Chemical Inhibition of the RORγt-dependent Transcriptional Network in Th17 cells [RNA-Seq 2]. Mus musculus

RORγt is a transcription factor required for T helper 17 (Th17) cell development. We identified three RORγt-specific inhibitors that suppress Th17 cell responses including Th17 cell-mediated autoimmune disease. We systemically characterized RORγt binding data in the presence and absence of drug with corresponding whole-transcriptome sequencing for wild-type and RORγt-deficient cells. RORγt is central in a densely interconnected regulatory network, acting both as a direct activator of genes important for Th17 cell differentiation and as a direct repressor of genes from other T-cell lineages. The three inhibitors identified here reversed both of these modes of action, but to varying extents and through distinct mechanisms. Whereas one inhibitor displaced RORγt from its target-loci, the two more potent inhibitors affected transcription predominantly without removing DNA-binding. Our work illustrates the power of a system-scale analysis of transcriptional regulation to characterize potential therapeutic compounds that inhibit pathogenic Th17 cells and suppress autoimmunity. Overall design: Transcriptional profiling of Th17 cells under chemical perturbations of RORγt, DMSO, and knockout of RORγt. It includes repeats for all the data in GSE56018, plus one additional condition.

视黄酸相关孤儿受体γt（RORγt）是辅助性T细胞17（T helper 17, Th17）细胞发育所必需的转录因子。本研究鉴定出三种RORγt特异性抑制剂，可抑制Th17细胞介导的免疫应答，包括Th17细胞引发的自身免疫疾病。我们针对野生型及RORγt缺陷型细胞，在药物存在与缺失两种条件下，对RORγt结合数据进行系统性表征，并同步获取了对应的全转录组测序结果。RORγt在高度互联的调控网络中处于核心地位：其既可直接激活Th17细胞分化所需的关键基因，又可直接抑制其他T细胞谱系相关基因的转录。本研究鉴定出的三种抑制剂均可逆转这两种调控模式，但作用程度与具体机制各不相同。其中一种抑制剂可将RORγt从其靶基因座位上解离，而另外两种活性更强的抑制剂则主要在不改变RORγt DNA结合能力的情况下调控转录过程。本研究证实了转录调控的系统级分析在表征可抑制致病性Th17细胞、缓解自身免疫疾病的潜在治疗性化合物方面的应用价值。实验整体设计：对经RORγt化学扰动、二甲基亚砜（Dimethyl Sulfoxide, DMSO）处理以及RORγt基因敲除的Th17细胞进行转录谱分析。本数据集包含GSE56018中所有数据的生物学重复，并新增了一组实验条件。