DNA氧化损伤修复基因microRNA结合位点的序列变异及其功能改变与年龄相关性白内障的关系

项目名称： DNA氧化损伤修复基因microRNA结合位点的序列变异及其功能改变与年龄相关性白内障的关系

项目编号： No.81470616

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： 管怀进

作者单位： 南通大学

项目金额： 73万元

中文摘要： 年龄相关性白内障（ARC）发病机制仍不清楚。我们之前的研究发现DNA氧化损伤修复基因启动子和编码区序列变异与ARC有关,但尚未探讨其致病机理。本课题假设可直接调节mRNA及蛋白质表达的氧化损伤修复基因3′UTR区microRNA结合位点的序列变异是导致ARC的分子机制。拟以人群样本和医院病例为对象（独立双样本），检测氧化损伤修复基因microRNA结合位点的序列改变及基因分型，分析其与ARC发病的关系；收集晶状体前囊膜上皮细胞分析相应基因的表达、细胞损伤与基因型和ARC的关系；转染不同基因型质粒到晶状体上皮细胞株证实基因型与基因表达的关系；转染microRNA的模拟物、阻遏物和对照质粒到晶状体上皮原代培养细胞，测试其对相应基因的调节及细胞损伤的影响。以上基因的功能调控和干预研究可进一步阐明ARC的发病机制，为设计以microRNA 为靶点的不同基因型ARC的个性化防治奠定基础。

中文关键词： 年龄相关性白内障；氧化损伤修复基因；3′非翻译区序列变异；微小RNA；功能改变

英文摘要： The pathogenesis of age-related cataract (ARC) remains unclear. Our previous study found that sequence variation of DNA oxidative damage repair gene in the promoter and coding region were associated with ARC, but not yet its pathogenesis. We assume that microRNA binding sequence variation of DNA oxidative damage repair gene 3′UTR region, which can regulate the expression of mRNA and protein directly, is the leading cause of ARC in molecular mechanisms. In this study, we will use samples from a population-based cohort and a hospital-based case-control to test the association of genotypes in microRNA bingding region of DNA oxidative damage repair genes with ARC. The surgery biopsy of lens anterior capsule will be used to measure the gene expression of the targeted genes and DNA lesion, and their relationship with the genotypes. Lens epithelium cell line by transfecting different SNP plasmids will further confirm the role of the genotypes in the gene regulation. Finally, we will establish primary culture lens epithelium cells, test their genotypes, transfect various microRNA mimics/inhibits/controls plasmid to detect the gene expression and DNA lesion. The functional regulation and intervention studies of the above genes may further elucidate the genetic pathogenesis of ARC. Most importantly, it will offer the proof of concept for microRNA intervention for ARC personalized prevention of different genotypes and therapy targeting ARC prevention.

英文关键词： Age-related cataract;Oxidative damage repair genes;3′UTR sequence variation;microRNA;Functional changes