YAP1调控卵泡膜-间质细胞雄激素合成在PCOS高雄发生中的作用机制

Polycystic ovary syndrome (PCOS) is a common ovarian dysfunction and endocrine metabolic disease affecting women of childbearing age, which is mainly characterized by signs of androgen excess, menstrual irregularitis and polycystic ovary, in which 60-80% PCOS are androgen excess. Our genome-wide associated study (GWAS) found that YAP1 is PCOS susceptibility gene. Our previous studies have shown that YAP1 specifically expressed in ovarian interstitial membranes cells (TICs); the testosterone level increased dramatically while the expression of YAP1 is upregulated in the preantral follicle and TICs, which indicated that Hippo pathway and YAP1 may function in the androgen synthesis in TICs. On the basis of our preliminary results, we use various experimental techniques, including RT-PCR, western blot, in situ hybridization, inmmunohistochemistry and viral infection to investigate that: 1) the differential expression of YAP1 and Hippo pathway in ovary tissue of PCOS patients and normal control; 2) the role and mechanism of YAP1 in androgen synthesis in TICs; 3) the treatment effect of anti-androgen drugs and corresponding inhibitors on PCOS androgen excess. This study will provide theoretical foundation and experimental evidence for the diagnosis, treatment and mechanism of PCOS.

多囊卵巢综合征（PCOS）是育龄期女性最常见生殖障碍与代谢紊乱性疾病，临床表现为高雄、月经稀发、卵巢多囊样改变等，其中60-80％PCOS患者雄激素水平过高。本课题组通过对PCOS患者进行全基因组关联分析发现Hippo通路靶因子YAP1为PCOS易感基因，且前期研究结果显示YAP1在小鼠卵泡膜-间质细胞（TICs）内高表达；上调小鼠窦前卵泡与TICs内YAP1表达后，其雄激素分泌水平显著上升，提示Hippo通路及YAP1参与TICs雄激素合成途径。本课题拟在此研究基础上，应用RT-PCR、蛋白印迹、原位杂交、免疫组化、病毒感染等多种实验方法，研究：1)YAP1及Hippo通路在PCOS患者与正常对照卵巢内表达差异；2)YAP1在TICs雄激素合成中的调控作用；3)抗雄药物及相应抑制剂对高雄的治疗作用机制，为PCOS的临床诊断治疗和发病机制提供理论基础与实验依据。

多囊卵巢综合征(PCOS)是育龄期女性最常见生殖障碍与代谢紊乱性疾病，临床表现为高雄、月经稀发、卵巢多囊样改变等，其中60-80%PCOS患者雄激素水平过高。本课题组通过对PCOS患者进行全基因组关联分析发现Hippo通路靶因子YAP1为PCOS易感基因，且本项目研究显示：(1) YAP1在小鼠卵巢内广泛表达，与小鼠卵泡膜-间质细胞、颗粒细胞内均高度表达，提示YAP1可能参与小鼠卵巢的生物学功能。(2) 构建可过表达YAP1病毒的腺病毒载体及阴性对照，以及可有效下调YAP1的小干扰RNA，分别感染或转染小鼠卵泡膜-间质细胞后，可见YAP1改变小鼠卵泡膜-间质细胞(TICs)增殖水平。(3) 改变小鼠窦前卵泡与TICs内YAP1表达后, 其雄激素分泌水平显著改变，且StAR与CYP17α1等雄激素合成关键酶类的RNA与蛋白表达水平相应发生改变；(4) 进一步研究发现，通过双荧光素酶报告基因系统检测可见YAP1增强了CYP17α1及StAR的启动子活性；(5) 同时ChIP assay实验表明YAP1通过结合StAR与CYP17α1启动子不同结合位点发挥调控作用。本项目应用RT-PCR、蛋白印迹、病毒感染、双荧光素酶报告基因系统及ChIP assay等多种实验方法，针对性研究YAP1分子对卵泡膜-间质细胞的增殖、雄激素合成与分泌功能的调控机制，为PCOS临床诊断治疗及发病机制提供了理论基础，并提供了新的治疗靶点。