人NOTCH2基因突变导致原发性卵巢功能不全的分子机制研究

原发性卵巢功能不全（POI）是女性不孕症中的一种严重疾病，表现为生殖力与卵泡储备显著降低。遗传基因突变是POI发病的原因之一，目前仍有大多数病例未找到致病原因。本课题前期针对显性遗传的POI家系母女患者进行全外显子组测序，定位到新的潜在致病基因NOTCH2及突变D1853H，并在100例散发POI样本中发现1个NOTCH2突变D1151N。我们根据前期研究推测NOTCH2基因突变可能通过影响颗粒细胞功能和发育进程进而导致POI，为验证假说本课题拟研究突变对NOTCH2下游靶基因转录的作用，对颗粒细胞分泌雌激素的影响以及对细胞凋亡的作用；然后利用人类胚胎干细胞体外诱导分化为颗粒细胞的体系结合基因编辑技术研究突变颗粒细胞发育进程中的受累情况，使用电镜对细胞器进行亚显微分析，并结合RNA测序手段研究NOTCH2突变致病的分子机理。通过本研究将有助于阐明与颗粒细胞功能和发育有关的POI致病机理。

Primary ovarian insufficiency (POI) is a severe type of women infertility due to the very low ovarian follicle reserve. Perturbations in genetic factors can cause or are associated with POI.We know very little about the molecular mechanism of POI despite enormous efforts to understand the genetic pathogenesis. Our stury performed whole-exome sequencing on two POI patients in a Chinese pedigree and identified a heterozygous mutation D1853H in NOTCH2 gene. Mutational analysis on 100 idiopathic POI patiens also found one heterozygous mutatoin D1151N. Further study will reveal the molecular mechanism of NOTCH2 mutations causing POI. We will perform molecular and cellular experiments to study the mutation's effect on estrogen secretion by granulosa cell, the downstream target genes and granulosa cell apoptosis. We will also use the platform of induction of granulosa cell from human embryonic stem cells combined with gene editing technology to study the mutation's effect on granulosa cell development.Electron microscope and RNA sequencing will be used to dissect the molecular pathology of POI. This study will aid us to elucidate the molecular mechanism of POI which is associated with granulosa cell function and development.