microRNAs及其靶基因遗传变异与湿性老年黄斑变性易感性的分子流行病学研究

湿性老年黄斑变性（AMD）是一种严重影响中老年人群生活质量的常见致盲眼病。课题组前期经生物信息学发现一些已知湿性AMD易感基因受miRNAs调控；采用测序获得55个miRNAs在湿性AMD模型细胞中差异表达，提示其可能参与该病发生。本项目通过Solexa测序绘制离体-在体miRNA和mRNA表达谱，结合生物信息学分析和双荧光素酶报告基因系统验证，获得候选miRNAs及其靶基因；采用病例对照设计，通过飞行时间质谱进行基因分型，分析候选miRNAs及其靶基因多态性与湿性AMD遗传易感性的关联，确定该病关键miRNAs及其靶基因；应用RNAi和过表达载体，在细胞及动物水平探讨关键miRNAs及其靶基因在湿性AMD发生中的调控机制。本研究从不同层面系统揭示关键miRNAs及其靶基因对罹患湿性AMD风险的影响及分子机制，为湿性AMD的早期诊断、个体化治疗和有效预防策略的建立提供理论依据和实验基础。

Wet age-related macular degeneration (AMD) is a common blinding eye disease which severely compromises life quality of middle aged and elderly people. Previously,we predicted some wet AMD susceptibility genes are regulated by microRNA (miRNAs), using bioinformatics approach; and then found that 55 miRNAs were different expressed between wet AMD model cells and controls using Solexa sequencing technology, which indicates miRNAs might be involved in the disease. In this project, we intend to study the expression profiles of miRNA and mRNA in vitro and in vivo using Solexa sequencing technology. In combination with the results of bioinformatics analysis and dual-luciferase reporter assay system, candidate miRNAs and their target genes are obtained. A population-based case-control study is conducted, and the relationships between polymorphisms of the candidate miRNAs and their target genes and genetic susceptibility of wet AMD are analyzed after genotyping using matrix-assisted laser desorption/ionization time of flight mass spectrometry; and the key miRNAs and their target genes are determined. After establishing cell and animal models, we discuss the regulatory mechanism of the key miRNAs and their target genes for wet AMD using technologies of RNAi and over expression. This study will reveal the influence and the molecular mechanism of the key miRNAs and their target genes on the risk of wet AMD from multi-aspect, so as to provide theoretical and experimental basis for early diagnosis and individualized treatment for wet AMD, and the establishment of effective prevention strategies.