假基因来源lncRNA PDIA3P1绑定JMJD2A调控DCN介导子痫前期滋养细胞表型的机制研究

Preeclampsia (PE) is known as a pregnancy-specifc disease. There is a close correlation between the functional phenotype changes of trophoblast and the pathogenesis of PE. Pseudogene PDIA3P1 was significantly down-regulated in PE tissues via high-through microarray detection. The earlier research revealed that silence of PDIA3P1 could change the normal functional phenotype of trophoblast. Transcriptome sequencing suggested that decorin (DCN), the key factor to regulate EVT functional phenotype, might be a candidate target gene of PDIA3P1. With the help of RNA-protein prediction and RIP experiments, we obtained that the Jumonji domain-containing protein 2A (JMJD2A), a histone demethylase, had binding sites with PDIA3P1, and the interaction of the two participated in the regulation of the expression of DCN. Therefore, we hypothesize that pseudogenes PDIA3P1 could regulate the expression of DCN by recruiting and binding JMJD2A to regulate the EVT phenotype in PE. This study intends to clarify the biological characteristics of PDIA3P1 at first. After that, using the cross over expression / silence strategy combined with experiments in vivo and in vitro, we propose to study the specific molecular mechanism of the regulation of DCN expression at the epigenetic level by pseudogenes PDIA3P1 through recruiting JMJD2A. Taking PDIA3P1 as the breakthrough point, the project shall further reveal the molecular mechanism details of PE complex pathological development, and may provide new clues and potential new targets for the prevention and treatment of PE.

子痫前期（PE）是一种妊娠期特发疾病。滋养细胞（EVT）功能表型的改变与PE发病密切相关。PDIA3P1是由芯片筛选、显著低表达于PE组织的假基因来源lncRNA。前期工作发现，沉默PDIA3P1可改变EVT功能表型；转录组测序提示调控EVT功能表型的关键因子核心蛋白聚糖（DCN）是PDIA3P1的下游靶基因；借助RNA蛋白预测及RIP实验，发现PDIA3P1与组蛋白去甲基化酶JMJD2A存在结合位点，且两者的相互作用参与调控DCN。故提出假基因PDIA3P1通过绑定JMJD2A调控DCN表达影响PE中EVT表型的科学假说。本研究拟阐明PDIA3P1的生物学特征；结合体内外多种环境，采用交叉过表达/沉默策略，研究假基因PDIA3P1通过JMJD2A在表观遗传水平调控DCN的分子机制。以PDIA3P1为切入点，将进一步揭示PE复杂病理发展的分子机制细节，并为PE的防治提供新线索和潜在新靶标。

子痫前期（PE）是妊娠期特发多系统多脏器受累的疾病，导致孕产妇及围产儿死亡的主要原因之一，但其具体发病机制尚不清楚。借助芯片筛选，我们发现一条显著低表达于PE组织的假基因来源lncRNA PDIA3P1。本研究在人胎盘组织标本、血液样本、滋养细胞系、动物模型等多个层次，采用芯片技术、转录组测序、生物信息学、qRT-PCR、western blotting、RNA干扰、核质分离、FISH、ChIP、流式细胞技术等手段，从分子水平、细胞水平、在体水平进行研究并得到研究结论：PDIA3P1可能通过募集并绑定JMJD2A蛋白，引起DCN启动子区H3组蛋白第4位赖氨酸去甲基化，使其转录降低，在表观遗传学沉默DCN表达。而在PE疾病中PDIA3P1低表达，其负性调控DCN的能力下降，改变DCN及下游相关信号通路的表达，使得EVT增殖及血管生成能力减弱、凋亡增多，从而造成子宫螺旋动脉重铸障碍。本研究将PDIA3P1引入PE发病机制研究中，并进行连续性的深入研究，从全新的角度系统阐述PDIA3P1在PE发生发展中的作用机制，为PE发病机理研究提供新方向，并为临床防治提供理论依据。