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介导过程调节基因表达。在我们的项目中，我们将利用CRISPR编辑技术在不同水平上分析表观遗传修饰因子对人类T细胞分化的影响：我们将在人类体外Treg诱导以及naïve T细胞中基因消融后的其他辅助性T细胞分化中研究溴域蛋白baz1b介导的染色质重塑对人类T细胞分化的影响。此外，我们将通过CRISPR编辑肿瘤源性T细胞中的表观遗传修饰因子抑制剂和BAZ1B，在一个具有单细胞RNA-seq读出的池筛选中。最后，我们将在naïve T细胞的聚合CRISPR筛选中，通过清除所有注释的表观遗传修饰因子（EpiFactors数据库），生成体外人类T辅助细胞分化的表观遗传修饰因子全球图谱，并评估其分化为Th1、Th2和Treg细胞的潜力。使用这些方法，我们将在功能上验证癌症治疗过程中表观遗传修饰因子的靶向性，并为未来的治疗方法确定新的候选基因。

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.