Epigenetic modulators in human T helper cell differentiation – identification and characterization with CRISPR screens (B16*)

表观遗传调节剂通过DNA甲基化、组蛋白修饰和不同的RNA介导的过程来调节基因表达。在我们的项目中，我们将使用CRISPR编辑来分析表观遗传修饰剂在不同水平上对人类T细胞分化的影响：布罗莫结构域蛋白BAZ 1B介导的染色质重塑对人类T细胞分化的影响将在人类体外Treg诱导以及幼稚T细胞中基因消融后的其他T辅助细胞分化中进行研究。此外，我们将在具有单细胞RNA-seq读数的合并筛选中通过肿瘤衍生的T细胞中的CRISPR编辑来消除表观遗传修饰剂的抑制剂以及BAZ 1B。最后，我们将通过在幼稚T细胞中的合并CRISPR筛选中消除所有注释的表观遗传修饰物（EpiFactors数据库）来生成离体人T辅助细胞分化中的表观遗传修饰物的全局图谱，并评估其分化为Th 1，Th 2和Treg细胞的潜力。使用这些方法，我们将在癌症治疗过程中功能验证表观遗传修饰剂的靶向作用，并为未来的治疗方法鉴定新的候选基因。

Epigenetic modulators regulate gene expression by DNA methylation, histone modifications and different RNA-mediated processes. In our project, we will use CRISPR editing to analyze the influence of epigenetic modifiers on human T cell differentiation on different levels: The effect of the bromodomain protein BAZ1B-mediated chromatin remodeling on human T cell differentiation will be investigated in human in vitro Treg induction as well as other T helper cell differentiation after ablation of the gene in naïve T cells. Furthermore, we will ablate inhibitors of epigenetic modifiers as well as BAZ1B by CRISPR editing in tumor-derived T cells in a pooled screen with single cell RNA-seq readout. Finally, we will generate a global map of epigenetic modifiers in ex vivo human T helper cell differentiation by ablating all annotated epigenetic modifiers (EpiFactors database) in a pooled CRISPR screen in naïve T cells and assess their potential to differentiate into Th1, Th2 and Treg cells. Using these approaches, we will functionally validate targeting of epigenetic modifiers during cancer therapy and identify novel candidate genes for future therapeutic approaches.