中药卷柏GPR54拮抗剂的发现及干预子宫内膜异位症的作用机理研究

GPR54具有重要生理功能，是妇科顽症子宫内膜异位症的潜在靶标。Treg细胞介导的免疫耐受在內异症中起重要作用，GPR54对Treg细胞有调控作用；但GPR54/Treg轴在內异症中的作用机制不明，迄今尚无天然小分子GPR54拮抗剂报道。课题组前期研究发现中药卷柏提取物对GPR54有拮抗作用，小鼠内异症模型显效，高通量筛选得到小分子拮抗剂，据此提出“中药卷柏GPR54拮抗剂调控免疫相关激素受体GPR54干预内异症”的科学假说。基于FLIPR和CLIP技术的高通量筛选平台导向分离，利用分子对接、质谱技术、定点突变和功能检验研究GPR54和拮抗剂相互作用机制，揭示GPR54结构和功能的关系；构建內异症患者ESCs-Treg细胞共培养体系、基因敲除和正常小鼠內异症模型，以GPR54拮抗剂为工具药物，研究GPR54在内异症中的病理机制。本研究预期可为内异症治疗提供新思路，为靶向药物设计奠定基础。

With important physiological functions, G protein-coupled receptor 54 (GPR54) is a potential target for endometriosis, a gynecologic persistent disease. Regulatory T cells (Treg) play a key role in the pathogenesis of endometriosis via the induction of immune tolerance, and GPR54 could regulate the Treg cells. However, natural non-peptidic GPR54 antagonist has not been reported as yet, and the mechanism of GPR54/Treg axis underlying endometriosis remains unclear. Our previous studies on Selaginella led to the discovery of natural GPR54 antagonists from Herba Selaginellae, possessing the inhibitory effect on endometriotic lesion formation in a mouse model. Therefore, we propose that GPR54 antagonists from Herba Selaginellae could produce a therapeutic effect on endometriosis via immunomodulation of GPR54. The main topics of this research are as follows: first, target-guided isolation of GPR54 antagonists based on FLIPR platform, and study on interaction mechanisms of GPR54 and its antagonists using the technology of molecular docking, targeted mutagenesis and ESI-Q-TOF-MS; GPR54 antagonists, as tool drugs, will be used to investigate the effect of GPR54 in the pathogenesis of endometriosis in vitro and in vivo on primary co-culture system of human endometrial cells and Treg cells, and normal/GPR54 gene knockout mouse model. This research aims to shed new light in treatment strategy for endometriosis, and laid the foundation for drug design in endometriosis.