Xia Li, Tian Enming, Yu Mingcheng, Liu Chenglong, Shen Lian, Huang Yafei, Wu Zhongen, Tian Jinlong, Yu Ker, Wang Yonghui, Xie Qiong, Zhu Di
Department of Pharmacology, School of Pharmacy, Fudan University, Shanghai, 201203, China.
Department of Pharmacology, School of Basic Medical Sciences, Fudan University, Shanghai, 200032, China.
J Exp Clin Cancer Res. 2022 Apr 23;41(1):155. doi: 10.1186/s13046-022-02289-2.
The overall response rate to checkpoint blockade remains unsatisfactory, partially due to the immune-suppressive tumor microenvironment. A retinoic acid-related orphan receptor γt (RORγt) agonist (LYC-55716) is currently used in clinical trials combined with anti-PD-1, but how the Th17 cell transcription factor RORγt enhances antitumor immunity of PD-1 in the tumor microenvironment remains elusive.
The expression of mRNA was analyzed using qPCR assays. Flow cytometry was used to sort and profile cells. Cell migration was analyzed using Transwell assays. Biacore was used to determine the binding affinity to the RORγt protein. The RORγt GAL4 cell-based reporter gene assay was used to measure activity in the RORγt driven luciferase reporter gene expression.
We designed a potent and selective small-molecule RORγt agonist (8-074) that shows robust antitumor efficacy in syngeneic tumor models and improves the efficacy of anti‑PD‑1 in a murine lung cancer model. RORγt agonist treatment increased intratumoral CD8 T cells, which were correlated with CXCL10 and monocyte-derived dendritic cells (MoDCs). In addition, the RORγt agonist promoted Type 17 T cell migration by upregulating CCL20 and CCR6 expression, and Type 17 T cell tumor infiltration. CCL20 induces MoDCs migration, and CXCL10 derived from MoDCs promotes CD8 T cell migration.
Our results revealed that the RORγt agonist improved the efficacy of anti-PD-1. The RORγt agonist increased the migration of MoDCs, which increased the local levels of CXCL10, thus promoting CD8 T cell tumor infiltration. Our findings provide the mechanistic insights implicating the RORγt agonist in immunotherapy and offer a strategy for targeting the RORγt agonist to improve PD-1 antibody efficacy in cancers.
检查点阻断的总体应答率仍不尽人意，部分原因是免疫抑制性肿瘤微环境。一种视黄酸相关孤儿受体γt（RORγt）激动剂（LYC-55716）目前正在与抗PD-1联合进行临床试验，但Th17细胞转录因子RORγt如何在肿瘤微环境中增强PD-1的抗肿瘤免疫力仍不清楚。
使用qPCR分析mRNA表达。流式细胞术用于分选和分析细胞。使用Transwell分析细胞迁移。Biacore用于测定与RORγt蛋白的结合亲和力。基于RORγt GAL4细胞的报告基因检测用于测量RORγt驱动的荧光素酶报告基因表达中的活性。
我们设计了一种强效且选择性的小分子RORγt激动剂（8-074），其在同基因肿瘤模型中显示出强大的抗肿瘤功效，并提高了小鼠肺癌模型中抗PD-1的疗效。RORγt激动剂治疗增加了肿瘤内CD8 T细胞，这与CXCL10和单核细胞衍生的树突状细胞（MoDCs）相关。此外，RORγt激动剂通过上调CCL20和CCR6表达促进17型T细胞迁移以及17型T细胞肿瘤浸润。CCL20诱导MoDCs迁移，而来自MoDCs的CXCL10促进CD8 T细胞迁移。
我们的结果表明RORγt激动剂提高了抗PD-1的疗效。RORγt激动剂增加了MoDCs的迁移，这增加了局部CXCL10水平，从而促进了CD8 T细胞肿瘤浸润。我们的研究结果提供了RORγt激动剂在免疫治疗中的作用机制见解，并提供了一种靶向RORγt激动剂以提高癌症中PD-1抗体疗效的策略。
