The Choriocapillaris in Aging and Macular Disease

描述（由申请人提供）：本项目旨在研究未被研究的问题，即绒毛膜毛细血管在早期年龄相关性黄斑变性（AMD）中的作用。提案中描述的初步数据表明，绒毛膜毛细血管内皮细胞是补体介导的AMD细胞裂解的靶标。这些细胞因损伤而丧失，和/或内皮细胞表型因损伤而改变，可能是这种致盲疾病发病机制的关键步骤。拟议的研究将处理这些问题。申请人提出，与绒毛膜内皮细胞损伤相关的事件可能导致灌注不足、内皮细胞活化、炎症、细胞外基质的异常合成或周转和/或新生血管形成，因此可能在AMD的发病机制中起重要作用。该项目的第一个具体目的是在大量供体眼睛中检查补体复合物与绒毛膜毛细血管在衰老和早期AMD中的关系，并将这种关系与同一只眼睛的绒毛膜毛细血管的形态特征联系起来。第二个特定目的是评估与AMD相关的内皮细胞活化，这是内皮细胞对补体介导的损伤和炎症的一种反应。我们将对分离的绒毛膜毛细血管进行定量分子分析，并对人供眼进行免疫组织化学研究，以探讨AMD中内皮细胞活化的问题。第三个具体目的是检查绒毛毛细血管细胞在体外的行为，特别是在补体介导的损伤反应。脉络膜的器官培养将在促进补体复合物形成的条件下进行处理，随后进行免疫组织化学和分子分析。内皮细胞培养将从人类供体的眼睛中制备，其长期目标是评估它们对补体介导的损伤和其他类型损伤的反应。这些文化的发展将为申请人在该领域的未来研究提供重要资源。这些实验有望为绒毛膜毛细血管在AMD病理生物学中的潜在作用提供新的见解。

DESCRIPTION (provided by applicant): This project is directed toward examining the understudied question, the role of the choriocapillaris in early age-related macular degeneration (AMD). Preliminary data described in the proposal suggest that choriocapillary endothelial cells are a target for complement-mediated cell lysis in AMD. The loss of these cells due to injury, and/or the change in endothelial cell phenotype as a consequence of injury, may be a crucial step in the pathogenesis of this blinding disorder. The proposed studies will address these issues. The applicant proposes that events associated with choriocapillaris endothelial cell injury are likely to lead to lack of perfusion, endothelial cell activation, inflammation, aberrant synthesis or turnover of the extracellular matrix, and/or neovascularization, and therefore may play a significant role in the pathogenesis of AMD. The first specific aim of this project is to examine the association of complement complexes with the choriocapillaris in aging and in early AMD in a large series of donor eyes, and to correlate this association with the morphometric characteristics of the choriocapillaris of the same eyes. The second specific aim is designed to assess endothelial cell activation in association with AMD, which is one response of endothelial cells to complement-mediated injury and inflammation. Quantitative molecular analyses on the isolated choriocapillaris and immunohistochemical studies on human donor eyes will be performed to examine the question of endothelial cell activation in AMD. The third specific aim is to examine the behavior of choriocapillaris cells in vitro, particularly in response to complement mediated injury. Organ cultures of the choroid will be treated with conditions that promote complement complex formation, followed by immunohistochemical and molecular analyses. Endothelial cell cultures will be prepared from human donor eyes, with the long-term goal of assessing their response to complement-mediated injury and other types of injury. The development of these cultures will represent an essential resource for future studies by the applicant in this field. These experiments are expected to provide novel insights into the potential role of the choriocapillaris in the pathobiology of AMD.