Drusen and AMD: sub-type isolation and characterization

描述（由申请人提供）：本申请的广泛、长期目标是从分子角度定义黄斑视网膜色素上皮（RPE）下软蛋白积累与发生年龄相关性黄斑变性（AMD）风险增加之间的联系。黄斑软性囊肿的存在是AMD发展的标志性危险因素。令人惊讶的是，人们对德鲁森的成分和起源知之甚少。为此目的，已经开发了一种新的方法，可以从供体眼组织中收集微克量的drusen。在分离时，可以鉴定和分离不同的毒株亚型，用于表征其分子组成的研究。老年性黄斑变性的诊断效用可比作血液中胆固醇水平的动脉粥样硬化。与血液中胆固醇的水平一样，drusen的存在和丰富程度表明了患者患这种疾病的风险程度。由于drusen与AMD的关系，了解不同drusen亚型的组成将为drusen发展的可能途径提供重要信息。在drusen中出现的新蛋白或常见的蛋白修饰，应该为治疗AMD的潜在药物靶点提供见解。目前的应用主要集中在利用这种drusen分离程序来确定正常和AMD组织中drusen亚型的分子组成、分布和细胞起源。三个具体目的是：(1)验证从供体眼组织中可以分离出不同亚群的假设。（硬的vs软的，中央凹的vs外围的，老的vs老的，球形的vs扁平的，琥珀色的vs透明的，不透明的vs粒状的，等等）。(2)通过光镜、组织化学、电镜、SDS/PAGE、Western blotting、质谱等方法，验证不同亚群结构特征反映不同分子组成的假设。(3)为了验证在drusen中存在的一些分子是新的并且在Bruch膜中没有发现的假设。

DESCRIPTION (provided by applicant): The broad, long-term objective of this application is to define in molecular terms the linkage between the accumulation of soft drusen below the retinal pigment epithelium (RPE) in the macula and the increased risk of developing age-related macular degeneration (AMD). The presence of soft drusen in the macula is the hallmark risk factor for developing AMD. Surprisingly little is known of the composition or origin of drusen. To this end a novel method for drusen isolation has been developed that allows the collection of microgram quantities of drusen from donor eye tissue. At the time of isolation, different drusen sub-types can be identified and separated for use in studies that will characterize their molecular composition. The diagnostic utility of drusen in AMD can be likened to that of blood levels of cholesterol in atherosclerosis. The presence and abundance of drusen, like the level of cholesterol in the blood, indicates the degree to which a patient is at risk for developing the disease. Because of the relationship of drusen and AMD, understanding the composition of different drusen sub-types will provide important information on possible pathways that are causally involved in drusen development. Novel proteins or common modifications of proteins present in drusen, should provide insight as to potential drug targets of therapeutic agents to treat AMD. The current application is focused on exploiting this drusen isolation procedure to define the molecular composition, distribution and cellular origin of drusen sub-types in normal and AMD tissues. The three specific aims are: (1) To test the hypothesis that different sub-populations of drusen can be isolated from donor eye tissue. (hard vs. soft, foveal vs. peripheral, old vs. older, spherical vs. flat, amber vs. clear, opaque vs. granular, etc.). (2) To test the hypothesis that different structural features of drusen sub-populations reflect different molecular composition (light microscopy, histochemistry, electron microscopy, SDS/PAGE, Western blotting, mass spectrometry). (3) To test the hypothesis that some of the molecules present in drusen are novel and are not found in Bruch's membrane.