenetic profiling provides evidence for oscillatory dynamics both in vitro and in vivo. These observations provide fresh insights into the emergence of epigenetic heterogeneity during early embryo development, indicating that dynamic changes in DNA methylation might influence early cell fate decisions.nnHighlightsO_LICo-expression of DNMT3s and TETs drive genome-scale oscillations of DNA methylationnC_LIO_LIOscillation amplitude is greatest at a CpG density characteristic of enhancersnC_LIO_LICell synchronisation reveals oscillation period and link with primary transcriptsnC_LIO_LIMultiomic single-cell profiling provides evidence for oscillatory dynamics in vivonC_LI
]]></description>
<dc:creator>Rulands, S.</dc:creator>
<dc:creator>Lee, H. J.</dc:creator>
<dc:creator>Clark, S. J.</dc:creator>
<dc:creator>Angermueller, C.</dc:creator>
<dc:creator>Smallwood, S. A.</dc:creator>
<dc:creator>Krueger, F.</dc:creator>
<dc:creator>Mohammed, H.</dc:creator>
<dc:creator>Dean, W.</dc:creator>
<dc:creator>Nichols, J.</dc:creator>
<dc:creator>Rugg-Gunn, P.</dc:creator>
<dc:creator>Kelsey, G.</dc:creator>
<dc:creator>Stegle, O.</dc:creator>
<dc:creator>Simons, B. D.</dc:creator>
<dc:creator>Reik, W.</dc:creator>
<dc:date>2018-06-05</dc:date>
<dc:identifier>doi:10.1101/338822</dc:identifier>
<dc:title><![CDATA[Genome-scale oscillations in DNA methylation during exit from pluripotency]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-06-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/431890v1?rss=1">
<title>
<![CDATA[
Dppa2 and Dppa4 directly regulate the Dux driven zygotic transcriptional programme 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/431890v1?rss=1"
</link>
<description><![CDATA[
The molecular regulation of zygotic genome activation (ZGA) in mammals remains poorly understood. Primed mouse embryonic stem cells contain a rare subset of "2C-like" cells that are epigenetically and transcriptionally similar to the two cell embryo and thus represent an ideal system for studying ZGA transcription regulation. Recently, the transcription factor Dux, expressed exclusively in the minor wave of ZGA, was described to activate many downstream ZGA transcripts. However, it remains unknown what upstream maternal factors initiate ZGA either in a Dux dependent or independent manner. Here we performed a candidate-based overexpression screen, identifying, amongst others, Developmental Pluripotency Associated 2 (Dppa2) and 4 (Dppa4) as positive regulators of 2C-like cells and ZGA transcription. In the germ line, promoter DNA demethylation coincides with upregulation of Dppa2 and Dppa4 which remain expressed until E7.5 when their promoters are remethylated. Furthermore, Dppa2 and Dppa4 are also expressed during iPSC reprogramming at the time 2C-like ZGA transcription transiently peaks. Through a combination of overexpression, knockdown, knockout and rescue experiments, together with transcriptional analyses, we show that Dppa2 and Dppa4 directly regulate the 2C-like cell population and associated transcripts, including Dux and the Zscan4 cluster. Importantly, we tease apart the molecular hierarchy in which the 2C-like transcriptional programme is initiated and stabilised. Dppa2 and Dppa4 require Dux to initiate 2C-like ZGA transcription, suggesting they act upstream by directly regulating Dux. Supporting this, ChIP-seq analysis revealed Dppa2 and Dppa4 bind to the Dux promoter and gene body and drive its expression. Zscan4c is also able to induce 2C-like cells in wild type cells, but, in contrast to Dux, can no longer do so in Dppa2/4 double knockout cells, suggesting it may act to stabilise rather than drive the transcriptional network. Our findings suggest a model in which Dppa2/4 binding to the Dux promoter leads to Dux upregulation and activation of the 2C-like transcriptional programme which is subsequently reinforced by Zscan4c.
]]></description>
<dc:creator>Eckersley-Maslin, M. A.</dc:creator>
<dc:creator>Alda-Catalinas, C.</dc:creator>
<dc:creator>Blotenburg, M.</dc:creator>
<dc:creator>Kreibich, E.</dc:creator>
<dc:creator>Krueger, C.</dc:creator>
<dc:creator>Reik, W.</dc:creator>
<dc:date>2018-10-01</dc:date>
<dc:identifier>doi:10.1101/431890</dc:identifier>
<dc:title><![CDATA[Dppa2 and Dppa4 directly regulate the Dux driven zygotic transcriptional programme]]></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/cgi/content/short/500900v1?rss=1">
<title>
<![CDATA[
Ageing affects DNA methylation drift and transcriptional cell-to-cell variability in muscle stem cells 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/500900v1?rss=1"
</link>
<description><![CDATA[
Age-related tissue alterations have been associated with a decline in stem cell number and function1. Although increased cell-to-cell variability in transcription or epigenetic marks has been proposed to be a major hallmark of ageing2-5, little is known about the molecular diversity of stem cells during ageing. Here, by combined single-cell transcriptome and DNA methylome profiling in mouse muscle stem cells, we show a striking global increase of uncoordinated transcriptional heterogeneity together with context-dependent alterations of DNA methylation with age. Importantly, promoters with increased methylation heterogeneity are associated with increased transcriptional heterogeneity of the genes they drive. Notably, old cells that change the most with age reveal alterations in the transcription of genes regulating cell-niche interactions. These results indicate that epigenetic drift, by accumulation of stochastic DNA methylation changes in promoters, is a substantial driver of the degradation of coherent transcriptional networks with consequent stem cell functional decline during ageing.
]]></description>
<dc:creator>Hernando-Herraez, I.</dc:creator>
<dc:creator>Evano, B.</dc:creator>
<dc:creator>Stubbs, T.</dc:creator>
<dc:creator>Commere, P.-H.</dc:creator>
<dc:creator>Clark, S.</dc:creator>
<dc:creator>Andrews, S.</dc:creator>
<dc:creator>Tajbakhsh, S.</dc:creator>
<dc:creator>Reik, W.</dc:creator>
<dc:date>2018-12-19</dc:date>
<dc:identifier>doi:10.1101/500900</dc:identifier>
<dc:title><![CDATA[Ageing affects DNA methylation drift and transcriptional cell-to-cell variability in muscle stem cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-12-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/502559v1?rss=1">
<title>
<![CDATA[
Microglia-dependent presynaptic disruption in an organotypic hippocampal slice culture model of neuroinflammation 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/502559v1?rss=1"
</link>
<description><![CDATA[
BackgroundSystemic inflammation, such as occurs during sepsis, bone fracture, infections or post-operative trauma, has been linked to synapse loss and cognitive decline in human patients and animal models. Organotypic hippocampal slice cultures (OHSCs) represent an underused tool in neuroinflammation; retaining much of the neuronal architecture, synaptic connections and diversity of cell types present in the hippocampus in vivo whilst providing convenient access to manipulate and sample the culture medium and observe cellular reactions as in other in vitro methods. Here we report the development of an OHSC model of synaptic disruption after aseptic inflammation and investigate the underlying mechanism.nnMethodsOHSCs were generated from P6-P9 C57BL/6, the APP transgenic TgCRND8 model, or wild-type littermate mice according to the interface method. Aseptic inflammation was induced via addition of lipopolysaccharide (LPS) and cultures were analysed for changes in synaptic proteins via western blot. qPCR and ELISA analysis of the slice tissue and culture medium respectively determined changes in gene expression and protein secretion. Microglia were selectively depleted using the toxin clodronate and the effect of IL1{beta} was assessed using a specific neutralising monoclonal antibody.nnResultsAddition of LPS caused a loss of the presynaptic protein synaptophysin via a mechanism dependent on microglia and involving IL1{beta}. Washout of LPS via medium exchange allows for partial recovery of synaptic protein levels after 2 weeks. TgCRND8 OHSCs do not show alterations in IL1{beta} expression at a timepoint where they exhibit spontaneous synaptophysin loss, and LPS does not alter levels of APP or A{beta} in wild-type OHSCs. This indicates that although synaptophysin loss is seen in both systems, there is likely to be distinct underlying pathogenic mechanisms between the neuroinflammatory and amyloid models.nnConclusionsWe report the development of an OHSC model of LPS-induced synaptophysin loss and demonstrate a key role for microglia and involvement of IL1{beta}. We propose that distinct molecular mechanisms lead to synaptophysin protein loss in LPS-exposed versus TgCRND8 OHSCs and provide a new experimental paradigm for assessing chronic changes in synaptic proteins, and synaptic plasticity, following acute inflammatory insults.
]]></description>
<dc:creator>Sheppard, O.</dc:creator>
<dc:creator>Coleman, M. P.</dc:creator>
<dc:creator>Durrant, C. S.</dc:creator>
<dc:date>2018-12-20</dc:date>
<dc:identifier>doi:10.1101/502559</dc:identifier>
<dc:title><![CDATA[Microglia-dependent presynaptic disruption in an organotypic hippocampal slice culture model of neuroinflammation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-12-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/519207v1?rss=1">
<title>
<![CDATA[
Single cell multi-omics profiling reveals a hierarchical epigenetic landscape during mammalian germ layer specification 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/519207v1?rss=1"
</link>
<description><![CDATA[
Formation of the three primary germ layers during gastrulation is an essential step in the establishment of the vertebrate body plan. Recent studies employing single cell RNA-sequencing have identified major transcriptional changes associated with germ layer specification. Global epigenetic reprogramming accompanies these changes, but the role of the epigenome in regulating early cell fate choice remains unresolved, and the coordination between different epigenetic layers is unclear. Here we describe the first single cell triple-omics map of chromatin accessibility, DNA methylation and RNA expression during the exit from pluripotency and the onset of gastrulation in mouse embryos. We find dynamic dependencies between the different molecular layers, with evidence for distinct modes of epigenetic regulation. The initial exit from pluripotency coincides with the establishment of a global repressive epigenetic landscape, followed by the emergence of local lineage-specific epigenetic patterns during gastrulation. Notably, cells committed to mesoderm and endoderm undergo widespread coordinated epigenetic rearrangements, driven by loss of methylation in enhancer marks and a concomitant increase of chromatin accessibility. In striking contrast, the epigenetic landscape of ectodermal cells is already established in the early epiblast. Hence, regulatory elements associated with each germ layer are either epigenetically primed or epigenetically remodelled prior to overt cell fate decisions during gastrulation, providing the molecular logic for a hierarchical emergence of the primary germ layers.nnHighlightsO_LIFirst map of mouse gastrulation using comprehensive single cell triple-omic analysis.nC_LIO_LIExit from pluripotency is associated with a global repressive epigenetic landscape, driven by a sharp gain of DNA methylation and a gradual decrease of chromatin accessibility.nC_LIO_LIDNA methylation and chromatin accessibility changes in enhancers, but not in promoters, are associated with germ layer formation.nC_LIO_LIMesoderm and endoderm enhancers become open and demethylated upon lineage commitment.nC_LIO_LIEctoderm enhancers are primed in the early epiblast and protected from the global repressive dynamics, supporting a default model of ectoderm commitment in vivo.nC_LI
]]></description>
<dc:creator>Argelaguet, R.</dc:creator>
<dc:creator>Mohammed, H.</dc:creator>
<dc:creator>Clark, S.</dc:creator>
<dc:creator>Stapel, C.</dc:creator>
<dc:creator>Krueger, C.</dc:creator>
<dc:creator>Kapourani, C. A.</dc:creator>
<dc:creator>Xiang, Y.</dc:creator>
<dc:creator>Hanna, C.</dc:creator>
<dc:creator>Smallwood, S.</dc:creator>
<dc:creator>Ibarra Soria, X.</dc:creator>
<dc:creator>Buettner, F.</dc:creator>
<dc:creator>Sanguinetti, G.</dc:creator>
<dc:creator>Krueger, F.</dc:creator>
<dc:creator>Xie, W.</dc:creator>
<dc:creator>Rugg-Gunn, P.</dc:creator>
<dc:creator>Kelsey, G.</dc:creator>
<dc:creator>Dean, W.</dc:creator>
<dc:creator>Nichols, J.</dc:creator>
<dc:creator>Stegle, O.</dc:creator>
<dc:creator>Marioni, J. C.</dc:creator>
<dc:creator>Reik, W.</dc:creator>
<dc:date>2019-01-13</dc:date>
<dc:identifier>doi:10.1101/519207</dc:identifier>
<dc:title><![CDATA[Single cell multi-omics profiling reveals a hierarchical epigenetic landscape during mammalian germ layer specification]]></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/cgi/content/short/526269v1?rss=1">
<title>
<![CDATA[
Chicdiff: a computational pipeline for detecting differential chromosomal interactions in Capture Hi-C data 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/526269v1?rss=1"
</link>
<description><![CDATA[
SummaryCapture Hi-C is a powerful approach for detecting chromosomal interactions involving, at least on one end, DNA regions of interest, such as gene promoters. We present Chicdiff, an R package for robust detection of differential interactions in Capture Hi-C data. ChiScdiff enhances a state-of-the-art differential testing approach for count data with bespoke normalisation and multiple testing procedures that account for specific statistical properties of Capture Hi-C. We validate Chicdiff on published Promoter Capture Hi-C data in human Monocytes and CD4+ T cells, identifying multitudes of cell type-specific interactions, and confirming the overall positive association between promoter interactions and gene expression. Chicdiff is implemented as an R package that is publicly available at https://github.com/RegulatoryGenomicsGroup/chicdiff.
]]></description>
<dc:creator>Cairns, J.</dc:creator>
<dc:creator>Orchard, W. R.</dc:creator>
<dc:creator>Malysheva, V.</dc:creator>
<dc:creator>Spivakov, M.</dc:creator>
<dc:date>2019-01-22</dc:date>
<dc:identifier>doi:10.1101/526269</dc:identifier>
<dc:title><![CDATA[Chicdiff: a computational pipeline for detecting differential chromosomal interactions in Capture Hi-C data]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-01-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/545830v1?rss=1">
<title>
<![CDATA[
Screening for genes that accelerate the epigenetic ageing clock in humans reveals a role for the H3K36 methyltransferase NSD1 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/545830v1?rss=1"
</link>
<description><![CDATA[
BackgroundEpigenetic clocks are mathematical models that predict the biological age of an individual using DNA methylation data, and which have emerged in the last few years as the most accurate biomarkers of the ageing process. However, little is known about the molecular mechanisms that control the rate of such clocks. Here, we have examined the human epigenetic clock in patients with a variety of developmental disorders, harbouring mutations in proteins of the epigenetic machinery.

ResultsUsing the Horvath epigenetic clock, we performed an unbiased screen for epigenetic age acceleration (EAA) in the blood of these patients. We demonstrate that loss-of-function mutations in the H3K36 histone methyltransferase NSD1, which cause Sotos syndrome, substantially accelerate epigenetic ageing. Furthermore, we show that the normal ageing process and Sotos syndrome share methylation changes and the genomic context in which they occur. Finally, we found that the Horvath clock CpG sites are characterised by a higher Shannon methylation entropy when compared with the rest of the genome, which is dramatically decreased in Sotos syndrome patients.

ConclusionsThese results suggest that the H3K36 methylation machinery is a key component of the epigenetic maintenance system in humans, which controls the rate of epigenetic ageing, and this role seems to be conserved in model organisms. Our observations provide novel insights into the mechanisms behind the epigenetic ageing clock and we expect will shed light on the different processes that erode the human epigenetic landscape during ageing.
]]></description>
<dc:creator>Martin-Herranz, D. E.</dc:creator>
<dc:creator>Aref-Eshghi, E.</dc:creator>
<dc:creator>Bonder, M. J.</dc:creator>
<dc:creator>Stubbs, T. M.</dc:creator>
<dc:creator>Stegle, O.</dc:creator>
<dc:creator>Sadikovic, B.</dc:creator>
<dc:creator>Reik, W.</dc:creator>
<dc:creator>Thornton, J. M.</dc:creator>
<dc:date>2019-02-10</dc:date>
<dc:identifier>doi:10.1101/545830</dc:identifier>
<dc:title><![CDATA[Screening for genes that accelerate the epigenetic ageing clock in humans reveals a role for the H3K36 methyltransferase NSD1]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-02-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/585489v1?rss=1">
<title>
<![CDATA[
Beta secretase 1-dependent amyloid precursor protein processing promotes excessive vascular sprouting through NOTCH3 signaling 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/585489v1?rss=1"
</link>
<description><![CDATA[
Amyloid beta peptides (A{beta}) proteins play a key role in vascular pathology in Alzheimers Disease (AD) including impairment of the blood brain barrier and aberrant angiogenesis. Although previous work has demonstrated a pro-angiogenic role of A{beta}, the exact mechanisms by which amyloid precursor protein (APP) processing and endothelial angiogenic signalling cascades interact in AD remain a largely unsolved problem. Here, we report that increased endothelial sprouting in human-APP transgenic mouse (TgCRND8) tissue is dependent on {beta}-secretase (BACE1) processing of APP. Higher levels of A{beta} processing in TgCRND8 tissue coincides with decreased NOTCH3/JAG1 signalling, over-production of endothelial filopodia and increased numbers of vascular pericytes. Using a novel in vitro approach to study sprouting angiogenesis in TgCRND8 organotypic brain slice cultures (OBSCs), we find that BACE1 inhibition normalises excessive endothelial filopodia formation and restores NOTCH3 signalling. These data present the first evidence for the potential of BACE1 inhibition as an effective therapeutic target for aberrant angiogenesis in AD.nnSignificanceIn this study, we show that targeting amyloid beta processing provides an opportunity to selectively target tip cell filopodia-driven angiogenesis and develop therapeutic targets for vascular dysfunction related to aberrant angiogenesis in AD. Our data provide the first evidence for a safe level of BACE1 inhibition that can normalize excess angiogenesis in AD, without inducing vascular deficits in healthy tissue. Our findings may pave the way for the development of new angiogenesis dependent therapeutic strategies in Alzheimers Disease.
]]></description>
<dc:creator>Durrant, C. S.</dc:creator>
<dc:creator>Ruscher, K.</dc:creator>
<dc:creator>Sheppard, O.</dc:creator>
<dc:creator>Coleman, M. P.</dc:creator>
<dc:creator>Özen, I.</dc:creator>
<dc:date>2019-03-21</dc:date>
<dc:identifier>doi:10.1101/585489</dc:identifier>
<dc:title><![CDATA[Beta secretase 1-dependent amyloid precursor protein processing promotes excessive vascular sprouting through NOTCH3 signaling]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-03-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/620062v1?rss=1">
<title>
<![CDATA[
Functional effects of variation in transcription factor binding highlight long-range gene regulation by epromoters 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/620062v1?rss=1"
</link>
<description><![CDATA[
Identifying DNA cis-regulatory modules (CRMs) that control the expression of specific genes is crucial for deciphering the logic of transcriptional control. Natural genetic variation can point to the possible gene regulatory function of specific sequences through their allelic associations with gene expression. However, comprehensive identification of causal regulatory sequences in brute-force association testing without incorporating prior knowledge is challenging due to limited statistical power and effects of linkage disequilibrium. Sequence variants affecting transcription factor (TF) binding at CRMs have a strong potential to influence gene regulatory function, which provides a motivation for prioritising such variants in association testing. Here, we generate an atlas of CRMs showing predicted allelic variation in TF binding affinity in human lymphoblastoid cell lines (LCLs) and test their association with the expression of their putative target genes inferred from Promoter Capture Hi-C and immediate linear proximity. We reveal over 1300 CRM TF-binding variants associated with target gene expression, the majority of them undetected with standard association testing. A large proportion of CRMs showing associations with the expression of genes they contact in 3D localise to the promoter regions of other genes, supporting the notion of  epromoters: dual-action CRMs with promoter and distal enhancer activity.
]]></description>
<dc:creator>Mitchelmore, J.</dc:creator>
<dc:creator>Grinberg, N.</dc:creator>
<dc:creator>Wallace, C.</dc:creator>
<dc:creator>Spivakov, M.</dc:creator>
<dc:date>2019-04-29</dc:date>
<dc:identifier>doi:10.1101/620062</dc:identifier>
<dc:title><![CDATA[Functional effects of variation in transcription factor binding highlight long-range gene regulation by epromoters]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-04-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/639807v1?rss=1">
<title>
<![CDATA[
An intergenic non-coding RNA promoter required for histone modifications in the human β-globin chromatin domain 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/639807v1?rss=1"
</link>
<description><![CDATA[
Transcriptome analyses show a surprisingly large proportion of the mammalian genome is transcribed; much more than can be accounted for by genes and introns alone. Most of this transcription is non-coding in nature and arises from intergenic regions, often overlapping known protein-coding genes in sense or antisense orientation. The functional relevance of this widespread transcription is unknown. Here we characterize a promoter responsible for initiation of an intergenic transcript located approximately 3.3 kb and 10.7 kb upstream of the adult-specific human {beta}-globin genes. Mutational analyses in {beta}-YAC transgenic mice show that alteration of intergenic promoter activity results in ablation of H3K4 di- and tri-methylation and H3 hyperacetylation extending over a 30 kb region immediately downstream of the initiation site, containing the adult {delta}- and {beta}-globin genes. This results in dramatically decreased expression of the adult genes through position effect variegation in which the vast majority of definitive erythroid cells harbor inactive adult globin genes. In contrast, expression of the neighboring {varepsilon}- and {gamma}-globin genes is completely normal in embryonic erythroid cells, indicating a developmentally specific variegation of the adult domain. Our results demonstrate a role for intergenic non-coding RNA transcription in the propagation of histone modifications over chromatin domains and epigenetic control of {beta}-like globin gene transcription during development.
]]></description>
<dc:creator>Debrand, E.</dc:creator>
<dc:creator>Chakalova, L.</dc:creator>
<dc:creator>Miles, J.</dc:creator>
<dc:creator>Dai, Y.-F.</dc:creator>
<dc:creator>Goyenechea, B.</dc:creator>
<dc:creator>Dye, S.</dc:creator>
<dc:creator>Osborne, C.</dc:creator>
<dc:creator>Harju-Baker, S.</dc:creator>
<dc:creator>Pink, R.</dc:creator>
<dc:creator>Caley, D.</dc:creator>
<dc:creator>Carter, D.</dc:creator>
<dc:creator>Peterson, K. R.</dc:creator>
<dc:creator>Fraser, P.</dc:creator>
<dc:date>2019-05-15</dc:date>
<dc:identifier>doi:10.1101/639807</dc:identifier>
<dc:title><![CDATA[An intergenic non-coding RNA promoter required for histone modifications in the human β-globin chromatin domain]]></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/cgi/content/short/641589v1?rss=1">
<title>
<![CDATA[
Follicular regulatory T cells can access the germinal centre independently of CXCR5 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/641589v1?rss=1"
</link>
<description><![CDATA[
The germinal centre (GC) response is critical for generating high-affinity humoral immunity and immunological memory, which forms the basis of successful immunisation. Control of the GC response is thought to require follicular regulatory T (Tfr) cells, a subset of suppressive Foxp3+ Treg cells located within GCs. Relatively little is known about the exact role of Tfr cells within the GC, and the mechanism/s through which they exert their suppressive function. A unique feature of Tfr cells is their reported CXCR5-dependent localisation to the GC. Here we show that the lack of CXCR5 on Foxp3+ regulatory T cells resulted in a reduced frequency, but not an absence of, GC-localised Tfr cells. This demonstrates that additional, CXCR5-independent mechanisms facilitate Treg cell homing to the GC.
]]></description>
<dc:creator>Vanderleyden, I.</dc:creator>
<dc:creator>Fra-Bido, S.</dc:creator>
<dc:creator>Innocentin, S.</dc:creator>
<dc:creator>Okkenhaug, H.</dc:creator>
<dc:creator>Evans-Bailey, N.</dc:creator>
<dc:creator>Pierson, W.</dc:creator>
<dc:creator>Linterman, M.</dc:creator>
<dc:date>2019-05-17</dc:date>
<dc:identifier>doi:10.1101/641589</dc:identifier>
<dc:title><![CDATA[Follicular regulatory T cells can access the germinal centre independently of CXCR5]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-05-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/741371v1?rss=1">
<title>
<![CDATA[
A single-cell transcriptomics CRISPR-activation screen identifies new epigenetic regulators of zygotic genome activation 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/741371v1?rss=1"
</link>
<description><![CDATA[
Zygotic genome activation (ZGA) is a crucial developmental milestone that remains poorly understood. This first essential transcriptional event in embryonic development coincides with extensive epigenetic reprogramming processes and is orchestrated, in part, by the interplay of transcriptional and epigenetic regulators. Here, we developed a novel high-throughput screening method that combines pooled CRISPR-activation (CRISPRa) with single-cell transcriptomics to systematically probe candidate regulators of ZGA. We screened 230 epigenetic and transcriptional regulators by upregulating their expression with CRISPRa in mouse embryonic stem cells (ESCs). Through single-cell RNA-sequencing (scRNA-seq) of CRISPRa-perturbed cells, we generated approximately 200,000 single-cell transcriptomes, each transduced with a unique short-guide RNA (sgRNA) targeting a specific candidate gene promoter. Using integrative dimensionality reduction of the perturbation scRNA-seq profiles, we characterized molecular signatures of ZGA and uncovered 44 factors that promote a ZGA-like response in ESCs, both in the coding and non-coding transcriptome. Upon upregulation of these factors, including the DNA binding protein Dppa2, the chromatin remodeller Smarca5 and the transcription factor Patz1, ESCs adopt an early embryonic-like state. Supporting their roles as ZGA regulators, Dppa2 and Smarca5 knock-out ESCs lose expression of ZGA genes, however, overexpression of Dppa2 in Smarca5 knock-out ESCs, but not vice versa, rescues ZGA-like expression, suggesting that Smarca5 regulates ZGA upstream and via Dppa2. Together, our single-cell transcriptomic profiling of CRISPRa-perturbed cells provides comprehensive system-level insights into the molecular mechanisms that orchestrate ZGA.nnHighlightsO_LIFirst large-scale screen combining pooled CRISPRa with scRNA-seq.nC_LIO_LIMulti-omics factor analysis identifies a ZGA-like signature for 44 of the candidate regulators.nC_LIO_LIDppa2, Smarca5 and Patz1 were validated as strong inducers of ZGA gene expression.nC_LIO_LISmarca5 regulates zygotic genome activation in a Dppa2-dependent manner.nC_LI
]]></description>
<dc:creator>Alda-Catalinas, C.</dc:creator>
<dc:creator>Bredikhin, D.</dc:creator>
<dc:creator>Hernando-Herraez, I.</dc:creator>
<dc:creator>Kubinyecz, O.</dc:creator>
<dc:creator>Santos, F.</dc:creator>
<dc:creator>Eckersley-Maslin, M. A.</dc:creator>
<dc:creator>Stegle, O.</dc:creator>
<dc:creator>Reik, W.</dc:creator>
<dc:date>2019-08-21</dc:date>
<dc:identifier>doi:10.1101/741371</dc:identifier>
<dc:title><![CDATA[A single-cell transcriptomics CRISPR-activation screen identifies new epigenetic regulators of zygotic genome activation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-08-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/744896v1?rss=1">
<title>
<![CDATA[
Transcription-induced formation of extrachromosomal DNA during yeast ageing 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/744896v1?rss=1"
</link>
<description><![CDATA[
Extrachromosomal circular DNA (eccDNA) facilitates adaptive evolution by allowing rapid and extensive gene copy number variation, and is implicated in the pathology of cancer and ageing. Here, we demonstrate that yeast aged under environmental copper accumulate high levels of eccDNA containing the copper resistance gene CUP1. Transcription of CUP1 causes CUP1 eccDNA accumulation, which occurs in the absence of phenotypic selection. We have developed a sensitive and quantitative eccDNA sequencing pipeline that reveals CUP1 eccDNA accumulation on copper exposure to be exquisitely site specific, with no other detectable changes across the eccDNA complement. eccDNA forms de novo from the CUP1 locus through processing of DNA double-strand breaks (DSBs) by Sae2 / Mre11 and Mus81, and genome-wide analyses show that other protein coding eccDNA species in aged yeast share a similar biogenesis pathway. Although abundant we find that CUP1 eccDNA does not replicate efficiently, and high copy numbers in aged cells arise through frequent formation events combined with asymmetric DNA segregation. The transcriptional stimulation of CUP1 eccDNA formation shows that age-linked genetic change varies with transcription pattern, resulting in gene copy number profiles tailored by environment.
]]></description>
<dc:creator>Hull, R.</dc:creator>
<dc:creator>King, M.</dc:creator>
<dc:creator>Pizza, G.</dc:creator>
<dc:creator>Krueger, F.</dc:creator>
<dc:creator>Vergara, X.</dc:creator>
<dc:creator>Houseley, J.</dc:creator>
<dc:date>2019-08-22</dc:date>
<dc:identifier>doi:10.1101/744896</dc:identifier>
<dc:title><![CDATA[Transcription-induced formation of extrachromosomal DNA during yeast ageing]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-08-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/370114v1?rss=1">
<title>
<![CDATA[
Dynamic modelling suggests differential mechanisms for initiation of non-selective autophagy and mitophagy 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/370114v1?rss=1"
</link>
<description><![CDATA[
During autophagy, the ULK complex nucleates autophagic precursors which give rise to autophagosomes. We analysed by live imaging and mathematical modelling translocation of ATG13 (part of ULK complex) to autophagic puncta in starvation-induced autophagy and ivermectin-induced mitophagy. In non-selective autophagy, the intensity and duration of ATG13 translocation approximated a normal distribution whereas wortmannin reduced this and shifted to a log-normal distribution. During mitophagy, multiple translocations of ATG13, with increasing time between peaks were observed. We hypothesised that these multiple translocations arise because engulfment of mitochondrial fragments requires successive nucleations of multiple phagophores on the same target, and a mathematical model based on this idea reproduced the oscillatory behaviour. Significantly, model and experimental data were also in agreement that the number of ATG13 translocations is directly proportional to the diameter of the targeted mitochondrial fragments. Our data provide novel insights into the early dynamics of selective and non-selective autophagy.
]]></description>
<dc:creator>Dalle Pezze, P.</dc:creator>
<dc:creator>Karanasios, E.</dc:creator>
<dc:creator>Kandia, V.</dc:creator>
<dc:creator>Walker, S.</dc:creator>
<dc:creator>Ktistakis, N.</dc:creator>
<dc:creator>Le Novere, N.</dc:creator>
<dc:date>2018-07-16</dc:date>
<dc:identifier>doi:10.1101/370114</dc:identifier>
<dc:title><![CDATA[Dynamic modelling suggests differential mechanisms for initiation of non-selective autophagy and mitophagy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2018-07-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/832873v1?rss=1">
<title>
<![CDATA[
Dppa2/4 target chromatin bivalency enabling multi-lineage commitment 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/832873v1?rss=1"
</link>
<description><![CDATA[
Bivalent chromatin marks developmental promoters in pluripotent cells, yet their targeting and precise impact on lineage commitment remains unclear. We uncover Developmental Pluripotency Associated 2 (Dppa2) and 4 (Dppa4) as epigenetic priming factors, establishing chromatin bivalency. Single-cell transcriptomics and differentiation assays reveal Dppa2/4 double knockout embryonic stem cells fail to exit pluripotency and differentiate efficiently. Dppa2/4 associate with COMPASS and Polycomb complexes and are required to recruit and maintain their binding at a subset of developmentally important bivalent promoters which are characterised by low expression and poised RNA polymerase. Consequently, upon Dppa2/4 knockout, these dependent promoters gain DNA methylation and are unable to be activated upon differentiation. Our findings uncover a novel targeting principle for bivalency to developmental promoters, poising them for future lineage specific activation.
]]></description>
<dc:creator>Eckersley-Maslin, M. A.</dc:creator>
<dc:creator>Parry, A. J.</dc:creator>
<dc:creator>Blotenburg, M.</dc:creator>
<dc:creator>Krueger, C.</dc:creator>
<dc:creator>Franklin, V. N. R.</dc:creator>
<dc:creator>Clark, S.</dc:creator>
<dc:creator>D'Santos, C. S.</dc:creator>
<dc:creator>Reik, W.</dc:creator>
<dc:date>2019-11-06</dc:date>
<dc:identifier>doi:10.1101/832873</dc:identifier>
<dc:title><![CDATA[Dppa2/4 target chromatin bivalency enabling multi-lineage commitment]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-11-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/769141v1?rss=1">
<title>
<![CDATA[
Polypyrimidine Tract Binding Proteins are essential for B cell development 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/769141v1?rss=1"
</link>
<description><![CDATA[
Polypyrimidine Tract Binding Protein 1 (PTBP1) is a RNA-binding protein (RBP) expressed throughout B cell development. Deletion of Ptbp1 in mouse pro-B cells results in upregulation of PTBP2 and normal B cell development. We show that PTBP2 compensates for PTBP1 in B cell ontogeny as deletion of both Ptbp1 and Ptbp2 results in a complete block at the pro-B cell stage and a lack of mature B cells. In pro-B cells PTBP1 ensures precise synchronisation of the activity of cyclin dependent kinases at distinct stages of the cell cycle, suppresses S-phase entry and promotes progression into mitosis. PTBP1 controls mRNA abundance and alternative splicing of important cell cycle regulators including CYCLIN-D2, c-MYC, p107 and CDC25B. Our results reveal a previously unrecognised mechanism mediated by a RBP that is essential for B cell ontogeny and integrates transcriptional and post-translational determinants of progression through the cell cycle.
]]></description>
<dc:creator>Monzon-Casanova, E.</dc:creator>
<dc:creator>Matheson, L. S.</dc:creator>
<dc:creator>Tabbada, K.</dc:creator>
<dc:creator>Zarnack, K.</dc:creator>
<dc:creator>Smith, C. W. J.</dc:creator>
<dc:creator>Turner, M.</dc:creator>
<dc:date>2019-09-14</dc:date>
<dc:identifier>doi:10.1101/769141</dc:identifier>
<dc:title><![CDATA[Polypyrimidine Tract Binding Proteins are essential for B cell development]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-09-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2019.12.13.875286v1?rss=1">
<title>
<![CDATA[
Network analysis of promoter interactions reveals the hierarchical differences in genome organisation between human pluripotent states 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2019.12.13.875286v1?rss=1"
</link>
<description><![CDATA[
SO_SCPCAPUMMARYC_SCPCAPA complex and poorly understood interplay between 3D genome organisation, transcription factors and chromatin state underpins cell identity. To gain a systems-level understanding of this interplay, we generated a high-resolution atlas of annotated chromatin interactions in naive and primed human pluripotent stem cells and developed a network-graph approach to examine the atlas at multiple spatial scales. Investigating chromatin interactions as a network uncovered highly connected hubs that changed substantially in interaction frequency and in transcriptional co-regulation between pluripotent states. Small hubs frequently merged to form larger networks in primed cells, often linked by newly-formed Polycomb-associated interactions. Importantly, we identified state-specific differences in enhancer activity and interactivity that corresponded with widespread reconfiguration of transcription factor binding and target gene expression. These findings provide multilayered insights into the gene regulatory control of human pluripotency and our systems-based network approach could be applied broadly to uncover new principles of 3D genome organisation.
]]></description>
<dc:creator>Chovanec, P.</dc:creator>
<dc:creator>Collier, A.</dc:creator>
<dc:creator>Krueger, C.</dc:creator>
<dc:creator>Varnai, C.</dc:creator>
<dc:creator>Schoenfelder, S.</dc:creator>
<dc:creator>Corcoran, A.</dc:creator>
<dc:creator>Rugg-Gunn, P.</dc:creator>
<dc:date>2019-12-13</dc:date>
<dc:identifier>doi:10.1101/2019.12.13.875286</dc:identifier>
<dc:title><![CDATA[Network analysis of promoter interactions reveals the hierarchical differences in genome organisation between human pluripotent states]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-12-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2019.12.20.882514v1?rss=1">
<title>
<![CDATA[
A neuronal thermostat controls membrane fluidity in C. elegans 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2019.12.20.882514v1?rss=1"
</link>
<description><![CDATA[
An organisms ability to adapt to heat can be key to its survival. Cells adapt to temperature shifts by adjusting lipid desaturation levels and the fluidity of membranes in a process that is thought to be controlled cell autonomously. We have discovered that subtle, step-wise increments in ambient temperature can lead to the conserved heat shock response being activated in head neurons of C. elegans. This response is exactly opposite to the expression of the lipid desaturase FAT-7 in the worms gut. We find that the over-expression of the master regulator of this response, Hsf-1, in head neurons, causes extensive fat remodeling to occur across tissues. These changes include a decrease in FAT-7 expression and a shift in the levels of unsaturated fatty acids in the plasma membrane. These shifts are in line with membrane fluidity requirements to survive in warmer temperatures. We have identified that the cGMP receptor, TAX-2/TAX-4, as well as TGF-{beta}/BMP signaling, as key players in the transmission of neuronal stress to peripheral tissues. This is the first study to suggest that a thermostat-based mechanism can centrally coordinate membrane fluidity in response to warm temperatures across tissues in multicellular animals.
]]></description>
<dc:creator>Casanueva, O.</dc:creator>
<dc:creator>Chauve, L.</dc:creator>
<dc:creator>Murdoch, S.</dc:creator>
<dc:creator>Masoudzadeh, F.</dc:creator>
<dc:creator>Hodge, F.</dc:creator>
<dc:creator>Lopez-Clavijo, A.</dc:creator>
<dc:creator>Okkenhaug, H.</dc:creator>
<dc:creator>Wingett, S.</dc:creator>
<dc:creator>Wakelam, M.</dc:creator>
<dc:creator>Kleigrewe, K.</dc:creator>
<dc:date>2019-12-20</dc:date>
<dc:identifier>doi:10.1101/2019.12.20.882514</dc:identifier>
<dc:title><![CDATA[A neuronal thermostat controls membrane fluidity in C. elegans]]></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/cgi/content/short/729657v1?rss=1">
<title>
<![CDATA[
Leptin resistance establishment in the ovary of diet-induced obese mice and its relationship with cumulus cells transcriptome 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/729657v1?rss=1"
</link>
<description><![CDATA[
Background/AimsObesity is associated with infertility, decreased ovarian performance and lipotoxicity. However, little is known about the aetiology of these reproductive impairments. Here, we hypothesise that the majority of changes in ovarian physiology in diet-induced obesity (DIO) are a consequence of transcriptional changes downstream of altered leptin signalling. Therefore, we investigated the extent to which leptin signalling is altered in the ovary upon obesity with particular emphasis on effects on cumulus cells (CCs), the intimate functional companions of the oocyte. Furthermore, we used the pharmacological hyperleptinemic (LEPT) mouse model to compare transcriptional profiles to DIO.

MethodsMice were subjected to DIO for 4 and 16 weeks (wk) and leptin treatment for 16 days, to study effects in the ovary in components of leptin signalling at the transcript and protein levels, using Western blot, Real-time PCR and immunostaining. Furthermore, we used low-cell RNA sequencing to characterise changes in the transcriptome of CCs in these models.

ResultsIn the DIO model, obesity led to establishment of ovarian leptin resistance after 16 wk high fat diet (HFD), as evidenced by increases in the feedback regulator suppressor of cytokine signalling 3 (SOCS3) and decreases in the positive effectors phosphorylation of tyrosine 985 of leptin receptor (ObRb-pTyr985) and Janus kinase 2 (pJAK2). Transcriptome analysis of the CCs revealed a complex response to DIO, with large numbers and distinct sets of genes deregulated at early and late stages of obesity; in addition, there was a striking correlation between body weight and global transcriptome profile of CCs. Further analysis indicated that the transcriptome profile in 4 wk HFD CCs resembled that of LEPT CCs, in the upregulation of cellular trafficking and impairment in cytoskeleton organisation. Conversely, after 16 wk HFD CCs showed expression changes indicative of augmented inflammatory responses, cell morphogenesis, and decreased metabolism and transport, mainly as a consequence of the physiological changes of obesity.

ConclusionObesity leads to ovarian leptin resistance and major time-dependent changes in gene expression in CCs, which in early obesity may be caused by increased leptin signalling in the ovary, whereas in late obesity are likely to be a consequence of metabolic changes taking place in the obese mother.
]]></description>
<dc:creator>Wolodko, K.</dc:creator>
<dc:creator>Walewska, E.</dc:creator>
<dc:creator>Castillo-Fernandez, J.</dc:creator>
<dc:creator>Kelsey, G. D.</dc:creator>
<dc:creator>Galvao, A.</dc:creator>
<dc:date>2019-08-15</dc:date>
<dc:identifier>doi:10.1101/729657</dc:identifier>
<dc:title><![CDATA[Leptin resistance establishment in the ovary of diet-induced obese mice and its relationship with cumulus cells transcriptome]]></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/cgi/content/short/2020.02.10.941989v1?rss=1">
<title>
<![CDATA[
Cohesin-dependent and independent mechanisms support chromosomal contacts between promoters and enhancers 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.02.10.941989v1?rss=1"
</link>
<description><![CDATA[
It is currently assumed that 3D chromosomal organisation plays a central role in transcriptional control. However, recent evidence shows that steady-state transcription of only a minority of genes is affected by depletion of architectural proteins such as cohesin and CTCF. Here, we have used Capture Hi-C to interrogate the dynamics of chromosomal contacts of all human gene promoters upon rapid architectural protein degradation. We show that promoter contacts lost in these conditions tend to be long-range, with at least one interaction partner localising in the vicinity of topologically associated domain (TAD) boundaries. In contrast, many shorter-range chromosomal contacts, particularly those that connect active promoters with each other and with active enhancers remain unaffected by cohesin and CTCF depletion. We demonstrate that the effects of cohesin depletion on nascent transcription can be explained by changes in the connectivity of their enhancers. Jointly, these results provide a mechanistic explanation to the limited, but consistent effects of cohesin and CTCF on steady-state transcription and point towards the existence of alternative enhancer-promoter pairing mechanisms that are independent of these proteins.
]]></description>
<dc:creator>Thiecke, M. J.</dc:creator>
<dc:creator>Wutz, G.</dc:creator>
<dc:creator>Muhar, M.</dc:creator>
<dc:creator>Tang, W.</dc:creator>
<dc:creator>Bevan, S.</dc:creator>
<dc:creator>Malysheva, V.</dc:creator>
<dc:creator>Stocsits, R. R.</dc:creator>
<dc:creator>Neumann, T.</dc:creator>
<dc:creator>Zuber, J.</dc:creator>
<dc:creator>Fraser, P.</dc:creator>
<dc:creator>Schoenfelder, S.</dc:creator>
<dc:creator>Peters, J.-M.</dc:creator>
<dc:creator>Spivakov, M.</dc:creator>
<dc:date>2020-02-11</dc:date>
<dc:identifier>doi:10.1101/2020.02.10.941989</dc:identifier>
<dc:title><![CDATA[Cohesin-dependent and independent mechanisms support chromosomal contacts between promoters and enhancers]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-02-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.02.25.964551v1?rss=1">
<title>
<![CDATA[
Causes of death in newborn C57BL/6J mice 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.02.25.964551v1?rss=1"
</link>
<description><![CDATA[
Neonatal mortality in wild-type laboratory mice is an overlooked welfare and financial problem in animal facilities around the world. Causes of death are often not reported and its causes remain unknown.

