PCP通路相关基因罕见变异及异常甲基化与胎儿无脑畸形的关联研究

项目名称： PCP通路相关基因罕见变异及异常甲基化与胎儿无脑畸形的关联研究

项目编号： No.81472987

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 医药、卫生

项目作者： 王琳琳

作者单位： 北京大学

项目金额： 85万元

中文摘要： 无脑畸形（anencephaly, AN）是一类具有高致死性的先天性神经管缺陷，是导致围产儿死亡的主要原因之一，主要表现为胎儿有少量的神经组织但头盖骨结构全部或部分缺失。AN病因复杂，尚不完全清楚。研究显示遗传因素占全部病因的50%以上。本项目采用基于核心家系的病例对照研究，病例为引产AN患儿，对照为引产无缺陷胎儿，对细胞平面极化通路(planar cell polarity, PCP)相关基因进行罕见变异和甲基化分析，以期发现与AN发生相关的新变异位点。同时，对新变异位点进行细胞水平和人群研究两方面的功能分析。细胞水平研究采用免疫共沉淀和酵母单杂交方法，分析目标基因新变异对蛋白质间或蛋白质-DNA间相互作用的影响；人群研究采用病例对照方法，检测病例和对照胎儿脑组织目标基因mRNA及蛋白的表达水平，分析表达差异性，揭示AN发生的分子遗传及表观遗传学机制。

中文关键词： 罕见变异；DNA甲基化；无脑畸形；PCP通路

英文摘要： Anencephaly (AN) is a kind of common congenital malformations with high rate of lethality that often lead to perinatal deaths. It is a condition that prevents the normal development of the brain and the bones of the skull. The etiology of AN remains poorly understood. Up to 50% of the variance in AN prevalence attributes to genetic factors. In the proposed research project, we aim to examine the association of AN with new rare variants and aberrant DNA methylation in planar cell polarity (PCP) pathway genes, by using case-control study based on nuclear family. Cases are fetuses terminated due to prenatal diagnosis of an AN and controls are fetuses terminated not for birth defects. Then, functional analysis will be conducted by using both human study and cell culture study. Co-Immunoprecipitation and yeast one-hybrid system will be performed to explore the effect of new variants on interaction between protein-protein and protein-DNA, respectively. In the human case-control study, protein and mRNA expression of candidate gene will be assayed in the brain tissues of cases and controls, and the differential expression will be determined. The proposed research project will partly discover the genetic and epigenetic mechanism in PCP pathway contributing to the development of AN.

英文关键词： rare variant;DNA methylation;anencephaly;PCP pathway