Birth and Death of Choriocapillaris.

脉络膜毛细血管的出生和死亡。

基本信息

批准号：
7627254
负责人：
Gerard Anthony Lutty
金额：
$ 35.83万
依托单位：
JOHNS HOPKINS UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2006
资助国家：
美国
起止时间：
2006-09-01 至 2011-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7627254
关键词：
Age Age related macular degeneration Alkaline Phosphatase Angiogenesis Inhibitors Angiopoietins Antibodies Apoptosis Area Atrophic Basement membrane Birth Blood Vessels Bruch&apos s basal membrane structure CCL2 gene CD34 gene Caspase Cell Adhesion Molecules Cell Death Cell Survival Cells Cessation of life Choroid Collagen Complement Complement Factor H Complex Cryoultramicrotomy Cytokeratin Development Embryonic Development Endostatins Endothelial Cells Equilibrium Erythropoietin Exudative age-related macular degeneration Eye Family Family member Fibroblast Growth Factor Functional disorder Gel Goals Growth Growth Factor Histocytochemistry Human IL6 gene IL8 gene Immunohistochemistry In Situ Nick-End Labeling In Vitro Inflammatory Intercellular adhesion molecule 1 Label Labyrinth fenestration Leukocyte Adhesion Molecules Leukocyte-Adhesion Receptors Leukocytes Maps Mediating Metabolic Neutrophil Infiltration P-Selectin PTPRC gene Photoreceptors Pregnancy Preparation Production Relative (related person)Research Personnel Role Series Small Interfering RNA Specimen Staging Stem Cell Factor Structure of retinal pigment epithelium Techniques Thrombospondin 1 Tissues Transmission Electron Microscopy Tube Up-Regulation Vascular Cell Adhesion Molecule-1 Vascular Endothelial Growth Factor Receptor Vascular Endothelial Growth Factors Western Blotting White Blood Cell Count procedure angiogenesis autocrine carboxylesterase cell type cytokine density fetal geographic atrophy improved inhibitor/antagonist member monocyte neutralizing antibody paracrine pigment epithelium-derived factor programs receptor spatial relationship

项目摘要

DESCRIPTION (provided by applicant): The retinal pigment epithelial cells (RPE) and the choriocapillaris (CC) have an intimate relationship in the eye. As nearest neighbors, separated only by Bruch's membrane, they rely on each other and assist each other in accomplishing their critical role of sustaining healthy photoreceptor cells. This proposal explores the relationship between these two cell types from the development or birth of choriocapillaris (5-23 weeks gestation) to the dysfunction or death of either cell type in two different forms of age-related macular degeneration (AMD), geographic atrophy (GA) and exudative AMD. The metabolic status and viability of CC will be evaluated using enzyme and immunohistochemistry (IH) and TUNEL labeling. RPE cell viability and function will be investigated with IH and TUNEL labeling. Growth factors (FGF's, VEGF, PIGF, angiopoietins, erythropoietin, stem cell factor, stromal derived factor-1) that the two cells types produce and use for themselves (autocrine) or their neighbor (paracrine), and their cognate receptors will be investigated by IH and bleaching of the tissue. Tube formation in vitro by fetal human endothelial cells in the presence or absence of fetal RPE will be assessed for paracrine and autocrine effects of growth factors using neutralizing antibodies and siRNA. The contribution of leukocytes and complement to CC death and dysfunction and to CNV in AMD will be investigated by IH to identify cell types, leukocyte numbers (nonspecific esterase, CD45, CD68), leukocyte adhesion molecules (ICAM-1, VCAM-1, P-selectin), and pro-inflammatory (TNFa, IL6, IL8, MCP-1) and angiogenic (FGF's, VEGF, PIGF, angiopoietins) cytokines. The spatial relationship between complement factor H, attack complex, leukocytes and CC viability will also be investigated. Using our alkaline phosphatase flat-embedding technique in which CC vascular density and percent of Bruch's membrane covered with RPE can be quantified, we have established and will continue to investigate the mutualistic symbiotic relationship between CC and RPE. This analysis will permit us to establish how this relationship is disrupted in GA and exudative AMD. The goal of this proposal is to better understand the relationship between these two neighboring cell types and elucidate important factors that could be targeted to maintain or improve this healthy relationship.

描述（由申请人提供）：视网膜色素上皮细胞（RPE）和脉络膜毛细血管（CC）在眼睛中具有密切的关系。作为最近的邻居，仅由布鲁赫膜隔开，它们相互依赖并相互协助，以完成维持健康感光细胞的关键作用。该提案探讨了这两种细胞类型之间的关系，从脉络膜毛细血管的发育或出生（妊娠 5-23 周）到两种不同形式的年龄相关性黄斑变性 (AMD)、地图样萎缩（妊娠 5-23 周）中任一细胞类型的功能障碍或死亡之间的关系。 GA) 和渗出性 AMD。 CC 的代谢状态和活力将使用酶和免疫组织化学 (IH) 以及 TUNEL 标记进行评估。 RPE 细胞活力和功能将通过 IH 和 TUNEL 标记进行研究。将研究两种细胞类型为自身（自分泌）或相邻细胞（旁分泌）产生和使用的生长因子（FGF、VEGF、PIGF、血管生成素、促红细胞生成素、干细胞因子、基质衍生因子-1）及其同源受体通过 IH 和组织漂白。在存在或不存在胎儿 RPE 的情况下，将使用中和抗体和 siRNA 评估生长因子的旁分泌和自分泌效应，评估胎儿人内皮细胞在体外的管形成。将通过 IH 研究白细胞和补体对 CC 死亡和功能障碍以及 AMD 中 CNV 的贡献，以确定细胞类型、白细胞数量（非特异性酯酶、CD45、CD68）、白细胞粘附分子（ICAM-1、VCAM-1、P -选择素）、促炎剂（TNFa、IL6、IL8、MCP-1）和血管生成剂（FGF、VEGF、PIGF、血管生成素）细胞因子。补体因子 H、攻击复合物、白细胞和 CC 活力之间的空间关系也将被研究。使用我们的碱性磷酸酶平面嵌入技术，可以量化 CC 血管密度和被 RPE 覆盖的布鲁赫膜的百分比，我们已经建立并将继续研究 CC 和 RPE 之间的互利共生关系。该分析将使我们能够确定这种关系在 GA 和渗出性 AMD 中是如何被破坏的。该提案的目标是更好地了解这两种相邻细胞类型之间的关系，并阐明可用于维持或改善这种健康关系的重要因素。