In this study, 324 newborn pups from two breeding colonies of healthy wildtype C57BL/6 mice underwent post-mortem analysis with special focus on obtaining proof of life after birth, evaluation of stomach contents and observation of congenital abnormalities that could compromise survival.

Based on a combination of lung morphology findings, outcome of lung float test, stomach contents and brown adipose tissue colouration, 21.6% of the pups found dead were considered stillbirths. Of the livebirths, only 3.2% were observed to have milk inside the stomach, indicating successful suckling. Congenital abnormalities were diagnosed only in a small fraction of the pups analysed. These results suggest that starvation was the most common cause of death, followed by stillbirth.
]]></description>
<dc:creator>Peneda, S. C.</dc:creator>
<dc:creator>Morello, G. M.</dc:creator>
<dc:creator>Lamas, S.</dc:creator>
<dc:creator>Olsson, A.</dc:creator>
<dc:creator>Gilbert, C.</dc:creator>
<dc:date>2020-02-26</dc:date>
<dc:identifier>doi:10.1101/2020.02.25.964551</dc:identifier>
<dc:title><![CDATA[Causes of death in newborn C57BL/6J mice]]></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/cgi/content/short/2020.02.25.953067v1?rss=1">
<title>
<![CDATA[
High laboratory mouse pre-weaning mortality associated with litter overlap, advanced mother age, small and large litters 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.02.25.953067v1?rss=1"
</link>
<description><![CDATA[
High and variable pre-weaning mortality is a persistent problem among the main mouse strains used in biomedical research. If a modest 15% mortality rate is assumed across all mouse strains used in the EU, approximately 1 million more pups must be produced yearly to compensate for those which die. A few environmental and social factors have been identified as affecting pup mortality, but optimizing these factors does not cease the problem. This study is the first large study to mine data records from 219,975 pups from two breeding facilities to determine the major risk factors associated with mouse pre-weaning mortality. It was hypothesized that litter overlap (i.e. the presence of older siblings in the cage when new pups are born), a recurrent social configuration in trio-housed mice, is associated with increased newborn mortality, along with high mother age, large litter size, as well as a high number and age of older siblings in the cage. The estimated probability of pup death was two to seven percentage points higher in cages with compared to those without litter overlap. Litter overlap was associated with an increase in percentage of litter losses of 19% and 103%, respectively, in the two breeding facilities. Increased number and age of older siblings, high mother age, small litter size (less than four pups born) and large litter size (over 11 pups born) were associated with increased probability of pup death. Results suggest that common social cage configurations at breeding facilities are dangerous for the survivability of young mouse pups. The underlying mechanisms and strategies to avoid these situations should be further investigated.
]]></description>
<dc:creator>Morello, G. M.</dc:creator>
<dc:creator>Hultgren, J.</dc:creator>
<dc:creator>Peneda, S. C.</dc:creator>
<dc:creator>Whiltshire, M.</dc:creator>
<dc:creator>Thomas, A.</dc:creator>
<dc:creator>Wardle-Jones, H.</dc:creator>
<dc:creator>Brajon, S.</dc:creator>
<dc:creator>Gilbert, C.</dc:creator>
<dc:creator>Olsson, A.</dc:creator>
<dc:date>2020-02-26</dc:date>
<dc:identifier>doi:10.1101/2020.02.25.953067</dc:identifier>
<dc:title><![CDATA[High laboratory mouse pre-weaning mortality associated with litter overlap, advanced mother age, small and large litters]]></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/cgi/content/short/2020.03.21.000042v1?rss=1">
<title>
<![CDATA[
IMPLICON: an ultra-deep sequencing method to uncover DNA methylation at imprinted regions 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.03.21.000042v1?rss=1"
</link>
<description><![CDATA[
Genomic imprinting is an epigenetic phenomenon leading to parental allele-specific expression. Dosage of imprinted genes is crucial for normal development and its dysregulation accounts for several human disorders. This unusual expression pattern is mostly dictated by differences in DNA methylation between parental alleles at specific regulatory elements known as imprinting control regions (ICRs). Although several approaches can be used for methylation inspection, we lack an easy and cost-effective method to simultaneously measure DNA methylation at multiple imprinted regions. Here, we present IMPLICON, a new high-throughput method measuring DNA methylation levels at imprinted regions with base-pair resolution and over 1000-fold coverage. We initially designed IMPLICON to look at ICRs in adult tissues of inbred mice. Then, we validated it in hybrid mice from reciprocal crosses for which we could discriminate methylation profiles in the two parental alleles. Lastly, we developed a human version of IMPLICON and detected imprinting errors in embryonic and induced pluripotent stem cells. We also provide rules and guidelines to adapt this method for investigating the DNA methylation landscape of any set of genomic regions. In summary, IMPLICON is a rapid, cost-effective and scalable method, which could become the gold standard in both imprinting research and diagnostics.
]]></description>
<dc:creator>Klobucar, T.</dc:creator>
<dc:creator>Kreibich, E.</dc:creator>
<dc:creator>Krueger, F.</dc:creator>
<dc:creator>Arez, M.</dc:creator>
<dc:creator>Polvora-Brandao, D.</dc:creator>
<dc:creator>von Meyenn, F.</dc:creator>
<dc:creator>da Rocha, S. T.</dc:creator>
<dc:creator>Eckersley-Maslin, M. A.</dc:creator>
<dc:date>2020-03-23</dc:date>
<dc:identifier>doi:10.1101/2020.03.21.000042</dc:identifier>
<dc:title><![CDATA[IMPLICON: an ultra-deep sequencing method to uncover DNA methylation at imprinted regions]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-03-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.03.31.007492v1?rss=1">
<title>
<![CDATA[
Specialisation and plasticity in a primitively social insect 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.03.31.007492v1?rss=1"
</link>
<description><![CDATA[
Biological systems not only have the remarkable capacity to build and maintain complex spatio-temporal structures in noisy environments, they can also rapidly break up and rebuild such structures. How such systems can simultaneously achieve both robust specialisation and plasticity is poorly understood. Here we use primitive societies of Polistes wasps as a model system where we experimentally perturb the social structure by removing the queen and follow the re-establishment of the social steady state over time. We combine a unique experimental strategy correlating time-resolved measurements across vastly different scales with a theoretical approach. We show that Polistes integrates antagonistic processes on multiple scales to distinguish between extrinsic and intrinsic perturbations and thereby achieve both robust specialisation and rapid plasticity. The long-term stability of the social structure relies on dynamic DNA methylation which controls transcriptional noise. Such dynamics provide a general principle of how both specialization and plasticity can be achieved in biological systems.

One Sentence SummaryA primitive social insect simultaneously achieves specialisation and plasticity by integrating antagonistic dynamics on different scales.

HighlightsO_LIWe employ a unique experimental approach correlating dynamics of societies, individuals, and epigenetic gene regulation
C_LIO_LIA social insect simultaneously achieves specialisation and plasticity by integrating antagonistic processes on different spatial scales
C_LIO_LIRegulation of population-level noise by DNA methylation ensures long-term stability of phenotypic specialisation
C_LI
]]></description>
<dc:creator>Patalano, S.</dc:creator>
<dc:creator>Alsina, A.</dc:creator>
<dc:creator>Gregorio-Rodriguez, C.</dc:creator>
<dc:creator>Bachman, M.</dc:creator>
<dc:creator>Dreier, S.</dc:creator>
<dc:creator>Hernando-Herraez, I.</dc:creator>
<dc:creator>Nana, P.</dc:creator>
<dc:creator>Balasubramanian, S.</dc:creator>
<dc:creator>Sumner, S.</dc:creator>
<dc:creator>Reik, W.</dc:creator>
<dc:creator>Rulands, S.</dc:creator>
<dc:date>2020-03-31</dc:date>
<dc:identifier>doi:10.1101/2020.03.31.007492</dc:identifier>
<dc:title><![CDATA[Specialisation and plasticity in a primitively social insect]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-03-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.05.07.082255v1?rss=1">
<title>
<![CDATA[
Intrinsic defects in lymph node stromal cells underpin poor germinal center responses during aging 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.05.07.082255v1?rss=1"
</link>
<description><![CDATA[
The failure to generate enduring humoral immunity after vaccination is a hallmark of advancing age. This can be attributed to a reduction in the germinal center response, which generates long-lived antibody-secreting cells that provide protection against (re)infection. Despite intensive investigation into the effect of age on the lymphoid compartment, the primary cellular defect that causes impaired germinal centers in aging has not been identified. Herein we demonstrate that aging reduces the capacity of germinal center-associated stromal cells to respond to vaccination. Heterochronic parabiosis and mathematical modeling demonstrate that a poor stromal cell response limits the size of the germinal center. This study reveals that age-associated defects in stromal cells are a significant barrier to efficacious vaccine responses in older individuals.
]]></description>
<dc:creator>Denton, A. E.</dc:creator>
<dc:creator>Silva-Cayetano, A.</dc:creator>
<dc:creator>Dooley, J.</dc:creator>
<dc:creator>Hill, D. L.</dc:creator>
<dc:creator>Carr, E. J.</dc:creator>
<dc:creator>Robert, P.</dc:creator>
<dc:creator>Meyer-Hermann, M.</dc:creator>
<dc:creator>Liston, A.</dc:creator>
<dc:creator>Linterman, M. A.</dc:creator>
<dc:date>2020-05-07</dc:date>
<dc:identifier>doi:10.1101/2020.05.07.082255</dc:identifier>
<dc:title><![CDATA[Intrinsic defects in lymph node stromal cells underpin poor germinal center responses during aging]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-05-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.05.14.096115v1?rss=1">
<title>
<![CDATA[
Non-canonical autophagy drives alternative ATG8 conjugation to phosphatidylserine 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.05.14.096115v1?rss=1"
</link>
<description><![CDATA[
Autophagy is a fundamental catabolic process essential for development, homeostasis and proper immune function 1. During autophagy, a cascade of ATG proteins target intracellular cargoes for lysosomal degradation and recycling 2. This pathway utilises a unique post-translational modification, the conjugation of ATG8 proteins to phosphatidylethanolamine (PE) at autophagosomes, which modulates cargo selection and maturation. ATG8 lipidation also occurs during non-canonical autophagy, a parallel pathway involving Single Membrane ATG8 Conjugation (SMAC) to endolysosomal compartments, which plays a key role in phagocytosis and other processes 3. It has been widely assumed that SMAC involves the same lipidation of ATG8 to PE, but this has yet to be formally tested. Here, we show that ATG8 undergoes alternative lipidation to phosphatidylserine (PS) during non-canonical autophagy/SMAC. Using mass spectrometry, we find that activation of SMAC, by pharmacological agents 4,5, or during non-canonical autophagy processes such as LC3-associated phagocytosis 6,7 and Influenza A virus infection 8, induces the covalent conjugation of ATG8 to PS, as well as PE. This alternative lipidation event is dependent on the ATG16L1 WD40 domain, and occurs at PS enriched endolysosomal membranes. Importantly, we find that the ATG8-PS and ATG8-PE adducts are differentially delipidated by isoforms of the ATG4 family, indicating significant molecular distinctions and mechanisms between these two species.

Together, these results provide an important new insight into autophagy signalling, revealing an alternative form of the hallmark ATG8-lipidation event, so widely used to define and assay autophagy. Furthermore, ATG8-PS lipidation provides a specific  molecular signature for non-canonical autophagy, uncovering a novel means of detecting and monitoring this emerging pathway.
]]></description>
<dc:creator>Durgan, J.</dc:creator>
<dc:creator>Lystad, A. H.</dc:creator>
<dc:creator>Sloan, K.</dc:creator>
<dc:creator>Carlsson, S. R.</dc:creator>
<dc:creator>Wilson, M. I.</dc:creator>
<dc:creator>Marcassa, E.</dc:creator>
<dc:creator>Ulferts, R.</dc:creator>
<dc:creator>Webster, J.</dc:creator>
<dc:creator>Lopez-Clavijo, A. F.</dc:creator>
<dc:creator>Wakelam, M. J.</dc:creator>
<dc:creator>Beale, R.</dc:creator>
<dc:creator>Simonsen, A.</dc:creator>
<dc:creator>Oxley, D.</dc:creator>
<dc:creator>Florey, O.</dc:creator>
<dc:date>2020-05-15</dc:date>
<dc:identifier>doi:10.1101/2020.05.14.096115</dc:identifier>
<dc:title><![CDATA[Non-canonical autophagy drives alternative ATG8 conjugation to phosphatidylserine]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-05-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.05.20.107219v1?rss=1">
<title>
<![CDATA[
Transcriptome and epigenome diversity and plasticity of muscle stem cells following transplantation 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.05.20.107219v1?rss=1"
</link>
<description><![CDATA[
Adult skeletal muscles are maintained during homeostasis and regenerated upon injury by muscle stem cells (MuSCs). A heterogeneity in self-renewal, differentiation and regeneration properties has been reported for MuSCs based on their anatomical location. Although MuSCs derived from extraocular muscles (EOM) have a higher regenerative capacity than those derived from limb muscles, the molecular determinants that govern these differences remain undefined. Here we show that EOM and limb MuSCs have distinct DNA methylation signatures associated with enhancers of location-specific genes, and that the EOM transcriptome is reprogrammed following transplantation into a limb muscle environment. Notably, EOM MuSCs expressed host-site specific positional Hox codes after engraftment and self-renewal within the host muscle. However, about 10% of EOM-specific genes showed engraftment-resistant expression, pointing to cell-intrinsic molecular determinants of the higher engraftment potential of EOM MuSCs. Our results underscore the molecular diversity of distinct MuSC populations and molecularly define their plasticity in response to microenvironmental cues. These findings provide insights into strategies designed to improve the functional capacity of MuSCs in the context of regenerative medicine.
]]></description>
<dc:creator>Evano, B.</dc:creator>
<dc:creator>Gill, D.</dc:creator>
<dc:creator>Hernando-Herraez, I.</dc:creator>
<dc:creator>Comai, G.</dc:creator>
<dc:creator>Stubbs, T.</dc:creator>
<dc:creator>Commere, P.-H.</dc:creator>
<dc:creator>Reik, W.</dc:creator>
<dc:creator>Tajbakhsh, S.</dc:creator>
<dc:date>2020-05-22</dc:date>
<dc:identifier>doi:10.1101/2020.05.20.107219</dc:identifier>
<dc:title><![CDATA[Transcriptome and epigenome diversity and plasticity of muscle stem cells following transplantation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-05-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.06.08.139642v1?rss=1">
<title>
<![CDATA[
Satellite repeat transcripts modulate heterochromatin condensates and safeguard chromosome stability in mouse embryonic stem cells 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.06.08.139642v1?rss=1"
</link>
<description><![CDATA[
Heterochromatin maintains genome integrity and function, and is organised into distinct nuclear domains. Some of these domains are proposed to form by phase separation through the accumulation of HP1. Mammalian heterochromatin contains noncoding major satellite repeats (MSR), which are highly transcribed in mouse embryonic stem cells (ESCs). Here, we report that MSR transcripts can drive the formation of HP1 droplets in vitro, and scaffold heterochromatin into dynamic condensates in ESCs, leading to the formation of large nuclear domains that are characteristic of pluripotent cells. Depleting MSR transcripts causes heterochromatin to transition into a more compact and static state. Unexpectedly, changing heterochromatins biophysical properties has severe consequences for ESCs, including chromosome instability and mitotic defects. These findings uncover an essential role for MSR transcripts in modulating the organisation and properties of heterochromatin to preserve genome stability. They also provide new insights into the processes that could regulate phase separation and the functional consequences of disrupting the properties of heterochromatin condensates.
]]></description>
<dc:creator>Novo, C. L.</dc:creator>
<dc:creator>Wong, E.</dc:creator>
<dc:creator>Hockings, C.</dc:creator>
<dc:creator>Poudel, C.</dc:creator>
<dc:creator>Sheekey, E.</dc:creator>
<dc:creator>Walker, S.</dc:creator>
<dc:creator>Kaminski Schierle, G. S.</dc:creator>
<dc:creator>Narlikar, G.</dc:creator>
<dc:creator>Rugg-Gunn, P.</dc:creator>
<dc:date>2020-06-08</dc:date>
<dc:identifier>doi:10.1101/2020.06.08.139642</dc:identifier>
<dc:title><![CDATA[Satellite repeat transcripts modulate heterochromatin condensates and safeguard chromosome stability in mouse embryonic stem cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-06-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.06.13.150037v1?rss=1">
<title>
<![CDATA[
Protein citrullination was introduced into animals by horizontal gene transfer from cyanobacteria 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.06.13.150037v1?rss=1"
</link>
<description><![CDATA[
Protein post-translational modifications (PTMs) add an enormous amount of sophistication to biological systems but their origins are largely unexplored. Citrullination, a key regulatory mechanism in human physiology and pathophysiology, is particularly enigmatic in an evolutionary context. The citrullinating enzymes peptidylarginine deiminases (PADIs) are ubiquitous across vertebrates but absent from yeast, worms and flies. Here, we map the surprising evolutionary trajectory of PADIs into the animal lineage. We present strong phylogenetic support for a clade encompassing animal and cyanobacterial PADIs that excludes fungal and other bacterial homologues. The animal and cyanobacterial PADIs share unique, functionally relevant synapomorphies that are absent from all other homologues. Molecular clock calculations and sequence divergence analyses using the fossil record estimate the last common ancestor of the cyanobacterial and animal PADIs to be approximately 1 billion years old, far younger than the 3.35-4.52 billion years known to separate bacterial and eukaryotic lineages. Under an assumption of vertical descent, PADI sequence change is anachronistically slow during this evolutionary time frame, even when compared to mitochondrial proteins, products of likely endosymbiont gene transfer and some of the most highly conserved proteins in life. The consilience of evidence indicates that PADIs were introduced from cyanobacteria into animals by horizontal gene transfer (HGT). The ancestral cyanobacterial protein is enzymatically active and can citrullinate eukaryotic proteins, suggesting that the PADI HGT event introduced a new catalytic capability into the regulatory repertoire of animals. This study reveals the unusual evolution of a pleiotropic protein modification with clear relevance in human physiology and disease.
]]></description>
<dc:creator>Cummings, T. F.</dc:creator>
<dc:creator>Gori, K.</dc:creator>
<dc:creator>Sanchez-Pulido, L.</dc:creator>
<dc:creator>Gavriilidis, G.</dc:creator>
<dc:creator>Moi, D.</dc:creator>
<dc:creator>Wilson, A. R.</dc:creator>
<dc:creator>Murchison, E. P.</dc:creator>
<dc:creator>Dessimoz, C.</dc:creator>
<dc:creator>Ponting, C.</dc:creator>
<dc:creator>Christophorou, M. A.</dc:creator>
<dc:date>2020-06-15</dc:date>
<dc:identifier>doi:10.1101/2020.06.13.150037</dc:identifier>
<dc:title><![CDATA[Protein citrullination was introduced into animals by horizontal gene transfer from cyanobacteria]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-06-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.07.23.217992v1?rss=1">
<title>
<![CDATA[
The global and promoter-centric 3D genome organization temporally resolved during a circadian cycle 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.07.23.217992v1?rss=1"
</link>
<description><![CDATA[
Circadian gene expression is essential for organisms to adjust cellular responses and anticipate daily changes in the environment. In addition to its physiological importance, the clock circuit represents an ideal, temporally resolved, system to study transcription regulation. Here, we analysed changes in spatial mouse liver chromatin conformation using genome-wide and promoter-capture Hi-C alongside daily oscillations in gene transcription in mouse liver. We found circadian topologically associated domains switched assignments to the transcriptionally active, open chromatin compartment and the inactive compartment at different hours of the day while their boundaries stably maintain their structure over time. Individual circadian gene promoters displayed maximal chromatin contacts at times of peak transcriptional output and the expression of circadian genes and contacted transcribed regulatory elements, or other circadian genes, was phase-coherent. Anchor sites of promoter chromatin loops were enriched in binding sites for liver nuclear receptors and transcription factors, some exclusively present in either rhythmic or stable contacts. The circadian 3D chromatin maps provided here identify the scales of chromatin conformation that parallel oscillatory gene expression and protein factors specifically associated with circadian or stable chromatin configurations.
]]></description>
<dc:creator>Furlan-Magaril, M.</dc:creator>
<dc:creator>Ando-Kuri, M.</dc:creator>
<dc:creator>Arzate-Mejia, R. G.</dc:creator>
<dc:creator>Morf, J.</dc:creator>
<dc:creator>Cairns, J.</dc:creator>
<dc:creator>Poot- Hernandez, C. A.</dc:creator>
<dc:creator>Andrews, S.</dc:creator>
<dc:creator>Varnai, C.</dc:creator>
<dc:creator>Virk, B.</dc:creator>
<dc:creator>Wingett, S. W.</dc:creator>
<dc:creator>Fraser, P.</dc:creator>
<dc:date>2020-07-24</dc:date>
<dc:identifier>doi:10.1101/2020.07.23.217992</dc:identifier>
<dc:title><![CDATA[The global and promoter-centric 3D genome organization temporally resolved during a circadian cycle]]></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/cgi/content/short/2020.06.29.177196v1?rss=1">
<title>
<![CDATA[
AutoSpill: a method for calculating spillover coefficients in high-parameter flow cytometry 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.06.29.177196v1?rss=1"
</link>
<description><![CDATA[
Compensating in classical flow cytometry or unmixing in spectral systems is an unavoidable challenge in the data analysis of fluorescence-based flow cytometry. In both cases, spillover coefficients are estimated for each fluorophore using single-color controls. This approach has remained essentially unchanged since its inception, and is increasingly limited in its ability to deal with high-parameter flow cytometry. Here, we present AutoSpill, a novel approach for calculating spillover coefficients or spectral signatures of fluorophores. The approach combines automated gating of cells, calculation of an initial spillover matrix based on robust linear regression, and iterative refinement to reduce error. Moreover, autofluorescence can be compensated out, by processing it as an endogenous dye in an unstained control. AutoSpill uses single-color controls and is compatible with common flow cytometry software, but it differs in two key aspects from current methods: (1) it is much less demanding in the preparation of controls, as it does not require the presence of well-defined positive and negative populations, and (2) it does not require manual tuning of the spillover matrix, as the algorithm iteratively computes the tuning, producing an optimal compensation matrix. Another algorithm, AutoSpread, complements this approach, providing a robust estimate of the Spillover Spreading Matrix (SSM), while avoiding the need for well-defined positive and negative populations. Together, AutoSpill and AutoSpread provide a superior solution to the problem of fluorophore spillover, allowing simpler and more robust workflows in high-parameter flow cytometry.
]]></description>
<dc:creator>Roca, C. P.</dc:creator>
<dc:creator>Burton, O. T.</dc:creator>
<dc:creator>Prezzemolo, T.</dc:creator>
<dc:creator>Whyte, C. E.</dc:creator>
<dc:creator>Halpert, R.</dc:creator>
<dc:creator>Kreft, Łukasz</dc:creator>
<dc:creator>Collier, J.</dc:creator>
<dc:creator>Botzki, A.</dc:creator>
<dc:creator>Spidlen, J.</dc:creator>
<dc:creator>Humblet-Baron, S.</dc:creator>
<dc:creator>Liston, A.</dc:creator>
<dc:date>2020-06-29</dc:date>
<dc:identifier>doi:10.1101/2020.06.29.177196</dc:identifier>
<dc:title><![CDATA[AutoSpill: a method for calculating spillover coefficients in high-parameter flow cytometry]]></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/cgi/content/short/2020.08.10.243931v1?rss=1">
<title>
<![CDATA[
Genome-wide analysis of DNA replication and DNA double strand breaks by TrAEL-seq 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.08.10.243931v1?rss=1"
</link>
<description><![CDATA[
Understanding the distribution of sites at which replication forks stall, and the ensuing fork processing events, requires genome-wide methods sensitive to both changes in replication fork structure and the formation of recombinogenic DNA ends. Here we describe Transferase-Activated End Ligation sequencing (TrAEL-seq), a method that captures single stranded DNA 3 ends genome-wide and with base pair resolution. TrAEL-seq labels DNA breaks, and profiles both stalled and processive replication forks in yeast and mammalian cells. Replication forks stalling at defined barriers and expressed genes are detected by TrAEL-seq with exceptional signal-to-noise, most likely through labelling of DNA 3 ends exposed during fork reversal. TrAEL-seq also labels unperturbed processive replication forks to yield maps of replication fork direction similar to those obtained by Okazaki fragment sequencing, however TrAEL-seq is performed on asynchronous populations of wild-type cells without incorporation of labels, cell sorting, or biochemical purification of replication intermediates, rendering TrAEL-seq simpler and more widely applicable than existing replication fork direction profiling methods. The specificity of TrAEL-seq for DNA 3 ends also allows accurate detection of double strand break sites after the initiation of DNA end resection, which we demonstrate by genome-wide mapping of meiotic double strand break hotspots in a dmc1{Delta} mutant. Overall, TrAEL-seq provides a flexible and robust methodology with high sensitivity and resolution for studying DNA replication and repair, which will be of significant use in determining mechanisms of genome instability.
]]></description>
<dc:creator>Kara, N.</dc:creator>
<dc:creator>Krueger, F.</dc:creator>
<dc:creator>Rugg-Gunn, P.</dc:creator>
<dc:creator>Houseley, J.</dc:creator>
<dc:date>2020-08-10</dc:date>
<dc:identifier>doi:10.1101/2020.08.10.243931</dc:identifier>
<dc:title><![CDATA[Genome-wide analysis of DNA replication and DNA double strand breaks by TrAEL-seq]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-08-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.08.16.250878v1?rss=1">
<title>
<![CDATA[
LipidFinder 2.0: advanced informatics pipeline for lipidomics discovery applications 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.08.16.250878v1?rss=1"
</link>
<description><![CDATA[
We present LipidFinder 2.0, incorporating four new modules that apply artefact filters, remove lipid and contaminant stacks, in-source fragments and salt clusters, and a new isotope deletion method which is significantly more sensitive than available open-access alternatives. We also incorporate a novel false discovery rate (FDR) method, utilizing a target-decoy strategy, which allows users to assess data quality. A renewed lipid profiling method is introduced which searches three different databases from LIPID MAPS and returns bulk lipid structures only, and a lipid category scatter plot with color blind friendly pallet. An API interface with XCMS Online is made available on LipidFinders online version. We show using real data that LipidFinder 2.0 provides a significant improvement over non-lipid metabolite filtering and lipid profiling, compared to available tools.

AvailabilityLipidFinder 2.0 is freely available at https://github.com/ODonnell-Lipidomics/LipidFinder and http://lipidmaps.org/resources/tools/lipidfinder.

Contactlipidfinder@cardiff.ac.uk

Supplementary informationSupplementary data are available at Bioinformatics online.
]]></description>
<dc:creator>Alvarez-Jarreta, J.</dc:creator>
<dc:creator>Rodrigues, P.</dc:creator>
<dc:creator>Fahy, E.</dc:creator>
<dc:creator>O'Connor, A.</dc:creator>
<dc:creator>Price, A.</dc:creator>
<dc:creator>Gaud, C.</dc:creator>
<dc:creator>Andrews, S.</dc:creator>
<dc:creator>Benton, P.</dc:creator>
<dc:creator>Siuzdak, G.</dc:creator>
<dc:creator>Hawksworth, J.</dc:creator>
<dc:creator>Valdivia, M.</dc:creator>
<dc:creator>Allen, S.</dc:creator>
<dc:creator>O'Donnell, V.</dc:creator>
<dc:date>2020-08-17</dc:date>
<dc:identifier>doi:10.1101/2020.08.16.250878</dc:identifier>
<dc:title><![CDATA[LipidFinder 2.0: advanced informatics pipeline for lipidomics discovery applications]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-08-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.09.24.298000v1?rss=1">
<title>
<![CDATA[
IL-7R signalling activates widespread VH and DH gene usage to drive antibody diversity in bone marrow B cells 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.09.24.298000v1?rss=1"
</link>
<description><![CDATA[
Generation of the primary antibody repertoire requires V(D)J recombination of hundreds of gene segments in the immunoglobulin heavy chain (Igh) locus. It has been proposed that interleukin-7 receptor (IL-7R) signalling is necessary for Igh recombination, but this has been challenging to partition from the receptors role in B cell survival and proliferation. By generating the first detailed description of the Igh repertoire of murine IL-7R-/- bone marrow B cells, we demonstrate that IL-7R signalling profoundly influences VH gene selection during VH-to-DJH recombination. We find skewing towards usage of 3 VH genes during de novo VH-to-DJH recombination that is more severe than the fetal liver (FL) B cell repertoire, and we now show a role for IL-7R signalling in DH-to-JH recombination. Transcriptome and accessibility analyses suggests reduced expression of B lineage-specific transcription factors (TFs) and their targets, and loss of DH and VH antisense transcription in IL-7R-/- B cells. These results refute models suggesting that IL-7R signalling is only required for survival and proliferation, and demonstrate a pivotal role in shaping the Igh repertoire by activating underpinning epigenetic mechanisms.
]]></description>
<dc:creator>Baizan Edge, A.</dc:creator>
<dc:creator>Stubbs, B. A.</dc:creator>
<dc:creator>Stubbington, M. J. T.</dc:creator>
<dc:creator>Bolland, D. J.</dc:creator>
<dc:creator>Tabbada, K.</dc:creator>
<dc:creator>Andrews, S.</dc:creator>
<dc:creator>Corcoran, A. E.</dc:creator>
<dc:date>2020-09-25</dc:date>
<dc:identifier>doi:10.1101/2020.09.24.298000</dc:identifier>
<dc:title><![CDATA[IL-7R signalling activates widespread VH and DH gene usage to drive antibody diversity in bone marrow B cells]]></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/cgi/content/short/2020.10.05.326082v1?rss=1">
<title>
<![CDATA[
Glyoxal fixation facilitates transcriptome analysis after antigen staining and cell sorting by flow cytometry 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.10.05.326082v1?rss=1"
</link>
<description><![CDATA[
A simple method for extraction of high quality RNA from cells that have been fixed, stained and sorted by flow cytometry would allow routine transcriptome analysis of highly purified cell populations and single cells. However, formaldehyde fixation impairs RNA extraction and inhibits RNA amplification. Here we show that good quality RNA can be readily extracted from stained and sorted mammalian cells if formaldehyde is replaced by glyoxal - a well-characterised fixative that is widely compatible with immunofluorescent staining methods. Although both formaldehyde and glyoxal efficiently form protein-protein crosslinks, glyoxal does not crosslink RNA to proteins nor form stable RNA adducts, ensuring that RNA remains accessible and amenable to enzymatic manipulation after glyoxal fixation. We find that RNA integrity is maintained through glyoxal fixation, permeabilisation with methanol or saponin, indirect immunofluorescent staining and flow sorting. RNA can then be extracted by standard methods and processed into RNA-seq libraries using commercial kits; mRNA abundances measured by poly(A)+ RNA-seq correlate well between freshly harvested cells and fixed, stained and sorted cells. We validate the applicability of this approach to flow cytometry by staining MCF-7 cells for the intracellular G2/M-specific antigen cyclin B1 (CCNB1), and show strong enrichment for G2/M-phase cells based on transcriptomic data. Switching to glyoxal fixation with RNA-compatible staining methods requires only minor adjustments of most existing staining and sorting protocols, and should facilitate routine transcriptomic analysis of sorted cells.
]]></description>
<dc:creator>Channathodiyil, P.</dc:creator>
<dc:creator>Houseley, J.</dc:creator>
<dc:date>2020-10-05</dc:date>
<dc:identifier>doi:10.1101/2020.10.05.326082</dc:identifier>
<dc:title><![CDATA[Glyoxal fixation facilitates transcriptome analysis after antigen staining and cell sorting by flow cytometry]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-10-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.10.27.357426v1?rss=1">
<title>
<![CDATA[
A booster dose enhances immunogenicity of the COVID-19 vaccine candidate ChAdOx1 nCoV-19 in aged mice 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.10.27.357426v1?rss=1"
</link>
<description><![CDATA[
The spread of SARS-CoV-2 has caused a global pandemic that has affected almost every aspect of human life. The development of an effective COVID-19 vaccine could limit the morbidity and mortality caused by infection, and may enable the relaxation of social distancing measures. Age is one of the most significant risk factors for poor health outcomes after SARS-CoV-2 infection, therefore it is desirable that any new vaccine candidates should elicit a robust immune response in older adults. Here, we test the immunogenicity of the adenoviral vectored vaccine ChAdOx1 nCoV-19 (AZD-1222) in aged mice. We find that a single dose of this vaccine induces cellular and humoral immunity in aged mice, but at a reduced magnitude than in younger adult mice. Furthermore, we report that a second dose enhances the immune response to this vaccine in aged mice, indicating that a primeboost strategy may be a rational approach to enhance immunogenicity in older persons.
]]></description>
<dc:creator>Silva-Cayetano, A.</dc:creator>
<dc:creator>Foster, W. S.</dc:creator>
<dc:creator>Innocentin, S.</dc:creator>
<dc:creator>Belij-Rammerstorfer, S.</dc:creator>
<dc:creator>Spencer, A. J.</dc:creator>
<dc:creator>Burton, O. T.</dc:creator>
<dc:creator>Fra-Bido, S.</dc:creator>
<dc:creator>Lee, J. L.</dc:creator>
<dc:creator>Thakur, N.</dc:creator>
<dc:creator>Conceicao, C.</dc:creator>
<dc:creator>Wright, D.</dc:creator>
<dc:creator>Barrett, J.</dc:creator>
<dc:creator>Evans-Bailey, N.</dc:creator>
<dc:creator>Noble, C.</dc:creator>
<dc:creator>Bailey, D.</dc:creator>
<dc:creator>Liston, A.</dc:creator>
<dc:creator>Gilbert, S. C.</dc:creator>
<dc:creator>Lambe, T.</dc:creator>
<dc:creator>Linterman, M. A.</dc:creator>
<dc:date>2020-10-27</dc:date>
<dc:identifier>doi:10.1101/2020.10.27.357426</dc:identifier>
<dc:title><![CDATA[A booster dose enhances immunogenicity of the COVID-19 vaccine candidate ChAdOx1 nCoV-19 in aged mice]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-10-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.11.20.391896v1?rss=1">
<title>
<![CDATA[
Highly multiplexed spatially resolved gene expression profiling of mouse organogenesis 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.11.20.391896v1?rss=1"
</link>
<description><![CDATA[
Transcriptional and epigenetic profiling of single-cells has advanced our knowledge of the molecular bases of gastrulation and early organogenesis. However, current approaches rely on dissociating cells from tissues, thereby losing the crucial spatial context that is necessary for understanding cell and tissue interactions during development. Here, we apply an image-based single-cell transcriptomics method, seqFISH, to simultaneously and precisely detect mRNA molecules for 387 selected target genes in 8-12 somite stage mouse embryo tissue sections. By integrating spatial context and highly multiplexed transcriptional measurements with two single-cell transcriptome atlases we accurately characterize cell types across the embryo and demonstrate how spatially-resolved expression of genes not profiled by seqFISH can be imputed. We use this high-resolution spatial map to characterize fundamental steps in the patterning of the midbrain-hindbrain boundary and the developing gut tube. Our spatial atlas uncovers axes of resolution that are not apparent from single-cell RNA sequencing data - for example, in the gut tube we observe early dorsal-ventral separation of esophageal and tracheal progenitor populations. In sum, by computationally integrating high-resolution spatially-resolved gene expression maps with single-cell genomics data, we provide a powerful new approach for studying how and when cell fate decisions are made during early mammalian development.
]]></description>
<dc:creator>Lohoff, T.</dc:creator>
<dc:creator>Ghazanfar, S.</dc:creator>
<dc:creator>Missarova, A.</dc:creator>
<dc:creator>Koulena, N.</dc:creator>
<dc:creator>Pierson, N.</dc:creator>
<dc:creator>Griffiths, J. A.</dc:creator>
<dc:creator>Bardot, E. S.</dc:creator>
<dc:creator>Eng, C.-H. L.</dc:creator>
<dc:creator>Tyser, R. C. V.</dc:creator>
<dc:creator>Argelaguet, R.</dc:creator>
<dc:creator>Guibentif, C.</dc:creator>
<dc:creator>Srinivas, S.</dc:creator>
<dc:creator>Briscoe, J.</dc:creator>
<dc:creator>Simons, B. D.</dc:creator>
<dc:creator>Hadjantonakis, A.-K.</dc:creator>
<dc:creator>Gottgens, B.</dc:creator>
<dc:creator>Reik, W.</dc:creator>
<dc:creator>Nichols, J.</dc:creator>
<dc:creator>Cai, L.</dc:creator>
<dc:creator>Marioni, J. C.</dc:creator>
<dc:date>2020-11-21</dc:date>
<dc:identifier>doi:10.1101/2020.11.20.391896</dc:identifier>
<dc:title><![CDATA[Highly multiplexed spatially resolved gene expression profiling of mouse organogenesis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-11-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.12.01.405902v1?rss=1">
<title>
<![CDATA[
BAP1/ASXL complex modulation regulates Epithelial-Mesenchymal Transition during trophoblast differentiation and invasion 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.12.01.405902v1?rss=1"
</link>
<description><![CDATA[
Normal function of the placenta depends on the earliest developmental stages when trophoblast cells differentiate and invade into the endometrium to establish the definitive maternal-fetal interface. Previously, we identified the ubiquitously expressed tumour suppressor BRCA1-associated protein 1 (BAP1) as a central factor of a novel molecular node controlling early mouse placentation. However, functional insights into how BAP1 regulates trophoblast biology are still missing. Using CRISPR/Cas9 knockout and overexpression technology, here we demonstrate that the downregulation of BAP1 protein is essential to trigger epithelial-mesenchymal transition (EMT) during trophoblast differentiation associated with a gain of invasiveness. This function, which is conserved in mouse and humans, is dependent on the binding of BAP1 binding to Additional sex comb-like (ASXL1/2/3) proteins to form the Polycomb repressive deubiquitinase (PR-DUB) complex. Our results reveal that the physiological modulation of BAP1 determines the invasive properties of trophoblast, delineating a new role of the BAP1 PR-DUB complex in regulating early placentation.
]]></description>
<dc:creator>Perez-Garcia, V.</dc:creator>
<dc:creator>Lopez-Jimenez, P.</dc:creator>
<dc:creator>Burton, G. J.</dc:creator>
<dc:creator>Moffett, A.</dc:creator>
<dc:creator>Turco, M. Y.</dc:creator>
<dc:creator>Hemberger, M.</dc:creator>
<dc:date>2020-12-02</dc:date>
<dc:identifier>doi:10.1101/2020.12.01.405902</dc:identifier>
<dc:title><![CDATA[BAP1/ASXL complex modulation regulates Epithelial-Mesenchymal Transition during trophoblast differentiation and invasion]]></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/cgi/content/short/2020.12.18.423431v1?rss=1">
<title>
<![CDATA[
Context-dependent effects of IL-2 rewire immunity into distinct cellular circuits 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.12.18.423431v1?rss=1"
</link>
<description><![CDATA[
Interleukin 2 (IL-2) is a key homeostatic cytokine, with potential therapeutic applications in both immunogenic and tolerogenic immune modulation. Clinical application has been hampered by pleiotropic functionality and wide-spread receptor expression, with unexpected adverse events during trials. To characterize the IL-2 homeostatic network, we developed a novel mouse strain allowing IL-2 production to be diverted. Rewiring of IL-2 production to diverse leukocyte sources allowed the identification of contextual influences over IL-2 impact. Network analysis identified a priority access for Tregs, and a competitive fitness cost induced among both Tregs and conventional CD4 T cells for IL-2 production. CD8 T cells and NK cells, by contrast, exhibited a preference for autocrine IL-2 production. IL-2 sourced from dendritic cells amplified the Treg circuit, while IL-2 produced by B cells induced two context-dependent circuits: dramatic expansion of CD8+ Tregs and ILC2 cells. The former was associated with an unexpected concentration of rare CD8+ Tregs in B cell zones, while the latter drove a downstream, IL-5-mediated, eosinophilic circuit. The source-specific effects demonstrate the contextual influence of IL-2 function and potentially explain unexpected adverse effects observed during clinical trials of exogenous IL-2. Targeted IL-2 production therefore has the potential to amplify or quench particular circuits in the IL-2 network, based on clinical desirability.

