AGE-induced Phenotype of the Retinal Pigment Epithelium

描述（由申请人提供）：虽然许多基因已经被 被假定参与年龄相关性黄斑变性（AMD）， 美国老年人失明的主要原因，一个全面的mRNA 视网膜色素上皮细胞（RPE）在体内的表型是目前 未知确定RPE mRNA表型可以识别基因 对AMD的发展至关重要，确定疾病的标志物，并导致 新的预防和治疗策略。布鲁赫膜基底沉积物 是在RPE退化之前发生的早期可识别的变化 并且是AMD的组织病理学标志物。最近，我们的实验室发现， 晚期糖基化终末产物（AGEs）的年龄依赖性积累， 布鲁赫膜和基底沉积物。AGEs是在衰老过程中形成的结构， 糖或其他前体之间的一系列非酶反应，以及 改变多种细胞类型表型的长寿蛋白质。我们 实验室最近也表明AGEs改变基因表达 参与RPE细胞中的基质调节、细胞极性和凋亡。我们 长期目标是确定健康人RPE的广泛mRNA表型 和AMD，这将反过来产生对AMD发病机制的见解。 具体来说，我们要定义表型的组成部分， 由AGEs控制。我们假设基底沉积物中AGEs的存在 负责RPE细胞表型的转换， 基质调节、RPE细胞极性和细胞凋亡的改变。到 测试这个假设，我们问3个问题：1。黄斑变性的mRNA表型 视网膜色素上皮细胞覆盖在基底层和正常布鲁赫膜上有什么不同？2. AGE修饰的基质是否在体外诱导RPE mRNA表型， 视网膜色素上皮细胞表达的表型的基础存款？3.是一 受体介导的AGE诱导的RPE细胞mRNA表型的亚群 晚期糖基化终末产物为了回答这些问题，我们将使用 激光捕获显微切割以从组织获得RPE细胞的纯样品 用于通过微阵列分析进行mRNA表型分析的样本。我们还将利用 体外AGE-基质系统，以确定哪些基因亚组受 年龄该项目希望建立一个广泛的RPE mRNA表型， 健康和AMD，并定义了一个子集的基因诱导的AGEs。

DESCRIPTION (provided by applicant): While a number of genes have been postulated to be involved in age-related macular degeneration (AMD), the leading cause of blindness among the elderly in the US, a comprehensive mRNA phenotype of the retinal pigment epithelium (RPE) in vivo is at present unknown. Determination of an RPE mRNA phenotype could identify the genes critical to the development of AMD, identify markers of disease, and lead to new preventative and treatment strategies. Basal deposits in Bruch's membrane are an early recognizable change that occur prior to degeneration of the RPE and are a histopathological marker for AMD. Recently, our laboratory identified an age-dependent accumulation of advanced glycation end products (AGEs) in Bruch's membrane and basal deposits. AGEs are structures formed during the series of nonenzymatic reactions between sugars or other precursors, and long-lived proteins that alter the phenotype of a variety of cell types. Our laboratory has also recently shown that AGEs alter the expression of genes involved in matrix regulation, cell polarity, and apoptosis in RPE cells. Our long term goal is to define an extensive mRNA phenotype of the RPE in health and AMD which will in turn, yield insights into the pathogenesis of AMD. Specifically, we want to define the component of that phenotype which is regulated by AGEs. We hypothesize that the presence of AGEs in basal deposits is responsible for a switch in the phenotype of RPE cells that is consistent with alterations in matrix regulation, RPE cell polarity, and apoptosis. To test this hypothesis we ask 3 questions: 1. Are the mRNA phenotypes of macular RPE cells overlying basal deposits and normal Bruch's membrane different? 2. Does AGE modified matrix induce an RPE mRNA phenotype in vitro that is a subset of the phenotype expressed by RPE cells overlying basal deposits? 3. Is a subset of the AGE induced mRNA phenotype of RPE cells mediated by the Receptor for Advanced Glycation End products? To answer these questions we will use laser capture microdissection to obtain pure samples of RPE cells from tissue specimens for mRNA phenotyping by microarray analysis. We will also utilize an in vitro AGE-matrix system to determine what subset of genes is regulated by AGEs. This project hopes to establish an extensive mRNA phenotype of the RPE in both health and AMD, and define a subset of genes induced by AGEs.