脂蛋白调节基因APOE在年龄相关性白内障发病中的作用研究

We have found that the abnormal serum lipid level was an independent risk factor of age-related cataract (ARC) in large population-based epidemiologic study. Furthermore, we also found that two SNPs mutation of the serum lipid protein regulating gene APOE significantly reduced the risk of ARC. In this study, we aim to deeply explore the effect of APOE gene in ARC development and its association with serum lipid protein. First, we plan to study the APOE gene polymorphism distribution and its effect on ARC development in a larger epidemiologic population by SNP-scan technique. Also, we will study the association of APOE polymorphism and serum lipid protein and their roles in ARC development using gene-environmental interaction model. Second, we will culture the human lens epithelia cells (LECs) and transfected the cells using APOE mutation plasmid. Then we will evaluate the expression of lipid protein in the medium and study the LECs apoptosis and viability using the ELISA, the quantitative RT-PCR, the luciferase experiment, the flow cytometry technique, et al. Third, we will confirm our findings in the clinical epidemiology. Briefly, in our research, we will deeply study the effect of APOE mutation in ARC development step by step through the population-based epidemiology, histocytology and clinical epidemiology. Hopefully our research will provide a new way and theoretical basis to the early prevention, control and treatment of ARC, as well as the new clinical drug target.

申请人前期已通过流行病学研究(样本量>1万人)发现了异常血脂水平是年龄相关性白内障(age related cataract, ARC)的重要独立危险因素之一,并通过基因学预实验发现了血脂蛋白调节基因APOE突变可显著降低ARC患病风险。为进一步探索APOE基因在ARC患病中的作用,申请人使用SNP-scan技术,在大规模流调人群中研究APOE基因多态性分布和对ARC患病的影响,并通过基因-环境交互作用分析APOE基因多态性和异常血脂水平在ARC患病中的调节关系。体外培养人晶状体上皮细胞(LECs),通过APOE基因突变转染细胞,利用ELISA、QRT-PCR、流式细胞等方法观察培养液中脂蛋白水平和LECs的活性,并在临床人群中加以验证。因此,本课题拟从人口流行病学、组织细胞学、临床流行病学三个层面分析分析APOE基因在调节ARC发病中的作用，研究结果将为ARC早期防控治疗提供理论依据。

申请人已通过流行病学研究(样本量>1万人)发现了异常血脂水平是年龄相关性白内障 (age related cataract, ARC)的重要独立危险因素之一，并通过基因学预实验发现了血脂蛋白调节基因APOE突变可显著降低ARC患病风险。为进一步探索APOE基因在ARC患病中的作用，申请人使用SNP-scan技术，在大规模流调人群中研究APOE基因突变的分布和对ARC患病的影响，并已完成大样本量(1616 ARC vs 1311 controls)APOE基因突变检测，发现 APOE rs7412 C>T，APOE rs440446 C>G 与 ARC 患病有密切关系。体外成功培养人晶状体上皮细胞(LECs)，成功构建转染质粒，通过APOE基因突变转染细胞，利用ELISA、QRT-PCR、流式细胞等方法观察培养液中脂蛋白水平和LECs的活性，并在临床人群中加以验证。因此，本课题从人口流行病学、组织细胞学、临床流行病学三个层面分析分析APOE基因在调节ARC发病中的作用，研究结果将为ARC早期防控治疗提供理论依据。

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