Homeostatic, repertoire and transcriptional relationships between colon T regulatory cell subsets

Foxp3+ regulatory T cells (Tregs) in the colon are key to promoting peaceful co-existence with symbiotic microbes. Differentiated in either thymic or peripheral locations, and modulated by microbes and other cellular influencers, colonic Treg subsets have been identified through key transcription factors (TF; Helios, Rorg, Gata3, cMaf), but their inter-relationships are unclear. Here, we perform genomic analysis of colonic lamina propria Tregs with conditional KOs of each of these TFs to better understand how each TF contributes to colonic Treg identity and function. Overall design: Mouse CD4+ regulatory Tregs from WT or Treg-specific conditional TF KO (Gata3, Ikzf2, cMaf, Rorc) littermate control mice were isolated from colonic lamina propria by FACS for scATAC-seq using 10x Genomics.

结肠内的Foxp3阳性调节性T细胞（Foxp3+ regulatory T cells, Tregs）是促进机体与共生微生物和谐共存的核心调控环节。这类细胞可在胸腺或外周组织中分化，并受微生物及其他细胞调控因子的调节；目前已通过关键转录因子（TF：Helios、Rorg、Gata3、cMaf）鉴定出多种结肠Treg亚群，但其相互关系仍不明确。
本研究针对上述各转录因子分别构建条件性敲除（KO）模型，对结肠固有层Treg开展基因组分析，以期更清晰地阐明每个转录因子对结肠Treg的细胞特性与功能的调控作用。
实验设计：从野生型（WT）小鼠或Treg特异性条件性转录因子敲除（Gata3、Ikzf2、cMaf、Rorc）同窝对照小鼠的结肠固有层中，通过荧光激活细胞分选术（FACS）分离小鼠CD4+调节性T细胞，采用10x Genomics平台进行scATAC-seq测序。