Graphical abstract O_FIG_DISPLAY_L [Figure 1] M_FIG_DISPLAY C_FIG_DISPLAY
]]></description>
<dc:creator>Whyte, C. E.</dc:creator>
<dc:creator>Singh, K.</dc:creator>
<dc:creator>Burton, O. T.</dc:creator>
<dc:creator>Aloulou, M.</dc:creator>
<dc:creator>Moudra, A.</dc:creator>
<dc:creator>Roca, C. P.</dc:creator>
<dc:creator>Naranjo, F. J.</dc:creator>
<dc:creator>Lombard-Vadnais, F.</dc:creator>
<dc:creator>Kouser, L.</dc:creator>
<dc:creator>Hochepied, T.</dc:creator>
<dc:creator>Halim, T. Y. F.</dc:creator>
<dc:creator>Schlenner, S.</dc:creator>
<dc:creator>Lesage, S.</dc:creator>
<dc:creator>Dooley, J.</dc:creator>
<dc:creator>Liston, A.</dc:creator>
<dc:date>2020-12-20</dc:date>
<dc:identifier>doi:10.1101/2020.12.18.423431</dc:identifier>
<dc:title><![CDATA[Context-dependent effects of IL-2 rewire immunity into distinct cellular circuits]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-12-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.01.15.426786v1?rss=1">
<title>
<![CDATA[
Multi-omic rejuvenation of human cells by maturation phase transient reprogramming 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.01.15.426786v1?rss=1"
</link>
<description><![CDATA[
Ageing is the gradual decline in organismal fitness that occurs over time leading to tissue dysfunction and disease. At the cellular level, ageing is associated with reduced function, altered gene expression and a perturbed epigenome. Somatic cell reprogramming, the process of converting somatic cells to induced pluripotent stem cells (iPSCs), can reverse these age-associated changes. However, during iPSC reprogramming somatic cell identity is lost, and can be difficult to reacquire as re-differentiated iPSCs often resemble foetal rather than mature adult cells. Recent work has demonstrated that the epigenome is already rejuvenated by the maturation phase of reprogramming, which suggests full iPSC reprogramming is not required to reverse ageing of somatic cells. Here we have developed the first "maturation phase transient reprogramming" (MPTR) method, where reprogramming factors are expressed until this rejuvenation point followed by withdrawal of their induction. Using dermal fibroblasts from middle age donors, we found that cells reacquire their fibroblast identity following MPTR, possibly as a result of persisting epigenetic memory at enhancers. Excitingly, our method substantially rejuvenated multiple cellular attributes including the transcriptome, which was rejuvenated by around 30 years as measured by a novel transcriptome clock. The epigenome, including H3K9me3 histone methylation levels and the DNA methylation ageing clock, was rejuvenated to a similar extent. The magnitude of rejuvenation instigated by MTPR is substantially greater than that achieved in previous transient reprogramming protocols. MPTR fibroblasts produced youthful levels of collagen proteins, suggesting functional rejuvenation. Overall, our work demonstrates that it is possible to separate rejuvenation from pluripotency reprogramming, which should facilitate the discovery of novel anti-ageing genes and therapies.

HighlightsO_LIWe developed a novel method by which human fibroblasts are reprogrammed until the maturation phase of iPSCs and are then returned to fibroblast identity
C_LIO_LIDNA methylation memory in fibroblast enhancers may allow recovery of cell identity when fibroblast gene expression programmes are already extinct
C_LIO_LIMolecular measures of ageing including transcriptome and DNA methylation clocks and H3K9me3 levels reveal robust and substantial rejuvenation
C_LIO_LIFunctional rejuvenation of fibroblasts by MPTR is suggested by reacquisition of youthful levels of collagen proteins
C_LI
]]></description>
<dc:creator>Gill, D.</dc:creator>
<dc:creator>Parry, A.</dc:creator>
<dc:creator>Santos, F.</dc:creator>
<dc:creator>Hernando-Herraez, I.</dc:creator>
<dc:creator>Stubbs, T. M.</dc:creator>
<dc:creator>Milagre, I.</dc:creator>
<dc:creator>Reik, W.</dc:creator>
<dc:date>2021-01-17</dc:date>
<dc:identifier>doi:10.1101/2021.01.15.426786</dc:identifier>
<dc:title><![CDATA[Multi-omic rejuvenation of human cells by maturation phase transient reprogramming]]></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/cgi/content/short/2021.01.21.426789v1?rss=1">
<title>
<![CDATA[
ORFLine: a bioinformatic pipeline to prioritise small open reading frames identifies candidate secreted small proteins from lymphocytes 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.01.21.426789v1?rss=1"
</link>
<description><![CDATA[
The annotation of small open reading frames (smORFs) of less than 100 codons (<300 nucleotides) is challenging due to the large number of such sequences in the genome. The recent development of next generation sequence and ribosome profiling enables identification of actively translated smORFs. In this study, we developed a computational pipeline, which we have named ORFLine, that stringently identifies smORFs and classifies them according to their position within transcripts. We identified a total of 5744 unique smORFs in datasets from mouse B and T lymphocytes and systematically characterized them using ORFLine. We further searched smORFs for the presence of a signal peptide, which predicted known secreted chemokines as well as novel micropeptides. Five novel micropeptides show evidence of secretion and are therefore candidate mediators of immunoregulatory functions.
]]></description>
<dc:creator>Hu, F.</dc:creator>
<dc:creator>Lu, J.</dc:creator>
<dc:creator>Munoz, M. D.</dc:creator>
<dc:creator>Saveliev, A.</dc:creator>
<dc:creator>Turner, M.</dc:creator>
<dc:date>2021-01-21</dc:date>
<dc:identifier>doi:10.1101/2021.01.21.426789</dc:identifier>
<dc:title><![CDATA[ORFLine: a bioinformatic pipeline to prioritise small open reading frames identifies candidate secreted small proteins from lymphocytes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-01-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.03.04.433911v1?rss=1">
<title>
<![CDATA[
Stimulation of adaptive gene amplification by origin firing under replication fork constraint 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.03.04.433911v1?rss=1"
</link>
<description><![CDATA[
Adaptive mutations can cause drug resistance in cancers and pathogens, and increase the tolerance of agricultural pests and diseases to chemical treatment. When and how adaptive mutations form is often hard to discern, but we have shown that adaptive copy number amplification of the copper resistance gene CUP1 occurs in response to environmental copper due to CUP1 transcriptional activation. Here we dissect the mechanism by which CUP1 transcription in budding yeast stimulates copy number variation (CNV). We show that transcriptionally stimulated CNV requires TREX-2 and Mediator, such that cells lacking TREX-2 or Mediator respond normally to copper but cannot acquire increased resistance. Mediator and TREX-2 cause replication stress by tethering transcribed loci to nuclear pores, a process known as gene gating, and transcription at the CUP1 locus causes a TREX-2-dependent accumulation of replication forks indicative of replication fork stalling. TREX-2-dependent CUP1 gene amplification occurs by a Rad52 and Rad51-mediated homologous recombination mechanism that is enhanced by histone H3K56 acetylation and repressed by Pol32, factors known to alter the frequency of template switching during break induced replication (BIR). CUP1 amplification is also critically dependent on late firing replication origins present in the CUP1 repeats, and mutations that remove or inactivate these origins strongly suppress the acquisition of copper resistance. We propose that replicative stress imposed by nuclear pore association causes replication bubbles from these origins to collapse soon after firing, leaving an epigenetic scar of H3K56 acetylation that promotes template switching during later break induced replication events. The capacity for inefficient replication origins to promote copy number variation renders certain genomic regions more fragile than others, and therefore more likely to undergo adaptive evolution through de novo gene amplification.
]]></description>
<dc:creator>Whale, A. J.</dc:creator>
<dc:creator>King, M.</dc:creator>
<dc:creator>Hull, R. M.</dc:creator>
<dc:creator>Krueger, F.</dc:creator>
<dc:creator>Houseley, J.</dc:creator>
<dc:date>2021-03-04</dc:date>
<dc:identifier>doi:10.1101/2021.03.04.433911</dc:identifier>
<dc:title><![CDATA[Stimulation of adaptive gene amplification by origin firing under replication fork constraint]]></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/cgi/content/short/2021.03.11.434836v1?rss=1">
<title>
<![CDATA[
Loss of SETD1B results in the redistribution of genomic H3K4me3 in the oocyte 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.03.11.434836v1?rss=1"
</link>
<description><![CDATA[
Histone 3 lysine 4 trimethylation (H3K4me3) is an epigenetic mark found at gene promoters and CpG islands. H3K4me3 is essential for mammalian development, yet mechanisms underlying its genomic targeting are poorly understood. H3K4me3 methyltransferases SETD1B and MLL2 are essential for oogenesis. We investigated changes in H3K4me3 in Setd1b conditional knockout (cKO) oocytes using ultra-low input ChIP-seq, with comparisons to DNA methylation and gene expression analyses. H3K4me3 was redistributed in Setd1b cKO oocytes showing losses at active gene promoters associated with downregulated gene expression. Remarkably, many regions also gained H3K4me3, in particular those that were DNA hypomethylated, transcriptionally inactive and CpG-rich, which are hallmarks of MLL2 targets. Consequently, loss of SETD1B disrupts the balance between MLL2 and de novo DNA methyltransferases in determining the epigenetic landscape during oogenesis. Our work reveals two distinct, complementary mechanisms of genomic targeting of H3K4me3 in oogenesis, with SETD1B linked to gene expression and MLL2 to CpG content.

Graphical AbstractIn oogenesis, SETD1B and CXXC1 target H3K4me3 to actively transcribed gene promoters, while MLL2 targets transcriptionally inactive regions based on underlying CpG composition (upper panel). When SETD1B is ablated, H3K4me3 is lost at a subset of active promoters, resulting in downregulation of transcription (lower panel). Loss of SETD1B alters the activity of MLL2, permitting MLL2 to deposit H3K4me3 at CpG-rich regions, many of which should otherwise be DNA methylated. Thus, it is evident that MLL2 and de novo DNMTs compete for genomic occupancy late in oogenesis, and loss of SETD1B disrupts the balance of these mechanisms.

O_FIG_DISPLAY_L [Figure 1] M_FIG_DISPLAY C_FIG_DISPLAY
]]></description>
<dc:creator>Hanna, C. W.</dc:creator>
<dc:creator>Huang, J.</dc:creator>
<dc:creator>Belton, C.</dc:creator>
<dc:creator>Reinhardt, S.</dc:creator>
<dc:creator>Dahl, A.</dc:creator>
<dc:creator>Andrews, S.</dc:creator>
<dc:creator>Stewart, A. F.</dc:creator>
<dc:creator>Kranz, A.</dc:creator>
<dc:creator>Kelsey, G.</dc:creator>
<dc:date>2021-03-11</dc:date>
<dc:identifier>doi:10.1101/2021.03.11.434836</dc:identifier>
<dc:title><![CDATA[Loss of SETD1B results in the redistribution of genomic H3K4me3 in the oocyte]]></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/cgi/content/short/2021.03.23.436572v1?rss=1">
<title>
<![CDATA[
DNA replication during acute MEK inhibition drives acquisition of resistance through amplification of the BRAF oncogene 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.03.23.436572v1?rss=1"
</link>
<description><![CDATA[
Mutations and gene amplifications that confer drug resistance emerge frequently during chemotherapy, but their mechanism and timing is poorly understood. Here, we investigate BRAFV600E amplification events that underlie resistance to the MEK inhibitor selumetinib (AZD6244/ARRY-142886) in COLO205 cells, a well-characterised model for reproducible emergence of drug resistance, and show that de novo amplification of BRAF is the primary path to resistance irrespective of pre-existing amplifications. Selumetinib causes long-term G1 arrest accompanied by reduced expression of DNA replication and repair genes, but cells stochastically re-enter the cell cycle during treatment despite continued repression of pERK1/2. Most DNA replication and repair genes are re-expressed as cells enter S and G2, however, mRNAs encoding a subset of factors important for error-free replication and chromosome segregation including TIPIN, PLK2 and PLK3 remain at low abundance. This suggests that DNA replication in drug is more error prone and provides an explanation for the DNA damage observed under long-term RAF-MEK-ERK1/2 pathway inhibition. To test the hypothesis that DNA replication in drug promotes de novo BRAF amplification, we exploited the combination of palbociclib and selumetinib. Combined treatment with selumetinib and a dose of palbociclib sufficient to reinforce G1 arrest in selumetinib-sensitive cells, but not to impair proliferation of resistant cells, delays the emergence of resistant colonies, meaning that escape from G1 arrest is critical in the formation of resistant clones. Our findings demonstrate that acquisition of MEK inhibitor resistance often occurs through de novo gene amplification and can be suppressed by impeding cell cycle entry in drug.
]]></description>
<dc:creator>Channathodiyil, P.</dc:creator>
<dc:creator>Segonds-Pichon, A.</dc:creator>
<dc:creator>Smith, P. D.</dc:creator>
<dc:creator>Cook, S. J.</dc:creator>
<dc:creator>Houseley, J.</dc:creator>
<dc:date>2021-03-23</dc:date>
<dc:identifier>doi:10.1101/2021.03.23.436572</dc:identifier>
<dc:title><![CDATA[DNA replication during acute MEK inhibition drives acquisition of resistance through amplification of the BRAF oncogene]]></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/cgi/content/short/2021.04.13.439513v1?rss=1">
<title>
<![CDATA[
Peptidylarginine deiminase IV (PADI4) is not essential for cell-autonomous HSC maintenance and normal haematopoiesis 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.04.13.439513v1?rss=1"
</link>
<description><![CDATA[
Peptidylarginine deiminases (PADIs, or PADs) are emerging as key regulators of human physiology and pathophysiology. The nuclear deiminase PADI4 regulates embryonic stem cell pluripotency, however its role in adult stem cells is unknown. PADI4 is expressed most highly in the bone marrow (BM), where it is found as part of a self-renewal-associated gene signature and shown to modulate the function of critical transcriptional regulators such as Tal1 and c-Myc, suggesting that it regulates haematopoietic development or regeneration. We investigated the functional significance of PADI4 in haematopoietic stem cell (HSC) biology and normal haematopoiesis. We employed two conditional mouse models of tissue-specific Padi4 ablation, where Padi4 was completely deleted either after the emergence of HSCs, or acutely in the BM of adult mice. We found that loss of PADI4 does not significantly affect HSC self-renewal or differentiation potential upon injury or serial transplantation, nor does it lead to exhaustion or premature ageing of HSCs. Thus, surprisingly, PADI4 is dispensable for cell-autonomous HSC maintenance, differentiation and haematopoietic regeneration. This work has important implications for the clinical use of PADI4 inhibitors as therapeutic agents in autoimmunity and cancer.

Key PointsO_LIPADI4 is dispensable for steady-state and post-transplantation haematopoiesis
C_LIO_LIHSCs do not require intrinsic PADI4 activity to respond to haematopoietic injury
C_LIO_LIPADI4 deficiency does not lead to premature HSC ageing or exhaustion
C_LI
]]></description>
<dc:creator>Young, C.</dc:creator>
<dc:creator>Russell, J. P.</dc:creator>
<dc:creator>Lawson, H.</dc:creator>
<dc:creator>Mapperley, C.</dc:creator>
<dc:creator>Kranc, K. R.</dc:creator>
<dc:creator>Christophorou, M. A.</dc:creator>
<dc:date>2021-04-14</dc:date>
<dc:identifier>doi:10.1101/2021.04.13.439513</dc:identifier>
<dc:title><![CDATA[Peptidylarginine deiminase IV (PADI4) is not essential for cell-autonomous HSC maintenance and normal haematopoiesis]]></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/cgi/content/short/2021.05.20.444917v1?rss=1">
<title>
<![CDATA[
V-ATPase is a universal regulator of LC3 associated phagocytosis and non-canonical autophagy 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.05.20.444917v1?rss=1"
</link>
<description><![CDATA[
Non-canonical autophagy is a key cellular pathway in immunity, cancer and neurodegeneration, characterised by Conjugation of ATG8 to endolysosomal Single-Membranes (CASM). CASM is activated by engulfment (endocytosis, phagocytosis), agonists (STING, TRPML1) and infection (influenza), dependent on the ATG16L1 WD40-domain, and specifically K490. However, the factor(s) associated with non-canonical ATG16L1 recruitment, and CASM induction, remain unknown. Here, we investigate a role for V-ATPase during non-canonical autophagy. We report that increased V0-V1 engagement is associated with, and sufficient for, CASM activation. Upon V0-V1 binding, V-ATPase directly recruits ATG16L1, via K490, during LC3-associated phagocytosis (LAP), STING- and drug-induced CASM, indicating a common mechanism. Furthermore, during LAP, key molecular players, including NADPH oxidase/ROS, converge on V-ATPase. Finally, we show that LAP is sensitive to Salmonella SopF, which disrupts the V-ATPase-ATG16L1 axis, and provide evidence that CASM contributes to the Salmonella host response. Together, these data identify V-ATPase as a universal regulator of CASM, and indicate that SopF evolved in part to evade non-canonical autophagy.
]]></description>
<dc:creator>Hooper, K. M.</dc:creator>
<dc:creator>Jacquin, E.</dc:creator>
<dc:creator>Li, T.</dc:creator>
<dc:creator>Goodwin, J. M.</dc:creator>
<dc:creator>Brumell, J. H.</dc:creator>
<dc:creator>Durgan, J.</dc:creator>
<dc:creator>Florey, O.</dc:creator>
<dc:date>2021-05-20</dc:date>
<dc:identifier>doi:10.1101/2021.05.20.444917</dc:identifier>
<dc:title><![CDATA[V-ATPase is a universal regulator of LC3 associated phagocytosis and non-canonical autophagy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-05-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.06.03.446738v1?rss=1">
<title>
<![CDATA[
The timing of differentiation and potency of CD8 effector function is set by RNA binding proteins 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.06.03.446738v1?rss=1"
</link>
<description><![CDATA[
CD8+ T cell differentiation into effector cells is initiated early after antigen encounter by signals from the T cell antigen receptor and costimulatory molecules. The molecular mechanisms that determine the timing and rate of differentiation however are not defined. Here we show that the RNA binding proteins (RBP) ZFP36 and ZFP36L1 limit the rate of differentiation of activated naive CD8+ T cells and the potency of the resulting cytotoxic lymphocytes. The RBP act in an early and short temporal window to enforce dependency on costimulation via CD28 for full T cell activation and effector differentiation by directly binding mRNA of NF-B, IRF8 and NOTCH1 transcription factors and cytokines, including IL2. Their absence in T cells, or the adoptive transfer of a small numbers of CD8+ T cells lacking the RBP, promotes resilience to influenza A virus infection without immunopathology. These findings highlight ZFP36 and ZFP36L1 as nodes for the integration of the early T cell activation signals determining the speed and quality of the CD8 response.
]]></description>
<dc:creator>Petkau, G.</dc:creator>
<dc:creator>Mitchell, T. J.</dc:creator>
<dc:creator>Chakraborty, K.</dc:creator>
<dc:creator>Bell, S. E.</dc:creator>
<dc:creator>DAngeli, V.</dc:creator>
<dc:creator>Matheson, L.</dc:creator>
<dc:creator>Turner, D. J.</dc:creator>
<dc:creator>Salerno, F.</dc:creator>
<dc:creator>Katsikis, P. D.</dc:creator>
<dc:creator>Turner, M.</dc:creator>
<dc:date>2021-06-03</dc:date>
<dc:identifier>doi:10.1101/2021.06.03.446738</dc:identifier>
<dc:title><![CDATA[The timing of differentiation and potency of CD8 effector function is set by RNA binding proteins]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-06-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.07.10.451887v1?rss=1">
<title>
<![CDATA[
TGFβ signalling is required to maintain pluripotency of human naïve pluripotent stem cells 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.07.10.451887v1?rss=1"
</link>
<description><![CDATA[
The signalling pathways that maintain primed human pluripotent stem cells (hPSCs) have been well characterised, revealing a critical role for TGF{beta}/Activin/Nodal signalling. In contrast, the signalling requirements of naive human pluripotency have not been fully established. Here, we demonstrate that TGF{beta} signalling is required to maintain naive hPSCs. The downstream effector proteins - SMAD2/3 - bind common sites in naive and primed hPSCs, including shared pluripotency genes. In naive hPSCs, SMAD2/3 additionally bind to active regulatory regions near to naive pluripotency genes. Inhibiting TGF{beta} signalling in naive hPSCs causes the downregulation of SMAD2/3-target genes and pluripotency exit. Single-cell analyses reveal that naive and primed hPSCs follow different transcriptional trajectories after inhibition of TGF{beta} signalling. Primed hPSCs differentiate into neuroectoderm cells, whereas naive hPSCs transition into trophectoderm. These results establish that there is a continuum for TGF{beta} pathway function in human pluripotency spanning a developmental window from naive to primed states.
]]></description>
<dc:creator>Osnato, A.</dc:creator>
<dc:creator>Brown, S.</dc:creator>
<dc:creator>Krueger, C.</dc:creator>
<dc:creator>Andrews, S.</dc:creator>
<dc:creator>Collier, A. J.</dc:creator>
<dc:creator>Nakanoh, S.</dc:creator>
<dc:creator>Quiroga Londono, M.</dc:creator>
<dc:creator>Wesley, B. T.</dc:creator>
<dc:creator>Muraro, D.</dc:creator>
<dc:creator>Brumm, S.</dc:creator>
<dc:creator>Niakan, K.</dc:creator>
<dc:creator>Vallier, L.</dc:creator>
<dc:creator>Ortmann, D.</dc:creator>
<dc:creator>Rugg-Gunn, P. J.</dc:creator>
<dc:date>2021-07-11</dc:date>
<dc:identifier>doi:10.1101/2021.07.10.451887</dc:identifier>
<dc:title><![CDATA[TGFβ signalling is required to maintain pluripotency of human naïve pluripotent stem cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-07-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.07.21.453193v1?rss=1">
<title>
<![CDATA[
The RNA m6A binding protein YTHDF2 promotes the B cell to plasma cell transition. 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.07.21.453193v1?rss=1"
</link>
<description><![CDATA[
To identify roles of RNA binding proteins (RBPs) in the differentiation of B cells to antibody-secreting plasma cells we performed a CRISPR/Cas9 knockout screen of 1213 mouse RBPs for their ability to affect proliferation and/or survival, and the emergence of differentiated CD138+ cells in vitro. We identified RBPs that promoted the appearance of CD138+ cells including CSDE1 and STRAP, as well as RBPs that inhibited CD138+ cell appearance such as EIF3 subunits EIF3K and EIF3L. Furthermore, we identified RBPs that share the property of recruiting the CCR4-NOT complex to their target transcripts have the potential to mediate opposing outcomes on B cell differentiation. One such RBP, the m6A binding protein YTHDF2 promotes the appearance of CD138+ cells in vitro. In chimeric mouse models YTHDF2-deficient B cells formed germinal centers in a cell-autonomous manner, however plasma cells failed to accumulate.
]]></description>
<dc:creator>Turner, D.</dc:creator>
<dc:creator>Saveliev, A.</dc:creator>
<dc:creator>Salerno, F.</dc:creator>
<dc:creator>Matheson, L. S.</dc:creator>
<dc:creator>Screen, M.</dc:creator>
<dc:creator>Lawson, H.</dc:creator>
<dc:creator>Wotherspoon, D.</dc:creator>
<dc:creator>Kranc, K.</dc:creator>
<dc:creator>Turner, M.</dc:creator>
<dc:date>2021-07-21</dc:date>
<dc:identifier>doi:10.1101/2021.07.21.453193</dc:identifier>
<dc:title><![CDATA[The RNA m6A binding protein YTHDF2 promotes the B cell to plasma cell transition.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-07-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.02.11.943357v1?rss=1">
<title>
<![CDATA[
Replicative aging is associated with loss of genetic heterogeneity from extrachromosomal circular DNA in Saccharomyces cerevisiae 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.02.11.943357v1?rss=1"
</link>
<description><![CDATA[
Circular DNA of chromosomal origin form from all parts of eukaryotic genomes. In yeast, circular rDNA accumulates as cells divide, contributing to replicative aging. However, little is known about how other chromosome-deri ved circles segregate and contribute to geneticvariation as cells age. We identified circular DNA across the genome of young S. cerevisiae populations and their aged descendants. Young cells had highly diverse circular DNA populations, but lost 94% of the different circular DNA after 20 divisions. Circles present in both young and old cells were characterized by replication origins and included circles from unique regions of the genome, rDNA circles and telomeric Y circles. The loss in genetic heterogeneity in aged cells was accompanied by massive accumulation of rDNA circles >95% of all circular DNA. We discovered circles had flexible inherence patterns. Glucose limited conditions selected for cells with glucose-transporter gene circles, [HXT6/7circle], and up to 50% of cells in a population carried them. [HXT6/7circle] cells were eventually substituted by cells carrying stable chromosomal HXT6 HXT6/7 HXT7 amplifications, suggesting circular DNA were intermediates in chromosomal amplifications. In conclusion, DNA circles can offer a flexible adaptive solution but cells lose genetic heterogeneity from circular DNA as they undergo replicative aging.
]]></description>
<dc:creator>Prada-Luengo, I.</dc:creator>
<dc:creator>Moller, H. D.</dc:creator>
<dc:creator>Henriksen, R. A.</dc:creator>
<dc:creator>Gao, Q.</dc:creator>
<dc:creator>Larsen, C. E.</dc:creator>
<dc:creator>Alizadeh, S.</dc:creator>
<dc:creator>Maretty, L.</dc:creator>
<dc:creator>Houseley, J.</dc:creator>
<dc:creator>Regenberg, B.</dc:creator>
<dc:date>2020-02-12</dc:date>
<dc:identifier>doi:10.1101/2020.02.11.943357</dc:identifier>
<dc:title><![CDATA[Replicative aging is associated with loss of genetic heterogeneity from extrachromosomal circular DNA in Saccharomyces cerevisiae]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-02-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.08.16.456479v1?rss=1">
<title>
<![CDATA[
Leptin Signalling in the Ovary of Diet-Induced Obese Mice Regulates Activation of Nod-Like Receptor Protein 3 Inflammasome 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.08.16.456479v1?rss=1"
</link>
<description><![CDATA[
1Obesity leads to ovarian dysfunction and the establishment of local leptin resistance. The aim of our study was to characterise levels of Nod-Like Receptor Protein 3 (NLRP3) inflammasome activation during obesity progression in the mouse ovaries and liver and test the putative role of leptin on its regulation. C57BL/6J mice were treated with equine chorionic gonadotropin (eCG) or human chorionic gonadotropin (hCG) for oestrous cycle synchronisation and ovaries collection. In diet-induced obesity (DIO) model, mice were fed chow diet (CD) or high fat diet (HFD) for 4 or 16 weeks (wk), whereas in hyperleptinemic model (LEPT), mice were injected with leptin for 16 days (16L) or saline (16C) and in the genetic obese leptin-deficient ob/ob (+/? and -/-) animals were fed CD for 4wk. Either ovaries and liver were collected, as well as cumulus cells (CCs) after superovulation from DIO and LEPT. In DIO protocol, protein expression of NLRP3 inflammasome components was increased in 4wk HFD, but decreased in 16wk HFD. Moreover LEPT and ob/ob models revealed NLRP3 and IL-1{beta} upregulation in 16L and downregulation in ob/ob. Transcriptome analysis of CC showed common genes between LEPT and 4wk HFD modulating NLRP3 inflammasome. Moreover analysis in the liver showed upregulation of NLRP3 protein only after 16wk HFD, but also the downregulation of NLRP3 protein in ob/ob-/-. We showed the link between leptin signalling and NLRP3 inflammasome activation in the ovary throughout obesity progression in mice, elucidating the molecular mechanisms underpinning ovarian failure in maternal obesity.
]]></description>
<dc:creator>Adamowski, M.</dc:creator>
<dc:creator>Wolodko, K.</dc:creator>
<dc:creator>Oliveira, J.</dc:creator>
<dc:creator>Castillo-Fernandez, J.</dc:creator>
<dc:creator>Murta, D.</dc:creator>
<dc:creator>Kelsey, G. D.</dc:creator>
<dc:creator>Galvao, A.</dc:creator>
<dc:date>2021-08-17</dc:date>
<dc:identifier>doi:10.1101/2021.08.16.456479</dc:identifier>
<dc:title><![CDATA[Leptin Signalling in the Ovary of Diet-Induced Obese Mice Regulates Activation of Nod-Like Receptor Protein 3 Inflammasome]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-08-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.08.27.457890v1?rss=1">
<title>
<![CDATA[
An improved iCLIP protocol 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.08.27.457890v1?rss=1"
</link>
<description><![CDATA[
Crosslinking and Immunoprecipitation (CLIP) is a powerful technique to obtain transcriptome-wide maps of in vivo protein-RNA interactions, which are important to understand the post-transcriptional mechanisms mediated by RNA binding proteins (RBPs). Many variant CLIP protocols have been developed to improve the efficiency and convenience of cDNA library preparation. Here we describe an improved individual nucleotide resolution CLIP protocol (iiCLIP), which can be completed within 4 days from UV crosslinking to libraries for sequencing. For benchmarking, we directly compared PTBP1 iiCLIP libraries with the iCLIP2 protocol produced under standardised conditions, and with public eCLIP and iCLIP PTBP1 data. We visualised enriched motifs surrounding the identified crosslink positions and RNA maps of these crosslinks around the alternative exons regulated by PTBP1. Notably, motif enrichment was higher in iiCLIP and iCLIP2 in comparison to public eCLIP and iCLIP, and we show how this impacts the specificity of RNA maps. In conclusion, iiCLIP is technically convenient and efficient, and enables production of highly specific datasets for identifying RBP binding sites.
]]></description>
<dc:creator>Lee, F. C. Y.</dc:creator>
<dc:creator>Chakrabarti, A. M.</dc:creator>
<dc:creator>Hänel, H.</dc:creator>
<dc:creator>Monzon-Casanova, E.</dc:creator>
<dc:creator>Hallegger, M.</dc:creator>
<dc:creator>Militti, C.</dc:creator>
<dc:creator>Capraro, F.</dc:creator>
<dc:creator>Sadee, C.</dc:creator>
<dc:creator>Toolan-Kerr, P.</dc:creator>
<dc:creator>Wilkins, O.</dc:creator>
<dc:creator>Turner, M.</dc:creator>
<dc:creator>König, J.</dc:creator>
<dc:creator>Sibley, C.</dc:creator>
<dc:creator>Ule, J.</dc:creator>
<dc:date>2021-08-27</dc:date>
<dc:identifier>doi:10.1101/2021.08.27.457890</dc:identifier>
<dc:title><![CDATA[An improved iCLIP protocol]]></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/cgi/content/short/2020.12.15.422926v1?rss=1">
<title>
<![CDATA[
Zfp36l1 and Zfp36l2 balances proliferation and differentiation in the developing retina 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.12.15.422926v1?rss=1"
</link>
<description><![CDATA[
Both transcriptional and post-transcriptional regulation of gene expression play significant roles in diverse biological processes, but little is known about how post-transcriptional regulation impacts retinal development. Here we report our study of the function of two members of the TTP (tristetraprolin) mRNA binding protein family, Zfp36l1 and Zfp36l2, in the developing retina. TTP proteins are highly conserved CCCH zinc finger proteins, which carry out their functions by promoting target mRNA decay and modulating translation. We found that Zfp36l1 and Zfp36l2 were expressed in retinal progenitor cells (RPCs) during development and Muller glial cells and photoreceptors in the mature retina. Our analysis of the mutant retinas showed that, whereas the single knockout retinas were largely normal, the double knockout (DKO) retina showed decreased RPC proliferation and increased differentiation of multiple retinal cell types. RNA-seq analysis confirmed the imbalance of proliferation and differentiation in the DKO retina. Gene ontology and in silico target gene analysis indicates that Zfp36l1 and Zfp36l2 exert their function by directly regulating multiple classes of proteins, including components of multiple signaling pathways such as the sonic hedgehog pathway and the Notch pathway, cell cycle regulators, and most interestingly transcription factors directly involved in retinal differentiation. These results reveal a new tier of gene regulation controlling retinal development.
]]></description>
<dc:creator>Mu, X.</dc:creator>
<dc:creator>Wu, F.</dc:creator>
<dc:creator>Kaczynski, T.</dc:creator>
<dc:creator>Matheson, L.</dc:creator>
<dc:creator>Liu, T.</dc:creator>
<dc:creator>Wang, J.</dc:creator>
<dc:creator>Turner, M.</dc:creator>
<dc:date>2020-12-15</dc:date>
<dc:identifier>doi:10.1101/2020.12.15.422926</dc:identifier>
<dc:title><![CDATA[Zfp36l1 and Zfp36l2 balances proliferation and differentiation in the developing retina]]></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/cgi/content/short/2021.02.22.432146v1?rss=1">
<title>
<![CDATA[
Human immunodeficiency reveals GIMAP5 as lymphocyte-specific regulator of senescence 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.02.22.432146v1?rss=1"
</link>
<description><![CDATA[
Elucidating the molecular basis of immunodeficiency diseases is a powerful approach to discovering new immunoregulatory pathways in humans. Here we report 10 affected individuals from 4 families with a new immunodeficiency disease comprising of severe progressive lymphopenia, autoimmunity, immunodeficiency, and liver disease due to recessive loss of function variants in "GTPase of immunity-associated proteins" protein 5 (GIMAP5). We show that the disease involves the progressive loss of naive T lymphocytes and a corresponding increase in antigen-experienced, but poorly functional and replicatively senescent T cells. In vivo treatment of Gimap5-deficient mice with rapamycin (an inhibitor of mTORC1) significantly restores the fraction of naive T lymphocytes. Furthermore, a GIMAP5-deficient human patient who was treated with rapamycin (sirolimus) showed a remarkable reduction in spleen/lymph node size. Together, these observations reveal that GIMAP5 plays a critical role in lymphocyte metabolism which is essential for senescence prevention and immune competence, suggesting that an inhibitor of mTORC1 could be a valuable clinical intervention in treating patients deficient for GIMAP5.
]]></description>
<dc:creator>Park, A. Y.</dc:creator>
<dc:creator>Leney-Greene, M.</dc:creator>
<dc:creator>Lynberg, M.</dc:creator>
<dc:creator>Xu, X.</dc:creator>
<dc:creator>Zheng, L.</dc:creator>
<dc:creator>Zhang, Y.</dc:creator>
<dc:creator>Matthews, H.</dc:creator>
<dc:creator>Chao, B.</dc:creator>
<dc:creator>Morawski, A.</dc:creator>
<dc:creator>Jiang, P.</dc:creator>
<dc:creator>Aluri, J.</dc:creator>
<dc:creator>Aydine, E. K.</dc:creator>
<dc:creator>Kiykim, A.</dc:creator>
<dc:creator>Pascall, J.</dc:creator>
<dc:creator>Barlan, I.</dc:creator>
<dc:creator>Sari, S.</dc:creator>
<dc:creator>Butcher, G.</dc:creator>
<dc:creator>Rao, V. K.</dc:creator>
<dc:creator>Lifton, R. P.</dc:creator>
<dc:creator>Baris, S.</dc:creator>
<dc:creator>Ozen, A.</dc:creator>
<dc:creator>Vilarinho, S.</dc:creator>
<dc:creator>Su, H.</dc:creator>
<dc:creator>Lenardo, M. J.</dc:creator>
<dc:date>2021-02-23</dc:date>
<dc:identifier>doi:10.1101/2021.02.22.432146</dc:identifier>
<dc:title><![CDATA[Human immunodeficiency reveals GIMAP5 as lymphocyte-specific regulator of senescence]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-02-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.09.24.461247v1?rss=1">
<title>
<![CDATA[
Solvent Precipitation SP3 (SP4) enhances recovery for proteomics sample preparation without magnetic beads 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.09.24.461247v1?rss=1"
</link>
<description><![CDATA[
Complete, reproducible extraction of protein material is essential for comprehensive and unbiased proteome analyses. A current gold standard is single-pot, solid-phase-enhanced sample preparation (SP3), in which organic solvent and magnetic beads are used to denature and capture proteins, with subsequently washes allowing contaminant removal. However, SP3 is dependent on effective protein immobilisation onto beads, risks losses during wash steps, and experiences a drop-off in protein recovery at higher protein inputs. Magnetic beads may also contaminate samples and instruments, and become costly for larger scale protein preparations. Here, we propose solvent precipitation SP3 (SP4) as an alternative to SP3, omitting magnetic beads and employing brief centrifugation--either with or without low-cost inert glass beads--as the means of aggregated protein capture. SP4 recovered equivalent or greater protein yields for 1-5000 {micro}g preparations and improved reproducibility (median protein R2 0.99 (SP4) vs. 0.97 (SP3)). Deep proteome profiling (n = 9,076) also demonstrated improved recovery by SP4 and a significant enrichment of membrane and low-solubility proteins vs. SP3. The effectiveness of SP4 was verified in three other labs, each confirming equivalent or improved proteome characterisation over SP3. This work suggests that protein precipitation is the primary mechanism of SP3, and reliance on magnetic beads presents protein losses, especially at higher concentrations and amongst hydrophobic proteins. SP4 represents an efficient and effective alternative to SP3, provides the option to omit beads entirely, and offers virtually unlimited scalability of input and volume--all whilst retaining the speed and universality of SP3.

BRIEFSolvent precipitation SP3 (SP4) captures aggregated protein for proteomics sample clean-up by omitting magnetic beads, instead employing brief centrifugation--with or without low-cost inert glass beads. SP4 offers improvements to protein yields, higher reproducibility, and greater recovery of membrane proteins, with verifications from three labs. Protein precipitation appears to be the primary mechanism of SP3, with reliance on magnetic beads presenting protein losses, especially at higher concentrations. SP4 presents an effective alternative to SP3 with improved scalability and equal speed and universality.

GRAPHICAL ABSTRACT

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]]></description>
<dc:creator>Johnston, H. E.</dc:creator>
<dc:creator>Yadav, K.</dc:creator>
<dc:creator>Kirkpatrick, J. M.</dc:creator>
<dc:creator>Biggs, G. S.</dc:creator>
<dc:creator>Oxley, D.</dc:creator>
<dc:creator>Kramer, H. B.</dc:creator>
<dc:creator>Samant, R. S.</dc:creator>
<dc:date>2021-09-24</dc:date>
<dc:identifier>doi:10.1101/2021.09.24.461247</dc:identifier>
<dc:title><![CDATA[Solvent Precipitation SP3 (SP4) enhances recovery for proteomics sample preparation without magnetic beads]]></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/cgi/content/short/2021.11.18.469000v1?rss=1">
<title>
<![CDATA[
The distinct effects of MEK and GSK3 inhibition upon the methylome and transcriptome of mouse embryonic stem cells 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.11.18.469000v1?rss=1"
</link>
<description><![CDATA[
Mouse embryonic stem cells (mESCs) were first cultured in vitro in serum-containing medium with leukaemia inhibitory factor, in which they exhibit heterogeneous expression of both pluripotency and some early differentiation markers. Over the last decade, however, it has become commonplace to grow mESCs with inhibitors of MEK and GSK3 signalling, which together elicit a more homogeneously  naive state of pluripotency. Whilst 2i/L-cultured mESCs have been shown to be globally hypomethylated, a comprehensive understanding of the distinct effects of these signalling inhibitors upon the DNA methylome is still lacking. Here we carried out whole genome bisulphite and RNA sequencing of mESCs grown with MEK or GSK3 inhibition alone, including different time points and concentrations of MEK inhibitor treatment. This demonstrated that MEK inhibition causes a dose-dependent impairment of maintenance methylation via loss of UHRF1 protein, as well as rapid impairment of de novo methylation. In contrast, GSK3 inhibition triggers impairment of de novo methylation alone, and consequent hypomethylation is enriched at enhancers with a 2i/L-specific chromatin signature and coincides with upregulation of nearby genes. Our study provides detailed insights into the epigenetic and transcriptional impacts of inhibiting MEK or GSK3 signalling in mouse pluripotent cells.

