Project B16 –Epigenetic modulators in human T helper cell differentiation – identification and characterization with CRISPR screens
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.