人类卵子成熟障碍新致病基因CDC20及ANAPC4的鉴定及致病机制研究

Oocyte maturation arrest is a genetic disease that seriously affects the reproductive health of women. Recently, our research group found the first gene TUBB8 whose mutation leads to human oocyte maturation arrest and illuminate the pathogenic mechanism. Previous studies show that TUBB8 mutations can only explain about 30% of the patients with oocyte maturation arrest while 70% remains unexplained, suggesting the existence of other genetic factors in the disease..In order to explore new genes related to human oocyte maturation arrest, we collected patient samples and found that mutations in meiosis key genes CDC20 and ANAPC4 resulted in human MI oocyte arrest through the methods of medical genetics. In future support of this project, we plan to identify novel mutations of CDC20 and ANAPC4 or new genes in the same signaling pathway. Phenotype will be confirmed using in vitro oocyte culture system and in vivo gene mutation mouse model. Gene function affected by mutation will be probed by protein interaction experiment and signaling pathway analysis applying molecular biology methods such as immunofluorescence and immunoblot. Finally, we will clarify the molecular mechanism of human oocyte maturation arrest resulted from CDC20 and ANAPC4 mutation which provides strong theoretical basis for precise diagnosis and intervention for patients with oocyte maturation arrest.

卵子成熟障碍是严重危害女性生殖健康的遗传疾病，此疾病的遗传原因及机制所知甚少。最近，我们课题组发现了第一个导致人类卵子成熟障碍的突变基因TUBB8并研究了致病机制，研究显示，TUBB8突变只可解释大约30%的卵子成熟障碍患者，仍有70%患者原因不明，提示还有其他致病基因在该疾病中发挥作用。.为寻找卵子成熟障碍的新致病基因，我们前期利用患者样本，通过医学遗传学方法及分析初步发现减数分裂过程中关键基因CDC20和ANAPC4在卵子MI期阻滞患者中存在突变。拟在本项目的资助下，进一步在疾病样本中鉴定CDC20和ANAPC4的新突变及该通路中的新致病基因；利用卵子体外培养及突变小鼠明确突变引起的表型；运用免疫荧光、免疫印迹等系列分子生物学方法研究相关基因突变的功能、相互作用及信号通路变化，阐明CDC20和ANAPC4导致卵子MI期阻滞的分子机制，最终为卵子成熟障碍患者的精确诊断及干预奠定理论基础。

卵子成熟障碍是严重危害女性生殖健康的遗传疾病，此疾病的遗传原因及机制所知甚少。本项目研究目标为根据前期鉴定的减数分裂过程中关键候选基因CDC20和ANAPC4，通过扩大样本寻找卵子成熟阻滞患者的新突变，并深入研究相关基因的致病机制。本项目完成了卵子成熟阻滞样本收集及相关基因突变筛查，鉴定出CDC20为卵子成熟阻滞新致病基因，并扩展了突变相关疾病表型包括卵子成熟阻滞、受精障碍及早期胚胎停育，研究了相关的分子机制，通过扩大样本筛查拓展了CDC20基因导致女性不孕的突变谱，探索了疾病的干预治疗方法。发表标注本项目SCI论文2篇，其中本领域高影响力期刊1篇，申请国内发明专利2项。研究成果将为相关疾病的机制认识、遗传咨询及分子治疗奠定基础。