HighlightsO_LIMEK inhibition causes dose-dependent impairment of maintenance methylation via loss of UHRF1 protein, as well as impairment of de novo methylation.
C_LIO_LIGSK3 inhibition triggers impairment of de novo methylation alone, which results in hypomethylation of enhancers and non-CGI promoters.
C_LIO_LIEnhancers that are hypomethylated following GSK3 inhibition are enriched for 2i/L-specific pluripotency factor binding, TET2 and H3K4me1.
C_LIO_LIEnhancer hypomethylation coincides with increased expression of nearby and Capture Hi-C linked genes.
C_LI
]]></description>
<dc:creator>Spindel, J.</dc:creator>
<dc:creator>Krueger, C.</dc:creator>
<dc:creator>Krueger, F.</dc:creator>
<dc:creator>Papachristou, E. K.</dc:creator>
<dc:creator>Kishore, K.</dc:creator>
<dc:creator>D'Santos, C. S.</dc:creator>
<dc:creator>Reik, W.</dc:creator>
<dc:date>2021-11-18</dc:date>
<dc:identifier>doi:10.1101/2021.11.18.469000</dc:identifier>
<dc:title><![CDATA[The distinct effects of MEK and GSK3 inhibition upon the methylome and transcriptome of mouse embryonic stem cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-11-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.02.08.430127v1?rss=1">
<title>
<![CDATA[
Nucleosomal Asymmetry Shapes Histone Mark Binding and Promotes Poising at Bivalent Domains 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.02.08.430127v1?rss=1"
</link>
<description><![CDATA[
Promoters of developmental genes in embryonic stem cells (ESCs) are marked by histone H3 lysine 4 trimethylation (H3K4me3) and H3K27me3 in an asymmetric nucleosomal conformation, with each sister histone H3 carrying only one mark. These bivalent domains are thought to poise genes for timely activation upon differentiation. Here we show that asymmetric bivalent nucleosomes recruit repressive H3K27me3 binders but fail to enrich activating H3K4me3 binders, despite presence of H3K4me3, thereby promoting a poised state. Strikingly, the bivalent mark combination further attracts chromatin proteins that are not recruited by each mark individually, including the histone acetyltransferase complex KAT6B (MORF). Knockout of KAT6B blocks neuronal differentiation, demonstrating that bivalency-specific readers are critical for proper ESC differentiation. These findings reveal how histone mark bivalency directly promotes establishment of a poised state at developmental genes, while highlighting how nucleosomal asymmetry is critical for histone mark readout and function.
]]></description>
<dc:creator>Bryan, E.</dc:creator>
<dc:creator>Warburton, M.</dc:creator>
<dc:creator>Webb, K. M.</dc:creator>
<dc:creator>McLaughlin, K. A.</dc:creator>
<dc:creator>Spanos, C.</dc:creator>
<dc:creator>Ambrosi, C.</dc:creator>
<dc:creator>Major, V.</dc:creator>
<dc:creator>Baubec, T.</dc:creator>
<dc:creator>Rappsilber, J.</dc:creator>
<dc:creator>Voigt, P.</dc:creator>
<dc:date>2021-02-08</dc:date>
<dc:identifier>doi:10.1101/2021.02.08.430127</dc:identifier>
<dc:title><![CDATA[Nucleosomal Asymmetry Shapes Histone Mark Binding and Promotes Poising at Bivalent Domains]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-02-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.07.21.214338v1?rss=1">
<title>
<![CDATA[
The TAZ2 domain of CBP/p300 directs acetylation towards H3K27 within chromatin 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.07.21.214338v1?rss=1"
</link>
<description><![CDATA[
The closely related acetyltransferases CBP and p300 are key regulators of gene expression in metazoans. CBP/p300 acetylate several specific lysine residues within nucleosomes, including histone H3 lysine 27 (H3K27), a hallmark of active enhancers and promoters. However, it has remained largely unclear how specificity of CBP/p300 towards H3K27 is achieved. Here we show that the TAZ2 domain of CBP is required for efficient acetylation of H3K27, while curbing activity towards other lysine residues within nucleosomes. We find that TAZ2 is a sequence-independent DNA binding module, promoting interaction between CBP and nucleosomes, thereby enhancing enzymatic activity and regulating substrate specificity of CBP. TAZ2 is further required to stabilize CBP binding to chromatin in mouse embryonic stem cells, facilitating specificity towards H3K27 and modulating gene expression. These findings reveal a crucial role of TAZ2 in regulating H3K27ac, while highlighting the importance of correct site-specific acetylation for proper regulation of gene expression.
]]></description>
<dc:creator>Sheahan, T. W.</dc:creator>
<dc:creator>Major, V.</dc:creator>
<dc:creator>Webb, K. M.</dc:creator>
<dc:creator>Bryan, E.</dc:creator>
<dc:creator>Voigt, P.</dc:creator>
<dc:date>2020-07-21</dc:date>
<dc:identifier>doi:10.1101/2020.07.21.214338</dc:identifier>
<dc:title><![CDATA[The TAZ2 domain of CBP/p300 directs acetylation towards H3K27 within chromatin]]></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/cgi/content/short/2022.04.27.489754v1?rss=1">
<title>
<![CDATA[
Molecular mechanism of Afadin substrate recruitment to the receptor phosphatase PTPRK via its pseudophosphatase domain 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.04.27.489754v1?rss=1"
</link>
<description><![CDATA[
Protein tyrosine phosphatase receptor-type kappa (PTPRK) is a transmembrane receptor that links extracellular homophilic interactions to intracellular catalytic activity. Previously we showed that PTPRK promotes cell-cell adhesion by selectively dephosphorylating several cell junction regulators including the protein Afadin (Fearnley et al., 2019). Here we demonstrate that Afadin is recruited for dephosphorylation by directly binding to the PTPRK D2 pseudophosphatase domain. We mapped this interaction to a putative coiled coil (CC) domain in Afadin that is separated by more than 100 amino acids from the substrate pTyr residue. We identify the residues that define PTP specificity, explaining how Afadin is selectively dephosphorylated by PTPRK yet not by the closely-related receptor tyrosine phosphatase PTPRM. Our work demonstrates that PTP substrate specificity can be determined by protein-protein interactions distal to the active site. This explains how PTPRK and other PTPs achieve substrate specificity despite a lack of specific sequence context at the substrate pTyr. Furthermore, by demonstrating that these interactions are phosphorylation-independent and mediated via binding to a non-catalytic domain, we highlight how receptor PTPs could function as intracellular scaffolds in addition to catalyzing protein dephosphorylation.
]]></description>
<dc:creator>Hay, I. M.</dc:creator>
<dc:creator>Mulholland, K. E.</dc:creator>
<dc:creator>Graham, S. C.</dc:creator>
<dc:creator>Sharpe, H.</dc:creator>
<dc:creator>Deane, J. E.</dc:creator>
<dc:date>2022-04-27</dc:date>
<dc:identifier>doi:10.1101/2022.04.27.489754</dc:identifier>
<dc:title><![CDATA[Molecular mechanism of Afadin substrate recruitment to the receptor phosphatase PTPRK via its pseudophosphatase domain]]></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/cgi/content/short/2022.07.14.500009v1?rss=1">
<title>
<![CDATA[
Senescence in yeast is associated with chromosome XII cleavage rather than ribosomal DNA circle accumulation 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.07.14.500009v1?rss=1"
</link>
<description><![CDATA[
The massive accumulation of extrachromosomal ribosomal DNA circles (ERCs) in yeast mother cells has been long cited as the primary driver of replicative ageing. ERCs arise through ribosomal DNA (rDNA) recombination and a wealth of genetic data connects rDNA instability events giving rise to ERCs with shortened lifespan and other ageing pathologies. However, we understand little about the molecular effects of ERC accumulation. Here we studied ageing in the presence and absence of ERCs, and unexpectedly found no evidence of gene expression differences that might indicate stress responses or metabolic feedback caused by ERCs. Neither did we observe any global change in the widespread disruption of gene expression that accompanies yeast ageing, altogether suggesting that ERCs are largely inert. Much of the differential gene expression that accompanies ageing in yeast was actually associated with markers of the Senescence Entry Point (SEP), showing that senescence rather than age underlies these changes. Cells passed the SEP irrespective of ERCs, but we found the SEP to be associated with copy number amplification of a region of chromosome XII between the rDNA and the telomere (ChrXIIr), which arises in aged cells due to rDNA instability but through a different mechanism to ERCs. Therefore, although rDNA copy number increases dramatically with age due to ERC accumulation, our findings implicate ChrXIIr, rather than ERCs, as the primary driver of senescence during budding yeast ageing.
]]></description>
<dc:creator>Zylstra, A.</dc:creator>
<dc:creator>Horkai, D.</dc:creator>
<dc:creator>Hadj-Moussa, H.</dc:creator>
<dc:creator>Piguet, B.</dc:creator>
<dc:creator>Houseley, J.</dc:creator>
<dc:date>2022-07-14</dc:date>
<dc:identifier>doi:10.1101/2022.07.14.500009</dc:identifier>
<dc:title><![CDATA[Senescence in yeast is associated with chromosome XII cleavage rather than ribosomal DNA circle accumulation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-07-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.07.19.500645v1?rss=1">
<title>
<![CDATA[
Dietary change without caloric restriction maintains a youthful profile in ageing yeast 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.07.19.500645v1?rss=1"
</link>
<description><![CDATA[
Caloric restriction increases lifespan and improves ageing health, but it is unknown whether these outcomes can be separated or achieved through less severe interventions. Here we show that an unrestricted galactose diet in early life minimises change during replicative ageing in budding yeast, irrespective of diet later in life. Lifespan and average mother cell division rate are comparable between glucose and galactose diets, but markers of senescence and the progressive dysregulation of gene expression observed on glucose are minimal on galactose, showing these to be associated rather than intrinsic aspects of the replicative ageing process. Respiration on galactose is critical for minimising hallmarks of ageing, and forced respiration during ageing on glucose by over-expression of the mitochondrial biogenesis factor Hap4 also has the same effect though only in a fraction of cells. This fraction maintains Hap4 activity to advanced age with low senescence and a youthful gene expression profile, whereas other cells in the same population lose Hap4 activity, undergo dramatic dysregulation of gene expression and accumulate fragments of chromosome XII (ChrXIIr), which are tightly associated with senescence. Our findings support the existence of two separable ageing trajectories in yeast. We propose that a complete shift to the healthy ageing mode can be achieved in wild-type cells through dietary change in early life without restriction.
]]></description>
<dc:creator>Horkai, D.</dc:creator>
<dc:creator>Houseley, J.</dc:creator>
<dc:date>2022-07-20</dc:date>
<dc:identifier>doi:10.1101/2022.07.19.500645</dc:identifier>
<dc:title><![CDATA[Dietary change without caloric restriction maintains a youthful profile in ageing yeast]]></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/cgi/content/short/2022.12.21.521417v1?rss=1">
<title>
<![CDATA[
Back to the Future: Unleashing your cytometers spectral potential 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.12.21.521417v1?rss=1"
</link>
<description><![CDATA[
With the recent growth in spectral flow cytometry many laboratories are investing in new spectral flow cytometers in order to maximise the information gathered about every cell. This study hypothesised that traditional cytometers already within many laboratories may be used as spectral cytometers and have shown using a range of different cytometers that data acquired may be unmixed after acquisition.
]]></description>
<dc:creator>Walker, R. V.</dc:creator>
<dc:creator>Hall, C.</dc:creator>
<dc:creator>Ibrahim, H.</dc:creator>
<dc:creator>Thompson, S.</dc:creator>
<dc:creator>Hobson, P.</dc:creator>
<dc:creator>Crofts, J.-A.</dc:creator>
<dc:creator>Nobes, P.</dc:creator>
<dc:creator>Lim, S.</dc:creator>
<dc:creator>Burpee, T.</dc:creator>
<dc:date>2022-12-22</dc:date>
<dc:identifier>doi:10.1101/2022.12.21.521417</dc:identifier>
<dc:title><![CDATA[Back to the Future: Unleashing your cytometers spectral potential]]></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/cgi/content/short/2022.12.22.521580v1?rss=1">
<title>
<![CDATA[
An integrated proteome and transcriptome of B cell maturation defines poised activation states of transitional and mature B cells 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.12.22.521580v1?rss=1"
</link>
<description><![CDATA[
During B cell maturation, transitional and mature B cells acquire cell-intrinsic features that determine their ability to exit quiescence and mount effective immune responses. We used label-free mass spectrometry to quantify the proteome of B cell subsets from the mouse spleen and map the differential expression of environmental sensing, transcription- and translation initiation-factors that define cellular identity and function. By comparing the full-length transcriptome and proteome within the same sample, we identified mRNAs linked to B cell activation and antibody secretion that are expressed without detectable protein. These "poised" mRNAs might enable rapid protein production through increased translation or protein stability. In addition, through interrogation of our proteomic dataset, we found that the translational repressor PDCD4 restrains the response of marginal zone B cells to a T-independent antigen. Our molecular characterization of B cell maturation is a valuable resource to further explore the mechanisms underpinning the specialised functions of B cell subsets.
]]></description>
<dc:creator>Salerno, F.</dc:creator>
<dc:creator>Howden, A. J.</dc:creator>
<dc:creator>Matheson, L. S.</dc:creator>
<dc:creator>Gizlenci, O.</dc:creator>
<dc:creator>Screen, M.</dc:creator>
<dc:creator>Lingel, H.</dc:creator>
<dc:creator>Brunner-Weinzierl, M.</dc:creator>
<dc:creator>Turner, M.</dc:creator>
<dc:date>2022-12-22</dc:date>
<dc:identifier>doi:10.1101/2022.12.22.521580</dc:identifier>
<dc:title><![CDATA[An integrated proteome and transcriptome of B cell maturation defines poised activation states of transitional and mature B cells]]></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/cgi/content/short/2023.03.22.533754v1?rss=1">
<title>
<![CDATA[
Lysosome damage triggers direct ATG8 conjugation and ATG2 engagement via CASM 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.03.22.533754v1?rss=1"
</link>
<description><![CDATA[
Cells harness multiple pathways to maintain lysosome integrity, a central homeostatic process. Damaged lysosomes can be repaired, or targeted for degradation by lysophagy, a selective autophagy process involving ATG8/LC3. Here, we describe a parallel ATG8/LC3 response to lysosome damage, mechanistically distinct from lysophagy. Using a comprehensive series of biochemical, pharmacological and genetic approaches, we show that lysosome damage induces rapid Conjugation of ATG8s to Single Membranes (CASM). ATG8 proteins are recruited directly onto damaged membranes, independently of ATG13/WIPI2, and conjugated to PS, as well as PE, a molecular hallmark of CASM. Lysosome damage drives V-ATPase V0-V1 association, and direct recruitment of ATG16L1, dependent on K490 (WD40-domain), and sensitive to Salmonella SopF. Lysosome damage-induced CASM is associated with the formation of dynamic LC3A-positive tubules, and promotes robust LC3A engagement with ATG2, a lipid transfer protein central to lysosome repair. Together, our data identify direct ATG8 conjugation as a rapid response to lysosome damage, with important links to lipid transfer and dynamics.
]]></description>
<dc:creator>Cross, J.</dc:creator>
<dc:creator>Durgan, J.</dc:creator>
<dc:creator>McEwan, D. G.</dc:creator>
<dc:creator>Florey, O.</dc:creator>
<dc:date>2023-03-22</dc:date>
<dc:identifier>doi:10.1101/2023.03.22.533754</dc:identifier>
<dc:title><![CDATA[Lysosome damage triggers direct ATG8 conjugation and ATG2 engagement via CASM]]></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/cgi/content/short/2023.05.11.539978v1?rss=1">
<title>
<![CDATA[
Zfp36l1 establishes the high affinity CD8 T cell response by directly linking TCR affinity to cytokine sensing 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.05.11.539978v1?rss=1"
</link>
<description><![CDATA[
How individual T cells compete for and respond to IL2 at the molecular level, and, as a consequence, how this shapes population dynamics and the selection of high affinity clones is still poorly understood. Here we describe how the RNA binding protein ZFP36L1, acts as a sensor of TCR affinity to promote clonal expansion of high affinity CD8 T cells. As part of an incoherent feed forward loop ZFP36L1 has a non-redundant role in suppressing negative regulators of cytokine signalling and mediating a selection mechanism based on competition for IL2. We suggest that ZFP36L1 acts as a sensor of antigen affinity and establishes dominance of high affinity T cells by installing a hierarchical response to IL2.
]]></description>
<dc:creator>Petkau, G.</dc:creator>
<dc:creator>Mitchell, T.</dc:creator>
<dc:creator>Jones Evans, M.</dc:creator>
<dc:creator>Matheson, L. S.</dc:creator>
<dc:creator>Salerno, F.</dc:creator>
<dc:creator>Turner, M.</dc:creator>
<dc:date>2023-05-13</dc:date>
<dc:identifier>doi:10.1101/2023.05.11.539978</dc:identifier>
<dc:title><![CDATA[Zfp36l1 establishes the high affinity CD8 T cell response by directly linking TCR affinity to cytokine sensing]]></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/cgi/content/short/2023.04.24.538044v1?rss=1">
<title>
<![CDATA[
B cells from aged mice do not have intrinsic defects in affinity maturation 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.04.24.538044v1?rss=1"
</link>
<description><![CDATA[
Affinity maturation, the progressive increase in serum antibody affinity after vaccination, is an essential process that contributes to an effective humoral response against vaccines and infections. Germinal centres (GCs) are key for affinity maturation, as they are where B cells undergo somatic hypermutation of their immunoglobulin genes in the dark zone, before going through positive selection in the light zone via interactions with T follicular helper cells and follicular dendritic cells. In aged mice, affinity maturation has been shown to be impaired, but whether B cell-intrinsic factors contribute to this defect remains unclear. In this study, we show that B cells from aged B cell receptor transgenic mice are able to become GC B cells, which are capable of receiving positive selection signals to a similar extent as B cells from young adult mice. Consistent with this, ageing also does not impact the ability of B cells to undergo somatic hypermutation and acquire affinity-enhancing mutations. Together, this shows that there are no B cell-intrinsic defects in affinity maturation with age when the B cell receptor repertoire is constant.
]]></description>
<dc:creator>Lee, J. L.</dc:creator>
<dc:creator>Innocentin, S.</dc:creator>
<dc:creator>Silva-Cayetano, A.</dc:creator>
<dc:creator>Guillaume, S.</dc:creator>
<dc:creator>Linterman, M. A.</dc:creator>
<dc:date>2023-04-25</dc:date>
<dc:identifier>doi:10.1101/2023.04.24.538044</dc:identifier>
<dc:title><![CDATA[B cells from aged mice do not have intrinsic defects in affinity maturation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-04-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.05.18.541123v1?rss=1">
<title>
<![CDATA[
PLEKHS1 drives PI3Ks and remodels pathway homeostasis in PTEN-null prostate 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.05.18.541123v1?rss=1"
</link>
<description><![CDATA[
The PIP3/PI3K network is a central regulator of metabolism and is frequently activated in cancer, commonly by loss of the PIP3/PI(3,4)P2-phosphatase, PTEN. Despite huge investment, the drivers of the PI3K network in normal tissues and how they adapt to overactivation are unclear.

We find that in healthy mouse prostate PI3K activity is driven by RTK/IRS signalling and constrained by pathway-feedback. In the absence of PTEN, the network is dramatically remodelled. A poorly understood, YXXM and PIP3/PI(3,4)P2-binding PH domain-containing, adaptor, PLEKHS1, became the dominant activator and was required to sustain PIP3, AKT-phosphorylation and growth in PTEN-null prostate. This was because PLEKHS1 evaded pathway-feedback and experienced enhanced PI3K and SRC-family kinase-dependent phosphorylation of Y258XXM, eliciting PI3K activation. hPLEKHS1-mRNA and activating-Y419-phosphorylation of hSRC correlated with PI3K-pathway activity in human prostate cancers. We propose that in PTEN-null cells, receptor-independent, SRC-dependent tyrosine-phosphorylation of PLEKHS1 creates positive-feedback that escapes homeostasis, drives PIP3- signalling and supports tumour progression.
]]></description>
<dc:creator>Chessa, T. A. M.</dc:creator>
<dc:creator>Jung, P.</dc:creator>
<dc:creator>Suire, S.</dc:creator>
<dc:creator>Anwar, A.</dc:creator>
<dc:creator>Anderson, K. E.</dc:creator>
<dc:creator>Barneda, D.</dc:creator>
<dc:creator>Kielkowska, A.</dc:creator>
<dc:creator>Sadiq, B. A.</dc:creator>
<dc:creator>Oxley, D.</dc:creator>
<dc:creator>Felisbino, S.</dc:creator>
<dc:creator>Spensberger, D.</dc:creator>
<dc:creator>Segonds-Pichon, A.</dc:creator>
<dc:creator>Wilson, M.</dc:creator>
<dc:creator>Walker, S.</dc:creator>
<dc:creator>Okkenhaug, H.</dc:creator>
<dc:creator>Cosulich, S.</dc:creator>
<dc:creator>Hawkins, P. T.</dc:creator>
<dc:creator>Stephens, L. R.</dc:creator>
<dc:date>2023-05-18</dc:date>
<dc:identifier>doi:10.1101/2023.05.18.541123</dc:identifier>
<dc:title><![CDATA[PLEKHS1 drives PI3Ks and remodels pathway homeostasis in PTEN-null prostate]]></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/cgi/content/short/2023.02.13.528037v1?rss=1">
<title>
<![CDATA[
Ubiquitination is a novel post-translational modification of VMP1 in autophagy of human tumor cells 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.02.13.528037v1?rss=1"
</link>
<description><![CDATA[
Autophagy is a tightly regulated catabolic process involved in the degradation and re-cycling of proteins and organelles. Ubiquitination plays an important role in the regulation of autophagy. Vacuole Membrane Protein 1 (VMP1) is an essential autophagy protein. The expression of VMP1 in pancreatic cancer stem cells carrying the activated Kirsten rat sarcoma viral oncogene homolog (KRAS) triggers autophagy and enables therapy resistance. Using biochemical and cellular approaches, we identified ubiquitination as a post-translational modification of VMP1 from the initial steps in autophagosome biogenesis. VMP1 remains ubiquitinated as part of the autophagosome membrane throughout autophagic flux until autolysosome formation. However, VMP1 is not degraded by autophagy, nor by the ubiquitin-proteasomal system. Mass spectrometry and immunoprecipitation showed that the cell division cycle protein cdt2 (Cdt2), the substrate recognition subunit of the E3 ligase complex associated with cancer, cullin-RING ubiquitin ligase complex 4 (CRL4), is a novel interactor of VMP1 and is involved in VMP1 ubiquitination. VMP1 ubiquitination decreases under the CRL inhibitor MLN4924 and increases with Cdt2 overexpression. Moreover, VMP1 recruitment and autophagosome formation is significantly affected by CRL inhibition. Our results indicate that ubiquitination is a novel post-translational modification of VMP1 during autophagy in human tumor cells. VMP1 ubiquitination may be of clinical relevance in tumor cell therapy resistance.

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]]></description>
<dc:creator>Renna, F. J.</dc:creator>
<dc:creator>Steinberg, J. H. E.</dc:creator>
<dc:creator>Manifava, M.</dc:creator>
<dc:creator>Gonzalez, C. D.</dc:creator>
<dc:creator>Tadic, M. S.</dc:creator>
<dc:creator>Orquera, T.</dc:creator>
<dc:creator>Vecino, C. V.</dc:creator>
<dc:creator>Ropolo, A.</dc:creator>
<dc:creator>Rossi, M.</dc:creator>
<dc:creator>Ktistakis, N. T.</dc:creator>
<dc:creator>Vaccaro, M. I.</dc:creator>
<dc:date>2023-02-13</dc:date>
<dc:identifier>doi:10.1101/2023.02.13.528037</dc:identifier>
<dc:title><![CDATA[Ubiquitination is a novel post-translational modification of VMP1 in autophagy of human tumor cells]]></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/cgi/content/short/2023.02.03.527081v1?rss=1">
<title>
<![CDATA[
MICA: A multi-omics method to predict gene regulatory networks in early human embryos 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.02.03.527081v1?rss=1"
</link>
<description><![CDATA[
Recent advances in single-cell-omics have been transformative to characterise cell types in challenging to study biological contexts. It is technically difficult to infer transcription factor-gene regulatory interactions from these datasets, especially in contexts with limited single-cell sample number such as the early human embryo. Here we systematically assessed the application of four different linear or non-linear gene regulatory network (GRN) prediction strategies to single-cell simulated and human embryo transcriptome datasets. We also compared how gene expression normalisation methods impact on regulatory network predictions, finding that TPM (transcripts per million reads) outperformed alternative methods. We identified more reproducible GRN inferences using a non-linear method based on mutual information (MI) applied to single-cell transcriptome datasets refined with chromatin accessibility (CA) (called MICA), compared to alternative network prediction methods tested. MICA captures complex non-monotonic dependencies and feedback loops. Using MICA, we generated the first GRN inferences in early human development. MICA predicted co-localisation of the AP-1 transcription factor subunit proto-oncogene JUND and the TFAP2C transcription factor AP-2{psi} in human preimplantation embryos. These interactions were supported by previous findings in other developmental and stem cell contexts. Overall, our comparative analysis of gene regulatory network prediction methods defines a pipeline that can be applied to single-cell multi-omics datasets in especially challenging contexts to infer interactions between transcription factor expression and target gene regulation.
]]></description>
<dc:creator>Alanis-Lobato, G.</dc:creator>
<dc:creator>Bartlett, T. E.</dc:creator>
<dc:creator>Huang, Q.</dc:creator>
<dc:creator>Simon, C.</dc:creator>
<dc:creator>McCarthy, A.</dc:creator>
<dc:creator>Elder, K.</dc:creator>
<dc:creator>Snell, P.</dc:creator>
<dc:creator>Christie, L.</dc:creator>
<dc:creator>Niakan, K.</dc:creator>
<dc:date>2023-02-03</dc:date>
<dc:identifier>doi:10.1101/2023.02.03.527081</dc:identifier>
<dc:title><![CDATA[MICA: A multi-omics method to predict gene regulatory networks in early human embryos]]></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/cgi/content/short/2023.01.30.526343v1?rss=1">
<title>
<![CDATA[
Single cell DNA methylation ageing in mouse blood 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.01.30.526343v1?rss=1"
</link>
<description><![CDATA[
Ageing is the accumulation of changes and overall decline of the function of cells, organs and organisms over time. At the molecular and cellular level, the concept of biological age has been established and biomarkers of biological age have been identified, notably epigenetic DNA-methylation based clocks. With the emergence of single-cell DNA methylation profiling methods, the possibility to study biological age of individual cells has been proposed, and a first proof-of-concept study, based on limited single cell datasets mostly from early developmental origin, indicated the feasibility and relevance of this approach to better understand organismal changes and cellular ageing heterogeneity.

Here we generated a large single-cell DNA methylation and matched transcriptome dataset from mouse peripheral blood samples, spanning a broad range of ages (10-101 weeks of age). We observed that the number of genes expressed increased at older ages, but gene specific changes were small. We next developed a robust single cell DNA methylation age predictor (scEpiAge), which can accurately predict age in a broad range of publicly available datasets, including very sparse data and it also predicts age in single cells. Interestingly, the DNA methylation age distribution is wider than technically expected in 19% of single cells, suggesting that epigenetic age heterogeneity is present in vivo and may relate to functional differences between cells. In addition, we observe differences in epigenetic ageing between the major blood cell types. Our work provides a foundation for better single-cell and sparse data epigenetic age predictors and highlights the significance of cellular heterogeneity during ageing.

Highlights- Model to estimate DNA methylation age in single cells
- Large multi-omics dataset of single cells from murine blood
- Epigenetic age deviations from chronological age are greater than technical expected from technical variability
- Number of genes expressed increases with chronological and epigenetic age
]]></description>
<dc:creator>Bonder, M. J.</dc:creator>
<dc:creator>Clark, S.</dc:creator>
<dc:creator>Krueger, F.</dc:creator>
<dc:creator>Luo, S.</dc:creator>
<dc:creator>Agostinho de Sousa, J.</dc:creator>
<dc:creator>Hashtroud, A. M.</dc:creator>
<dc:creator>Stubbs, T. M.</dc:creator>
<dc:creator>Stark, A.-K.</dc:creator>
<dc:creator>Rulands, S.</dc:creator>
<dc:creator>Stegle, O.</dc:creator>
<dc:creator>Reik, W.</dc:creator>
<dc:creator>von Meyenn, F.</dc:creator>
<dc:date>2023-02-01</dc:date>
<dc:identifier>doi:10.1101/2023.01.30.526343</dc:identifier>
<dc:title><![CDATA[Single cell DNA methylation ageing in mouse blood]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-02-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.01.26.524308v1?rss=1">
<title>
<![CDATA[
The LCLAT1/LYCAT acyltransferase supports EGF-mediated phosphatidylinositol-3,4,5-trisphosphate and Akt signalling 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.01.26.524308v1?rss=1"
</link>
<description><![CDATA[
Receptor tyrosine kinases such as epidermal growth factor receptor (EGFR) stimulate phosphoinositide 3-kinases (PI3Ks) to convert phosphatidylinositol-4,5-bisphosophate [PtdIns(4,5)P2] into phosphatidylinositol-3,4,5-trisphosphate [PtdIns(3,4,5)P3]. PtdIns(3,4,5)P3 then remodels actin and gene expression, and boosts cell survival and proliferation. PtdIns(3,4,5)P3 partly achieves these functions by triggering activation of the kinase Akt, which phosphorylates targets like Tsc2 and GSK3{beta}. Consequently, unchecked upregulation of PtdIns(3,4,5)P3-Akt signalling promotes tumour progression. Interestingly, 50-70% of PtdIns and PtdInsPs have stearate and arachidonate at sn-1 and sn-2 positions of glycerol, respectively, forming a species known as 38:4-PtdIns/PtdInsPs. LCLAT1 and MBOAT7 acyltransferases partly enrich PtdIns in this acyl format. We previously showed that disruption of LCLAT1 lowered PtdIns(4,5)P2 levels and perturbed endocytosis and endocytic trafficking. However, the role of LCLAT1 in receptor tyrosine kinase and PtdIns(3,4,5)P3 signaling was not explored. Here, we show that LCLAT1 silencing in MDA-MB-231 and ARPE-19 cells abated the levels of PtdIns(3,4,5)P3 in response to EGF signalling. Importantly, LCLAT1-silenced cells were also impaired for EGF-driven and insulin-driven Akt activation and downstream signalling. Thus, our work provides first evidence that the LCLAT1 acyltransferase is required for receptor tyrosine kinase signalling.
]]></description>
<dc:creator>Chan, V.</dc:creator>
<dc:creator>Bone, L.</dc:creator>
<dc:creator>Anderson, K. E.</dc:creator>
<dc:creator>Zhang, K.</dc:creator>
<dc:creator>Orofiamma, L.</dc:creator>
<dc:creator>Awadeh, Y.</dc:creator>
<dc:creator>Lee, D. K. C.</dc:creator>
<dc:creator>Fu, N. J.</dc:creator>
<dc:creator>Chow, J. T. S.</dc:creator>
<dc:creator>Salmena, L.</dc:creator>
<dc:creator>Stephens, L. R.</dc:creator>
<dc:creator>Hawkins, P. T.</dc:creator>
<dc:creator>Antonescu, C. N.</dc:creator>
<dc:creator>Botelho, R. J.</dc:creator>
<dc:date>2023-01-27</dc:date>
<dc:identifier>doi:10.1101/2023.01.26.524308</dc:identifier>
<dc:title><![CDATA[The LCLAT1/LYCAT acyltransferase supports EGF-mediated phosphatidylinositol-3,4,5-trisphosphate and Akt signalling]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-01-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.01.11.523441v1?rss=1">
<title>
<![CDATA[
Dynamic DNA methylation turnover at the exit of pluripotency epigenetically primes gene regulatory elements for hematopoietic lineage specification 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.01.11.523441v1?rss=1"
</link>
<description><![CDATA[
Epigenetic mechanisms govern developmental cell fate decisions, but how DNA methylation coordinates with chromatin structure and three-dimensional DNA folding to enact cell-type specific gene expression programmes remains poorly understood. Here, we use mouse embryonic stem and epiblast-like cells deficient for 5-methyl cytosine or its oxidative derivatives (5-hydroxy-, 5-formyl- and 5-carboxy-cytosine) to dissect the gene regulatory mechanisms that control cell lineage specification at the exit of pluripotency. Genetic ablation of either DNA methyltransferase (Dnmt) or Ten-eleven-translocation (Tet) activity yielded largely distinct sets of dysregulated genes, revealing divergent transcriptional defects upon perturbation of individual branches of the DNA cytosine methylation cycle. Unexpectedly, we found that disrupting DNA methylation or oxidation interferes with key enhancer features, including chromatin accessibility, enhancer-characteristic histone modifications, and long-range chromatin interactions with putative target genes. In addition to affecting transcription of select genes in pluripotent stem cells, we observe impaired enhancer priming, including a loss of three-dimensional interactions, at regulatory elements associated with key lineage-specifying genes that are required later in development, as we demonstrate for the key hematopoietic genes Klf1 and Lyl1. Consistently, we observe impaired transcriptional activation of blood genes during embryoid body differentiation of knockout cells. Our findings identify a novel role for the dynamic turnover of DNA methylation at the exit of pluripotency to establish and maintain chromatin states that epigenetically prime enhancers for later activation during developmental cell diversification.

HighlightsO_LIWe perform a detailed epigenetic characterisation of the mouse embryonic stem cell (ESC) to epiblast-like cell (EpiLC) transition in wild type, Tet triple-knockout (TKO) and Dnmt TKO lines and develop a novel clustering approach to interrogate the data.
C_LIO_LITet TKO reduces H3K4me1 and H3K27ac levels across enhancer elements upon pluripotency exit whilst Dnmt TKO affects only H3K4me1 levels, suggesting a novel role for oxidative derivatives in H3K4me1 deposition.
C_LIO_LITet TKO and Dnmt TKO affect enhancer priming in EpiLCs which is associated with failure to upregulate hematopoietic genes upon differentiation.
C_LIO_LILong-range chromosomal interactions between primed enhancers and their target genes are weakened in both Dnmt and Tet TKO.
C_LI
]]></description>
<dc:creator>Parry, A. J.</dc:creator>
<dc:creator>Krueger, C.</dc:creator>
<dc:creator>Lohoff, T.</dc:creator>
<dc:creator>Wingett, S.</dc:creator>
<dc:creator>Schoenfelder, S.</dc:creator>
<dc:creator>Reik, W.</dc:creator>
<dc:date>2023-01-11</dc:date>
<dc:identifier>doi:10.1101/2023.01.11.523441</dc:identifier>
<dc:title><![CDATA[Dynamic DNA methylation turnover at the exit of pluripotency epigenetically primes gene regulatory elements for hematopoietic lineage specification]]></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/cgi/content/short/2022.12.28.522149v1?rss=1">
<title>
<![CDATA[
Dynamic protein deacetylation is a limited carbon source for acetyl-CoA-dependent metabolism 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.12.28.522149v1?rss=1"
</link>
<description><![CDATA[
The ability of cells to store and rapidly mobilize energy reserves in response to nutrient availability is essential for survival. Breakdown of carbon stores produces acetyl-coenzyme-A (acetyl-CoA), which fuels various metabolic pathways and is also the acyl donor for protein lysine acetylation. Notably, histone acetylation is sensitive to acetyl-CoA availability and nutrient replete conditions induce a substantial accumulation of acetylation on histones. Deacetylation releases acetate, which can be recycled to acetyl-CoA, suggesting that deacetylation could be mobilized as an acetyl-CoA source to feed downstream metabolic processes under nutrient depletion. While the notion of histones as a metabolic reservoir has been frequently proposed, experimental evidence has been lacking. Therefore, to test this concept directly, we developed an experimental system to trace deacetylation-derived acetate and its incorporation into acetyl-CoA, using 13C2-acetate in ATP citrate lyase-deficient fibroblasts (Acly-/- MEFs), which are primarily dependent on acetate for protein acetylation. We find that dynamic protein deacetylation in Acly-/- MEFs contributes carbons to acetyl-CoA and proximal downstream metabolites. However, there is no significant effect on acyl-CoA pool sizes, and even at maximal acetylation, deacetylation transiently supplies approximately 9% of cellular acetyl-CoA. Together, our data reveal that although protein acetylation is dynamic and sensitive to nutrient availability, its potential for maintaining cellular acetyl-CoA-dependent metabolic pathways is limited compared to cellular demand.
]]></description>
<dc:creator>Soaita, I.</dc:creator>
<dc:creator>Megill, E.</dc:creator>
<dc:creator>Kantner, D.</dc:creator>
<dc:creator>Chatoff, A.</dc:creator>
<dc:creator>Arany, Z.</dc:creator>
<dc:creator>Snyder, N. W.</dc:creator>
<dc:creator>Wellen, K. E.</dc:creator>
<dc:creator>Trefely, S. W.</dc:creator>
<dc:date>2022-12-29</dc:date>
<dc:identifier>doi:10.1101/2022.12.28.522149</dc:identifier>
<dc:title><![CDATA[Dynamic protein deacetylation is a limited carbon source for acetyl-CoA-dependent metabolism]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-12-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.11.03.467112v1?rss=1">
<title>
<![CDATA[
Time-dependent regulation of cytokine production by RNA binding proteins defines T cell effector function 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.11.03.467112v1?rss=1"
</link>
<description><![CDATA[
Potent T cell responses against infections and malignancies depend on the release of effector molecules, such as pro-inflammatory cytokines. Because effector molecules can be toxic, their production is tightly regulated through post-transcriptional events at 3 Untranslated Regions (3UTRs). RNA binding proteins (RBPs) were shown to be key regulators herein. With an RNA aptamer-based capture assay from human T cells, we identified >130 RBPs interacting with the IFNG, TNF and IL2 3UTRs in human T cells. T cell activation altered RBP-RNA interactions, revealing that RBP-target mRNA interactions rapidly respond to stimulation. Furthermore, we uncovered the intricate and time-dependent regulation of cytokine production by RBPs: whereas HuR supports early cytokine production, ZFP36L1, ATXN2L and ZC3HAV1 dampen and shorten the production duration, each at different time points. Strikingly, even though ZFP36L1 deletion did not phenotypically rescue T cell dysfunction in tumors, the increased production of cytokines and cytotoxic molecules resulted in superior anti-tumoral T cell responses in vivo. Our findings thus show that identifying RBP-RNA interactions reveals key modulators of T cell responses in health and disease.
]]></description>
<dc:creator>Popovic, B.</dc:creator>
<dc:creator>Guislain, A.</dc:creator>
<dc:creator>Engels, S.</dc:creator>
<dc:creator>Nicolet, B.</dc:creator>
<dc:creator>Jurgens, A. P.</dc:creator>
<dc:creator>Paravinja, N.</dc:creator>
<dc:creator>Freen-van Heeren, J. J.</dc:creator>
<dc:creator>van Alphen, F. P.</dc:creator>
<dc:creator>van den Biggelaar, M.</dc:creator>
<dc:creator>Salerno, F.</dc:creator>
<dc:creator>Wolkers, M.</dc:creator>
<dc:date>2021-11-03</dc:date>
<dc:identifier>doi:10.1101/2021.11.03.467112</dc:identifier>
<dc:title><![CDATA[Time-dependent regulation of cytokine production by RNA binding proteins defines T cell effector function]]></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/cgi/content/short/2022.06.15.496239v1?rss=1">
<title>
<![CDATA[
Decoding gene regulation in the mouse embryo using single-cell multi-omics 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.06.15.496239v1?rss=1"
</link>
<description><![CDATA[
Following gastrulation, the three primary germ layers develop into the major organs in a process known as organogenesis. Single-cell RNA sequencing has enabled the profiling of the gene expression dynamics of these cell fate decisions, yet a comprehensive map of the interplay between transcription factors and cis-regulatory elements is lacking, as are the underlying gene regulatory networks. Here we generate a multi-omics atlas of mouse early organogenesis by simultaneously profiling gene expression and chromatin accessibility from tens of thousands of single cells. We develop a computational method to leverage the multimodal readouts to predict transcription factor binding events in cis-regulatory elements, which we then use to infer gene regulatory networks that underpin lineage commitment events. Finally, we show that these models can be used to generate in silico predictions of the effect of transcription factor perturbations. We validate this experimentally by showing that Brachyury is essential for the differentiation of neuromesodermal progenitors to somitic mesoderm fate by priming cis-regulatory elements. The data set can be interactively explored at https://www.bioinformatics.babraham.ac.uk/shiny/shiny_multiome_organogenesis/
]]></description>
<dc:creator>Argelaguet, R.</dc:creator>
<dc:creator>Lohoff, T.</dc:creator>
<dc:creator>Li, J. G.</dc:creator>
<dc:creator>Nakhuda, A.</dc:creator>
<dc:creator>Drage, D.</dc:creator>
<dc:creator>Krueger, F.</dc:creator>
<dc:creator>Velten, L.</dc:creator>
<dc:creator>Clark, S. J.</dc:creator>
<dc:creator>Reik, W.</dc:creator>
<dc:date>2022-06-15</dc:date>
<dc:identifier>doi:10.1101/2022.06.15.496239</dc:identifier>
<dc:title><![CDATA[Decoding gene regulation in the mouse embryo using single-cell multi-omics]]></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/cgi/content/short/2022.10.19.512842v1?rss=1">
<title>
<![CDATA[
High-resolution promoter interaction analysis in Type 3 Innate Lymphoid Cells implicates Batten Disease gene CLN3 in Crohn's Disease aetiology 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.10.19.512842v1?rss=1"
</link>
<description><![CDATA[
Innate lymphoid cells (ILCs) are rare, tissue-resident innate lymphocytes that functionally mirror CD4+ T helper cell lineages but lack antigen receptors. Type 3 ILCs (ILC3s) are enriched in the gut, airways, and mucosal lymphoid tissues, where they regulate inflammation and promote barrier integrity. To define the regulatory architecture of primary human ILC3s, we map promoter-anchored chromosomal contacts using high-resolution, low-input Promoter Capture Hi-C (PCHi-C) in these cells alongside CD4+ T cells. By combining statistical detection with a PCHi-C-adapted Activity-by-Contact approach, we link promoters to distal regulatory elements, identifying hundreds of ILC3-specific contacts. We use these maps to connect genome-wide association study (GWAS) risk variants for Crohns disease to target genes using multiCOGS, a Bayesian framework that integrates PCHi-C with summary-statistic imputation and multivariate fine-mapping. This analysis highlights both known and unanticipated candidates, including CLN3, a causal gene for the neurodevelopmental Batten disease. Using a mouse ILC3-like cell line, we show that Cln3 is downregulated upon cytokine stimulation, and Cln3 overexpression alters stimulation-induced transcriptional programmes and cytokine secretion. Extending this approach, we generate a catalogue of ILC3-linked risk genes for five additional autoimmune conditions and show that they are enriched for regulators of the ILC3 inflammatory response identified in a CRISPR interference screen. Together, these findings illuminate long-range gene control in ILC3s and prioritise known and newly implicated autoimmune risk genes with potential roles in this clinically important cell type.
]]></description>
<dc:creator>Malysheva, V.</dc:creator>
<dc:creator>Ray-Jones, H.</dc:creator>
<dc:creator>Cazares, T. A.</dc:creator>
<dc:creator>Clay, O.</dc:creator>
<dc:creator>Ohayon, D.</dc:creator>
<dc:creator>Artemov, P.</dc:creator>
<dc:creator>Wayman, J. A.</dc:creator>
<dc:creator>Della Rosa, M.</dc:creator>
<dc:creator>Petitjean, C.</dc:creator>
<dc:creator>Ellaway, J. I. J.</dc:creator>
<dc:creator>Orchard, W.</dc:creator>
<dc:creator>Chen, X.</dc:creator>
<dc:creator>Parameswaran, S.</dc:creator>
<dc:creator>Nagano, T.</dc:creator>
<dc:creator>Fraser, P.</dc:creator>
<dc:creator>Schoenfelder, S.</dc:creator>
<dc:creator>Weirauch, M.</dc:creator>
<dc:creator>Kottyan, L. C.</dc:creator>
<dc:creator>Smith, D. F.</dc:creator>
<dc:creator>Powell, N.</dc:creator>
<dc:creator>Weimer, J.</dc:creator>
<dc:creator>Wallace, C.</dc:creator>
<dc:creator>Miraldi, E. R.</dc:creator>
<dc:creator>Waggoner, S.</dc:creator>
<dc:creator>Spivakov, M.</dc:creator>
<dc:date>2022-10-21</dc:date>
<dc:identifier>doi:10.1101/2022.10.19.512842</dc:identifier>
<dc:title><![CDATA[High-resolution promoter interaction analysis in Type 3 Innate Lymphoid Cells implicates Batten Disease gene CLN3 in Crohn's Disease aetiology]]></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/cgi/content/short/2022.07.04.498412v1?rss=1">
<title>
<![CDATA[
FOXA2 controls the antioxidant response in FH-deficient cells independent of NRF2 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.07.04.498412v1?rss=1"
</link>
<description><![CDATA[
Hereditary Leiomyomatosis and renal cell cancer (HLRCC) is a cancer syndrome caused by inactivating germline mutations in fumarate hydratase (FH) and subsequent accumulation of fumarate. Fumarate accumulation leads to the activation of an anti-oxidant response via nuclear translocation of the transcription factor NRF2. The activation of the anti-oxidant response is key for cellular survival in FH-deficient cells, yet the extent to which chromatin remodelling shapes the anti-oxidant response is currently unknown. Here, we explored the global effects of FH loss on the chromatin landscape to identify transcription factor networks involved in the highly remodelled chromatin landscape of FH-deficient cells. We identify FOXA2 as a key transcription factor which directly regulates anti-oxidant response genes and subsequent metabolic rewiring. Moreover, we also find that FOXA2 regulates anti-oxidant genes independent of the canonical anti-oxidant regulator NRF2. The identification of FOXA2 as an anti-oxidant regulator provides new insights into the molecular mechanisms behind cell responses to fumarate accumulation, and potentially provides new avenues for therapeutic intervention for HLRCC.
]]></description>
<dc:creator>Rogerson, C.</dc:creator>
<dc:creator>Sciacovelli, M.</dc:creator>
<dc:creator>Maddalena, L. A.</dc:creator>
<dc:creator>Valcarcel-Jimenez, L.</dc:creator>
<dc:creator>Schmidt, C.</dc:creator>
<dc:creator>Yang, M.</dc:creator>
<dc:creator>Ivanova, E.</dc:creator>
<dc:creator>Kent, J.</dc:creator>
<dc:creator>Cheeseman, D.</dc:creator>
<dc:creator>Carroll, J. S.</dc:creator>
<dc:creator>Kelsey, G.</dc:creator>
<dc:creator>Frezza, C.</dc:creator>
<dc:date>2022-07-04</dc:date>
<dc:identifier>doi:10.1101/2022.07.04.498412</dc:identifier>
<dc:title><![CDATA[FOXA2 controls the antioxidant response in FH-deficient cells independent of NRF2]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-07-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.09.27.509783v1?rss=1">
<title>
<![CDATA[
Inter-gastruloid heterogeneity revealed by single cell transcriptomics time course: implications for organoid based perturbation studies 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.09.27.509783v1?rss=1"
</link>
<description><![CDATA[
Recent advances in organoid and genome editing technologies are allowing for perturbation experiments at an unprecedented scale. However, before doing such experiments it is important to understand the gene expression profile in each of the organoids cells, as well as how much heterogeneity there is between individual organoids. Here we characterise an organoid model of mouse gastrulation called gastruloids using single cell RNA-sequencing of individual organoids at half-day intervals between day 3 and day 5 of differentiation (roughly corresponding to E6.5-E8.75 in vivo). Our study reveals multiple differentiation trajectories that have hitherto not been characterised in gastruloids. Intriguingly, we observe that individual gastruloids displayed a strong bias towards producing either mesodermal (largely somitic) or ectodermal (specifically neural) cell types. This bifurcation is already seen at the earliest sampled time point, and is characterised by increased activity of WNT-associated pathways in mesodermally-biased gastruloids as compared to neurally-biased gastruloids. Notably, at day 5, mesodermal gastruloids show an increase in the proportion of neural cells, while neural gastruloids do not produce notably more mesodermal cells. This is in line with previous studies on how the balance between these cell types is regulated. We demonstrate using in silico simulations that without proper understanding of the inter-organoid heterogeneity, perturbation experiments have either very high false positive or negative rates, depending on the statistical model used. Thus in future studies, modelling of inter-organoid heterogeneity will be crucial when designing organoid-based perturbation studies.

