PLAGL1印迹控制区异常甲基化影响辅助生殖胚胎及子代长期发育的机制研究

The long-term health of the offspring born by assisted reproductive technology (ART) has received considerable public attention. At present, the most obvious internal relation between ART and genetic risks lies in the changes of newborns’ genomic imprinting. However, the underlying mechanism is still unclear. The applicant has found that abnormal DNA reprogramming existed in preimplantation blastocysts in ART, and has identified 33 human imprinting control region (ICR) including PLAGL1. On this basis, this project will take advantage of the placenta and embryo tissues from the abortion and reduction in ART. Focusing on PLAGL1, which is an important control component of an imprinting gene network (IGN), this project will explore the DNA methylation and expression of PLAGL1 and its ICR in ART, select their potential ART sensitive CpG sites and discuss their influence on pregnancy outcome consulting clinical manifestation. Meanwhile, this project will use epigenetic editing technology, explore the effects of PLAGL1 ICR’s abnormal DNA methylation on the methylation and expression of PLAGA1 and the other imprinting gene in IGN， as well as elucidating the effects on embryo development. In conclusion, from the perspective of ICR, this project will replenish the mechanism of embryonic heteroplasia and birth defect, and provide theoretical basis for improving ART genetic security.

辅助生殖子代的长期健康受到社会的普遍关注。目前辅助生殖与遗传风险之间最明显的联系为新生儿基因组印记的改变，但其机制尚不明确。申请人前期发现辅助生殖中囊胚存在异常的DNA甲基化重编程，并鉴定出PLAGL1等33个人类印迹基因控制区（ICR）。在此基础上，本课题将进一步针对印迹基因网络的重要调控组分PLAGL1，对该基因及其ICR在辅助生殖及自然受孕子代临床样本中的甲基化及表达谱进行分析，结合临床表型探讨辅助生殖中基因印迹发生异常的机制，以及ICR对胚胎发育的影响。同时，本课题将利用表观遗传编辑技术，在杂合小鼠胚胎中探究早期胚胎PLAGL1的ICR异常DNA甲基化对PLAGL1等印记基因的甲基化状态以及基因表达的影响，阐明PLAGL1印记控制区的异常重编程对胚胎发育的影响。综上，本课题将从ICR的角度，补充辅助生殖中胚胎发育异常以及出生缺陷的发生机制，为提高辅助生殖的遗传安全性提供理论依据。

辅助生殖子代的长期健康受到社会的普遍关注。目前辅助生殖与遗传风险之间最明显的联系为新生儿基因组印记的改变，但其机制尚不明确。申请人前期发现辅助生殖中囊胚存在异常的DNA甲基化重编程，并鉴定出PLAGL1等33个人类印迹基因控制区（ICR）。在此基础上，本课题将进一步针对印迹基因网络的重要调控组分PLAGL1，利用基因编辑技术，在杂合小鼠胚胎中探究早期胚胎PLAGL1的ICR异常对PLAGL1等印记基因的甲基化状态以及基因表达的影响，阐明PLAGL1印记控制区的异常重编程对胚胎发育的影响。综上，本课题将从ICR的角度，补充辅助生殖中胚胎发育异常以及出生缺陷的发生机制，为提高辅助生殖的遗传安全性提供理论依据。