Choriocapillaris Activation in Macular Degeneration

产品说明：视网膜相关性黄斑变性（AMD）是西方世界无法治愈的失明的主要原因，仅在美国就有数百万人受到影响。随着人口老龄化，AMD对社会的负面影响将增加。不幸的是，AMD的发病机制目前知之甚少。由于AMD的流行及其对视力的严重损害，评估AMD的疾病机制是国家眼科研究所的使命的一部分，并在2004年国家眼科和视力研究计划中得到强调。 在这个项目中，我们将研究脉络膜毛细血管激活分子在AMD中的作用。鉴于炎症和补体系统在AMD中的作用，我们建议探讨脉络膜毛细血管内皮细胞的激活可能是这种疾病的发病机制中的中心事件的假设。 该项目的第一个具体目标是检查正常眼睛和AMD眼睛中内皮细胞活化的程度和性质。将对正常眼、早期AMD眼和晚期AMD眼进行持续的分子和组织学研究。 在第二个具体目标中，我们将确定内皮细胞是否在暴露于可能发生在AMD中的微环境损伤后在培养物中被激活。将用补体复合物（使用我们开发的方法）和补体副产物（过敏毒素）挑战内皮细胞培养物。将使用微阵列、免疫细胞化学、真实的时间PCR和Western印迹来研究这些细胞的活化反应。 在第三个具体目标中，我们将使用生物信息学驱动的方法来检验编码内皮激活分子的基因中的突变或多态性与AMD相关的假设。将在30个候选基因中筛选单核苷酸多态性（每年资助6个）。将使用免疫组织化学和人眼睛的蛋白质印迹法对遗传发现进行随访，以检查特定基因型的可能功能后果。 总结：组成血管的内皮细胞可能在AMD中发挥重要作用。这些细胞促进炎症的一种方式是吸引白色血细胞进入发炎组织。内皮细胞通过表达在内皮“活化”过程中丰度增加的细胞表面蛋白来发挥这一作用。我们推测，在这种疾病中，眼内内皮细胞活化是导致组织损伤和异常血管生长的原因。涉及遗传学、计算机工程、生物化学、细胞培养和显微镜的跨学科实验有望提供关于脉络膜毛细血管激活在AMD发展中的潜在作用的新信息。

DESCRIPTION: Age-related macular degeneration (AMD) is the leading cause of untreatable blindness in the Western world, with millions affected in the U.S. alone. As the population ages, the negative impact of AMD on society will increase. Unfortunately, the pathogenesis of AMD is currently poorly understood. Because of the prevalence of AMD, and its severe toll on vision, evaluating disease mechanisms in AMD is part of the mission of the National Eye Institute and is highlighted in the 2004 National Plan for Eye and Vision Research. In this project we will examine the role of choriocapillaris activation molecules in AMD. In view of the role of inflammation and the complement system in AMD, we propose to explore the hypothesis that activation of choriocapillaris endothelial cells may be a central event in the pathogenesis of this disease. The first specific aim of this project is to examine the extent and nature of endothelial cell activation in normal eyes and eyes with AMD. Ongoing molecular and histological studies will be performed on normal eyes, early AMD eyes, and advanced AMD eyes. In the second specific aim we will determine whether endothelial cells become activated in culture following exposure to microenvironmental insults that are likely to occur in AMD. Endothelial cell cultures will be challenged with complement complexes (using methodology we have developed) and complement byproducts (anaphylatoxins). The activation response of these cells will be studied using microarrays, immunocytochemistry, real time PCR and Western blotting. In the third specific aim we will test the hypothesis that mutations or polymorphisms in the genes encoding endothelial activation molecules are associated with AMD using a bioinformatics-driven approach. Single nucleotide polymorphisms will be screened in 30 candidate genes (6 per year of the grant). Genetic findings will be followed up using immunohistochemistry and Western blotting of human eyes to examine the possible functional consequences of specific genotypes. Summary: The endothelial cells that comprise blood vessels are likely to play an important role in AMD. One way that these cells promote inflammation is by attracting white blood cells into the inflamed tissue. Endothelial cells perform this role through expression of cell surface proteins that increase in abundance during endothelial "activation". We hypothesize that endothelial cell activation in the eye is responsible for the tissue injury and abnormal blood vessel growth in this disease. Interdisciplinary experiments involving genetics, computer engineering, biochemistry, cell culture and microscopy are expected to provide new information about the potential role of choriocapillaris activation in the development of AMD.