HighlightsO_LIA single cell RNA-sequencing time course of day 3 to day 5 mouse gastruloids reveals multiple mesodermal and neural differentiation trajectories hitherto uncharacterised in gastruloids
C_LIO_LISingle gastruloid, single cell RNA-sequencing of mouse gastruloids reveals that gastruloids are either mesodermally- or neurally-biased
C_LIO_LIThe two classes of gastruloid arise from differences in response strength to the WNT-agonist chiron
C_LIO_LIAt day 5, mesodermal gastruloids start making more neural cells, while neural gastruloids do not make more mesodermal cells, aligning with previously studied in vivo feedback loops
C_LIO_LIWe show using simulations that understanding interorganoid heterogeneity is a crucial consideration in the design and analysis of well-powered organoid-based perturbation studies
C_LI
]]></description>
<dc:creator>Rosen, L. U.</dc:creator>
<dc:creator>Stapel, L. C.</dc:creator>
<dc:creator>Argelaguet, R.</dc:creator>
<dc:creator>Barker, C. G.</dc:creator>
<dc:creator>Yang, A.</dc:creator>
<dc:creator>Reik, W.</dc:creator>
<dc:creator>Marioni, J. C.</dc:creator>
<dc:date>2022-09-28</dc:date>
<dc:identifier>doi:10.1101/2022.09.27.509783</dc:identifier>
<dc:title><![CDATA[Inter-gastruloid heterogeneity revealed by single cell transcriptomics time course: implications for organoid based perturbation studies]]></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/cgi/content/short/2021.12.20.473460v1?rss=1">
<title>
<![CDATA[
GLP is critical for oogenesis exhibiting a G9A-independent role in transcriptional repression 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.12.20.473460v1?rss=1"
</link>
<description><![CDATA[
GLP (EHMT1) is a multifunctional protein, best known for its role as an H3K9me1 and H3K9me2 methyltransferase through its reportedly obligatory dimerization with G9A (EHMT2). Here, we investigate the role of GLP in the oocyte in comparison to G9A using oocyte-specific conditional knockout mouse models (G9a cKO, Glp cKO, G9a-Glp cDKO). Loss of GLP in Glp cKO and G9a-Glp cDKO oocytes re-capitulated meiotic defects observed in the G9a cKO; however, there was a significant impairment in oocyte maturation and developmental competence in Glp cKO and G9a-Glp cDKO oocytes beyond that observed in the G9a cKO. Consequently, loss of GLP in oogenesis results upon fertilisation in mid-gestation embryonic lethality. To assess the molecular functions of GLP and G9A, we applied a multi-omics approach, supported by immunofluorescence, to identify changes in epigenomic, transcriptomic and proteomic signatures in cKO oocytes. H3K9me2 was equally depleted in all cKO oocytes, whereas H3K9me1 was decreased only upon loss of GLP. The transcriptome, DNA methylome and proteome were markedly more affected in G9a-Glp cDKO than G9a cKO oocytes, with transcriptional de-repression associated with increased protein abundance and gains in genic DNA methylation in G9a-Glp cDKO oocytes. Together, our findings suggest that GLP contributes to transcriptional repression in the oocyte, independent of G9A, and is critical for oogenesis and oocyte developmental competence.
]]></description>
<dc:creator>Demond, H.</dc:creator>
<dc:creator>Hanna, C. W.</dc:creator>
<dc:creator>Castillo-Fernandez, J.</dc:creator>
<dc:creator>Santos, F.</dc:creator>
<dc:creator>Papachristou, E. K.</dc:creator>
<dc:creator>Segonds-Pichon, A.</dc:creator>
<dc:creator>Kishore, K.</dc:creator>
<dc:creator>D'Santos, C. S.</dc:creator>
<dc:creator>Kelsey, G.</dc:creator>
<dc:date>2021-12-21</dc:date>
<dc:identifier>doi:10.1101/2021.12.20.473460</dc:identifier>
<dc:title><![CDATA[GLP is critical for oogenesis exhibiting a G9A-independent role in transcriptional repression]]></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/cgi/content/short/2022.06.23.497309v1?rss=1">
<title>
<![CDATA[
Structural determinants of homophilic adhesion by receptor tyrosine phosphatases 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.06.23.497309v1?rss=1"
</link>
<description><![CDATA[
The type IIB receptor protein tyrosine phosphatases (R2B RPTPs) are cell surface transmembrane proteins that engage in cell adhesion via their extracellular domains (ECDs) and cell signaling via their cytoplasmic phosphatase domains. The ECDs of R2B RPTPs form stable, homophilic, trans interactions between adjacent cell membranes. Previous work has demonstrated how one family member, PTPRM, forms homodimers; however, the determinants of homophilic specificity remain unknown. We have solved the X-ray crystal structure of the membrane-distal, N-terminal domains of PTPRK that form a head-to-tail dimer consistent with intermembrane adhesion. Comparison with the PTPRM structure demonstrates inter-domain conformational differences that may define homophilic specificity. Using small-angle X-ray scattering we determined the solution structures of the full-length ECDs of PTPRM and PTPRK, identifying that both are rigid, extended molecules that differ in their overall long-range conformation. Furthermore, we identify one residue, W351, within the interaction interface that differs between PTPRM and PTPRK and show that mutation to glycine, the equivalent residue in PTPRM, abolishes PTPRK dimer formation in vitro. This comparison of two members of the receptor tyrosine phosphatase family suggest that homophilic specificity is driven by a combination of shape complementarity and specific but limited sequence differences.

SIGNIFICANCE STATEMENTCell-cell contacts are dynamically regulated, in part, by the actions of tyrosine kinases and phosphatases. The R2B family of receptor tyrosine phosphatases combine an adhesive extracellular domain with intracellular catalytic domains that bind and dephosphorylate key cell adhesion and polarity proteins. Previous work demonstrated that the extracellular domains form head-to-tail homodimers but, as the interface was composed of residues conserved across the family, homophilic specificity determinants remained unclear. We have used a range of structural techniques including X-ray crystallography, small angle X-ray scattering and AlphaFold modelling to demonstrate that, despite their similarity, two members of the R2B family possess significant differences in their overall shape. Our results support that a combination of subtle shape and sequence variations may determine homophilic binding.
]]></description>
<dc:creator>Hay, I. M.</dc:creator>
<dc:creator>Shamin, M.</dc:creator>
<dc:creator>Caroe, E. R.</dc:creator>
<dc:creator>Mohammed, A. S. A.</dc:creator>
<dc:creator>Svergun, D. I.</dc:creator>
<dc:creator>Jeffries, C. M.</dc:creator>
<dc:creator>Graham, S. C.</dc:creator>
<dc:creator>Sharpe, H. J.</dc:creator>
<dc:creator>Deane, J. E.</dc:creator>
<dc:date>2022-06-23</dc:date>
<dc:identifier>doi:10.1101/2022.06.23.497309</dc:identifier>
<dc:title><![CDATA[Structural determinants of homophilic adhesion by receptor tyrosine phosphatases]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-06-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.08.18.504441v1?rss=1">
<title>
<![CDATA[
NANOGP1, a tandem duplicate of NANOG, exhibits partial functional conservation in human naive pluripotent stem cells 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.08.18.504441v1?rss=1"
</link>
<description><![CDATA[
Gene duplication events are important drivers of evolution by providing genetic material for new gene functions. They also create opportunities for diverse developmental strategies to emerge between species. To study the contribution of duplicated genes to human early development, we examined the evolution and function of NANOGP1, a tandem duplicate of the key transcription factor NANOG. We found that NANOGP1 and NANOG have overlapping but distinct expression profiles, with high NANOGP1 expression restricted to early epiblast cells and naive-state pluripotent stem cells. Sequence analysis and epitope-tagging of the endogenous locus revealed that NANOGP1 is protein-coding with an intact homeobox domain. NANOGP1 has been retained only in great apes, whereas Old World monkeys have disabled the gene in different ways including point mutations in the homeodomain. NANOGP1 is a strong inducer of naive pluripotency; however, unlike NANOG, it is not required to maintain the undifferentiated status of human naive pluripotent cells. By retaining expression, sequence and partial functional conservation with its ancestral copy, NANOGP1 exemplifies how gene duplication and subfunctionalisation can contribute to transcription factor activity in human pluripotency and development.

Summary statementEstablishing that NANOGP1 has retained partial functional conservation with its ancestral copy NANOG sheds light on the role of gene duplication and subfunctionalisation in human pluripotency and development.
]]></description>
<dc:creator>Maskalenka, K.</dc:creator>
<dc:creator>Alagoz, G.</dc:creator>
<dc:creator>Krueger, F.</dc:creator>
<dc:creator>Wright, J.</dc:creator>
<dc:creator>Rostovskaya, M.</dc:creator>
<dc:creator>Nakhuda, A.</dc:creator>
<dc:creator>Bendall, A.</dc:creator>
<dc:creator>Krueger, C.</dc:creator>
<dc:creator>Walker, S.</dc:creator>
<dc:creator>Scally, A.</dc:creator>
<dc:creator>Rugg-Gunn, P.</dc:creator>
<dc:date>2022-08-19</dc:date>
<dc:identifier>doi:10.1101/2022.08.18.504441</dc:identifier>
<dc:title><![CDATA[NANOGP1, a tandem duplicate of NANOG, exhibits partial functional conservation in human naive pluripotent stem cells]]></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/cgi/content/short/2022.05.23.492985v1?rss=1">
<title>
<![CDATA[
Native size exclusion chromatography-based mass spectrometry (SEC-MS) identifies novel components of the Heat Shock Protein 90-dependent proteome 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.05.23.492985v1?rss=1"
</link>
<description><![CDATA[
The molecular chaperone heat shock protein 90 (HSP90) works in concert with co-chaperones to stabilize its client proteins, which include multiple drivers of oncogenesis and malignant progression. Pharmacologic inhibitors of HSP90 have been observed to exert a wide range of effects on the proteome, including depletion of client proteins, induction of heat shock proteins, dissociation of co-chaperones from HSP90, disruption of client protein signaling networks, and recruitment of the protein ubiquitylation and degradation machinery--suggesting widespread remodeling of cellular protein complexes. However, proteomics studies to date have focused on inhibitor-induced changes in total protein levels, often overlooking protein complex alterations. Here, we use size-exclusion chromatography in combination with mass spectrometry (SEC-MS) to characterize the changes in native protein complexes following treatment with the HSP90 inhibitor tanespimycin (17-AAG) in the HT29 colon adenocarcinoma cell line. After confirming the signature cellular response to HSP90 inhibition (e.g., induction of heat shock proteins, decreased total levels of client proteins), we were surprised to find only modest perturbations to the global distribution of protein elution profiles in inhibitor-treated cells. Similarly, co-chaperones that co-eluted with HSP90 displayed no clear difference between control and treated conditions. However, two distinct analysis strategies identified multiple inhibitor-induced changes, including several known components of the HSP90 proteome, as well as numerous proteins and protein complexes with no previous links to HSP90. We present this dataset as a resource for the HSP90, proteostasis, and cancer communities (https://www.bioinformatics.babraham.ac.uk/shiny/HSP90/SEC-MS/), laying the groundwork for future mechanistic and therapeutic studies related to HSP90 pharmacology. Data are available via ProteomeXchange with identifier PXD033459.
]]></description>
<dc:creator>Samant, R. S.</dc:creator>
<dc:creator>Batista, S.</dc:creator>
<dc:creator>Larance, M.</dc:creator>
<dc:creator>Ozer, B.</dc:creator>
<dc:creator>Milton, C. I.</dc:creator>
<dc:creator>Bludau, I.</dc:creator>
<dc:creator>Biggins, L.</dc:creator>
<dc:creator>Andrews, S.</dc:creator>
<dc:creator>Hervieu, A.</dc:creator>
<dc:creator>Johnston, H. E.</dc:creator>
<dc:creator>Al-Lazikhani, B.</dc:creator>
<dc:creator>Lamond, A. I.</dc:creator>
<dc:creator>Clarke, P. A.</dc:creator>
<dc:creator>Workman, P.</dc:creator>
<dc:date>2022-05-23</dc:date>
<dc:identifier>doi:10.1101/2022.05.23.492985</dc:identifier>
<dc:title><![CDATA[Native size exclusion chromatography-based mass spectrometry (SEC-MS) identifies novel components of the Heat Shock Protein 90-dependent proteome]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-05-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.04.26.489502v1?rss=1">
<title>
<![CDATA[
Widespread allele-specific topological domains in the human genome are not confined to imprinted gene clusters. 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.04.26.489502v1?rss=1"
</link>
<description><![CDATA[
BackgroundThere is widespread interest in the three-dimensional chromatin conformation of the genome and its impact on gene expression. However, these studies frequently do not consider parent-of-origin differences, such as genomic imprinting, which result in monoallelic expression. In addition, genome-wide allele-specific chromatin conformation associations have not been extensively explored. There are few accessible bioinformatic workflows for investigating allelic conformation differences and these require pre-phased haplotypes which are not widely available.

ResultsWe assembled a bioinformatic pipeline, "HiCFlow", which performs haplotype assembly and visualisation of parental chromatin architecture. We benchmarked the pipeline using prototype haplotype phased Hi-C data from GM12878 cells at three disease associated imprinted gene clusters. Using RC-HiC (Region Capture Hi-C) and Hi-C data from further human cell lines (1-7HB2, IMR-90, and H1-hESCs) we were able to robustly identify the known stable allele-specific interactions at the H19/IGF2 locus. Other imprinted loci (DLK1 and SNRPN) were more variable and there was no "canonical imprinted 3D structure", but we could detect allele-specific differences in A/B compartmentalisation. Genome-wide, when TADs were unbiasedly ranked according to their allele-specific contact frequencies, a set of "allele-specific TADs" (ASTADs) could be defined. These occurred in genomic regions of high sequence variation. In addition to imprinted genes, ASTADs were also enriched for allele-specific expressed (ASE) genes. We found loci in ASTADs that have not previously been identified as ASE such as the bitter taste receptors (TAS2Rs).

ConclusionsThis study highlights the widespread differences in chromatin conformation between heterozygous loci and provides a new framework for understanding ASE.
]]></description>
<dc:creator>Richer, S.</dc:creator>
<dc:creator>Yuan, T.</dc:creator>
<dc:creator>Schoenfelder, S.</dc:creator>
<dc:creator>Hurst, L.</dc:creator>
<dc:creator>Murrell, A.</dc:creator>
<dc:creator>Pisignano, G.</dc:creator>
<dc:date>2022-04-27</dc:date>
<dc:identifier>doi:10.1101/2022.04.26.489502</dc:identifier>
<dc:title><![CDATA[Widespread allele-specific topological domains in the human genome are not confined to imprinted gene clusters.]]></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/cgi/content/short/2022.04.07.487453v1?rss=1">
<title>
<![CDATA[
Chemical conversion of human conventional Pluripotent Stem Cells to Trophoblast Stem Cells 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.04.07.487453v1?rss=1"
</link>
<description><![CDATA[
In human embryos, naive pluripotent cells of the inner cell mass generate epiblast, primitive endoderm and Trophectoderm (TE) lineage, whence trophoblast cells derive. In vitro, naive pluripotent stem cells (PSCs) retain this potential and can generate trophoblast stem cells (TSCs), while conventional PSCs form amnion-like cells and lack the competence to generate TSCs. Transient histone deacetylase and MEK inhibitions with LIF stimulation can be used to chemically reset conventional to naive PSCs. Here we report that chemical resetting induced expression of both naive and TSC markers and of placental imprinted genes. A modified chemical resetting protocol allowed for the fast and efficient conversion of conventional PSCs into TSCs, entailing shutdown of pluripotency genes and full activation of the trophoblast master regulators, without induction of amnion markers. Chemical resetting generates a responsive intermediate state, in which conventional PSCs rapidly acquire competence to form TSCs without the need of stabilisation and expansion in a naive state. The efficiency and rapidity of our system will be useful for the study of cell fate transitions, and to generate models of placental disorders.
]]></description>
<dc:creator>Zorzan, I.</dc:creator>
<dc:creator>Betto, R. M.</dc:creator>
<dc:creator>Rossignoli, G.</dc:creator>
<dc:creator>Arboit, M.</dc:creator>
<dc:creator>Drusin, A.</dc:creator>
<dc:creator>Martini, P.</dc:creator>
<dc:creator>Martello, G.</dc:creator>
<dc:date>2022-04-08</dc:date>
<dc:identifier>doi:10.1101/2022.04.07.487453</dc:identifier>
<dc:title><![CDATA[Chemical conversion of human conventional Pluripotent Stem Cells to Trophoblast Stem Cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-04-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.03.11.483903v1?rss=1">
<title>
<![CDATA[
The AppNL-G-F mouse model of Alzheimer's disease is refractory to regulatory T cell treatment 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.03.11.483903v1?rss=1"
</link>
<description><![CDATA[
BackgroundAlzheimers Disease is a neurodegenerative disease with a neuroinflammatory component. Due to the multifunctional capacity of regulatory T cells to prevent and reverse inflammation, regulatory T cells have been proposed as a potential therapeutic in Alzheimers Disease, either as a direct cell therapy or through the use of IL2 as a biologic to expand the endogenous population.

MethodsHere we characterize the longitudinal immunological changes occurring in T cells in the AppNL-G-F mouse model of Alzheimers disease.

ResultsAge-dependent immunological changes, in both the brain and periphery, occurred in parallel in both AppNL-G-F mice and control AppNL mice. As the endogenous IL2 axis was disturbed with age, we sought to determine the effect of IL2 supplementation on disease progression. Using a genetic model of IL2 provision in the periphery or in the brain, we found that expanding regulatory T cells in either location was unable to alter the progression of key pathological events or behavioral changes.

ConclusionThese results suggest that either the AppNL-G-F mouse model does not recapitulate key regulatory T cell-dependent process of Alzheimers disease, or that regulatory T cell therapy is not a promising candidate for APP-mutation-driven Alzheimers disease.
]]></description>
<dc:creator>Yshii, L.</dc:creator>
<dc:creator>Mascali, L.</dc:creator>
<dc:creator>Kouser, L.</dc:creator>
<dc:creator>Lemaitre, P.</dc:creator>
<dc:creator>Marino, M.</dc:creator>
<dc:creator>Dooley, J.</dc:creator>
<dc:creator>Burton, O.</dc:creator>
<dc:creator>Haughton, J.</dc:creator>
<dc:creator>Callaerts-Vegh, Z.</dc:creator>
<dc:creator>de Strooper, B.</dc:creator>
<dc:creator>Holt, M.</dc:creator>
<dc:creator>Pasciuto, E.</dc:creator>
<dc:creator>Liston, A.</dc:creator>
<dc:date>2022-03-14</dc:date>
<dc:identifier>doi:10.1101/2022.03.11.483903</dc:identifier>
<dc:title><![CDATA[The AppNL-G-F mouse model of Alzheimer's disease is refractory to regulatory T cell treatment]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-03-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.03.03.482829v1?rss=1">
<title>
<![CDATA[
Polypyrimidine Tract Binding Protein 1 regulates the activation of mouse CD8 T cells 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.03.03.482829v1?rss=1"
</link>
<description><![CDATA[
We show that the RNA-binding protein Polypyrimidine Tract Binding Protein 1 (PTBP1) is dispensable for the development of naive mouse CD8 T cells, but is necessary for the optimal expansion and production of effector molecules by antigen-specific CD8 T cells in vivo. PTBP1 has an essential role in regulating the early events following activation of the naive CD8 T cell leading to IL-2 and TNF production. It is also required to protect activated CD8 T cells from apoptosis. PTBP1 controls alternative splicing of over 400 genes in naive CD8 T cells in addition to regulating the abundance of [~]200 mRNAs. PTBP1 is required for the nuclear accumulation of c-Fos, NFATc2 and NFATc3, but not NFATc1. This selective effect on NFAT proteins correlates with PTBP1-promoted expression of the shorter A{beta}1 isoform and exon 13 skipped A{beta}2 isoform of the catalytic A-subunit of calcineurin phosphatase. These findings reveal a crucial role for PTBP1 in regulating CD8 T cell activation.
]]></description>
<dc:creator>D'Angeli, V.</dc:creator>
<dc:creator>Monzon-Casanova, E.</dc:creator>
<dc:creator>Matheson, L. S.</dc:creator>
<dc:creator>Gizlenci, O.</dc:creator>
<dc:creator>Petkau, G.</dc:creator>
<dc:creator>Gooding, C.</dc:creator>
<dc:creator>Berrens, R. V.</dc:creator>
<dc:creator>Smith, C. W. J.</dc:creator>
<dc:creator>Turner, M.</dc:creator>
<dc:date>2022-03-04</dc:date>
<dc:identifier>doi:10.1101/2022.03.03.482829</dc:identifier>
<dc:title><![CDATA[Polypyrimidine Tract Binding Protein 1 regulates the activation of mouse CD8 T cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-03-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.03.01.482519v1?rss=1">
<title>
<![CDATA[
Molecular and cognitive signatures of ageing partially restored through synthetic delivery of IL2 to the brain 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.03.01.482519v1?rss=1"
</link>
<description><![CDATA[
Cognitive decline is a common pathological outcome during aging, with an ill-defined cellular or molecular basis. Among the cellular changes observed with age are alterations to neuronal plasticity, changes in the glial compartment and the decline of the neurogenic niche. In the recent years, the concept of inflammaging, defined as a low-grade inflammation increasing with age, has emerged as a nexus for age-related diseases. This increase of basal inflammation is also observed in the central nervous system. While not classically considered a neurological cell type, infiltrating T cells increase in the brain with age, and may be responsible for amplification of inflammatory cascades and disruptions to the neurogenic niche. Recently, a small resident population of regulatory T cells has been identified in the brain, and the capacity of IL2-mediated expansion of this population to counter neuroinflammatory disease has been demonstrated. Here we test a brain-specific IL2 delivery system for the prevention of neurological decline in aging mice. We identify the molecular hallmarks of aging in the brain glial compartments, and identify partial restoration of this signature through IL2 treatment. At a behavioral level, brain IL2 delivery prevented the age-induced defect in spatial learning, without improving the general decline in motor skill or arousal. These results identify immune modulation as a potential path to preserving cognitive function for healthy ageing.
]]></description>
<dc:creator>Lemaitre, P.</dc:creator>
<dc:creator>Tareen, S.</dc:creator>
<dc:creator>Pasciuto, E.</dc:creator>
<dc:creator>Mascali, L.</dc:creator>
<dc:creator>Martirosyan, A.</dc:creator>
<dc:creator>Callaerts-Vegh, Z.</dc:creator>
<dc:creator>Dooley, J.</dc:creator>
<dc:creator>Holt, M.</dc:creator>
<dc:creator>Yshii, L.</dc:creator>
<dc:creator>Liston, A.</dc:creator>
<dc:date>2022-03-04</dc:date>
<dc:identifier>doi:10.1101/2022.03.01.482519</dc:identifier>
<dc:title><![CDATA[Molecular and cognitive signatures of ageing partially restored through synthetic delivery of IL2 to the brain]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-03-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.02.28.482297v1?rss=1">
<title>
<![CDATA[
Astrocyte-targeted gene delivery of interleukin 2 specifically increases brain-resident regulatory T cell numbers and protects against pathological neuroinflammation 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.02.28.482297v1?rss=1"
</link>
<description><![CDATA[
The ability of immune-modulating biologics to prevent and reverse pathology has transformed recent clinical practice. Full utility in the neuroinflammation space, however, requires identification of both effective targets for local immune-modulation and a delivery system capable of crossing the blood-brain-barrier. The recent identification and characterization of a small population of regulatory T cells (Tregs) resident in the brain presents one such potential therapeutic target. Here we identified brain IL2 levels as a limiting factor for brain-resident Tregs. We developed a gene-delivery approach for astrocytes, with a small-molecule on-switch to allow temporal control, and enhanced production in reactive astrocytes to spatially-direct delivery to inflammatory sites. Mice with brain-specific IL2 delivery were protected from traumatic brain injury, stroke and multiple sclerosis models, without impacting the peripheral immune system. These results validate brain-specific IL2 gene-delivery as effective protection against neuroinflammation, and provide a versatile platform for delivery of diverse biologics to neuroinflammatory patients.
]]></description>
<dc:creator>Yshii, L.</dc:creator>
<dc:creator>Pasciuto, E.</dc:creator>
<dc:creator>Bielefeld, P.</dc:creator>
<dc:creator>Mascali, L.</dc:creator>
<dc:creator>Lemaitre, P.</dc:creator>
<dc:creator>Marino, M.</dc:creator>
<dc:creator>Dooley, J.</dc:creator>
<dc:creator>Kouser, L.</dc:creator>
<dc:creator>Verschoren, S.</dc:creator>
<dc:creator>Lagou, V.</dc:creator>
<dc:creator>Kemps, H.</dc:creator>
<dc:creator>Gervois, P.</dc:creator>
<dc:creator>de Boer, A.</dc:creator>
<dc:creator>Burton, O.</dc:creator>
<dc:creator>Wahis, J.</dc:creator>
<dc:creator>Tareen, S.</dc:creator>
<dc:creator>Roca, C.</dc:creator>
<dc:creator>Singh, K.</dc:creator>
<dc:creator>Whyte, C.</dc:creator>
<dc:creator>Kerstens, A.</dc:creator>
<dc:creator>Callaerts-Vegh, Z.</dc:creator>
<dc:creator>Poovathingal, S.</dc:creator>
<dc:creator>Prezzemolo, T.</dc:creator>
<dc:creator>Wierda, K.</dc:creator>
<dc:creator>Dashwood, A.</dc:creator>
<dc:creator>Xie, J.</dc:creator>
<dc:creator>Van Wonterghem, E.</dc:creator>
<dc:creator>Creemers, E.</dc:creator>
<dc:creator>Aloulou, M.</dc:creator>
<dc:creator>Gsell, W.</dc:creator>
<dc:creator>Abiega, O.</dc:creator>
<dc:creator>Munck, S.</dc:creator>
<dc:creator>Vandenbroucke, R.</dc:creator>
<dc:creator>Lemmens, R.</dc:creator>
<dc:creator>De Strooper, B.</dc:creator>
<dc:creator>Van Den Bosch, L.</dc:creator>
<dc:creator>Himmelreich, U.</dc:creator>
<dc:creator>Fitzsimons, C. P.</dc:creator>
<dc:creator>Holt, M.</dc:creator>
<dc:creator>Liston, A.</dc:creator>
<dc:date>2022-03-02</dc:date>
<dc:identifier>doi:10.1101/2022.02.28.482297</dc:identifier>
<dc:title><![CDATA[Astrocyte-targeted gene delivery of interleukin 2 specifically increases brain-resident regulatory T cell numbers and protects against pathological neuroinflammation]]></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/cgi/content/short/2022.01.20.477063v1?rss=1">
<title>
<![CDATA[
Multi-omic rejuvenation of naturally aged tissues by a single cycle of transient reprogramming 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.01.20.477063v1?rss=1"
</link>
<description><![CDATA[
The expression of the pluripotency factors OCT4, SOX2, KLF4 and MYC (OSKM) can convert somatic differentiated cells into pluripotent stem cells in a process known as reprogramming. Notably, cycles of brief OSKM expression do not change cell identity but can reverse markers of aging in cells and extend longevity in progeroid mice. However, little is known about the mechanisms involved. Here, we have studied changes in the DNA methylome, transcriptome and metabolome in naturally aged mice subject to a single period of transient OSKM expression. We found that this is sufficient to reverse DNA methylation changes that occur upon aging in the pancreas, liver, spleen and blood. Similarly, we observed reversion of transcriptional changes, especially regarding biological processes known to change during aging. Finally, some serum metabolites altered with aging were also restored to young levels upon transient reprogramming. These observations indicate that a single period of OSKM expression can drive epigenetic, transcriptomic and metabolomic changes towards a younger configuration in multiple tissues and in the serum.
]]></description>
<dc:creator>Chondronasiou, D.</dc:creator>
<dc:creator>Gill, D.</dc:creator>
<dc:creator>Mosteiro, L.</dc:creator>
<dc:creator>Urdinguio, R. G.</dc:creator>
<dc:creator>Berenguer, A.</dc:creator>
<dc:creator>Aguilera, M.</dc:creator>
<dc:creator>Durand, S.</dc:creator>
<dc:creator>Aprahamian, F.</dc:creator>
<dc:creator>Nirmalathasan, N.</dc:creator>
<dc:creator>Abad, M.</dc:creator>
<dc:creator>Martin-Herranz, D. E.</dc:creator>
<dc:creator>Stephan-Otto Attolini, C.</dc:creator>
<dc:creator>Prats, N.</dc:creator>
<dc:creator>Kroemer, G.</dc:creator>
<dc:creator>Fraga, M. F.</dc:creator>
<dc:creator>Reik, W.</dc:creator>
<dc:creator>Serrano, M.</dc:creator>
<dc:date>2022-01-21</dc:date>
<dc:identifier>doi:10.1101/2022.01.20.477063</dc:identifier>
<dc:title><![CDATA[Multi-omic rejuvenation of naturally aged tissues by a single cycle of transient reprogramming]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-01-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.01.14.476154v1?rss=1">
<title>
<![CDATA[
Plasma cell maintenance and antibody secretion are under the control of Sec22b-mediated regulation of organelle dynamics 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.01.14.476154v1?rss=1"
</link>
<description><![CDATA[
Despite the essential role of plasma cells in health and disease, the cellular mechanisms controlling their survival and secretory capacity are still poorly understood. Here, we identified the SNARE Sec22b as a unique and critical regulator of plasma cell maintenance and function. In absence of Sec22b, plasma cells were barely detectable and serum antibody titres were dramatically reduced. Accordingly, Sec22b deficient mice fail to mount a protective immune response. At the mechanistic level, we demonstrated that Sec22b is indispensable for efficient antibody secretion but also for plasma cell fitness through the regulation of the morphology of the endoplasmic reticulum and mitochondria. Altogether, our results unveil a critical role for Sec22b-mediated regulation of plasma cell biology through the control of organelle dynamics.
]]></description>
<dc:creator>Bonaud, A.</dc:creator>
<dc:creator>Gargowitsch, L.</dc:creator>
<dc:creator>Gilbert, S.</dc:creator>
<dc:creator>Rajan, E.</dc:creator>
<dc:creator>Canales-Herrerias, P.</dc:creator>
<dc:creator>Stockholm, D.</dc:creator>
<dc:creator>Rahman, N.</dc:creator>
<dc:creator>Collins, M.</dc:creator>
<dc:creator>Hill, D. L.</dc:creator>
<dc:creator>Alloatti, A.</dc:creator>
<dc:creator>Alouche, N.</dc:creator>
<dc:creator>Balor, S.</dc:creator>
<dc:creator>Soldan, V.</dc:creator>
<dc:creator>Gillet, D.</dc:creator>
<dc:creator>Barbier, J.</dc:creator>
<dc:creator>Bachelerie, F.</dc:creator>
<dc:creator>Smith, K. G. C.</dc:creator>
<dc:creator>Bruhns, P.</dc:creator>
<dc:creator>Amigorena, S.</dc:creator>
<dc:creator>Balabanian, K.</dc:creator>
<dc:creator>Linterman, M. A.</dc:creator>
<dc:creator>Peden, A.</dc:creator>
<dc:creator>Espeli, M.</dc:creator>
<dc:date>2022-01-15</dc:date>
<dc:identifier>doi:10.1101/2022.01.14.476154</dc:identifier>
<dc:title><![CDATA[Plasma cell maintenance and antibody secretion are under the control of Sec22b-mediated regulation of organelle dynamics]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-01-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.12.29.474486v1?rss=1">
<title>
<![CDATA[
Pronounced sequence specificity of the TET enzyme catalytic domain guides its cellular function 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.12.29.474486v1?rss=1"
</link>
<description><![CDATA[
TET (ten-eleven translocation) enzymes catalyze the oxidation of 5-methylcytosine bases in DNA, thus driving active and passive DNA demethylation. Here, we report that the catalytic cores of mammalian TET enzymes favor CpGs embedded within bHLH and bZIP transcription factor binding sites, with 250-fold preference in vitro. Crystal structures and molecular dynamics calculations show that sequence preference is caused by intra-substrate interactions and CpG flanking sequence indirectly affecting enzyme conformation. TET sequence preferences are physiologically relevant as they explain the rates of DNA demethylation in TET-rescue experiments in culture and in vivo within the zygote and germline. Most and least favorable TET motifs represent DNA sites that are bound by methylation-sensitive immediate-early transcription factors and OCT4, respectively, illuminating TET function in transcriptional responses and pluripotency support.

One-Sentence SummaryThe catalytic domains of the enzymes that facilitate passive and drive active DNA demethylation have intrinsic sequence preferences that target DNA demethylation to bHLH and bZIP transcription factor binding sites.
]]></description>
<dc:creator>Ravichandran, M.</dc:creator>
<dc:creator>Rafalski, D.</dc:creator>
<dc:creator>Ortega-Recalde, O.</dc:creator>
<dc:creator>Davies, C. I.</dc:creator>
<dc:creator>Glanfield, C. R.</dc:creator>
<dc:creator>Kotter, A.</dc:creator>
<dc:creator>Misztal, K.</dc:creator>
<dc:creator>Wang, A. H.</dc:creator>
<dc:creator>Wojciechowski, M.</dc:creator>
<dc:creator>Razew, M.</dc:creator>
<dc:creator>Mayyas, I. M.</dc:creator>
<dc:creator>Kardailsky, O.</dc:creator>
<dc:creator>Schwarz, U.</dc:creator>
<dc:creator>Zembrzycki, K.</dc:creator>
<dc:creator>Morison, I. M.</dc:creator>
<dc:creator>Helm, M.</dc:creator>
<dc:creator>Weichenhan, D.</dc:creator>
<dc:creator>Jurkowska, R. Z.</dc:creator>
<dc:creator>Krueger, F.</dc:creator>
<dc:creator>Plass, C.</dc:creator>
<dc:creator>Zacharias, M.</dc:creator>
<dc:creator>Hore, T. A.</dc:creator>
<dc:creator>Bochtler, M.</dc:creator>
<dc:creator>Jurkowski, T. P.</dc:creator>
<dc:date>2021-12-30</dc:date>
<dc:identifier>doi:10.1101/2021.12.29.474486</dc:identifier>
<dc:title><![CDATA[Pronounced sequence specificity of the TET enzyme catalytic domain guides its cellular function]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-12-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.12.20.473453v1?rss=1">
<title>
<![CDATA[
Single-Cell Atlas of Common Variable Immunodeficiency reveals germinal center-associated epigenetic dysregulation in B cell responses 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.12.20.473453v1?rss=1"
</link>
<description><![CDATA[
Common variable immunodeficiency (CVID), the most prevalent symptomatic primary immunodeficiency, is characterized by impaired terminal B-cell differentiation and defective antibody responses. Incomplete genetic penetrance and a wide range of phenotypic expressivity in CVID suggest the participation of additional pathogenic mechanisms. Monozygotic (MZ) twins discordant for CVID are uniquely valuable for studying the contribution of epigenetics to the disease. We used single-cell epigenomics and transcriptomics to create a cell census of naive-to-memory B cell differentiation in a pair of CVID-discordant MZ twins. Our analysis identifies DNA methylation, chromatin accessibility and transcriptional defects in memory B cells that mirror defective cell-cell communication defects following activation. These findings were validated in a cohort of CVID patients and healthy donors. Our findings provide a comprehensive multi-omics map of alterations in naive-to-memory B-cell transition in CVID and reveal links between the epigenome and immune cell cross-talk. Our resource, publicly available at the Human Cell Atlas, paves the way for future diagnosis and treatments of CVID patients.
]]></description>
<dc:creator>Rodriguez-Ubreva, J.</dc:creator>
<dc:creator>Arutyunyan, A.</dc:creator>
<dc:creator>Bonder, M. J.</dc:creator>
<dc:creator>Del Pino-Molina, L.</dc:creator>
<dc:creator>Clark, S.</dc:creator>
<dc:creator>de la Calle-Fabregat, C.</dc:creator>
<dc:creator>Garcia-Alonso, L.</dc:creator>
<dc:creator>Handfield, L.-F.</dc:creator>
<dc:creator>Ciudad, L.</dc:creator>
<dc:creator>Andres-Leon, E.</dc:creator>
<dc:creator>Krueger, F.</dc:creator>
<dc:creator>Catala-Moll, F.</dc:creator>
<dc:creator>Rodriguez-Cortez, V. C.</dc:creator>
<dc:creator>Polanski, K.</dc:creator>
<dc:creator>Mamanova, L.</dc:creator>
<dc:creator>van Dongen, S.</dc:creator>
<dc:creator>Kiselev, V. Y.</dc:creator>
<dc:creator>Martinez-Saavedra, M. T.</dc:creator>
<dc:creator>Heyn, H.</dc:creator>
<dc:creator>Martin, J.</dc:creator>
<dc:creator>Warnatz, K.</dc:creator>
<dc:creator>Lopez-Granados, E.</dc:creator>
<dc:creator>Rodriguez-Gallego, C.</dc:creator>
<dc:creator>Stegle, O.</dc:creator>
<dc:creator>Kelsey, G. D.</dc:creator>
<dc:creator>Vento-Tormo, R.</dc:creator>
<dc:creator>Ballestar, E.</dc:creator>
<dc:date>2021-12-21</dc:date>
<dc:identifier>doi:10.1101/2021.12.20.473453</dc:identifier>
<dc:title><![CDATA[Single-Cell Atlas of Common Variable Immunodeficiency reveals germinal center-associated epigenetic dysregulation in B cell responses]]></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/cgi/content/short/2020.08.06.236307v1?rss=1">
<title>
<![CDATA[
Genome wide efficiency profiling reveals modulation of maintenance and de novo methylation by Tets 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.08.06.236307v1?rss=1"
</link>
<description><![CDATA[
A precise understanding of DNA methylation dynamics on a genome wide scale is of great importance for the comprehensive investigation of a variety of biological processes such as reprogramming of somatic cells to iPSCs, cell differentiation and also cancer development. To date, a complex integration of multiple and distinct genome wide data sets is required to derive the global activity of DNA modifying enzymes. We present GwEEP - Genome-wide Epigenetic Efficiency Profiling as a versatile approach to infer dynamic efficiency changes of DNA modifying enzymes at base pair resolution on a genome wide scale. GwEEP relies on genome wide oxidative Hairpin Bisulfite sequencing (HPoxBS) data sets, which are translated by a sophisticated hidden Markov model into quantitative enzyme efficiencies with reported confidence around the estimates. GwEEP in its present form predicts de novo and maintenance methylation efficiencies of Dnmts, as well as the hydroxylation efficiency of Tets but its purposefully flexible design allows to capture further oxidation processes such as formylation and carboxylation given available data in the future. Applied to a well characterized ES cell model, GwEEP precisely predicts the complex epigenetic changes following a Serum-to-2i shift i.e., (i) instant reduction in maintenance efficiency (ii) gradually decreasing de novo methylation efficiency and (iii) increasing Tet efficiencies. In addition, a complementary analysis of Tet triple knock-out ES cells confirms the previous hypothesized mutual interference of Dnmts and Tets. GwEEP is applicable to a wide range of biological samples including cell lines, but also tissues and primary cell types.

