女性不孕新致病基因AAAS的鉴定和分子机制研究

早期胚胎停育是导致女性不孕及反复试管婴儿失败的常见原因之一，但其遗传因素和分子机制大多未知。此前，申请人及其所在团队成员成功鉴定出导致人类早期胚胎停育的致病基因PADI6、NLRP2、NLRP5和REC114，但仍有近90%的患者原因不明。因此，本项目的科学问题是：导致早期胚胎停育的遗传因素还有哪些？是否存在新的机制？.为回答以上问题，申请人前期利用全外显子组测序结合生物信息分析，鉴定出导致早期胚胎停育的新致病基因AAAS。初步研究发现，突变导致AAAS蛋白细胞定位异常。后续拟在本项目的支持下，扩大患者样本数，寻找AAAS新突变，建立突变谱与表型谱；同时，构建AAAS点突变和敲除鼠模型，通过体外受精，显微注射以及免疫共沉淀等方法，系统研究AAAS在早期胚胎发育中的生理功能和突变导致早期胚胎停育的分子机制。该研究将为部分胚胎停育患者明确病因，同时为临床诊断和遗传咨询提供靶点。

Early embryonic arrest is the important causes of female infertility. However, the genetic factors and molecular mechanism are largely unknown. In previous studies, our group has successfully identified PADI6, NLRP2, NLRP5 and REC114 responsible for early embryonic arrest via the approach of medical genetics, but about 90% of patients still could not find the genetic pathogenesis So, this project aiming is: What other genetic factors lead to early embryonic arrest? Is there a new mechanism? .for answering these questions, we analyzed whole exome sequencing data and found that patients diagnosed with early embryonic arrest carried recessive mutations of AAAS which is a member of nuclear pore complex. We found that mutations could lead proteins mislocalized in primary research. With the support of this project, we will expand the samples, screen AAAS mutations and establish the mutational and phenotypic spectrum. By way of constructing point mutation and knock-out mouse models, on the other hand, we will investigate the molecular mechanism of AAAS in early embryonic development, and AAAS mutations leading to early embryonic arrest by multiple methods including in-vitro fertilization, microinjection and Co-Immunoprecipitation, etc. Finally, we hope to explain the pathogenesis for some patients diagnosed with early embryonic arrest and give the target of clinical diagnosis and genetic counseling for them in future.