Project description:B cell differentiation is tightly regulated through coordinated changes in metabolism, division, expression of transcription factors, and epigenetic programming mediated by histone modifying enzymes. In this study, we examined the role of an epigenetic writer, the histone H3K9 mono and dimethyltransferse G9a, in regulating the transcriptional programs during B-cell development and plasma cell (PC) formation. Utilizing a B-cell specific G9afl/flCd19Cre/+ conditional mouse model we performed RNA-seq on naive and activated B cells and plasma cells that formed in response to LPS.
Project description:B cell differentiation is tightly regulated through coordinated changes in metabolism, division, expression of transcription factors, and epigenetic programming mediated by histone modifying enzymes. In this study, we examined the role of an epigenetic writer, the histone H3K9 mono and dimethyltransferse G9a, in regulating the chromatin accessibility programs during B-cell development and plasma cell (PC) formation. Utilizing a B-cell specific G9afl/flCd19Cre/+ conditional mouse model we performed ATAC-seq on naive and activated B cells and plasma cells that formed in response to LPS.
Project description:As methyltransferase G9a has been implicated in immune cell differentiation we have assessed the role of G9a enzymatic inhibitors (UNC0642) in the differentiation of human naïve T-cells under different stimulation conditions.
Project description:As methyltransferase G9a has been implicated in immune cell differentiation we have assessed the role of G9a enzymatic inhibitors (UNC0642) in the differentiation of human naïve T-cells under different stimulation conditions.
Project description:Posttranslational modifications of histone N-terminal tails influence the status of chromatin and eventually control the transcriptional outcome of a particular gene. As a histone H3K9 methyltransferase (HMTase) in higher eukaryotes, G9a-mediated transcriptional repression is the major epigenetic silencing machinery. UHRF1 (ubiquitin-like with PHD and ring finger domains I) binds to hemi-methylated DNA and plays essential role in maintenance of DNA methylation by recruiting DNMT1. Here, we provide evidence that UHRF1 is transcriptionally downregulated by H3K9 HMTase G9a. We found that increased expression of G9a along with transcription factor YY1 specifically represses UHRF1 transcription. We uncovered showed that G9a regulates UHRF1-mediated H3K23 ubiquitylation and proper DNA replication maintenance by FACS analysis and propose that H3K9 HMTase G9a is a specific epigenetic regulator of UHRF1.
Project description:Elucidating mechanisms of T cell development can guide in vitro T cell differentiation from Pluripotent Stem Cells (iPSCs) and inform off-the-shelf T cell-based immunotherapies. Using a stroma-free human iPSC-T cell differentiation platform, we screened for epigenetic modulators that govern T cell specification and identified the H3K9-directed histone methyltransferases G9a/GLP as repressors of T cell fate. We show that G9a/GLP controls the cell fate decision between myeloid and lymphoid lineages in hematopoietic stem and progenitor cells (HSPCs). Inhibition of G9a/GLP promotes the production of lymphoid cells during zebrafish embryonic hematopoiesis, demonstrating the evolutionary conservation of G9a/GLP function. Importantly, chemical-induced epigenetic reprogramming via G9a/GLP inhibition facilitates the generation of robust iPSC-T cells that bear transcriptional similarity to peripheral blood αβ T cells. When engineered to express Chimeric Antigen Receptors, the epigenetically engineered iPSC-T cells exhibit enhanced antitumor activity both in vitro and in a xenograft mouse model.
Project description:Elucidating mechanisms of T cell development can guide in vitro T cell differentiation from Pluripotent Stem Cells (iPSCs) and inform off-the-shelf T cell-based immunotherapies. Using a stroma-free human iPSC-T cell differentiation platform, we screened for epigenetic modulators that govern T cell specification and identified the H3K9-directed histone methyltransferases G9a/GLP as repressors of T cell fate. We show that G9a/GLP controls the cell fate decision between myeloid and lymphoid lineages in hematopoietic stem and progenitor cells (HSPCs). Inhibition of G9a/GLP promotes the production of lymphoid cells during zebrafish embryonic hematopoiesis, demonstrating the evolutionary conservation of G9a/GLP function. Importantly, chemical-induced epigenetic reprogramming via G9a/GLP inhibition facilitates the generation of robust iPSC-T cells that bear transcriptional similarity to peripheral blood αβ T cells. When engineered to express Chimeric Antigen Receptors, the epigenetically engineered iPSC-T cells exhibit enhanced antitumor activity both in vitro and in a xenograft mouse model.