MOTIVATIONDynamic changes of DNA methylation patterns are a common phenomenon in epigenetics. Although a stable DNA methylation profile is essential for cell identity, developmental processes require the rearrangement of 5-methylcytosine in the genome. Stable methylation patterns are the result of balanced Dnmts and Tets activities, while methylome transformation results from a coordinated change in Dnmt and Tet efficiencies. Such transformations occur on a global scale, for example during the reprogramming of maternal and paternal methylation patterns and the establishment of novel cell type specific methylomes during embryonic development in vivo, but also in vitro during (re)programming of induced pluripotent stem cells, as well as somatic cells. In addition, local (de)methylation events are key for gene regulation during cell differentiation. A detailed understanding of Dnmt and Tet cooperation is essential for understanding natural epigenetic adaptation as well as optimization of in vitro (re)programming protocols. For this purpose, we developed a pipeline for quantitative and precise estimation of Dnmt and Tet activity. Using only double strand methylation information, GwEEP infers accurate maintenance and de novo methylation efficiency of Dnmts, as well as hydroxylation efficiency of Tets at single base resolution. Thus, we believe GwEEP provides a powerful tool for the investigation of methylome rearrangements in various systems.
]]></description>
<dc:creator>Giehr, P.</dc:creator>
<dc:creator>Kyriakopoulos, C.</dc:creator>
<dc:creator>Karl Nordström, K.</dc:creator>
<dc:creator>Salhab, A.</dc:creator>
<dc:creator>Müller, F.</dc:creator>
<dc:creator>von Meyenn, F.</dc:creator>
<dc:creator>Ficz, G.</dc:creator>
<dc:creator>Reik, W.</dc:creator>
<dc:creator>Wolf, V.</dc:creator>
<dc:creator>Walter, J.</dc:creator>
<dc:date>2020-08-07</dc:date>
<dc:identifier>doi:10.1101/2020.08.06.236307</dc:identifier>
<dc:title><![CDATA[Genome wide efficiency profiling reveals modulation of maintenance and de novo methylation by Tets]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-08-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.10.28.466259v1?rss=1">
<title>
<![CDATA[
Modelling human zygotic genome activation in 8C-like cells in vitro 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.10.28.466259v1?rss=1"
</link>
<description><![CDATA[
The remodelling of the epigenome and transcriptome of the fertilised oocyte to establish totipotency in the zygote and developing embryo is one of the most critical processes in mammalian embryogenesis. Zygotic or embryonic genome activation (ZGA, EGA) in the 2-cell embryo in mouse, and the 8-cell embryo in humans, constitutes the first major wave of transcription. Failure to initiate ZGA leads to developmental defects, and contributes to the high attrition rates of human pre-implantation embryos. Due to limitations in cell numbers and experimental tractability, the mechanisms that regulate human embryonic genome activation in the totipotent embryo remain poorly understood. Here we report the discovery of human 8-cell like cells (8CLCs) specifically among naive embryonic stem cells, but not primed pluripotent cells. 8CLCs express ZGA marker genes such as ZSCAN4, LEUTX and DUXA and their transcriptome closely resembles that of the 8-cell human embryo. 8-cell like cells reactivate 8-cell stage specific transposable elements such as HERVL and MLT2A1 and are characterized by upregulation of the DNA methylation regulator DPPA3. 8CLCs show reduced SOX2 protein, and can be identified based on expression of the novel ZGA-associated protein markers TPRX1 and H3.Y in vitro. Overexpression of the transcription factor DUX4. as well as spliceosome inhibition increase ZGA-like transcription and enhance TPRX1+ 8CLCs formation. Excitingly, the in vitro identified 8CLC marker proteins TPRX1 and H3.Y are also expressed in 8-cell human embryos at the time of genome activation and may thus be relevant in vivo. The discovery of 8CLCs provides a unique opportunity to model and manipulate human ZGA-like transcriptional programs in vitro, and might provide critical functional insights into one of the earliest events in human embryogenesis in vivo.

HighlightsO_LIZGA markers and transposable elements are expressed in 8CLCs among naive human stem cells
C_LIO_LIThe transcription factor DUX4 and spliceosome inhibition induce ZGA-like transcription
C_LIO_LI8CLC marker proteins TPRX1 and H3.Y are expressed in nuclei of 8-cell human embryos
C_LIO_LI8CLCs serve as a novel in vitro model for human ZGA
C_LI
]]></description>
<dc:creator>Taubenschmid-Stowers, J.</dc:creator>
<dc:creator>Rostovskaya, M.</dc:creator>
<dc:creator>Santos, F.</dc:creator>
<dc:creator>Ljung, S.</dc:creator>
<dc:creator>Argelaguet, R.</dc:creator>
<dc:creator>Krueger, F.</dc:creator>
<dc:creator>Nichols, J.</dc:creator>
<dc:creator>Reik, W.</dc:creator>
<dc:date>2021-10-29</dc:date>
<dc:identifier>doi:10.1101/2021.10.28.466259</dc:identifier>
<dc:title><![CDATA[Modelling human zygotic genome activation in 8C-like cells in vitro]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-10-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.09.13.460183v1?rss=1">
<title>
<![CDATA[
Maternal Dppa2 and Dppa4 are dispensable for zygotic genome activation but important for offspring survival 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.09.13.460183v1?rss=1"
</link>
<description><![CDATA[
Zygotic Genome Activation (ZGA) represents the initiation of transcription following fertilisation. Despite its importance in shifting developmental control from primarily maternal stores in the oocyte to the embryo proper, we know little of the molecular events that initiate ZGA in vivo. Recent in vitro studies in mouse embryonic stem cells (ESCs) have revealed Developmental Pluripotency Associated 2 and 4 (Dppa2/4) as key regulators of ZGA-associated transcription. However, their roles in initiating ZGA in vivo remain unexplored. We reveal Dppa2/4 proteins are present in the nucleus at all stages of preimplantation development and associate with mitotic chromatin. We generated single and double maternal knockout mouse models to deplete maternal stores of Dppa2/4. Importantly, while fertile, Dppa2/4 maternal knockout mice had reduced litter sizes, indicating decreased offspring survival. Immunofluorescence and transcriptome analyses of 2-cell embryos revealed while ZGA took place there were subtle defects in embryos lacking maternal Dppa2/4. Strikingly, heterozygous offspring that inherited the null allele maternally had higher preweaning lethality than those that inherited the null allele paternally. Together our results show that while Dppa2/4 are dispensable for ZGA transcription, maternal stores have an important role in offspring survival, potentially via epigenetic priming of developmental genes.
]]></description>
<dc:creator>Kubinyecz, O.</dc:creator>
<dc:creator>Santos, F.</dc:creator>
<dc:creator>Drage, D.</dc:creator>
<dc:creator>Reik, W.</dc:creator>
<dc:creator>Eckersley-Maslin, M. A.</dc:creator>
<dc:date>2021-09-14</dc:date>
<dc:identifier>doi:10.1101/2021.09.13.460183</dc:identifier>
<dc:title><![CDATA[Maternal Dppa2 and Dppa4 are dispensable for zygotic genome activation but important for offspring survival]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-09-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.07.09.451752v1?rss=1">
<title>
<![CDATA[
Single-cell bisulfite sequencing of spermatozoa from lean and obese humans reveals potential for the transmission of epimutations 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.07.09.451752v1?rss=1"
</link>
<description><![CDATA[
Epigenetic marks in gametes modulate developmental programming after fertilization. Spermatozoa from obese men exhibit distinct epigenetic signatures compared to lean men, however, whether epigenetic differences are concentrated in a sub-population of spermatozoa or spread across the ejaculate population is unknown. Here, by using whole-genome single-cell bisulfite sequencing on 87 motile spermatozoa from 8 individuals (4 lean and 4 obese), we found that spermatozoa within single ejaculates are highly heterogeneous and contain subsets of spermatozoa with marked imprinting defects. Comparing lean and obese subjects, we discovered methylation differences across two large CpG dense regions located near PPM1D and LINC01237. These findings confirm that sperm DNA methylation is altered in human obesity and indicate that single ejaculates contain subpopulations of spermatozoa carrying distinct DNA methylation patterns. Distinct epigenetic patterns of spermatozoa within an ejaculate may result in different intergenerational effects and therefore influence strategies aiming to prevent epigenetic-related disorders in the offspring.
]]></description>
<dc:creator>Barres, R.</dc:creator>
<dc:creator>Andersen, E.</dc:creator>
<dc:creator>Reik, W.</dc:creator>
<dc:creator>Clark, S.</dc:creator>
<dc:creator>Ingerslev, L.</dc:creator>
<dc:creator>Lundell, L.</dc:creator>
<dc:date>2021-07-10</dc:date>
<dc:identifier>doi:10.1101/2021.07.09.451752</dc:identifier>
<dc:title><![CDATA[Single-cell bisulfite sequencing of spermatozoa from lean and obese humans reveals potential for the transmission of epimutations]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-07-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.02.22.432209v1?rss=1">
<title>
<![CDATA[
GABARAP membrane conjugation sequesters the FLCN-FNIP tumor suppressor complex to activate TFEB and lysosomal biogenesis 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.02.22.432209v1?rss=1"
</link>
<description><![CDATA[
Adaptive changes in lysosomal capacity are driven by the transcription factors TFEB and TFE3 in response to increased autophagic flux and endolysosomal stress, yet the molecular details of their activation are unclear. LC3 and GABARAP members of the ATG8 protein family are required for selective autophagy and sensing perturbation within the endolysosomal system. Here we show that during single membrane ATG8 conjugation (SMAC), Parkin-dependent mitophagy, and Salmonella-induced xenophagy, the membrane conjugation of GABARAP, but not LC3, is required for activation of TFEB/TFE3 to control lysosomal homeostasis and capacity. GABARAP directly binds to a novel LC3-interacting motif (LIR) in the FLCN/FNIP tumor suppressor complex with picomolar affinity and regulates its relocalization to these GABARAP-conjugated membrane compartments. This disrupts the regulation of RagC/D by the FLCN/FNIP GAP complex, resulting in impaired mTOR-dependent phosphorylation of TFEB without changing mTOR activity towards other substrates. Thus, the GABARAP-FLCN/FNIP-TFEB axis serves as a universal molecular sensor that coordinates lysosomal homeostasis with perturbations and cargo flux within the autophagy-lysosomal network.
]]></description>
<dc:creator>Goodwin, J.</dc:creator>
<dc:creator>Walkup, W. G.</dc:creator>
<dc:creator>Hooper, K.</dc:creator>
<dc:creator>Li, T.</dc:creator>
<dc:creator>Kishi-Itakura, C.</dc:creator>
<dc:creator>Ng, A.</dc:creator>
<dc:creator>Lehmberg, T.</dc:creator>
<dc:creator>Jha, A.</dc:creator>
<dc:creator>Kommineni, S.</dc:creator>
<dc:creator>Fletcher, K.</dc:creator>
<dc:creator>Garcia-Fortanet, J.</dc:creator>
<dc:creator>Fan, Y.</dc:creator>
<dc:creator>Tang, Q.</dc:creator>
<dc:creator>Wei, M.</dc:creator>
<dc:creator>Agrawal, A.</dc:creator>
<dc:creator>Budhe, S. R.</dc:creator>
<dc:creator>Rouduri, S. R.</dc:creator>
<dc:creator>Baird, D.</dc:creator>
<dc:creator>Saunders, J.</dc:creator>
<dc:creator>Kiselar, J.</dc:creator>
<dc:creator>Chance, M. R.</dc:creator>
<dc:creator>Ballabio, A.</dc:creator>
<dc:creator>Appleton, B. A.</dc:creator>
<dc:creator>Brumell, J. H.</dc:creator>
<dc:creator>Florey, O.</dc:creator>
<dc:creator>Murphy, L. O.</dc:creator>
<dc:date>2021-02-22</dc:date>
<dc:identifier>doi:10.1101/2021.02.22.432209</dc:identifier>
<dc:title><![CDATA[GABARAP membrane conjugation sequesters the FLCN-FNIP tumor suppressor complex to activate TFEB and lysosomal biogenesis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-02-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.02.03.429496v1?rss=1">
<title>
<![CDATA[
Three-dimensional interactions between integrated HPV genomes and cellular chromatin dysregulate host gene expression in early cervical carcinogenesis 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.02.03.429496v1?rss=1"
</link>
<description><![CDATA[
Development of cervical cancer is directly associated with integration of human papillomavirus (HPV) genomes into host chromosomes and subsequent modulation of HPV oncogene expression, which correlates with multi-layered epigenetic changes at the integrated HPV genomes. However, the process of integration itself and dysregulation of host gene expression at sites of integration in our model of HPV16 integrant clone natural selection has remained enigmatic. We now show, using a state-of-the-art  HPV integrated site capture (HISC) technique, that integration likely occurs through microhomology-mediated repair (MHMR) mechanisms via either a direct process, resulting in host sequence deletion (in our case, partially homozygously) or via a  looping mechanism by which flanking host regions become amplified. Furthermore, using our  HPV16-specific Region Capture Hi-C technique, we have determined that three-dimensional (3D) interactions between the integrated virus genome and host chromosomes, both at short- (<500 kbp) and long-range (>500 kbp), appear to drive host gene dysregulation through the disruption of local host:host 3D interactions known as topologically associating domains (TADs). This mechanism of HPV-induced host gene expression modulation indicates that integration of virus genomes near to or within a  cancer-causing gene is not essential to influence such genes within an entire TAD and that these modifications to 3D interactions could have a major role in selection of HPV integrants at the early stage of cervical neoplastic progression.
]]></description>
<dc:creator>Groves, I. J.</dc:creator>
<dc:creator>Drane, E. L.</dc:creator>
<dc:creator>Michalski, M.</dc:creator>
<dc:creator>Monahan, J. M.</dc:creator>
<dc:creator>Scarpini, C. G.</dc:creator>
<dc:creator>Smith, S. P.</dc:creator>
<dc:creator>Bussotti, G.</dc:creator>
<dc:creator>Varnai, C.</dc:creator>
<dc:creator>Schoenfelder, S.</dc:creator>
<dc:creator>Fraser, P.</dc:creator>
<dc:creator>Enright, A. J.</dc:creator>
<dc:creator>Coleman, N.</dc:creator>
<dc:date>2021-02-03</dc:date>
<dc:identifier>doi:10.1101/2021.02.03.429496</dc:identifier>
<dc:title><![CDATA[Three-dimensional interactions between integrated HPV genomes and cellular chromatin dysregulate host gene expression in early cervical carcinogenesis]]></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/cgi/content/short/2021.02.03.429569v1?rss=1">
<title>
<![CDATA[
Inhibiting glycogen synthase kinase 3 suppresses TDP-43-mediated neurotoxicity in a caspase-dependant manner 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.02.03.429569v1?rss=1"
</link>
<description><![CDATA[
Amyotrophic lateral sclerosis-frontotemporal dementia (ALS-FTD) is a progressive and ultimately fatal disease spectrum characterised by 43-kDa TAR DNA-binding protein (TDP-43) pathology. Current disease modifying drugs have modest effects and novel therapies are sorely needed. We previously showed that deletion of glycogen synthase kinase-3 (GSK3) suppresses TDP-43-mediated motor neuron degeneration in Drosophila. Here, we investigated the potential of GSK3 inhibition to ameliorate TDP43-mediated toxicity in mammalian neurons. Expression of TDP-43 was found to both activate GSK3 and promote caspase mediated cleavage of TDP-43. Inhibition of GSK3 reduced the abundance of full-length and cleaved TDP-43 in rodent neurons expressing wild-type or disease-associated mutant TDP-43 and also ameliorated neurotoxicity. Our results suggest that TDP-43 turnover is promoted by GSK3 inhibition in a caspase-dependent manner, and that targeting GSK3 activity could have therapeutic value.
]]></description>
<dc:creator>White, M. A.</dc:creator>
<dc:creator>Massenzio, F.</dc:creator>
<dc:creator>Li, X.</dc:creator>
<dc:creator>Coleman, M. P.</dc:creator>
<dc:creator>Barmada, S. J.</dc:creator>
<dc:creator>Sreedharan, J.</dc:creator>
<dc:date>2021-02-03</dc:date>
<dc:identifier>doi:10.1101/2021.02.03.429569</dc:identifier>
<dc:title><![CDATA[Inhibiting glycogen synthase kinase 3 suppresses TDP-43-mediated neurotoxicity in a caspase-dependant manner]]></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/cgi/content/short/2020.07.07.192427v1?rss=1">
<title>
<![CDATA[
Enhancer priming by H3K4 methylation safeguards germline competence 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.07.07.192427v1?rss=1"
</link>
<description><![CDATA[
Germline specification in mammals occurs through an inductive process whereby competent cells in the post-implantation epiblast differentiate into primordial germ cells (PGC). The intrinsic factors that endow epiblast cells with the competence to respond to germline inductive signals remain unknown. Single-cell RNA sequencing across multiple stages of an in vitro PGC-like cells (PGCLC) differentiation system shows that PGCLC genes initially expressed in the naive pluripotent stage become homogeneously dismantled in germline competent epiblast like-cells (EpiLC). In contrast, the decommissioning of enhancers associated with these germline genes is incomplete. Namely, a subset of these enhancers partly retain H3K4me1, accumulate less heterochromatic marks and remain accessible and responsive to transcriptional activators. Subsequently, as in vitro germline competence is lost, these enhancers get further decommissioned and lose their responsiveness to transcriptional activators. Importantly, using H3K4me1 deficient cells, we show that the loss of this histone modification reduces the germline competence of EpiLC and decreases PGCLC differentiation efficiency. Our work suggests that, although H3K4me1 might not be essential for enhancer function, it can facilitate the (re)activation of enhancers and the establishment of gene expression programs during specific developmental transitions.
]]></description>
<dc:creator>Bleckwehl, T.</dc:creator>
<dc:creator>Schaaf, K.</dc:creator>
<dc:creator>Crispatzu, G.</dc:creator>
<dc:creator>Respuela, P.</dc:creator>
<dc:creator>Bartusel, M.</dc:creator>
<dc:creator>Benson, L.</dc:creator>
<dc:creator>Clark, S. J.</dc:creator>
<dc:creator>Dorighi, K. M.</dc:creator>
<dc:creator>Barral, A.</dc:creator>
<dc:creator>Laugsch, M.</dc:creator>
<dc:creator>Manzanares, M.</dc:creator>
<dc:creator>Wysocka, J.</dc:creator>
<dc:creator>Reik, W.</dc:creator>
<dc:creator>Rada-Iglesias, A.</dc:creator>
<dc:date>2020-07-07</dc:date>
<dc:identifier>doi:10.1101/2020.07.07.192427</dc:identifier>
<dc:title><![CDATA[Enhancer priming by H3K4 methylation safeguards germline competence]]></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/cgi/content/short/2021.01.12.426343v1?rss=1">
<title>
<![CDATA[
Conserved and unique transcriptional features of pharyngeal arches in the skate (Leucoraja erinacea) and evolution of the jaw 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.01.12.426343v1?rss=1"
</link>
<description><![CDATA[
The origin of the jaw is a long-standing problem in vertebrate evolutionary biology. Classical hypotheses of serial homology propose that the upper and lower jaw evolved through modifications of dorsal and ventral gill arch skeletal elements, respectively. If the jaw and gill arches are derived members of a primitive branchial series, we predict that they would share common developmental patterning mechanisms. Using candidate and RNAseq/differential gene expression analyses, we find broad conservation of dorsoventral patterning mechanisms within the developing mandibular, hyoid and gill arches of a cartilaginous fish, the skate (Leucoraja erinacea). Shared features include expression of genes encoding members of the ventralising BMP and endothelin signalling pathways and their effectors, the joint markers bapx1 and gdf5 and pro-chondrogenic transcription factors barx1 and gsc, and the dorsalising transcription factor pou3f3. Additionally, we find that mesenchymal expression of eya1/six1 is an ancestral feature of the mandibular arch of jawed vertebrates, while differences in notch signalling distinguish the mandibular and gill arches in skate. Comparative transcriptomic analyses of mandibular and gill arch tissues reveal additional genes differentially expressed along the dorsoventral axis of the pharyngeal arches, including scamp5 as a novel marker of the dorsal mandibular arch, as well as distinct transcriptional features of mandibular and gill arch muscle progenitors and developing gill buds. Taken together, our findings reveal conserved patterning mechanisms in the pharyngeal arches of jawed vertebrates, consistent with serial homology of their skeletal derivatives, as well as unique transcriptional features that may underpin distinct jaw and gill arch morphologies.
]]></description>
<dc:creator>Hirschberger, C.</dc:creator>
<dc:creator>Sleight, V. A.</dc:creator>
<dc:creator>Criswell, K. E.</dc:creator>
<dc:creator>Clark, S.</dc:creator>
<dc:creator>Gillis, J. A.</dc:creator>
<dc:date>2021-01-12</dc:date>
<dc:identifier>doi:10.1101/2021.01.12.426343</dc:identifier>
<dc:title><![CDATA[Conserved and unique transcriptional features of pharyngeal arches in the skate (Leucoraja erinacea) and evolution of the jaw]]></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/cgi/content/short/2020.12.30.424823v1?rss=1">
<title>
<![CDATA[
Inference of emergent spatio-temporal processes from single-cell sequencing reveals feedback between de novo DNA methylation and chromatin condensates 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.12.30.424823v1?rss=1"
</link>
<description><![CDATA[
Recent breakthroughs in single-cell genomics allow probing molecular states of cells with unprecedented detail along the sequence of the DNA. Biological function relies, however, on emergent processes in the three-dimensional space of the nucleus, such as droplet formation through phase separation. Here, we use single-cell multi-omics sequencing to develop a theoretical framework to rigorously map epigenome profiling along the DNA sequence onto a description of the emergent spatial dynamics in the nucleus. Drawing on scNMT-seq multi-omics sequencing in vitro and in vivo we exemplify our approach in the context of exit from pluripotency and global de novo methylation of the genome. We show how DNA methylation patterns of the embryonic genome are established through the interplay between spatially correlated DNA methylation and topological changes to the DNA. This feedback leads to the predicted formation of 30-40nm sized condensates of methylated DNA and determines genome-scale DNA methylation rates. We verify these findings with orthogonal single cell multi-omics data that combine the methylome with HiC measurements. Notably, this scale of chromatin organization has recently been described by super-resolution microscopy. Using this framework, we identify local methylation correlations in gene bodies that precede transcriptional changes at the exit from pluripotency. Our work provides a general framework of how mechanistic insights into emergent processes underlying cell fate decisions can be gained by the combination of single-cell multi-omics and methods from theoretical physics that have not been applied in the context of genomics before.

HighlightsO_LIWe develop methodology to infer collective spatio-temporal processes in the physical space of the nucleus from single-cell methylome sequencing experiments.
C_LIO_LIWe show that DNA methylation relies on a feedback between de novo methylation and nanoscale changes in DNA topology, leading to the formation of methylation condensates.
C_LIO_LIChromatin condensates at this scale have recently been described by high-resolution microscopy but have remained without mechanistic explanation.
C_LIO_LIUsing this framework, we identify changes in the distribution of DNA methylation marks in gene bodies that precede gene silencing at the exit from pluripotency.
C_LI
]]></description>
<dc:creator>Olmeda, F.</dc:creator>
<dc:creator>Lohoff, T.</dc:creator>
<dc:creator>Clark, S.</dc:creator>
<dc:creator>Benson, L.</dc:creator>
<dc:creator>Krueger, F.</dc:creator>
<dc:creator>Reik, W.</dc:creator>
<dc:creator>Rulands, S.</dc:creator>
<dc:date>2021-01-01</dc:date>
<dc:identifier>doi:10.1101/2020.12.30.424823</dc:identifier>
<dc:title><![CDATA[Inference of emergent spatio-temporal processes from single-cell sequencing reveals feedback between de novo DNA methylation and chromatin condensates]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-01-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.11.20.370973v1?rss=1">
<title>
<![CDATA[
Sex of donor cell and reprogramming conditions predict the extent and nature of imprinting defects in mouse iPSCs 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.11.20.370973v1?rss=1"
</link>
<description><![CDATA[
Reprogramming of somatic cells into induced Pluripotent Stem Cells (iPSCs) is a major leap towards personalized approaches to disease modelling and cell-replacement therapies. However, we still lack the ability to fully control the epigenetic status of iPSCs, which is a major hurdle for their downstream applications. A sensible indicator for epigenetic fidelity is genomic imprinting, a phenomenon dependent on DNA methylation, which is frequently perturbed in iPSCs by yet unidentified reasons. By using a secondary reprogramming system with murine hybrid donor cells, we conducted a thorough imprinting analysis using IMPLICON in multiple female and male iPSCs generated under different culture conditions. Our results show that imprinting defects are remarkably common in mouse iPSCs causing dysregulation of the typical monoallelic expression of imprinted genes. Interestingly, the nature of imprinting defects depends on the sex of the donor cell and their respective response to culture conditions. Under serum-free conditions, male iPSCs show global hypomethylation at imprinted regions, whereas in serum conditions show focal hypermethylation at specific loci. In contrast, female iPSCs always exhibit hypomethylation defects regardless of culture conditions. These imprinting defects are more severe than the global changes in DNA methylation, highlighting the sensitivity of imprinting loci to current iPSC generation protocols. Our results reveal clear predictors underlying different types of imprinting defects in mouse iPSCs. This knowledge is essential to devise novel reprogramming strategies aiming at generating epigenetically faithful iPSCs.
]]></description>
<dc:creator>Arez, M.</dc:creator>
<dc:creator>Eckersley-Maslin, M.</dc:creator>
<dc:creator>Klobucar, T.</dc:creator>
<dc:creator>von Gilsa Lopes, J.</dc:creator>
<dc:creator>Krueger, F. A.</dc:creator>
<dc:creator>Raposo, A.</dc:creator>
<dc:creator>Gendrel, A.-V.</dc:creator>
<dc:creator>Bernardes de Jesus, B.</dc:creator>
<dc:creator>Teixeira da Rocha, S.</dc:creator>
<dc:date>2020-11-20</dc:date>
<dc:identifier>doi:10.1101/2020.11.20.370973</dc:identifier>
<dc:title><![CDATA[Sex of donor cell and reprogramming conditions predict the extent and nature of imprinting defects in mouse iPSCs]]></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/cgi/content/short/2020.11.18.388306v1?rss=1">
<title>
<![CDATA[
Subtractive CRISPR screen identifies factors involved in non-canonical LC3 lipidation. 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.11.18.388306v1?rss=1"
</link>
<description><![CDATA[
Although commonly associated with autophagosomes, LC3 can also be recruited to membranes in a variety of non-canonical contexts. These include responses to ionophores such as the M2 proton channel of influenza A virus. LC3 is attached to membranes by covalent lipidation that depends on recruitment of the ATG5-12-16L1 complex. Non-canonical LC3 lipidation requires the C-terminal WD40 domain of ATG16L1 that is dispensable for canonical autophagy. We devised a subtractive CRISPR knock-out screening strategy to investigate the requirements for non-canonical LC3-lipidation. This correctly identified the enzyme complexes directly responsible for LC3-lipidation. We additionally identified the RALGAP complex as important for M2-induced, but not ionophore drug induced LC3 lipidation. In contrast, we identified ATG4D as responsible for LC3 recycling in M2-induced and basal LC3-lipidation. Identification of a vacuolar ATPase subunit in the screen suggested a common mechanism for non-canonical LC3 recruitment. Influenza-induced and ionophore drug induced LC3-lipidation leads to association of the vacuolar ATPase and ATG16L1 and can be antagonised by Salmonella SopF. LC3 recruitment to erroneously neutral compartments may therefore represent a response to damage caused by diverse invasive pathogens.
]]></description>
<dc:creator>Ulferts, R.</dc:creator>
<dc:creator>Marcassa, E.</dc:creator>
<dc:creator>Timimi, L.</dc:creator>
<dc:creator>Lee, L. C.</dc:creator>
<dc:creator>Daley, A.</dc:creator>
<dc:creator>Montaner, B.</dc:creator>
<dc:creator>Turner, S. D.</dc:creator>
<dc:creator>Florey, O.</dc:creator>
<dc:creator>Baillie, J. K.</dc:creator>
<dc:creator>Beale, R.</dc:creator>
<dc:date>2020-11-18</dc:date>
<dc:identifier>doi:10.1101/2020.11.18.388306</dc:identifier>
<dc:title><![CDATA[Subtractive CRISPR screen identifies factors involved in non-canonical LC3 lipidation.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-11-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.10.16.341446v1?rss=1">
<title>
<![CDATA[
Histone modifications form a cell-type-specific chromosomal bar code that modulates and maintains patterns of gene expression through the cell cycle 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.10.16.341446v1?rss=1"
</link>
<description><![CDATA[
BackgroundChromatin configuration influences gene expression in eukaryotes at multiple levels, from individual nucleosomes to chromatin domains several Mb long. Post-translational modifications (PTM) of core histones seem to be involved in chromatin structural transitions, but how remains unclear.

To explore this, we used ChIP-seq and two cell types, HeLa and lymphoblastoid (LCL) to define how changes in chromatin packaging through the cell cycle influence the distributions of three transcription-associated histone modifications, H3K9ac, H3K4me3 and H3K27me3.

ResultsChromosome regions (bands) of 10-50Mb, detectable by immunofluorescence microscopy of metaphase (M) chromosomes, are also present in G1 and G2. We show that they comprise 1-5Mb sub-bands that differ between HeLa and LCL but remain consistent through the cell cycle. The same sub-bands are defined by H3K9ac and H3K4me3, while H3K27me3 spreads more widely.

We found little change between cell cycle phases, whether compared by 5Kb rolling windows or when analysis was restricted to functional elements such as transcription start sites and topologically associating domains.

Only a small number of genes showed cell-cycle related changes: at genes encoding proteins involved in mitosis, H3K9 became highly acetylated in G2M, possibly because of ongoing transcription.

ConclusionsModified histone isoforms H3K9ac, H3K4me3 and H3K27me3 exhibit a characteristic genomic distribution at resolutions of 1Mb and below that differs between HeLa and lymphoblastoid cells but remains remarkably consistent through the cell cycle. We suggest that this cell-type-specific chromosomal bar-code is part of a homeostatic mechanism by which cells retain their characteristic gene expression patterns, and hence their identity, through multiple mitoses.
]]></description>
<dc:creator>Halsall, J. A.</dc:creator>
<dc:creator>Andrews, S.</dc:creator>
<dc:creator>Krueger, F.</dc:creator>
<dc:creator>Rutledge, C.</dc:creator>
<dc:creator>Ficz, G.</dc:creator>
<dc:creator>Reik, W.</dc:creator>
<dc:creator>Turner, B. M.</dc:creator>
<dc:date>2020-10-16</dc:date>
<dc:identifier>doi:10.1101/2020.10.16.341446</dc:identifier>
<dc:title><![CDATA[Histone modifications form a cell-type-specific chromosomal bar code that modulates and maintains patterns of gene expression through the cell cycle]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-10-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.07.28.224832v1?rss=1">
<title>
<![CDATA[
MSK phosphorylation of H3S28 is required for immediate early gene induction and cardiac hypertrophy 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.07.28.224832v1?rss=1"
</link>
<description><![CDATA[
Heart failure is a leading cause of death that develops subsequent to deleterious hypertrophic cardiac remodelling. MAPK pathways play a key role in coordinating the induction of gene expression during hypertrophy. Induction of the immediate early gene (IEG) response is a necessary and early event in this process. How MAPK and IEG expression are coupled during cardiac hypertrophy is not yet resolved. Here, in vitro, in rodent models and in human samples, we demonstrate that MAPK-stimulated IEG induction depends on the Mitogen and Stress activated protein Kinase (MSK) and its phosphorylation of histone H3 at serine 28 (pH3S28). pH3S28 in IEG promoters in turn recruits Brg1, a BAF60 ATP-dependent chromatin remodelling complex component, initiating gene expression. Without MSK activity and IEG induction, the hypertrophic response is suppressed. These studies provide new mechanistic insights and highlight the role of signalling to the epigenome in gene expression regulation during cardiac hypertrophy.

Brief summary one sentenceMSK1/2 phosphorylation of Histone 3 Serine 28 couples MAPK signalling with chromatin remodelling and immediate early gene expression to induce pro-hypertrophic cardiac transcriptional responses.
]]></description>
<dc:creator>Robinson, E. L.</dc:creator>
<dc:creator>Drawnel, F. M.</dc:creator>
<dc:creator>Mehdi, S.</dc:creator>
<dc:creator>Archer, C. R.</dc:creator>
<dc:creator>Liu, W.</dc:creator>
<dc:creator>Okkenhaug, H.</dc:creator>
<dc:creator>Alkass, K.</dc:creator>
<dc:creator>Aronsen, J. M.</dc:creator>
<dc:creator>Nagaraju, C. K.</dc:creator>
<dc:creator>Sjaastad, I.</dc:creator>
<dc:creator>Sipido, K. R.</dc:creator>
<dc:creator>Bergmann, O.</dc:creator>
<dc:creator>Arthur, J. S. C.</dc:creator>
<dc:creator>Wang, X.</dc:creator>
<dc:creator>Roderick, H. L.</dc:creator>
<dc:date>2020-07-29</dc:date>
<dc:identifier>doi:10.1101/2020.07.28.224832</dc:identifier>
<dc:title><![CDATA[MSK phosphorylation of H3S28 is required for immediate early gene induction and cardiac hypertrophy]]></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/cgi/content/short/2020.08.07.241034v1?rss=1">
<title>
<![CDATA[
Epigenetic change induced by in utero dietary challenge provokes phenotypic variability across multiple generations of mice 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.08.07.241034v1?rss=1"
</link>
<description><![CDATA[
Transmission of epigenetic information between generations occurs in nematodes, flies and plants, mediated by specialised small RNA pathways, histone H3K9me3, H3K27me3, H4K16ac and DNA methylation1-3. In higher vertebrates, epidemiological and experimental evidence supports similar trans-generational effects4,5 although the mechanisms that underpin these are incompletely understood6-9. We generated a luciferase reporter knock-in mouse for the imprinted Dlk1 locus, to visualise and track epigenetic fidelity across generations. We showed that exposure to high-fat diet (HFD) in pregnancy provokes sustained re-expression of the normally silent maternal Dlk1 allele in offspring, coincident with increased DNA methylation at the Dlk1 sDMR. Interestingly, maternal Dlk1 mis-expression was also evident in the next generation (F2), exclusively in animals derived from F1-exposed females. Oocytes from these females showed altered microRNA and gene expression, without any major changes in underlying DNA methylation, and correctly imprinted Dlk1 expression resumed in subsequent generations (F3 onwards). Our results reveal how canonical and non-canonical imprinting mechanisms enable the foetal epigenome to adapt to in utero challenge to modulate the properties of two successive generations of offspring.
]]></description>
<dc:creator>Van de Pette, M.</dc:creator>
<dc:creator>Galvao, A.</dc:creator>
<dc:creator>Millership, S. J.</dc:creator>
<dc:creator>To, W.</dc:creator>
<dc:creator>Dimond, A.</dc:creator>
<dc:creator>Prodani, C.</dc:creator>
<dc:creator>McNamara, G.</dc:creator>
<dc:creator>Bruno, L.</dc:creator>
<dc:creator>Sardini, A.</dc:creator>
<dc:creator>Webster, Z.</dc:creator>
<dc:creator>McGinty, J.</dc:creator>
<dc:creator>French, P. M.</dc:creator>
<dc:creator>Uren, A.</dc:creator>
<dc:creator>Castillo-Fernandez, J.</dc:creator>
<dc:creator>John, R. M.</dc:creator>
<dc:creator>Ferguson-Smith, A.</dc:creator>
<dc:creator>Merkenschlager, M.</dc:creator>
<dc:creator>Kelsey, G. D.</dc:creator>
<dc:creator>Fisher, A. G.</dc:creator>
<dc:date>2020-08-07</dc:date>
<dc:identifier>doi:10.1101/2020.08.07.241034</dc:identifier>
<dc:title><![CDATA[Epigenetic change induced by in utero dietary challenge provokes phenotypic variability across multiple generations of mice]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-08-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.02.09.579641v1?rss=1">
<title>
<![CDATA[
Regulation of IFNgamma production by ZFP36L2 in T cells is context-dependent 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.02.09.579641v1?rss=1"
</link>
<description><![CDATA[
CD8+ T cells kill target cells by releasing cytotoxic molecules and pro-inflammatory cytokines, such as TNF and IFN{gamma}. The magnitude and duration of cytokine production is defined by post-transcriptional regulation, and a critical regulator herein are RNA-binding proteins (RBPs). Although the functional importance of RBPs in regulating cytokine production is established, the kinetics and mode of action through which RBPs control cytokine production is not well understood. Previously, we showed that the RBP ZFP36L2 blocks translation of pre-formed cytokine encoding mRNA in quiescent memory T cells. Here, we uncover that ZFP36L2 regulates cytokine production in a context-dependent manner. T cell-specific deletion of ZFP36L2 (CD4-cre) had no effect on T cell development, or on cytokine production during early time points (2-6h) of T cell activation. In contrast, ZFP36L2 specifically dampened the production of IFN{gamma} during prolonged T cell activation (20-48h). ZFP36L2 deficiency also resulted in increased production of IFN{gamma} production in tumour-infiltrating T cells that are chronically exposed to antigen. Mechanistically, ZFP36L2 regulates IFN{gamma} production at late time points of activation by destabilizing Ifng mRNA in an AU-rich element-dependent manner. Together, our results reveal that ZFP36L2 employs different regulatory nodules in effector and memory T cells to regulate cytokine production.
]]></description>
<dc:creator>Zandhuis, N. D.</dc:creator>
<dc:creator>Guislain, A.</dc:creator>
<dc:creator>Popalzij, A.</dc:creator>
<dc:creator>Engels, S.</dc:creator>
<dc:creator>Popovic, B.</dc:creator>
<dc:creator>Turner, M.</dc:creator>
<dc:creator>Wolkers, M. C.</dc:creator>
<dc:date>2024-02-12</dc:date>
<dc:identifier>doi:10.1101/2024.02.09.579641</dc:identifier>
<dc:title><![CDATA[Regulation of IFNgamma production by ZFP36L2 in T cells is context-dependent]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-02-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.08.19.608560v1?rss=1">
<title>
<![CDATA[
The consequences of mismatched buffers in spectral cell sorting 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.08.19.608560v1?rss=1"
</link>
<description><![CDATA[
Although spectral flow cytometry has become a ubiquitous tool for cell analysis, the use of spectral cytometry on cell sorters requires additional considerations arising from the unique requirements of sorting workflows. Here, we show that care should be taken when ascertaining the purity of a sort on a spectral cell sorter, as the mismatch of buffers used for initial sample suspension and the buffers used for sort collection can affect the unmixing of the data, potentially giving rise to erroneous purity check results.
]]></description>
<dc:creator>Dapaah, R. A. S.</dc:creator>
<dc:creator>Ferrer Font, L.</dc:creator>
<dc:creator>Shi, X.</dc:creator>
<dc:creator>Hall, C.</dc:creator>
<dc:creator>Thompson, S.</dc:creator>
<dc:creator>Catharina Costa, L.</dc:creator>
<dc:creator>Mage, P. L.</dc:creator>
<dc:creator>Tyznik, A. J.</dc:creator>
<dc:creator>Lundsten, K.</dc:creator>
<dc:creator>Walker, R. V.</dc:creator>
<dc:date>2024-08-19</dc:date>
<dc:identifier>doi:10.1101/2024.08.19.608560</dc:identifier>
<dc:title><![CDATA[The consequences of mismatched buffers in spectral cell sorting]]></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/cgi/content/short/2024.08.01.604391v1?rss=1">
<title>
<![CDATA[
Protein degradation shapes developmental tempo in mouse and human neural progenitors 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.08.01.604391v1?rss=1"
</link>
<description><![CDATA[
The pace of embryonic development differs markedly across mammalian species, yet the molecular mechanisms underlying these tempo differences remain largely unknown. Here, we systematically compared protein dynamics in mouse and human neural progenitors (NPs) and examined how protein stability influences developmental timing. We find that mouse NPs exhibit faster protein production and degradation than human NPs. Human NPs display broadly increased protein half-lives, independent of cellular compartment or protein function, and this difference persists in post-mitotic neurons. Consistent with this, proteasomal activity is lower in human embryonic spinal cord and stem cell-derived neural progenitors than mouse, correlating with reduced expression of proteasome-associated proteins. Functionally, accelerating the degradation of the key transcriptional repressor IRX3 in mouse NPs speeds the activation of its target gene, providing causal evidence that protein turnover modulates developmental tempo. These results reveal that species-specific regulation of protein degradation shapes the timing of neural development and suggest that evolutionary tuning of proteasomal activity contributes to differences in embryonic developmental pace.
]]></description>
<dc:creator>Nakanoh, S.</dc:creator>
<dc:creator>Stamataki, D.</dc:creator>
<dc:creator>Garcia-Perez, L.</dc:creator>
<dc:creator>Azzi, C.</dc:creator>
<dc:creator>Carr, H. L.</dc:creator>
<dc:creator>Pokhilko, A.</dc:creator>
<dc:creator>Howell, S.</dc:creator>
<dc:creator>Yu, L.</dc:creator>
<dc:creator>Skehel, M.</dc:creator>
<dc:creator>Oxley, D.</dc:creator>
<dc:creator>Andrews, S.</dc:creator>
<dc:creator>Briscoe, J.</dc:creator>
<dc:creator>Rayon, T.</dc:creator>
<dc:date>2024-08-01</dc:date>
<dc:identifier>doi:10.1101/2024.08.01.604391</dc:identifier>
<dc:title><![CDATA[Protein degradation shapes developmental tempo in mouse and human neural progenitors]]></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/cgi/content/short/2024.08.15.607884v1?rss=1">
<title>
<![CDATA[
RAF inhibitors activate the integrated stress response by direct activation of GCN2 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.08.15.607884v1?rss=1"
</link>
<description><![CDATA[
Paradoxical RAF activation by chemical RAF inhibitors (RAFi) is a well-understood  on-target biological and clinical response. In this study, we show that a range of RAFi drive ERK1/2-independent activation of the Unfolded Protein Response (UPR), including expression of ATF4 and CHOP, that required the translation initiation factor eIF2. RAFi-induced ATF4 and CHOP expression was not reversed by inhibition of PERK, a known upstream activator of the eIF2-dependent Integrated Stress Response (ISR). Rather, we found that RAFi exposure activated GCN2, an alternate eIF2 kinase, leading to eIF2-dependent (and ERK1/2-independent) ATF4 and CHOP expression. The GCN2 kinase inhibitor A-92, GCN2 RNAi, GCN2 knock-out or ISRIB (an eIF2 antagonist) all reversed RAFi-induced expression of ATF4 and CHOP indicating that RAFi require GCN2 to activate the ISR. RAFi also activated full-length recombinant GCN2 in vitro and in cells, generating a characteristic  bell-shaped concentration-response curve, reminiscent of RAFi-driven paradoxical activation of WT RAF dimers. Activation of the ISR by RAFi was abolished by GCN2 kinase dead mutations and M802A or M802G gatekeeper mutations, suggesting that RAFi bind directly to the GCN2 kinase domain; this was supported by mechanistic structural models of RAFi interaction with GCN2. Since the ISR is a critical pathway for determining cell survival or death, our observations may be relevant to the clinical use of RAFi, where paradoxical GCN2 activation may be a previously unappreciated off-target effect that may modulate tumour cell responses.
]]></description>
<dc:creator>Gilley, R.</dc:creator>
<dc:creator>Kidger, A. M.</dc:creator>
<dc:creator>Neill, G.</dc:creator>
<dc:creator>Severson, P.</dc:creator>
<dc:creator>Byrne, D. P.</dc:creator>
<dc:creator>Kenneth, N. S.</dc:creator>
<dc:creator>Bollag, G.</dc:creator>
<dc:creator>Zhang, C.</dc:creator>
<dc:creator>de Oliveira, T. M.</dc:creator>
<dc:creator>Eyers, P.</dc:creator>
<dc:creator>Bayliss, R.</dc:creator>
<dc:creator>Masson, G. R.</dc:creator>
<dc:creator>Cook, S. J.</dc:creator>
<dc:date>2024-08-16</dc:date>
<dc:identifier>doi:10.1101/2024.08.15.607884</dc:identifier>
<dc:title><![CDATA[RAF inhibitors activate the integrated stress response by direct activation of GCN2]]></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/cgi/content/short/2024.08.13.607790v1?rss=1">
<title>
<![CDATA[
Eomes directs the formation of spatially and functionally diverse extra-embryonic hematovascular tissues 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.08.13.607790v1?rss=1"
</link>
<description><![CDATA[
During mouse gastrulation, extraembryonic mesoderm (ExEM) contributes to the extraembryonic yolk sac (YS) and allantois, both of which are essential for successful gestation. Although the genetic networks coordinating intra-embryonic mesodermal subtype specification are well-studied, the mechanisms driving ExEM diversification are poorly understood. Here, we reveal that embryoid body in vitro differentiation generates two distinct lineages of mesodermal cells matching YS and allantois respectively. Combining in vitro models with in vivo chimeric embryo analysis, we discover that Eomesodermin (Eomes) regulates the formation of a subset of YS-fated ExEM but is dispensable for allantois formation. Furthermore, simultaneous disruption of Eomes and T impedes the specification of any YS or allantois mesoderm, indicating compensatory roles for T during allantois formation when Eomes is disrupted. Our study highlights previously unrecognized functional and mechanistic diversity in ExEM diversification and endothelial development and introduces a tractable EB model to dissect the signaling pathways and transcriptional networks driving the formation of key extraembryonic tissues.
]]></description>
<dc:creator>Theeuwes, B.</dc:creator>
<dc:creator>Harland, L. T.</dc:creator>
<dc:creator>Bisia, A.</dc:creator>
<dc:creator>Costello, I.</dc:creator>
<dc:creator>Ton, M.-L.</dc:creator>
<dc:creator>Lohoff, T.</dc:creator>
<dc:creator>Clark, S. J.</dc:creator>
<dc:creator>Argelaguet, R.</dc:creator>
<dc:creator>Wilson, N. K.</dc:creator>
<dc:creator>Reik, W.</dc:creator>
<dc:creator>Bikoff, E.</dc:creator>
<dc:creator>Robertson, E. J.</dc:creator>
<dc:creator>Gottgens, B.</dc:creator>
<dc:date>2024-08-16</dc:date>
<dc:identifier>doi:10.1101/2024.08.13.607790</dc:identifier>
<dc:title><![CDATA[Eomes directs the formation of spatially and functionally diverse extra-embryonic hematovascular tissues]]></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/cgi/content/short/2023.08.14.553196v1?rss=1">
<title>
<![CDATA[
The tissue-resident regulatory T cell pool is shaped by transient multi-tissue migration and a conserved residency program 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.08.14.553196v1?rss=1"
</link>
<description><![CDATA[
The tissues are the site of many of the most important immunological reactions, yet the immunology of the tissues has remained relatively opaque. Recent studies have identified Foxp3+ regulatory T cells (Tregs) in several non-lymphoid tissues. These tissue-resident populations have been ascribed unique characteristics based on comparisons to lymphoid Tregs. Here we performed a systematic analysis of the Treg population residing in non-lymphoid organs throughout the body, revealing shared phenotypes, transient residency and common molecular dependencies. Further, tissue Tregs from different non-lymphoid organs shared T cell receptor (TCR) sequences, with functional capacity to drive multi-tissue Treg entry. Finally, tissue Tregs extracted from non-lymphoid organs were tissue-agnostic on re-entry, without homing preference for their tissue of origin. Together these results demonstrate that the tissue-resident Treg pool in most non-lymphoid organs, other than the gut, is largely constituted by broadly self-reactive Tregs, characterised by transient multi-tissue migration and a common residency program.
]]></description>
<dc:creator>Burton, O.</dc:creator>
<dc:creator>Bricard, O.</dc:creator>
<dc:creator>Tareen, S.</dc:creator>
<dc:creator>Gergelits, V.</dc:creator>
<dc:creator>Andrews, S.</dc:creator>
<dc:creator>Roca, C. P.</dc:creator>
<dc:creator>Whyte, C.</dc:creator>
<dc:creator>Junius, S.</dc:creator>
<dc:creator>Brajic, A.</dc:creator>
<dc:creator>Pasciuto, E.</dc:creator>
<dc:creator>Ali, M.</dc:creator>
<dc:creator>Lemaitre, P.</dc:creator>
<dc:creator>Schlenner, S. M.</dc:creator>
<dc:creator>Ishigame, H.</dc:creator>
<dc:creator>Brown, B.</dc:creator>
<dc:creator>Dooley, J.</dc:creator>
<dc:creator>Liston, A.</dc:creator>
<dc:date>2023-08-14</dc:date>
<dc:identifier>doi:10.1101/2023.08.14.553196</dc:identifier>
<dc:title><![CDATA[The tissue-resident regulatory T cell pool is shaped by transient multi-tissue migration and a conserved residency program]]></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/cgi/content/short/2023.12.19.572318v1?rss=1">
<title>
<![CDATA[
Single-cell analysis reveals lasting immunological consequences of influenza infection and respiratory immunisation in the pig lung 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.12.19.572318v1?rss=1"
</link>
<description><![CDATA[
The pig is a natural host for influenza viruses and integrally involved in virus evolution through interspecies transmissions between humans and swine. Swine have many physiological, anatomical, and immunological similarities to humans, and are an excellent model for human influenza. Here, we employed single RNA-sequencing (scRNA-seq) and flow cytometry to characterize the major leucocyte subsets in bronchoalveolar lavage (BAL), twenty-one days after H1N1pdm09 infection or respiratory immunization with an adenoviral vector vaccine expressing haemagglutinin and nucleoprotein with or without IL-1{beta}. Mapping scRNA-seq clusters from BAL onto those previously described in peripheral blood facilitated annotation and highlighted differences between tissue resident and circulating immune cells. ScRNA-seq data and functional assays revealed lasting impacts of immune challenge on BAL populations. First, mucosal administration of IL-1{beta} reduced the number of functionally active Treg. Second, influenza infection upregulated IFI6 in BAL cells, decreasing their susceptibility to virus replication in vitro. Our data provides a reference map of porcine BAL cells and reveals lasting immunological consequences of influenza infection and respiratory immunisation in a highly relevant large animal model for respiratory virus infection.

