应用人类iPS细胞模型研究Xq26-q28区关键基因缺失导致原发性卵巢功能不全的分子机制

项目名称： 应用人类iPS细胞模型研究Xq26-q28区关键基因缺失导致原发性卵巢功能不全的分子机制

项目编号： No.81471432

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 医药、卫生

项目作者： 谭跃球

作者单位： 中南大学

项目金额： 73万元

中文摘要： 原发性卵巢功能不全（POI）是一类常见的严重损害女性生殖健康的疾病，Xq26-q28杂合性缺失（缺失型）是重要病因之一，但分子机制不清楚，一批逃逸X失活基因的表达下降可能起重要作用。前期的研究发现，诱导性多能干细胞（iPSC）可作为POI疾病模型，缺失型POI患者iPSC诱导成原始生殖细胞（PGC特化）能力下降。本研究拟以构建的iPSC为模型进一步研究：比较正常和缺失型POI的iPSC诱导的PGC的分化、增殖及凋亡等生物学特征；应用单细胞转录组学技术比较正常和POI的iPSC诱导的PGC的表达谱，筛选逃逸X失活的关键基因；结合上述两方面的结果，挑选1-2个关键基因，应用RNAi和过表达技术来探讨所筛选基因与POI的关系，并在特发性POI患者中验证所筛选基因是否为致病基因。本研究将突破缺乏有效模型研究缺失型POI发生分子机制的瓶颈，为最终实现通过靶向基因治疗恢复这些患者的卵巢功能提供基础。

中文关键词： 原发性卵巢功能不全；诱导性多能干细胞；原始生殖细胞；X染色体关键区；特化

英文摘要： Primary ovarian insufficiency (POI) is a type of common disorder which leads to serious deterioation to the reproductive health of the females. Heterozygous deletion of Xq26-q28 is one of the important causes, but the molecular mechanism is still unclear. We have evidence to believe that the decreased expression level of genes escaping X inactivation might play a role in the POI etiology. In our previous studies, we found that induced pluripotent stem cells (iPSC) could be used as a model for POI mechanism research and the ability of primordial germ cells differentiation (PGC specialization) in iPSCs from POI patients with Xq26-q28 deletion was decreased compared to those from females with a normal karyotype. In this study, we propose to investigate the mechanisms of POI in three aspects using the established iPSC model: first, we will compare the biological characters of the induced PGCs between the normal and POI derived iPSCs; second, we will compare the gene expression profile of the induced PGCs between the normal and POI derived iPSCs using single cell RNA transcriptome technique and screen for the critical genes which escape the X inactivation; third, by combining the results of the first two components, we will select one or two critical genes and study whether the ability of PGC specification is contributed by these genes using RNAi and over expression techniques, and confirm whether the deletion of these genes is present in the idiopathic POI patients. This study will overcome the bottleneck of lacking an experimental model in studying the molecular mechanism of of POI and provide the useful data for treating POI via recovering ovarian function in POI patients using targeted gene therapy.

英文关键词： Primary ovarian insufficiency;Induced pluripotent stem cells;Primordial germ cells;X chromosome critical region;Specification