The First Affiliated Hospital, Chongqing Medical University, Chongqing Key Laboratory of Ophthalmology, Chongqing, P. R. China.
Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou, 510530, China.
Cell Death Dis. 2020 Aug 22;11(8):697. doi: 10.1038/s41419-020-02891-2.
Th17 cells, a lymphocyte subpopulation that is characterized by the expression of the transcription factor "retinoic acid receptor-related orphan receptor gamma-t" (RORγt), plays an important role in the pathogenesis of autoimmune disease. The current study was set up to discover novel and non-steroidal small-molecule inverse agonists of RORγt and to determine their effects on autoimmune disease. Structure-based virtual screening (SBVS) was used to find compounds targeting RORγt. Flow cytometry was used to detect the Th17 cell differentiation. Inverse agonists were intraperitoneally administered to mice undergoing experimental autoimmune uveitis (EAU), experimental autoimmune encephalomyelitis (EAE) or type 1 diabetes. The effects of the inverse agonists were evaluated by clinical or histopathological scoring. Among 1.3 million compounds screened, CQMU151 and CQMU152 were found to inhibit Th17 cell differentiation without affecting the differentiation of Th1 and Treg lineages (both P = 0.001). These compounds also reduced the severity of EAU (P = 0.01 and 0.013) and functional studies showed that they reduced the number of Th17 cell and the expression of IL-17(Th17), but not IFN-γ(Th1) and TGF-β(Treg) in mouse retinas. Further studies showed that these compounds may reduce the expression of p-STAT3 by reducing the positive feedback loop of IL-17/IL-6/STAT3. These compounds also reduced the impaired blood-retinal barrier function by upregulating the expression of tight junction proteins. These compounds were also found to reduce the severity of EAE and type 1 diabetes. Our results showed that RORγt inverse agonists may inhibit the development of autoimmune diseases and may provide new clues for the treatment of Th17-mediated immune diseases.
Th17 细胞是一种淋巴细胞亚群，其特征在于表达转录因子“视黄酸受体相关孤儿受体 γt”（RORγt），在自身免疫性疾病的发病机制中发挥重要作用。本研究旨在发现新型非甾体小分子 RORγt 反向激动剂，并确定它们对自身免疫性疾病的影响。基于结构的虚拟筛选（SBVS）用于寻找针对 RORγt 的化合物。流式细胞术用于检测 Th17 细胞分化。反向激动剂经腹腔注射用于实验性自身免疫性葡萄膜炎（EAU）、实验性自身免疫性脑脊髓炎（EAE）或 1 型糖尿病小鼠。通过临床或组织病理学评分评估反向激动剂的作用。在筛选的 130 万种化合物中，发现 CQMU151 和 CQMU152 可抑制 Th17 细胞分化，而不影响 Th1 和 Treg 谱系的分化（均 P = 0.001）。这些化合物还降低了 EAU 的严重程度（P = 0.01 和 0.013），功能研究表明，它们减少了 Th17 细胞的数量和 IL-17（Th17）的表达，但不影响 IFN-γ（Th1）和 TGF-β（Treg）在小鼠视网膜中的表达。进一步的研究表明，这些化合物可能通过降低 IL-17/IL-6/STAT3 的正反馈环来降低 p-STAT3 的表达。这些化合物还通过上调紧密连接蛋白的表达来减少受损的血视网膜屏障功能。这些化合物还降低了 EAE 和 1 型糖尿病的严重程度。我们的结果表明，RORγt 反向激动剂可能抑制自身免疫性疾病的发展，并为治疗 Th17 介导的免疫性疾病提供新的线索。