Author SummaryPigs and humans have a similar anatomy and physiology. In humans, cells from lung-washes are used to study immune responses and it was shown that these cells are crucial in protection against respiratory diseases such as influenza and COVID-19. To better understand lung immunity, we compared genes expressed in cells of pig lung-wash to white blood cells, providing an atlas for future studies of immunity in the lung. We also tested a vaccine given to the lung containing IL-1{beta}, a strong immune activator that protects mice against influenza virus infection. However, although IL-1{beta} increased pig immune responses it did not protect pigs against infection. We also showed that the number of immune cells that dampen immune responses (regulatory T cells) is reduced. In addition, we demonstrated increased expression of a protein, IFI6, 21 days after infection showing that while immune cells in the lung have common properties, the invading organisms influence them significantly. Our study elucidates why some vaccines fail despite inducing powerful immune responses, emphasizes the need for caution when applying results from small animals like mice to humans, and indicates the importance of the pig as a model to study disease in humans and livestock.
]]></description>
<dc:creator>Muir, A.</dc:creator>
<dc:creator>Paudyal, B.</dc:creator>
<dc:creator>Schmidt, S.</dc:creator>
<dc:creator>Sedaghat-Rostani, E.</dc:creator>
<dc:creator>Chakravarti, S.</dc:creator>
<dc:creator>Villanueva Hernandez, S.</dc:creator>
<dc:creator>Moffat, K.</dc:creator>
<dc:creator>Polo, N.</dc:creator>
<dc:creator>Angelopoulos, N.</dc:creator>
<dc:creator>Schmidt, A.</dc:creator>
<dc:creator>Tenbusch, M.</dc:creator>
<dc:creator>Freimanis, G.</dc:creator>
<dc:creator>Gerner, W.</dc:creator>
<dc:creator>Richard, A. C.</dc:creator>
<dc:creator>Tchilian, E.</dc:creator>
<dc:date>2023-12-19</dc:date>
<dc:identifier>doi:10.1101/2023.12.19.572318</dc:identifier>
<dc:title><![CDATA[Single-cell analysis reveals lasting immunological consequences of influenza infection and respiratory immunisation in the pig lung]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-12-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.08.14.606984v1?rss=1">
<title>
<![CDATA[
Paradoxical Activation of GCN2 by ATP-competitive inhibitors via allosteric activation and autophosphorylation 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.08.14.606984v1?rss=1"
</link>
<description><![CDATA[
Recently it has been found that General Control Non-derepressible 2 (GCN2) can be activated by an array of small molecule ATP-competitive inhibitors, including clinically relevant compounds such as Ponatinib, and compounds specifically designed to be GCN2 inhibitors, such as GCN2iB. Furthermore, we recently showed that GCN2 can be activated in cells by clinically approved small molecule RAF inhibitors. GCN2 is a drug target, specifically in cancers such as mesothelioma, and a better understanding of this paradoxical activation is required to develop drugs which truly inhibit the enzyme. Using biochemical assays and structural mass spectrometry, we present a model for how GCN2 is activated by these compounds by promoting an active conformation in the HisRS domain while competitively inhibiting the kinase domain. This conformation promotes activating phosphorylation of GCN2, potentially through phosphorylation of other activated GCN2 molecules which are not bound to compound. Together this model suggests that efforts to inhibit GCN2 would benefit from exploring allosteric routes rather than targeting the ATP-binding pocket of the kinase domain.
]]></description>
<dc:creator>Masson, G. R.</dc:creator>
<dc:creator>Neill, G.</dc:creator>
<dc:creator>Vinciauskaite, V.</dc:creator>
<dc:creator>Paul, M.</dc:creator>
<dc:creator>Gilley, R.</dc:creator>
<dc:creator>Cook, S. J.</dc:creator>
<dc:date>2024-08-14</dc:date>
<dc:identifier>doi:10.1101/2024.08.14.606984</dc:identifier>
<dc:title><![CDATA[Paradoxical Activation of GCN2 by ATP-competitive inhibitors via allosteric activation and autophosphorylation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-08-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.07.10.547212v1?rss=1">
<title>
<![CDATA[
Investigating the effects of radiation, T cell depletion, and bone marrow transplantation on murine gut microbiota 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.07.10.547212v1?rss=1"
</link>
<description><![CDATA[
Microbiome research has gained much attention in recent years as the importance of gut microbiota in regulating host health becomes increasingly evident. However, the impact of radiation on the microbiota in the murine bone marrow transplantation model is still poorly understood. In this paper, we present the major conclusions of our investigation into the effects of radiation and subsequent bone marrow transplantation with or without T cell depletion of the donor bone-marrow on the microbiota of the ileum and cecum. Our findings show that radiation has different effects on the microbiota of the two intestinal regions, with the cecum showing increased interindividual variation, suggesting an impaired ability of the host to regulate microbial symbionts, consistent with the Anna Karenina principle. Additionally, we observed changes in the ileum composition, including an increase in bacterial taxa that are important modulators of host health, such as Akkermansia and Faecalibaculum.

In contrast, radiation in the cecum was associated with an increased abundance of several common commensal taxa in the gut, including Lachnospiraceae and Bacteroides. Finally, we found that high doses of radiation had more substantial effects on the caecal microbiota of the T-cell-depleted group than that of the non-T-cell-depleted group. Overall, our results contribute to a better understanding of the complex relationship between radiation and the gut microbiota in the context of bone marrow transplantation and highlight the importance of considering different intestinal regions when studying microbiome responses to environmental stressors.
]]></description>
<dc:creator>Kreisinger, J.</dc:creator>
<dc:creator>Dooley, J.</dc:creator>
<dc:creator>Singh, K.</dc:creator>
<dc:creator>Cizkova, D.</dc:creator>
<dc:creator>Schmiedova, L.</dc:creator>
<dc:creator>Bendova, B.</dc:creator>
<dc:creator>Liston, A.</dc:creator>
<dc:creator>Moudra, A.</dc:creator>
<dc:date>2023-07-10</dc:date>
<dc:identifier>doi:10.1101/2023.07.10.547212</dc:identifier>
<dc:title><![CDATA[Investigating the effects of radiation, T cell depletion, and bone marrow transplantation on murine gut microbiota]]></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/cgi/content/short/2024.06.28.601066v1?rss=1">
<title>
<![CDATA[
Phospholipase Cgamma regulates lacrimal gland branching by competing with PI3K in phosphoinositide metabolism 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.06.28.601066v1?rss=1"
</link>
<description><![CDATA[
Although the regulation of branching morphogenesis by spatially distributed cues is well established, the role of intracellular signaling in determining the branching pattern remains poorly understood. In this study, we investigated the regulation and function of phospholipase C gamma (PLC{gamma}) in Fibroblast Growth Factor (FGF) signaling in lacrimal gland development. We showed that deletion of PLC{gamma}1 in the lacrimal gland epithelium leads to ectopic branching and acinar hyperplasia, which was phenocopied by either mutating the PLC{gamma}1 binding site on Fgfr2 or disabling any of its SH2 domains. PLC{gamma}1 inactivation did not change the level of Fgfr2 or affect MAPK signaling, but instead led to sustained AKT phosphorylation due to increased PIP3 production. Consistent with this, PLC{gamma}1 mutant phenotype can be reproduced by elevation of PI3K signaling in Pten knockout and attenuated by blocking AKT signaling. This study demonstrated that PLC{gamma} modulates PI3K signaling by shifting phosphoinositide metabolism, revealing an important role of signaling dynamics in conjunction with spatial cues in shaping branching morphogenesis.
]]></description>
<dc:creator>Wang, Q.</dc:creator>
<dc:creator>Tao, C.</dc:creator>
<dc:creator>Wu, Y.</dc:creator>
<dc:creator>Anderson, K. E.</dc:creator>
<dc:creator>Hannan, A.</dc:creator>
<dc:creator>Lin, C.-s.</dc:creator>
<dc:creator>Hawkins, P. T.</dc:creator>
<dc:creator>Stephens, L.</dc:creator>
<dc:creator>Zhang, X.</dc:creator>
<dc:date>2024-07-02</dc:date>
<dc:identifier>doi:10.1101/2024.06.28.601066</dc:identifier>
<dc:title><![CDATA[Phospholipase Cgamma regulates lacrimal gland branching by competing with PI3K in phosphoinositide metabolism]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-07-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.09.29.560184v1?rss=1">
<title>
<![CDATA[
Stability of Genomic Imprinting and X-Chromosome Inactivation in the Aging Brain 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.09.29.560184v1?rss=1"
</link>
<description><![CDATA[
Epigenetic drift is a hallmark of aging that contributes to the irreversible decline in organismal fitness ultimately leading to aging-related diseases. Epigenetic modifications regulate the cellular memory of the epigenetic processes of genomic imprinting and X-chromosome inactivation to ensure monoallelic expression of imprinted and X-linked genes. Whether epigenetic drift affects maintenance of genomic imprinting and X-chromosome inactivation has not been comprehensively studied. Here, we investigate the allele-specific transcriptional and epigenetic signatures of the aging brain, by comparing juvenile and old hybrid mice obtained from C57BL/6J (BL6) & CAST/EiJ (CAST) reciprocal crosses, with an emphasis on the hippocampus (HCP). We confirm that the aged HCP shows an increase of DNA hydroxymethylation, a sign of epigenetic drift, and a typical aging transcriptional signature. Genomic imprinting was found to be largely unaffected with stable parent-of-origin-specific DNA methylation in HCP, but also other brain regions such as the cerebellum (CB), nucleus accumbens, hypothalamus and prefrontal cortex. Consistently, transcriptomics analysis confirmed unaltered imprinting expression in the aged HCP. An exception are three novel non-coding transcripts (B230209E15Rik, Ube2nl and A330076H08Rik) at the Prader-Willi syndrome/Angelman syndrome (PWS/AS) imprinted locus which lose strict monoallelic expression during aging. Like imprinting, X-chromosome inactivation was remarkably stable with no signs of aging-driven skewing or relaxation of monoallelic expression of X-linked genes. Our study provides a valuable resource for evaluating monoallelic expression in the aging brain and reveals that, despite epigenetic drift during aging, genomic imprinting and X-chromosome inactivation remain predominantly stable throughout the process of physiological aging in the mouse brain.
]]></description>
<dc:creator>Mancino, S.</dc:creator>
<dc:creator>Seneviratne, J.</dc:creator>
<dc:creator>Mupo, A.</dc:creator>
<dc:creator>Krueger, F.</dc:creator>
<dc:creator>Oxley, D.</dc:creator>
<dc:creator>Eckersley-Maslin, M.</dc:creator>
<dc:creator>da Rocha, S. T.</dc:creator>
<dc:date>2023-09-30</dc:date>
<dc:identifier>doi:10.1101/2023.09.29.560184</dc:identifier>
<dc:title><![CDATA[Stability of Genomic Imprinting and X-Chromosome Inactivation in the Aging Brain]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-09-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.10.07.561361v1?rss=1">
<title>
<![CDATA[
Whole Genome Methylation Sequencing via Enzymatic Conversion (EM-seq): Protocol, Data Processing and Analysis 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.10.07.561361v1?rss=1"
</link>
<description><![CDATA[
Whole genome bisulfite sequencing (WGBS) has been the gold standard technique for base resolution analysis of DNA methylation for the last 15 years. It has been, however, associated with technical biases, which lead to overall overestimation of global and regional methylation values, and significant artifacts in extreme cytosine-rich DNA sequence contexts. Enzymatic conversion of cytosine is the newest approach, set to replace entirely the use of the damaging bisulfite conversion of DNA. The EM-seq technique utilises TET2, T4-BGT and APOBEC in a two-step conversion process, where the modified cytosines are first protected by oxidation and glucosylation, followed by deamination of all unmodified cytosines to uracil. As a result, EM-seq is degradation-free and bias-free, requires low DNA input, and produces high library yields with longer reads, little batch variation, less duplication, uniform genomic coverage, accurate methylation over a larger number of captured CpGs, and no sequence-specific artifacts.
]]></description>
<dc:creator>Olova, N. N.</dc:creator>
<dc:creator>Andrews, S.</dc:creator>
<dc:date>2023-10-10</dc:date>
<dc:identifier>doi:10.1101/2023.10.07.561361</dc:identifier>
<dc:title><![CDATA[Whole Genome Methylation Sequencing via Enzymatic Conversion (EM-seq): Protocol, Data Processing and Analysis]]></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/cgi/content/short/2024.10.22.619613v1?rss=1">
<title>
<![CDATA[
A distinctive PI(4,5)P2 compartment forms during entosis and related engulfment processes 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.10.22.619613v1?rss=1"
</link>
<description><![CDATA[
Entosis is a form of cell cannibalism prevalent in human tumours. During entosis, one live and viable epithelial cell is completely internalised by another, then housed inside a large, single-membrane, endolysosomal vacuole in the host cytosol. The composition and maturation of this specialised, macroendocytic compartment has not been fully defined, but ultimately, the inner cell is killed and digested by host lysosomes. Here, we investigate the molecular characteristics and maturation profile of the entotic vacuole. Like phagosomes and macropinosomes, this vacuole undergoes a series of phospholipid modifications, but its maturation profile bears distinctive dynamics. While PI(4,5)P2 is lost rapidly during phagosome maturation, entosis yields an unusual, intracellular PI(4,5)P2-positive compartment, that can persist for hours, suggesting vacuole maturation is uncoupled from membrane scission. Loss of PI(4,5)P2 is eventually triggered, a requisite step for lysosomal killing of the internalised cell. More broadly, PI(4,5)P2-positive vacuoles also form during T-cell engulfment by thymic nurse cells, dependent on ROCK activity, suggesting this distinctive compartment represents a shared feature of entosis-like cell engulfments.
]]></description>
<dc:creator>Durgan, J.</dc:creator>
<dc:creator>Sloan, K.</dc:creator>
<dc:creator>Domart, M.-C.</dc:creator>
<dc:creator>Collinson, L.</dc:creator>
<dc:creator>Florey, O.</dc:creator>
<dc:date>2024-10-22</dc:date>
<dc:identifier>doi:10.1101/2024.10.22.619613</dc:identifier>
<dc:title><![CDATA[A distinctive PI(4,5)P2 compartment forms during entosis and related engulfment processes]]></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/cgi/content/short/2023.10.29.564595v1?rss=1">
<title>
<![CDATA[
The N-terminal region of DNMT3A combines multiple chromatin reading motifs to guide recruitment. 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.10.29.564595v1?rss=1"
</link>
<description><![CDATA[
DNA methyltransferase 3A (DNMT3A) plays a critical role in establishing and maintaining DNA methylation patterns. However, the mechanisms underlying DNMT3A recruitment to and function within different chromatin environments remain unclear. Using a combination of biochemical and structural approaches we find that DNMT3A interacts using multiple interfaces with chromatin; directly binding generic nucleosome features as well as site-specific post-translational histone modifications. The N-terminal region, unique to the DNMT3A1 isoform, is essential for these interactions and stabilises H3K36me2-nucleosome recruitment. Intriguingly, in the same region critical for nucleosome binding we also map a ubiquitylation-dependent recruitment motif (UDR). The UDR binds specifically to ubiquitylated H2AK119, explaining the previously observed recruitment to Polycomb-occupied heterochromatin. A cryo-EM structure of DNMT3A1-DNMT3L with a modified nucleosome reveals that the UDR interacts with the nucleosome surface including the acidic patch. Previously unexplained disease-associated mutations are present in the UDR and ablate nucleosome interactions. This leads to an increased understanding of how DNMT3A1 recruitment occurs in the genome and highlights the importance of multivalent binding of DNMT3A to histone modifications and the nucleosome.
]]></description>
<dc:creator>Wapenaar, H.</dc:creator>
<dc:creator>Clifford, G.</dc:creator>
<dc:creator>Rolls, W.</dc:creator>
<dc:creator>Burdett, H.</dc:creator>
<dc:creator>Zhang, Y.</dc:creator>
<dc:creator>Deak, G.</dc:creator>
<dc:creator>Zou, J.</dc:creator>
<dc:creator>Taylor, M. R.</dc:creator>
<dc:creator>Mills, J.</dc:creator>
<dc:creator>Watson, J. A.</dc:creator>
<dc:creator>Kumar, D.</dc:creator>
<dc:creator>Das, A.</dc:creator>
<dc:creator>Valsakumar, D.</dc:creator>
<dc:creator>Bramham, J.</dc:creator>
<dc:creator>Voigt, P.</dc:creator>
<dc:creator>Wilson, M. D.</dc:creator>
<dc:date>2023-10-29</dc:date>
<dc:identifier>doi:10.1101/2023.10.29.564595</dc:identifier>
<dc:title><![CDATA[The N-terminal region of DNMT3A combines multiple chromatin reading motifs to guide recruitment.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.07.11.548598v1?rss=1">
<title>
<![CDATA[
Chromatin remodeling of histone H3 variants underlies epigenetic inheritance of DNA methylation 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.07.11.548598v1?rss=1"
</link>
<description><![CDATA[
Epigenetic inheritance refers to the faithful replication of DNA methylation and histone modification independent of DNA sequence. Nucleosomes block access to DNA methyltransferases, unless they are remodeled by DECREASE IN DNA METHYLATION1 (DDM1Lsh/HELLS), a Snf2-like master regulator of epigenetic inheritance. We show that DDM1 activity results in replacement of the transcriptional histone variant H3.3 for the replicative variant H3.1 during the cell cycle. In ddm1 mutants, DNA methylation can be restored by loss of the H3.3 chaperone HIRA, while the H3.1 chaperone CAF-1 becomes essential. The single-particle cryo-EM structure at 3.2 [A] of DDM1 with a variant nucleosome reveals direct engagement at SHL2 with histone H3.3 at or near variant residues required for assembly, as well as with the deacetylated H4 tail. An N-terminal autoinhibitory domain binds H2A variants to allow remodeling, while a disulfide bond in the helicase domain is essential for activity in vivo and in vitro. We show that differential remodeling of H3 and H2A variants in vitro reflects preferential deposition in vivo. DDM1 co-localizes with H3.1 and H3.3 during the cell cycle, and with the DNA methyltransferase MET1Dnmt1. DDM1 localization to the chromosome is blocked by H4K16 acetylation, which accumulates at DDM1 targets in ddm1 mutants, as does the sperm cell specific H3.3 variant MGH3 in pollen, which acts as a placeholder nucleosome in the germline and contributes to epigenetic inheritance.
]]></description>
<dc:creator>Lee, S. C.</dc:creator>
<dc:creator>Adams, D. W.</dc:creator>
<dc:creator>Ipsaro, J. J.</dc:creator>
<dc:creator>Cahn, J.</dc:creator>
<dc:creator>Lynn, J.</dc:creator>
<dc:creator>Kim, H.-S.</dc:creator>
<dc:creator>Berube, B.</dc:creator>
<dc:creator>Major, V.</dc:creator>
<dc:creator>Calarco, J. P.</dc:creator>
<dc:creator>LeBlanc, C.</dc:creator>
<dc:creator>Bhattacharjee, S.</dc:creator>
<dc:creator>Ramu, U.</dc:creator>
<dc:creator>Grimanelli, D.</dc:creator>
<dc:creator>Jacob, Y.</dc:creator>
<dc:creator>Voigt, P.</dc:creator>
<dc:creator>Joshua-Tor, L.</dc:creator>
<dc:creator>Martienssen, R. A.</dc:creator>
<dc:date>2023-07-11</dc:date>
<dc:identifier>doi:10.1101/2023.07.11.548598</dc:identifier>
<dc:title><![CDATA[Chromatin remodeling of histone H3 variants underlies epigenetic inheritance of DNA methylation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-07-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.05.04.442453v1?rss=1">
<title>
<![CDATA[
Histone marks are drivers of the splicing changes necessary for an epithelial-to-mesenchymal transition 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.05.04.442453v1?rss=1"
</link>
<description><![CDATA[
Cell differentiation and reprogramming depend on coordinated changes in specific alternative splicing events. How these cell type-specific splicing patterns are dynamically modified in response to a stimulus remains elusive. Taking advantage of the epithelial-to-mesenchymal transition (EMT), a reversible cell reprogramming intimately involved in cancer cell invasiveness and metastasis, we found a strong correlation between changes in the alternative splicing of key exons for EMT, such as at the Fgfr2 and Cnntd1 loci, and changes in the enrichment levels of specific histone modifications, namely H3K27ac and H3K27me3. Localised CRISPR epigenome editing of these exon-specific histone marks was sufficient to induce changes in splicing capable of recapitulating important aspects of EMT, such as a motile and invasive cell phenotype. Whereas, impairment of the changes in H3K27 marks observed during EMT, using histone deacetylase inhibitors, repressed inclusion of the mesenchymal isoform despite an EMT induction, supporting a driving effect for H3K27 modifications in establishing the new cell type-specific splicing patterns necessary for EMT cell reprogramming. Finally, H3K27 marks were shown to impact splicing by modulating recruitment of the splicing factor PTB to its RNA binding sites, suggesting a direct link between chromatin modifications and the splicing machinery. Taken together, these results prove the causal role of H3K27 marks in driving the dynamic splicing changes necessary for induction of important aspects of EMT. They also prove that chromatin-mediated splicing changes are sufficient to impact the cells phenotype, which expands the cells toolkit to adapt and respond to diverse stimuli, such as EMT induction.
]]></description>
<dc:creator>Segelle, A.</dc:creator>
<dc:creator>Nunez-Alvarez, Y.</dc:creator>
<dc:creator>Oldfield, A. J.</dc:creator>
<dc:creator>Webb, K. M.</dc:creator>
<dc:creator>Voigt, P.</dc:creator>
<dc:creator>Luco, R. F.</dc:creator>
<dc:date>2021-05-04</dc:date>
<dc:identifier>doi:10.1101/2021.05.04.442453</dc:identifier>
<dc:title><![CDATA[Histone marks are drivers of the splicing changes necessary for an epithelial-to-mesenchymal transition]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-05-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.02.26.432832v1?rss=1">
<title>
<![CDATA[
H3K36 methylation and DNA-binding are critical for Ioc4 recruitment and Isw1b remodeller function 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.02.26.432832v1?rss=1"
</link>
<description><![CDATA[
The Isw1b chromatin-remodelling complex is specifically recruited to gene bodies to help retain pre-existing histones during transcription by RNA polymerase II. Recruitment is dependent on H3K36 methylation and the Isw1b subunit Ioc4, which contains an N-terminal PWWP domain. Here, we present the crystal structure of the Ioc4-PWWP domain, including a detailed functional characterization of the domain on its own as well as in the context of full-length Ioc4 and the Isw1b remodeller. The Ioc4-PWWP domain preferentially binds H3K36me3-containing nucleosomes. Its ability to bind DNA is required for nucleosome binding. It is also furthered by the unique insertion motif present in Ioc4-PWWP. The ability to bind H3K36me3 and DNA promote the interaction of full-length Ioc4 with nucleosomes in vitro and they are necessary for its recruitment to gene bodies in vivo. Furthermore, a fully functional Ioc4-PWWP domain promotes efficient remodelling by Isw1b and the maintenance of ordered chromatin in vivo, thereby preventing the production of non-coding RNAs.
]]></description>
<dc:creator>Li, J.</dc:creator>
<dc:creator>Bergmann, L.</dc:creator>
<dc:creator>Webb, K. M.</dc:creator>
<dc:creator>Gogol, M. M.</dc:creator>
<dc:creator>Voigt, P.</dc:creator>
<dc:creator>Liu, Y.</dc:creator>
<dc:creator>Liang, H.</dc:creator>
<dc:creator>Smolle, M. M.</dc:creator>
<dc:date>2021-02-26</dc:date>
<dc:identifier>doi:10.1101/2021.02.26.432832</dc:identifier>
<dc:title><![CDATA[H3K36 methylation and DNA-binding are critical for Ioc4 recruitment and Isw1b remodeller function]]></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/cgi/content/short/2021.03.04.433942v1?rss=1">
<title>
<![CDATA[
The memory B cell response to influenza vaccination is impaired in older persons 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.03.04.433942v1?rss=1"
</link>
<description><![CDATA[
Influenza imparts an age-related increase in mortality and morbidity. The most effective countermeasure is vaccination; however, vaccines offer modest protection in older adults. To investigate how ageing impacts the memory B cell response we tracked haemagglutinin specific B cells by indexed flow sorting and single cell RNA sequencing in twenty healthy adults administered the trivalent influenza vaccine. We found age-related skewing in the memory B cell compartment six weeks after vaccination, with younger adults developing haemagglutinin specific memory B cells with an FCRL5+ "atypical" phenotype, showing evidence of somatic hypermutation and positive selection, which happened to a lesser extent in older persons. We confirmed the germinal center ancestry of these FCRL5+ "atypical" memory B cells using scRNASeq from fine needle aspirates of influenza responding human lymph nodes and paired blood samples. Together, this study shows that the aged human germinal center reaction and memory B cell response following vaccination is defective.

SummaryImmune responses to vaccination wane with age. Using single cell RNA sequencing of influenza vaccine specific B cells, this study delineates changes in B cell memory generation, antibody mutation and their subsequent selection in older persons.
]]></description>
<dc:creator>Carr, E. J.</dc:creator>
<dc:creator>Wheatley, A. K.</dc:creator>
<dc:creator>Hill, D. L.</dc:creator>
<dc:creator>Linterman, M. A.</dc:creator>
<dc:date>2021-03-04</dc:date>
<dc:identifier>doi:10.1101/2021.03.04.433942</dc:identifier>
<dc:title><![CDATA[The memory B cell response to influenza vaccination is impaired in older persons]]></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/cgi/content/short/2023.08.04.551986v1?rss=1">
<title>
<![CDATA[
The Fork Protection Complex protects long replicons from DNA damage at the cost of genome instability induced by DNA topological stress. 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.08.04.551986v1?rss=1"
</link>
<description><![CDATA[
Tof1/Timeless protects eukaryotic cells from DNA replication stress as part of the Fork Protection Complex (FPC). Tof1 supports rapid DNA replication, fork pausing, and resolution of DNA topological stress. Here, we show that disruption of FPC function through loss of either Tof1 or Mrc1 results in DNA damage in long replicons. Despite increasing DNA damage in long replicons, loss of either Tof1 or Mrc1 concurrently reduces DNA damage in regions prone to damage caused by DNA topological stress, indicating that the rapid replication promoted by the FPC fosters completing DNA replication at the cost of increased vulnerability to DNA topological stress. Supporting this we find that a tof1 mutation that selectively inhibits DNA topological stress resolution increases DNA damage in contexts prone to DNA topological stress. Our data indicates that the FPC balances rapid replication with recruitment of topoisomerase I to resolve the topological stress generated by increased DNA unwinding.
]]></description>
<dc:creator>Keszthelyi, A.</dc:creator>
<dc:creator>Whale, A.</dc:creator>
<dc:creator>Houseley, J.</dc:creator>
<dc:creator>Baxter, J.</dc:creator>
<dc:date>2023-08-04</dc:date>
<dc:identifier>doi:10.1101/2023.08.04.551986</dc:identifier>
<dc:title><![CDATA[The Fork Protection Complex protects long replicons from DNA damage at the cost of genome instability induced by DNA topological stress.]]></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/cgi/content/short/2024.01.09.574858v1?rss=1">
<title>
<![CDATA[
p110α-dependent hepatocyte signaling is critical for liver gene expression and its rewiring in MASLD 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.01.09.574858v1?rss=1"
</link>
<description><![CDATA[
Insulin and other growth factors are key regulators of liver gene expression, including in metabolic diseases. Most of the phosphoinositide 3-kinase (PI3K) activity induced by insulin is dependent on PI3K. We used mice lacking p110, the catalytic subunit of PI3K, to investigate its role in the regulation of liver gene expression in health and in Metabolic dysfunction-Associated Steatotic Liver Disease (MASLD). The absence of hepatocyte PI3K signaling promoted glucose intolerance in lean mice and significantly regulated liver gene expression, including insulin-sensitive genes, in ad libitum feeding. Some of the defective regulation of gene expression in response to hepatocyte-restricted insulin receptor deletion was related to PI3K signaling. In addition, though PI3K deletion in hepatocytes promoted insulin resistance, it was protective against steatotic liver disease in diet-induced obesity. In the absence of hepatocyte PI3K, the effect of diet-induced obesity on liver gene expression was significantly altered, with changes in rhythmic gene expression in liver. Altogether, this study highlights the specific role of p110 in the control of liver gene expression in physiology and in the metabolic rewiring that occurs during MASLD.
]]></description>
<dc:creator>Regnier, M.</dc:creator>
<dc:creator>Polizzi, A.</dc:creator>
<dc:creator>Fougeray, T.</dc:creator>
<dc:creator>Fougerat, A.</dc:creator>
<dc:creator>Perrier, P.</dc:creator>
<dc:creator>Anderson, K.</dc:creator>
<dc:creator>Lippi, Y.</dc:creator>
<dc:creator>Smati, S.</dc:creator>
<dc:creator>Lukowicz, C.</dc:creator>
<dc:creator>Lasserre, F.</dc:creator>
<dc:creator>Fouche, E.</dc:creator>
<dc:creator>Huillet, M.</dc:creator>
<dc:creator>Rives, C.</dc:creator>
<dc:creator>Tramunt, B.</dc:creator>
<dc:creator>Naylies, C.</dc:creator>
<dc:creator>Garcia, G.</dc:creator>
<dc:creator>Rousseau-Bacquie, E.</dc:creator>
<dc:creator>Bertrand-Michel, J.</dc:creator>
<dc:creator>Canlet, C.</dc:creator>
<dc:creator>Chevolleau-Mege, S.</dc:creator>
<dc:creator>Debrauwer, L.</dc:creator>
<dc:creator>Heymes, C.</dc:creator>
<dc:creator>Burcelin, R.</dc:creator>
<dc:creator>Levade, T.</dc:creator>
<dc:creator>Gourdy, P.</dc:creator>
<dc:creator>Wahli, W.</dc:creator>
<dc:creator>Blum, Y.</dc:creator>
<dc:creator>Gamet-Payrastre, L.</dc:creator>
<dc:creator>Ellero-Simatos, S.</dc:creator>
<dc:creator>Guillermet-Guibert, J.</dc:creator>
<dc:creator>Hawkins, P.</dc:creator>
<dc:creator>Stephens, L.</dc:creator>
<dc:creator>Montagner, A.</dc:creator>
<dc:creator>Loiseau, N.</dc:creator>
<dc:creator>Guillou, H.</dc:creator>
<dc:date>2024-01-10</dc:date>
<dc:identifier>doi:10.1101/2024.01.09.574858</dc:identifier>
<dc:title><![CDATA[p110α-dependent hepatocyte signaling is critical for liver gene expression and its rewiring in MASLD]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-01-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.02.26.580534v1?rss=1">
<title>
<![CDATA[
A targetable PREX2/RAC1/PI3Kβ signalling axis confers resistance to clinically relevant therapeutic approaches in melanoma 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.02.26.580534v1?rss=1"
</link>
<description><![CDATA[
Metastatic melanoma remains a major clinical challenge. Large-scale genomic sequencing of melanoma has identified bona fide activating mutations in RAC1, with mutations of its upstream regulator, the RAC-GEF PREX2, also commonly detected. Crucially, RAC1 mutations are associated with resistance to BRAF-targeting therapies. Despite the role of its homologue PREX1 in melanomagenesis, and evidence that some truncating PREX2 mutations drive increased RAC1 activity, no hotspot mutations have been identified, and the impact of PREX2 mutation remains contentious. Here, we use genetically engineered mouse models and patient-derived BRAFV600E-driven melanoma cell lines to dissect the role of PREX2 in melanomagenesis and response to therapy. We show that while PREX2 is dispensable for the initiation and progression of melanoma, its loss confers sensitivity to clinically relevant therapeutics. Importantly, genetic and pharmacological targeting of the RAC1 effector kinase PI3K{beta} phenocopies PREX2 loss, sensitizing our model systems to therapy. Our data reveal a druggable PREX2/RAC1/PI3K{beta} signalling axis in BRAF-mutant melanoma that could be exploited clinically.

