Age-Related Macular Degeneration: Genetic Variations and Animal Model

老年性黄斑变性（老年性黄斑变性，AMD）是世界上老年人群中导致不可逆中枢性视力丧失的主要原因。各种研究表明AMD有显著的遗传成分。目前的证据支持基因变异导致疾病易感性的假设。2003年，我们启动了这个项目，招募了晚期AMD患者和视网膜正常的年龄控制个体。迄今为止，已有419人被纳入研究，并收集了60例AMD的组织病理学病例。我们还收到并分析了来自澳大利亚蓝山眼科研究的835份DNA样本和来自AREDS历史NEI项目的534份DNA样本。我们比较了AMD和对照组之间候选基因内单核苷酸多态性（snp）的等位基因频率，并通过体外和/或体内实验对这些snp进行了功能研究。通过这种方法，我们已经确定了AMD的遗传危险因素以及这些基因变异在疾病发病机制中的可能作用。基于上述方法获得的信息，已经产生了一种基因工程动物作为AMD模型。在2009财年，我们完成了以下工作：(1)建立了成熟的数据挖掘平台，进行大规模数据分析；(2)完成Htra1、TLR3、TLR4 SNP-AMD相关性的研究，并在Ophthalmology、IOVS上发表论文2篇；(3)完成衣原体感染与AMD snp相互作用的研究，并在BJO上发表；(4)进一步表征了AMD小鼠模型Ccl2/Cx3cr1双敲除（DKO）小鼠的视网膜病变，并报道了Htra2/Omi参与该模型的视网膜病变；(4)增加了长链omega-3脂肪酸对DKO小鼠视网膜病变的影响实验，并在AJP上发表论文；(5)开始了各种化合物和基因治疗在体外和体内（DKO）模型的测试；(6)为NIH和制药公司以外的研究人员创建了7份材料转移协议（MTA）和1份合作研究与开发协议（CRADA），以便将我们的DKO模型用于机械和治疗目的。

Age-related macular degeneration (AMD) is the leading cause of irreversible central visual loss in the aged population in the world. Various studies suggest that AMD has a significant genetic component. Current evidence supports the hypothesis that gene variation causes a predisposition to the disease. In 2003, we initiated this project by recruiting advanced AMD patients and age-control individuals with normal retinas. Up to date, 419 individuals have been enrolled and 60 histopathological cases with AMD have been collected. We also received and analyzed 835 DNA samples from the Blue Mountain Eye Study in Australia and 534 DNA samples from a historical NEI project of AREDS. We are comparing the allelic frequencies of single nucleotide polymorphisms (SNPs) within candidate genes between AMD and control subjects followed by the functional studies of these SNPs by in vitro and/or in vivo experiments. Through this approach, we have identified genetic risk factors of AMD and the possible roles of these gene variations in the pathogenesis of the disease. Based on the information obtained from the above approches, a genetically engineered animal has been generated to act as the AMD model. In FY2009, we have accomplished the following: (1) established a sophisticated data mining platform for large scale data analysis; (2) completed the study on Htra1, TLR3 and TLR4 SNP-AMD association and published two papers in Ophthalmology and IOVS; (3) Completed the the study of the interation of Chlamydia infection and AMD SNPs and published the result in BJO; (4)further characterized the retinal lesions in Ccl2/Cx3cr1 double knock-out (DKO) mice - a murine model of AMD, and reported the involvement of Htra2/Omi in the retina lesion of the model; (4) Added the experiments on the test of long-chain omega-3 fatty acid on the retinal lesions of the DKO mice and published the paper in AJP, (5) started to test various compounds and gene therapy in vitro and in vivo (DKO) models; (6) creared 7 Material Transfer Agreetments (MTA), and 1 Collaborative Research and Development Agreement(CRADA) for researchers other than NIH and pharmaceutical company to use our DKO model for mechanistic and therapeutic purposes.