人卵母细胞和早期胚胎发育异常的分子机制及干预研究

About 10-15% couples are facing reproductive failure worldwide, which has been deveoped as a major issue affecting our society but at least to our family. In China, the infertile couples are over 50 millions. Although artificial reproductive technology (ART) has been quickly developed and many infertile couples have been benifited from it, recurrent abortion even with ART remains very high. It is well known that this high recurrent abortion is closely related with germ cell developmental defect and early embryo developmental failure. However, the molecular insights of this developmental failure remains largely unexplored. In this study, we will first collect samples from these patients with recurrent abortion caused by germ cell or early embryo developmental failure. Then through whole genome exome sequencing and bioinformatics analysis, the candidate gene mutations can be potentially identified. By further using CRISPR-Cas9 gene editing technology, we can successfully generate mouse models with the specific mutations obtained through sequencing. All these efforts are expected to disclose the molecular mechanism of the reproductive failure caused by either germ cell or early embryo developmantal failure. Furthermore, we can further rescue this developmental abnormalities. Overall, we have built-up very strong gene knock-in mouse platform, as well as the high-thorughput sequencing bioinformatic analysis technology. We believe that through this study, we can identify many critical gene mutations causing both germ cell and early embryo developmental failure. Most importantly, we will put our efforts to rescue these defects.

全球大约有10～15％的育龄夫妇面临不孕不育的困境，给家庭和社会带来了很大影响。我国不孕不育患者已超过5000万。随着生殖医学的发展，辅助生殖技术（ART）已成为治疗不孕不育症的重要手段之一。但是，目前辅助生殖反复失败的发生率非常高，其中与配子及早期胚胎发育异常密切相关。本项目将在之前研究基础上广泛收集人卵母细胞和早期胚胎发育异常的患者，利用全基因组外显子测序并结合生物信息学分析及基因功能验证筛选出致病基因。通过实验室成熟的CRISPR/Cas9技术高效构建携带致病基因突变的小鼠疾病模型，对患者和小鼠疾病模型的研究将揭示致病基因突变导致卵子及胚胎发育异常的分子机制。项目还将对此类疾病的诊断方法进行探讨，结合致病基因的分子机理探索有效安全的干预方法，为此类疾病的治疗提供实验基础。本项目的实施将为由于卵子和早期胚胎异常导致的不孕不育患者的临床诊断和治疗提供指导，并最终提高辅助生殖治疗的成功率。

配子和胚胎发育对生殖健康至关重要，在国家自然科学基金委重点项目的资助下，本项目围绕配子和早期胚胎发育的调控机制，圆满完成了各项既定研究目标，主要成果包括：1）广泛收集了卵子和早期胚胎发育异常的临床样本，通过全外显子测序及生物信息学方法筛选出多个致病基因；2）利用CRISPR/Cas9等技术建立了多个人卵子及早期胚胎发育异常的动物疾病模型；3）结合胚胎显微操作技术和微量细胞测序的方法揭示多种导致人和小鼠早期胚胎发育异常的表观遗传机制；4）通过对小鼠模型及临床样品的研究发现高龄男性精子tsRNA改变与子代神经系统疾病相关，衰老引起tsRNAs变化可将焦虑代际遗传给雄性子代。相应研究成果发表在Nature Cell Biology、Cell Stem Cell、Nature Communication等国际知名学术期刊上共计20余篇。研究中发现的致病基因突变及一些治疗方法，已经应用于临床不孕不育患者的辅助生殖诊治。本项目相应的研究成果及方法已申报2项发明专利。通过本项目的实施，培养硕士研究生2名，博士研究生毕业5名。其中国家奖学金2人，校级奖学金3人，上海市优秀毕业生1人，校级优秀学生3人。培养团队青年教师获得国家、省部级人才项目9项，所培养的博士生获得国家博士后创新人才支持计划项目3项。在项目实施过程中负责人以第一完成人分别获得2019年教育部自然科学一等奖和2020年度国家自然科学奖二等奖。