Statement of SignificanceMetastatic melanoma remains both a clinical problem, and an opportunity for therapeutic benefit. Co-targeting of the MAPK pathway and the PREX2/RAC1/PI3K{beta} has remarkable efficacy and outperforms monotherapy MAPK targeting in vivo.
]]></description>
<dc:creator>Ford, C. A.</dc:creator>
<dc:creator>Koludrovic, D.</dc:creator>
<dc:creator>Centeno, P. P.</dc:creator>
<dc:creator>Foth, M.</dc:creator>
<dc:creator>Tsonou, E.</dc:creator>
<dc:creator>Vlahov, N.</dc:creator>
<dc:creator>Sphyris, N.</dc:creator>
<dc:creator>Gilroy, K.</dc:creator>
<dc:creator>Bull, C.</dc:creator>
<dc:creator>Nixon, C.</dc:creator>
<dc:creator>Serrels, B.</dc:creator>
<dc:creator>Munro, A. F.</dc:creator>
<dc:creator>Dawson, J. C.</dc:creator>
<dc:creator>Carragher, N. O.</dc:creator>
<dc:creator>Pavet, V.</dc:creator>
<dc:creator>Hornigold, D. C.</dc:creator>
<dc:creator>Dunne, P. D.</dc:creator>
<dc:creator>Downward, J.</dc:creator>
<dc:creator>Welch, H. C. E.</dc:creator>
<dc:creator>Barry, S. T.</dc:creator>
<dc:creator>Sansom, O. J.</dc:creator>
<dc:creator>Campbell, A. D.</dc:creator>
<dc:date>2024-02-28</dc:date>
<dc:identifier>doi:10.1101/2024.02.26.580534</dc:identifier>
<dc:title><![CDATA[A targetable PREX2/RAC1/PI3Kβ signalling axis confers resistance to clinically relevant therapeutic approaches in melanoma]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-02-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.01.16.524218v1?rss=1">
<title>
<![CDATA[
Dbx2, an aging-related homeobox gene, inhibits the proliferation of adult neural progenitors 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.01.16.524218v1?rss=1"
</link>
<description><![CDATA[
The subventricular zone (SVZ) of the adult mouse brain contains quiescent neural stem cells, which can be activated (aNSCs) to generate transit amplifying progenitors (TAPs), neuroblasts (NBs) and newborn neurons. Neurogenesis declines during aging, as the aged SVZ niche causes transcriptomic changes that promote NSC quiescence and decrease proliferating neural/stem progenitor cells (NSPCs). The transcription factors mediating these changes, however, remain unclear. We previously found that the homeobox gene Dbx2 is upregulated in aged SVZ NSPCs and inhibits NSPC culture growth. Here, we report that Dbx2 is repressed by Epidermal Growth Factor Receptor signaling, which promotes NSPC proliferation and decreases in the aged SVZ. We show that Dbx2 inhibits NSPC proliferation by hindering the G2/M transition and elucidate the transcriptomic networks modulated by Dbx2, highlighting its role in the downregulation of the cell cycle molecular pathways. Accordingly, Dbx2 function is negatively correlated with the transcriptional signatures of proliferative NSPCs (aNSCs, TAPs and early NBs). These results point to Dbx2 as a molecular node relaying the anti-neurogenic input of the aged niche to the NSPC transcriptome.
]]></description>
<dc:creator>Giuliani, A.</dc:creator>
<dc:creator>Licursi, V.</dc:creator>
<dc:creator>Nisi, P. S.</dc:creator>
<dc:creator>Fiore, M.</dc:creator>
<dc:creator>Biagioni, S.</dc:creator>
<dc:creator>Negri, R.</dc:creator>
<dc:creator>Rugg-Gunn, P.</dc:creator>
<dc:creator>Cacci, E.</dc:creator>
<dc:creator>Lupo, G.</dc:creator>
<dc:date>2023-01-19</dc:date>
<dc:identifier>doi:10.1101/2023.01.16.524218</dc:identifier>
<dc:title><![CDATA[Dbx2, an aging-related homeobox gene, inhibits the proliferation of adult neural progenitors]]></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/cgi/content/short/2023.05.29.541316v1?rss=1">
<title>
<![CDATA[
Delay of human early development via in vitro diapause 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.05.29.541316v1?rss=1"
</link>
<description><![CDATA[
Many mammals can control the timing of gestation and birth by pausing embryonic development at the blastocyst stage. It is unknown whether the capacity to pause development is conserved, in general across mammals, and more specifically in humans. Activity of the growth regulating mTOR pathway governs developmental pausing in the mouse (1). Here we show a stage-specific capacity to delay the progression of human development via mTOR inhibition. In this context, human blastoids and pluripotent stem cells in naive and naive-like, but not primed, states can be induced to enter a dormant state, which is reversible at the functional and molecular level. Comparative analysis of mouse and human naive cells longitudinal response to mTORi revealed distinct temporal dynamics and metabolic requirements of dormancy in each species. Mouse and human blastocysts show similar tissue-specific patterns of mTOR pathway activity, suggesting that the mTOR pathway may be a conserved regulator of blastocyst development and timing in both species. Our results raise the possibility that the developmental timing of the human embryo may be controllable, with implications for reproductive therapies.
]]></description>
<dc:creator>Bulut-Karslioglu, A.</dc:creator>
<dc:creator>Iyer, D.</dc:creator>
<dc:creator>van der Weijden, V.</dc:creator>
<dc:creator>Heidari Khoei, H.</dc:creator>
<dc:creator>McCarthy, A.</dc:creator>
<dc:creator>Rayon, T.</dc:creator>
<dc:creator>Simon, C.</dc:creator>
<dc:creator>Dunkel, I.</dc:creator>
<dc:creator>Wamaitha, S.</dc:creator>
<dc:creator>Schulz, E.</dc:creator>
<dc:creator>Niakan, K.</dc:creator>
<dc:creator>Rivron, N. C.</dc:creator>
<dc:date>2023-05-29</dc:date>
<dc:identifier>doi:10.1101/2023.05.29.541316</dc:identifier>
<dc:title><![CDATA[Delay of human early development via in vitro diapause]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-05-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.10.19.618135v1?rss=1">
<title>
<![CDATA[
KRAS G12 mutant alleles differentially control glutamine metabolism via FOXO1 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.10.19.618135v1?rss=1"
</link>
<description><![CDATA[
Mutations in KRAS, particularly at codon 12, are frequent in adenocarcinomas of the colon, lungs and pancreas, driving carcinogenesis by altering cell signalling and reprogramming metabolism. However, the specific mechanisms by which different KRAS G12 alleles initiate distinctive patterns of metabolic reprogramming is unclear. Using isogenic panels of colorectal cell lines harbouring the G12A, G12C, G12D and G12V heterozygous mutations and employing transcriptomics, metabolomics, and extensive biochemical validation, we demonstrate distinctive features of each allele. We also demonstrate that cells harbouring the common G12D and G12V oncogenic mutations significantly alter glutamine metabolism and nitrogen recycling through FOXO1-mediated regulation compared to parental lines. Moreover, with a combination of small molecule inhibitors targeting glutamine and glutamate metabolism, we also identify a common vulnerability that eliminates mutant cells selectively. These results highlight a previously unreported mutant-specific effect of KRAS alleles on metabolism and signalling that could be potentially harnessed for cancer therapy.

In briefBer et al. reveal common and distinct features of oncogenic KRAS mutant isogenic cell lines. Mutant lines upregulate different nitrogen-recycling metabolic pathways and show sensitivity to combinatorial drugging of glutamine metabolism.



O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=200 SRC="FIGDIR/small/618135v1_ufig1.gif" ALT="Figure 1">
View larger version (101K):
org.highwire.dtl.DTLVardef@1459558org.highwire.dtl.DTLVardef@14fcd43org.highwire.dtl.DTLVardef@2d22ccorg.highwire.dtl.DTLVardef@147b068_HPS_FORMAT_FIGEXP  M_FIG C_FIG HighlightsO_LIDifferent KRAS mutations trigger distinct metabolic phenotypes in colorectal cell lines
C_LIO_LIGlutamine metabolism and nitrogen recycling are upregulated in KRAS mutant cells
C_LIO_LIFOXO1 is a key regulator of KRAS-induced rewiring of glutamine and nitrogen metabolism
C_LIO_LIInhibition of glutamine synthetase and glutaminase selectively kills mutant KRAS lines
C_LI
]]></description>
<dc:creator>Ber, S.</dc:creator>
<dc:creator>Yang, M.</dc:creator>
<dc:creator>Sciacovelli, M.</dc:creator>
<dc:creator>Samarajiwa, S.</dc:creator>
<dc:creator>Patel, K.</dc:creator>
<dc:creator>Nikitopoulou, E.</dc:creator>
<dc:creator>Howitt, A.</dc:creator>
<dc:creator>Cook, S. J.</dc:creator>
<dc:creator>Venkitaraman, A. R.</dc:creator>
<dc:creator>Frezza, C.</dc:creator>
<dc:creator>Esposito, A.</dc:creator>
<dc:date>2024-10-22</dc:date>
<dc:identifier>doi:10.1101/2024.10.19.618135</dc:identifier>
<dc:title><![CDATA[KRAS G12 mutant alleles differentially control glutamine metabolism via FOXO1]]></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/cgi/content/short/2023.10.09.561545v1?rss=1">
<title>
<![CDATA[
A maternal-effect Padi6 variant results in abnormal nuclear localization of DNMT1 and failure of epigenetic reprogramming and zygotic genome activation in mouse embryos 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.10.09.561545v1?rss=1"
</link>
<description><![CDATA[
PADI6 belongs to the multi-protein sub-cortical maternal complex (SCMC) that is present specifically in mammalian oocytes and early embryos. Maternal inactivation of SCMC genes generally results in early embryo lethality. In humans, variants in a subset of SCMC genes have been found in the healthy mothers of children affected by genomic imprinting disorders and characterized by multi-locus imprinting disturbances (MLID). However, how the SCMC controls the DNA methylation required to regulate imprinting remains poorly defined. To address this issue, we generated a mouse line carrying a Padi6 missense variant that had been identified in the mother of two sisters affected by Beckwith-Wiedemann syndrome and MLID. We found that if homozygous in female mice this variant resulted in interruption of embryo development at the 2-cell stage. Single-cell DNA methylation and RNA analyses demonstrated genomic hypermethylation, down-regulation of zygotic genome activation (ZGA) genes and up-regulation of maternal decay genes in 2-cell embryos from homozygous females. In addition, immunofluorescence analysis showed abnormal localization of DNMT1 and UHRF1 in mutant oocytes and zygotes. Taken together, this study demonstrates that PADI6 controls the subcellular localization of DNMT1 that is necessary for pre-implantation epigenetic reprogramming and ZGA.
]]></description>
<dc:creator>Giaccari, C.</dc:creator>
<dc:creator>Cecere, F.</dc:creator>
<dc:creator>Argenziano, L.</dc:creator>
<dc:creator>Galvao, A.</dc:creator>
<dc:creator>Acampora, D.</dc:creator>
<dc:creator>Rossi, G.</dc:creator>
<dc:creator>Hay Mele, B.</dc:creator>
<dc:creator>Cubellis, M. V.</dc:creator>
<dc:creator>Cerrato, F.</dc:creator>
<dc:creator>Andrews, S.</dc:creator>
<dc:creator>Cecconi, S.</dc:creator>
<dc:creator>Kelsey, G.</dc:creator>
<dc:creator>Riccio, A.</dc:creator>
<dc:date>2023-10-10</dc:date>
<dc:identifier>doi:10.1101/2023.10.09.561545</dc:identifier>
<dc:title><![CDATA[A maternal-effect Padi6 variant results in abnormal nuclear localization of DNMT1 and failure of epigenetic reprogramming and zygotic genome activation in mouse embryos]]></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/cgi/content/short/2023.07.31.551250v1?rss=1">
<title>
<![CDATA[
MiR-155 epigenetically counteracts hypoxia for mitochondrial fitness during germinal center positive selection 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.07.31.551250v1?rss=1"
</link>
<description><![CDATA[
To increase antibody affinity against pathogens, positively selected GC-B cells initiate cell division in the light zone (LZ) of germinal centres (GCs). Among those, higher-affinity clones migrate to the dark zone (DZ) and vigorously proliferate by relying on oxidative phosphorylation (OXPHOS). However, it remains unknown how positively selected GC-B cells adapt their metabolism for cell division in the glycolysis-dominant, cell cycle arrest-inducing, hypoxic LZ microenvironment. Here, we show that microRNA (miR)-155 mediates metabolic reprogramming during positive selection to protect high-affinity clones. Transcriptome examination and mass spectrometry analysis revealed that miR-155 regulates H3K36me2 levels by directly repressing hypoxia-induced histone lysine demethylase, Kdm2a. This is indispensable for enhancing OXPHOS through optimizing the expression of vital nuclear mitochondrial genes under hypoxia. The miR-155-Kdm2a interaction is crucial to prevent excessive production of reactive oxygen species and apoptosis. Thus, miR-155-mediated epigenetic regulation promotes mitochondrial fitness in high-affinity clones, ensuring their expansion and consequently affinity maturation.
]]></description>
<dc:creator>Nakagawa, R.</dc:creator>
<dc:creator>Llorian-Sopena, M.</dc:creator>
<dc:creator>Varsani-brown, S.</dc:creator>
<dc:creator>Chakravarty, P.</dc:creator>
<dc:creator>Camarillo, J. M.</dc:creator>
<dc:creator>Barry, D.</dc:creator>
<dc:creator>George, R.</dc:creator>
<dc:creator>Blackledge, N. P.</dc:creator>
<dc:creator>Duddy, G.</dc:creator>
<dc:creator>Klose, R.</dc:creator>
<dc:creator>Turner, M.</dc:creator>
<dc:creator>Calado, D. P.</dc:creator>
<dc:date>2023-08-02</dc:date>
<dc:identifier>doi:10.1101/2023.07.31.551250</dc:identifier>
<dc:title><![CDATA[MiR-155 epigenetically counteracts hypoxia for mitochondrial fitness during germinal center positive selection]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-08-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.12.22.521572v1?rss=1">
<title>
<![CDATA[
A quantitative and site-specific atlas of the in vivo citrullinome reveals widespread existence of citrullination 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.12.22.521572v1?rss=1"
</link>
<description><![CDATA[
Citrullination is the conversion of peptidyl-arginine into the non-coded amino acid citrulline. Despite its importance in physiology and disease, global identification of citrullinated proteins and precise modification sites has remained challenging. Here, we employed quantitative mass spectrometry-based proteomics to generate a comprehensive atlas of citrullination sites in a physiologically relevant cell type. Collectively, we identified 14.056 citrullination sites within 4.008 proteins and quantified their regulation upon inhibition of the citrullinating enzyme PADI4. Using this rich dataset, we uncover general mechanistic and cell biological principles of citrullination function, while providing site-specific and quantitative information on thousands of PAD4 substrates within cells. Our findings include signature histone marks and numerous modifications on transcriptional regulators and chromatin-related signaling effectors. Additionally, we identify precise citrullination sites on an extensive list of known autoantigens. Collectively, we describe systems attributes of the human citrullinome and provide a resource framework for understanding citrullinaiton at the mechanistic level.
]]></description>
<dc:creator>Rebak, A.</dc:creator>
<dc:creator>Hendriks, I. A.</dc:creator>
<dc:creator>Buch-Larsen, S. C.</dc:creator>
<dc:creator>Elsborg, J. D.</dc:creator>
<dc:creator>Kirsch, R.</dc:creator>
<dc:creator>Doncheva, N. T.</dc:creator>
<dc:creator>Jensen, L. J.</dc:creator>
<dc:creator>Christophorou, M. A.</dc:creator>
<dc:creator>Nielsen, M. L.</dc:creator>
<dc:date>2022-12-22</dc:date>
<dc:identifier>doi:10.1101/2022.12.22.521572</dc:identifier>
<dc:title><![CDATA[A quantitative and site-specific atlas of the in vivo citrullinome reveals widespread existence of citrullination]]></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/cgi/content/short/2022.11.02.514842v1?rss=1">
<title>
<![CDATA[
Intracellular calcium elevations drive the nucleation of FIP200- and ATG13-containing pre-autophagosomal structures that become omegasomes 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.11.02.514842v1?rss=1"
</link>
<description><![CDATA[
Ca2+ modulates autophagy at multiple steps including the induction and maturation of autophagosomes, but the magnitude and spatiotemporal properties of this calcium signal and its ultimate effect on the autophagic machinery are unclear. Focusing on the induction step leading to omegasome formation, we report that low but sustained elevations in cytosolic calcium levels induce omegasome formation but treatments that only transiently elevate calcium do not. The calcium-induced structures are early intermediates that mature into omegasomes but do not constitute full autophagosomes because they are partially devoid of late autophagy proteins ATG16 and LC3. In addition to omegasomes, all four components of the ULK complex (ULK1, FIP200, ATG13, ATG101) respond to calcium modulation: they translocate to early autophagy puncta in complete medium upon calcium elevation, and are inhibited from translocation during starvation by calcium chelation with BAPTA-2 AM. The principal early step affected by calcium lies downstream of mTORC1 inactivation and upstream of VPS34 activation, coinciding biochemically with phosphorylation of ATG13 at serine 318, which is known to require ULK1 activity. However, although the calcium-mediated step requires ATG9, FIP200 and ATG13, it does not require ULK1/2, suggesting that calcium does not directly regulate ULK1 activity but rather it regulates the mechanism by which the ULK complex components ATG13 and FIP200, together with ATG9, nucleate pre-autophagosomal precursors. This calcium-induced nucleation is sufficient to drive autophagy induction up to the omegasome step, but not beyond it.
]]></description>
<dc:creator>Smith, M.</dc:creator>
<dc:creator>Schoenfelder, P.</dc:creator>
<dc:creator>Manifava, M.</dc:creator>
<dc:creator>Polson, H.</dc:creator>
<dc:creator>Tooze, S.</dc:creator>
<dc:creator>Roderick, L.</dc:creator>
<dc:creator>Ktistakis, N.</dc:creator>
<dc:date>2022-11-02</dc:date>
<dc:identifier>doi:10.1101/2022.11.02.514842</dc:identifier>
<dc:title><![CDATA[Intracellular calcium elevations drive the nucleation of FIP200- and ATG13-containing pre-autophagosomal structures that become omegasomes]]></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/cgi/content/short/2024.02.15.580302v1?rss=1">
<title>
<![CDATA[
Tissue-resident regulatory T cells exert dualistic anti-tumour and pro-repair function in the exocrine pancreas 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.02.15.580302v1?rss=1"
</link>
<description><![CDATA[
Regulatory T cells are fundamentally important for maintaining immune homeostasis, and their potent immune-suppressive roles make them attractive immunotherapeutic targets in cancer. Recent work suggests potential functions of tissue-resident Tregs (trTregs) in tissue-repair and epithelial cell homeostasis. Here, we describe a rare population of trTreg in the exocrine pancreas. We show that these cells share common features of trTregs, including expression of the IL-33 receptor ST2 and production of the epithelial growth factor Amphiregulin, and display an oligoclonal T cell receptor repertoire. Using a mouse model of acute pancreatitis, we show that pancreatic Tregs rapidly expand upon release of IL-33 by fibroblasts. Moreover, depletion of Tregs after initiation of pancreatic injury impairs the regeneration of the exocrine parenchyma. This effect is due, in part, through a direct effect of Tregs on acinar cell proliferation. Finally, we show that transient Treg depletion in established orthotopic pancreatic tumours leads to tumour rejection yet provokes long-lasting damages to surrounding exocrine parenchyma. In all, our results demonstrate the tissue-repair capacity of pancreatic Tregs, and highlight a dualistic role of these cells in the pancreatic tumour ecosystem, with their harmful immune-suppressive function in the tumour coupled to a beneficial tissue-repair function in the surrounding tissue.
]]></description>
<dc:creator>Stockis, J.</dc:creator>
<dc:creator>Yip, T.</dc:creator>
<dc:creator>Raghunathan, S.</dc:creator>
<dc:creator>Garcia, C.</dc:creator>
<dc:creator>Lee, S.</dc:creator>
<dc:creator>Simpson, C.</dc:creator>
<dc:creator>Silvain, S.</dc:creator>
<dc:creator>Schuijs, M. J.</dc:creator>
<dc:creator>Araos, J.</dc:creator>
<dc:creator>Png, S.</dc:creator>
<dc:creator>Raddi, G.</dc:creator>
<dc:creator>Bricard, O.</dc:creator>
<dc:creator>So, T. Y.</dc:creator>
<dc:creator>Mack, S.</dc:creator>
<dc:creator>Papadopoulos, P.</dc:creator>
<dc:creator>Sawle, A.</dc:creator>
<dc:creator>Jodrell, D. I.</dc:creator>
<dc:creator>McKenzie, A. N. J.</dc:creator>
<dc:creator>Thaventhiran, J.</dc:creator>
<dc:creator>Clatworthy, M.</dc:creator>
<dc:creator>Biffi, G.</dc:creator>
<dc:creator>Saeb-Parsy, K.</dc:creator>
<dc:creator>Liston, A.</dc:creator>
<dc:creator>Halim, T. Y. F.</dc:creator>
<dc:date>2024-02-19</dc:date>
<dc:identifier>doi:10.1101/2024.02.15.580302</dc:identifier>
<dc:title><![CDATA[Tissue-resident regulatory T cells exert dualistic anti-tumour and pro-repair function in the exocrine pancreas]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-02-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.01.25.525562v1?rss=1">
<title>
<![CDATA[
Ageing impairs the regenerative capacity of regulatory T cells in central nervous system remyelination 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.01.25.525562v1?rss=1"
</link>
<description><![CDATA[
Myelin regeneration (remyelination) is essential to prevent neurodegeneration in demyelinating diseases such as Multiple Sclerosis, however, its efficiency declines with age. Regulatory T cells (Treg) recently emerged as critical players in tissue regeneration, including remyelination. However, the effect of ageing on Treg-mediated regenerative processes is poorly understood. Here, we show that expansion of aged Treg does not rescue age-associated remyelination impairment due to an intrinsically diminished capacity of aged Treg to promote oligodendrocyte differentiation and myelination. This decline in regenerative Treg functions can be rescued by a young environment. We identified Melanoma Cell Adhesion Molecule 1 (MCAM1) and Integrin alpha 2 (ITGA2) as novel candidates of Treg-mediated oligodendrocyte differentiation that decrease with age. Our findings demonstrate that ageing limits the neuroregenerative capacity of Treg, likely limiting their remyelinating therapeutic potential in aged patients, and describe two novel mechanisms implicated in Treg-driven remyelination that may be targetable to overcome this limitation.
]]></description>
<dc:creator>de la Fuente, A. G.</dc:creator>
<dc:creator>Dittmer, M.</dc:creator>
<dc:creator>Heesbeen, E.</dc:creator>
<dc:creator>de la Vega-Gallardo, N.</dc:creator>
<dc:creator>White, J. A.</dc:creator>
<dc:creator>Young, A.</dc:creator>
<dc:creator>Mayne, K.</dc:creator>
<dc:creator>Falconer, J.</dc:creator>
<dc:creator>McMurran, C. E.</dc:creator>
<dc:creator>Innayatulah, M.</dc:creator>
<dc:creator>Tiwari, V.</dc:creator>
<dc:creator>Penalva, R.</dc:creator>
<dc:creator>Ingram, R. J.</dc:creator>
<dc:creator>Dombrowski, Y.</dc:creator>
<dc:creator>Fitzgerald, D. C.</dc:creator>
<dc:date>2023-01-25</dc:date>
<dc:identifier>doi:10.1101/2023.01.25.525562</dc:identifier>
<dc:title><![CDATA[Ageing impairs the regenerative capacity of regulatory T cells in central nervous system remyelination]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-01-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.12.01.569004v1?rss=1">
<title>
<![CDATA[
Protein tyrosine phosphatase receptor kappa regulates glycolysis and de novo lipogenesis to promote hepatocyte metabolic reprogramming in obesity 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.12.01.569004v1?rss=1"
</link>
<description><![CDATA[
Fat accumulation, de novo lipogenesis, and glycolysis are key drivers of hepatocyte reprogramming and the consequent metabolic dysfunction-associated steatotic liver disease (MASLD). Here we report that obesity leads to dysregulated expression of hepatic protein-tyrosine phosphatases (PTPs). PTPRK was found to be increased in steatotic hepatocytes in both humans and mice, and positively correlated with PPAR{gamma}-induced lipogenic signalling. High-fat-fed PTPRK knockout mice displayed reduced weight gain and hepatic fat accumulation. Phosphoproteomic analysis in primary hepatocytes and hepatic metabolomics identified fructose-1,6-bisphosphatase 1 and glycolysis as PTPRK targets in metabolic reprogramming. Silencing PTPRK in hepatoma cell lines resulted in reduced colony-forming ability and PTPRK knockout mice developed smaller tumours after diethylnitrosamine-induced hepatocarcinogenesis. Our study defines a novel role for PTPRK in regulating hepatic glycolysis, lipid metabolism, and tumour development. PTPRK inhibition may provide therapeutic possibilities in obesity-associated liver diseases.

HighlightsO_LIHepatic receptor-type PTPs are increased in MASLD
C_LIO_LIPTPRK is expressed in hepatocytes and upregulated in obesity
C_LIO_LIPTPRK deficiency reduces body fat mass and liver steatosis in diet-induced obesity
C_LIO_LIPTPRK regulates hepatic glycolysis and lipogenesis, promoting tumorigenesis
C_LI
]]></description>
<dc:creator>Gilglioni, E. H.</dc:creator>
<dc:creator>Li, A.</dc:creator>
<dc:creator>Saint-Pierre Wijckmans, W.</dc:creator>
<dc:creator>Shen, T.-K.</dc:creator>
<dc:creator>Perez-Chavez, I.</dc:creator>
<dc:creator>Hovhannisyan, G.</dc:creator>
<dc:creator>Lisjak, M.</dc:creator>
<dc:creator>Negueruela, J.</dc:creator>
<dc:creator>Vandenbempt, V.</dc:creator>
<dc:creator>Bauza-Martinez, J.</dc:creator>
<dc:creator>Herranz, J. M.</dc:creator>
<dc:creator>Ezerina, D.</dc:creator>
<dc:creator>Demine, S.</dc:creator>
<dc:creator>Feng, Z.</dc:creator>
<dc:creator>Vignane, T.</dc:creator>
<dc:creator>Otero-Sanchez, L.</dc:creator>
<dc:creator>Lambertucci, F.</dc:creator>
<dc:creator>Prasnicka, A.</dc:creator>
<dc:creator>Deviere, J.</dc:creator>
<dc:creator>Hay, D. C.</dc:creator>
<dc:creator>Encinar, J. A. N.</dc:creator>
<dc:creator>Singh, S. P.</dc:creator>
<dc:creator>Messens, J.</dc:creator>
<dc:creator>Filipovic, M. R.</dc:creator>
<dc:creator>Sharpe, H. J.</dc:creator>
<dc:creator>Trepo, E.</dc:creator>
<dc:creator>Wu, W.</dc:creator>
<dc:creator>Gurzov, E. N.</dc:creator>
<dc:date>2023-12-01</dc:date>
<dc:identifier>doi:10.1101/2023.12.01.569004</dc:identifier>
<dc:title><![CDATA[Protein tyrosine phosphatase receptor kappa regulates glycolysis and de novo lipogenesis to promote hepatocyte metabolic reprogramming in obesity]]></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/cgi/content/short/2024.12.01.626220v1?rss=1">
<title>
<![CDATA[
RNA-binding proteins control the G2-M checkpoint of the germinal centre B cell 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.12.01.626220v1?rss=1"
</link>
<description><![CDATA[
How germinal centre (GC) B cells undergo rapid cell division while maintaining genome stability is poorly understood. Here, we show that the RNA-binding proteins ZFP36L1 and ZFP36L2 act downstream of antigen-sensing and protect GC B cells from replication stress by controlling a cell cycle-related RNA post-transcriptional regulon. ZFP36L1 and ZFP36L2 safeguard faithful completion of mitosis by restraining the expression of CDK1 and cyclin B1, whilst controlling their activity through regulation of a p21-mediated negative feedback loop. In the absence of ZFP36L1 and ZFP36L2, GC B cells arrest in G2-M and die by apoptosis, resulting in curtailed GC responses. This is associated with stalling of the DNA replication fork at active replication initiation zones, which causes replication stress and increased activity of the ATR/CHK1 DNA damage response. Our findings reveal that gene regulation by RNA-binding proteins is essential for a functional G2-M checkpoint to operate in GC B cells.
]]></description>
<dc:creator>Salerno, F.</dc:creator>
<dc:creator>Whale, A.</dc:creator>
<dc:creator>Matheson, L. S.</dc:creator>
<dc:creator>Foster, W.</dc:creator>
<dc:creator>Stammers, M.</dc:creator>
<dc:creator>Bell, S. E.</dc:creator>
<dc:creator>Hidson, D. J.</dc:creator>
<dc:creator>Linterman, M. A.</dc:creator>
<dc:creator>Houseley, J. A.</dc:creator>
<dc:creator>Turner, M.</dc:creator>
<dc:date>2024-12-05</dc:date>
<dc:identifier>doi:10.1101/2024.12.01.626220</dc:identifier>
<dc:title><![CDATA[RNA-binding proteins control the G2-M checkpoint of the germinal centre B cell]]></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/cgi/content/short/2025.01.10.632317v1?rss=1">
<title>
<![CDATA[
RNA helicase DDX1 regulates germinal centre selection and affinity maturation by promoting tRNA ligase activity 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.01.10.632317v1?rss=1"
</link>
<description><![CDATA[
Clonal expansion of antigen-specific B-cells defines effective germinal centre responses and is key for the generation of high-affinity antibodies. While positive selection in germinal centres has been associated with anabolic metabolism and cell growth, the downstream drivers of B-cell proliferation are not well understood. Here we report that the RNA helicase DDX1 is required for germinal centre maturation and accrual of dark-zone cellularity. Upon interaction with T-follicular helper cells, DDX1-deficient B-cells upregulate c-MYC but do not clonally expand. We show that positive selection is coupled with an increase in mRNA translation, that is dependent on DDX1. DDX1 endows B-cells with the protein biosynthetic capability that is required for rapid cell proliferation. It does so by modulating the activity of the tRNA ligase complex and tRNA splicing. Our data reveal that mRNA translation efficiency is a key determinant of B-cell fitness during germinal centre responses.
]]></description>
<dc:creator>Kimber, R.</dc:creator>
<dc:creator>Ingelsfield, S.</dc:creator>
<dc:creator>Screen, M.</dc:creator>
<dc:creator>Salerno, F.</dc:creator>
<dc:creator>Matheson, L. S.</dc:creator>
<dc:creator>Okkenhaug, H.</dc:creator>
<dc:creator>Whale, A.</dc:creator>
<dc:creator>Assalaarachchi, J.</dc:creator>
<dc:creator>Stammers, M.</dc:creator>
<dc:creator>Akdeniz, D.</dc:creator>
<dc:creator>Andrews, S.</dc:creator>
<dc:creator>Ribeiro de Almeida, C.</dc:creator>
<dc:date>2025-01-14</dc:date>
<dc:identifier>doi:10.1101/2025.01.10.632317</dc:identifier>
<dc:title><![CDATA[RNA helicase DDX1 regulates germinal centre selection and affinity maturation by promoting tRNA ligase activity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-01-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.03.27.645766v1?rss=1">
<title>
<![CDATA[
Re-engineering of acetyl coenzyme A metabolism prevents senescence in budding yeast 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.03.27.645766v1?rss=1"
</link>
<description><![CDATA[
Although lifespan has long been the focus of ageing research, preventing functional decline late in life is a more pressing societal need. Here, we investigate the basis of senescence and declining fitness during replicative ageing in budding yeast, and describe a metabolic perturbation that preserves late-life fitness even on an unrestricted glucose diet. We show that senescence can be prevented by constitutive activation of AMPK, though only for approximately half the ageing population, and use genetic and functional assays to link this heterogeneous response with differences in cytosolic acetyl coenzyme A (Acetyl-CoA) metabolism. In one class of ageing cell, AMPK activity maintains fitness late-in-life through pathways that transport cytosolic Acetyl-CoA into mitochondria, but AMPK also inhibits fatty acid synthesis which leads to lipid starvation in the other class of ageing cell. Therefore, AMPK activity has both positive and negative effects, but we show that constitutive AMPK activity uncoupled from fatty acid synthesis inhibition (the A2A mutant) suppresses senescence and maintains fitness in both classes of ageing cell. We further implicate lipid starvation and excess acetyl coenzyme A availability as major drivers of senescence in replicatively aged wild-type yeast. Our work shows that ageing is not intrinsically associated with declining fitness, at least in yeast, and that re-engineering highly conserved metabolic pathways allows fitness to be preserved very late in life.

Graphical abstract

O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=151 SRC="FIGDIR/small/645766v2_ufig1.gif" ALT="Figure 1">
View larger version (74K):
org.highwire.dtl.DTLVardef@173a897org.highwire.dtl.DTLVardef@4166c3org.highwire.dtl.DTLVardef@1c1c1aeorg.highwire.dtl.DTLVardef@19295c5_HPS_FORMAT_FIGEXP  M_FIG C_FIG
]]></description>
<dc:creator>Hadj-Moussa, H.</dc:creator>
<dc:creator>Ulusan, M.</dc:creator>
<dc:creator>Horkai, D.</dc:creator>
<dc:creator>Mirza, M. K. A.</dc:creator>
<dc:creator>Houseley, J.</dc:creator>
<dc:date>2025-03-27</dc:date>
<dc:identifier>doi:10.1101/2025.03.27.645766</dc:identifier>
<dc:title><![CDATA[Re-engineering of acetyl coenzyme A metabolism prevents senescence in budding yeast]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-03-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.04.27.650842v1?rss=1">
<title>
<![CDATA[
Pervanadate-induced oxidation relieves autoinhibition of SRC protein tyrosine kinase 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.04.27.650842v1?rss=1"
</link>
<description><![CDATA[
Dynamic regulation of protein tyrosine phosphorylation (pTyr) by phosphatases (PTPs) and kinases enables cells to sense and respond to environmental changes. The widely used chemical probe Pervanadate (PV) induces accumulation of high levels of pTyr in cells, an effect primarily attributed to its properties as a PTP inhibitor. This led to the assertion that PTPs are the master gatekeeper of intracellular pTyr homeostasis. Here, we use diverse approaches to reveal that PV disrupts cellular redox homeostasis and directly activates SRC family tyrosine kinases via oxidation of specific cysteine residues. Using mass spectrometry and biophysical approaches, we show that oxidation activates SRC by disrupting autoinhibition and altering phosphopeptide binding by its SH2 domain. We further establish that redox-sensitive cysteine residues are essential for SRC to promote cellular overgrowth. Our findings call for a re-evaluation of PV-based experiments and provide compelling evidence that oxidation is a crucial mechanism in controlling the oncogenic properties of SRC.
]]></description>
<dc:creator>Mulholland, K. E.</dc:creator>
<dc:creator>Bourguet, M.</dc:creator>
<dc:creator>Cheng, N.</dc:creator>
<dc:creator>Rahman, O.</dc:creator>
<dc:creator>Ezerina, D.</dc:creator>
<dc:creator>Daly, L. A.</dc:creator>
<dc:creator>Lai, T.</dc:creator>
<dc:creator>Casanova, S. A.</dc:creator>
<dc:creator>Creixell, P.</dc:creator>
<dc:creator>Eyers, C.</dc:creator>
<dc:creator>Eyers, P.</dc:creator>
<dc:creator>Messens, J.</dc:creator>
<dc:creator>Byrne, D.</dc:creator>
<dc:creator>Sharpe, H.</dc:creator>
<dc:date>2025-04-27</dc:date>
<dc:identifier>doi:10.1101/2025.04.27.650842</dc:identifier>
<dc:title><![CDATA[Pervanadate-induced oxidation relieves autoinhibition of SRC protein tyrosine kinase]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-04-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.06.09.658559v1?rss=1">
<title>
<![CDATA[
TrAEL-seq captures DNA replication dynamics in mammalian cells 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.06.09.658559v1?rss=1"
</link>
<description><![CDATA[
Precise DNA replication is critical to the maintenance of genome stability, and the DNA replication machinery is a focal point of many current and upcoming chemotherapeutics. TrAEL-seq is a robust method for profiling DNA replication genome-wide that works in unsynchronised cells and does not require treatment with drugs or nucleotide analogues. Here, we provide an updated method for TrAEL-seq including multiplexing of up to 6 samples that dramatically improves sample quality and throughput, and we validate TrAEL-seq in multiple mammalian cell lines. The updated protocol is straightforward and robust yet provides excellent resolution comparable to OK-seq in mammalian cell samples. High resolution replication profiles can be obtained across large panels of samples and in dynamic systems, for example during the progressive onset of oncogene induced senescence. In addition to mapping zones where replication initiates and terminates, TrAEL-seq is sensitive to replication fork speed, revealing effects of both transcription and proximity to replication Initiation Zones on fork progression. Although forks move more slowly through transcribed regions, this does not have a significant impact on the broader dynamics of replication fork progression, which is dominated by rapid fork movement in long replication regions (>1Mb). Short and long replication regions are not intrinsically different, and instead replication forks accelerate across the first [~]1 Mb of travel such that forks progress faster in the middle of regions lying between widely spaced Initiation Zones. We propose that this is a natural consequence of fewer replication forks being active later in S phase when these distal regions are replicated and there being less competition for replication factors.
]]></description>
<dc:creator>Kara, N.</dc:creator>
<dc:creator>Biggins, L.</dc:creator>
<dc:creator>Grinkevich, V.</dc:creator>
<dc:creator>Whale, A.</dc:creator>
<dc:creator>Garran Garcia, P.</dc:creator>
<dc:creator>Srinivasan, J.</dc:creator>
<dc:creator>Rugg-Gunn, P.</dc:creator>
<dc:creator>Andrews, S.</dc:creator>
<dc:creator>Parry, A.</dc:creator>
<dc:creator>Robinson, H. M. R.</dc:creator>
<dc:creator>Houseley, J.</dc:creator>
<dc:date>2025-06-11</dc:date>
<dc:identifier>doi:10.1101/2025.06.09.658559</dc:identifier>
<dc:title><![CDATA[TrAEL-seq captures DNA replication dynamics in mammalian cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-06-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.09.09.675154v1?rss=1">
<title>
<![CDATA[
A post-transcriptional regulatory checkpoint controls the response of tumor-infiltrating cytotoxic CD4+ T cells to immunotherapy 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.09.09.675154v1?rss=1"
</link>
<description><![CDATA[
Acquisition of cytotoxic activity in CD4+ T cells (TCTX) can promote potent anti-tumor activity thus holding promise as a therapeutic approach. However, how this activity is regulated remains poorly understood. Here, we demonstrate that tumor-infiltrating CD4+ TCTX activity is restrained by a post-transcriptional regulatory checkpoint. In untreated tumors, CD4+ TCTX exist in a poised state, characterized by abundant Gzmb mRNA but limited Granzyme B (GzmB) protein. Differentiation into poised TCTX is regulated by the Blimp-1-Bcl6 axis and requires type-I interferon signaling. Treatment with anti-CTLA-4 or anti-LAG-3 plus anti-PD-1 removed the block to GzmB protein production by repressing expression of the post-transcriptional regulator Zfp36l1. Constitutive Zfp36l1 expression abrogated the effects of anti-CTLA-4 while deletion of Zfp36l1 and its paralog Zfp36 triggered GzmB protein production and promoted tumor control. These data identify ZFP36/ZFP36L1 as a key post-transcriptional regulatory checkpoint of CD4+ TCTX activity and a potential immunotherapy target in cancer.
]]></description>
<dc:creator>Vila de Mucha, M.</dc:creator>
<dc:creator>Hlond, J.</dc:creator>
<dc:creator>Costoya, C.</dc:creator>
<dc:creator>Lau, C.-I.</dc:creator>
<dc:creator>Sledzinska, A.</dc:creator>
<dc:creator>Uddin, I.</dc:creator>
<dc:creator>Navarrete, M.</dc:creator>
<dc:creator>Shah, M.</dc:creator>
<dc:creator>Mastrokalos, G.</dc:creator>
<dc:creator>Karagianni, D.</dc:creator>
<dc:creator>Bell, S. E.</dc:creator>
<dc:creator>Nattress, C.</dc:creator>
<dc:creator>Beattie, G.</dc:creator>
<dc:creator>Tape, C. J.</dc:creator>
<dc:creator>Turner, M.</dc:creator>
<dc:creator>Quezada, S. A.</dc:creator>
<dc:creator>Jenner, R. G.</dc:creator>
<dc:date>2025-09-12</dc:date>
<dc:identifier>doi:10.1101/2025.09.09.675154</dc:identifier>
<dc:title><![CDATA[A post-transcriptional regulatory checkpoint controls the response of tumor-infiltrating cytotoxic CD4+ T cells to immunotherapy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-09-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.12.09.627617v1?rss=1">
<title>
<![CDATA[
H3.1K27M-induced misregulation of the TSK/TONSL-H3.1 pathway causes genomic instability 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.12.09.627617v1?rss=1"
</link>
<description><![CDATA[
The oncomutation lysine 27-to-methionine in histone H3 (H3K27M) is frequently identified in tumors of patients with diffuse midline glioma-H3K27 altered (DMG-H3K27a). H3K27M inhibits the deposition of the histone mark H3K27me3, which affects the maintenance of transcriptional programs and cell identity. Cells expressing H3K27M are also characterized by defects in genome integrity, but the mechanisms linking expression of the oncohistone to DNA damage remain mostly unknown. In this study, we demonstrate that expression of H3.1K27M in the model plant Arabidopsis thaliana interferes with post-replicative chromatin maturation mediated by the H3.1K27 methyltransferases ATXR5 and ATXR6. As a result, H3.1 variants on nascent chromatin remain unmethylated at K27 (H3.1K27me0), leading to ectopic activity of TONSOKU (TSK), which induces DNA damage and genomic alterations. Elimination of TSK activity suppresses the genome stability defects associated with H3.1K27M expression, while inactivation of specific DNA repair pathways prevents survival of H3.1K27M-expressing plants. Overall, our results suggest that H3.1K27M disrupts the chromatin-based mechanisms regulating TSK/TONSL activity, which causes genomic instability and may contribute to the etiology of DMG-H3K27a.
]]></description>
<dc:creator>Yuan, W.</dc:creator>
<dc:creator>Huang, Y.-C.</dc:creator>
<dc:creator>LeBlanc, C.</dc:creator>
<dc:creator>Poulet, A.</dc:creator>
<dc:creator>Valsakumar, D.</dc:creator>
<dc:creator>van Wolfswinkel, J. C.</dc:creator>
<dc:creator>Voigt, P.</dc:creator>
<dc:creator>Jacob, Y.</dc:creator>
<dc:date>2024-12-11</dc:date>
<dc:identifier>doi:10.1101/2024.12.09.627617</dc:identifier>
<dc:title><![CDATA[H3.1K27M-induced misregulation of the TSK/TONSL-H3.1 pathway causes genomic instability]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-12-11</prism:publicationDate>
<prism:section></prism:section>
</item>
</rdf:RDF>
