Birth and Death of Choriocapillaris.

脉络膜毛细血管的出生和死亡。

• 批准号: 7454181
• 负责人: Gerard Anthony Lutty
• 金额: $ 35.11万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 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/7454181
• 关键词: Age; Age related macular degeneration; Alkaline Phosphatase; Angiogenesis Inhibitors; Angiopoietins; Antibodies; Apoptosis; Area; Atrophic; Basement membrane; Birth; Blood Vessels; Bruch' 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; Geographic Locations; 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 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; Week; Western Blotting; White Blood Cell Count procedure; angiogenesis; autocrine; carboxylesterase; cell type; cytokine; density; fetal; improved; inhibitor/antagonist; member; monocyte; neutralizing antibody; paracrine; pigment epithelium-derived factor; programs; receptor; spatial relationship

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，它们只被布鲁赫膜分开，它们相互依赖，相互帮助，完成维持健康感光细胞的关键作用。该提案探索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），地图状萎缩（GA）和渗出性AMD中任一细胞类型的功能障碍或死亡。将使用酶和免疫组织化学（IH）和TUNEL标记评价CC的代谢状态和活力。将用IH和TUNEL标记研究RPE细胞活力和功能。将通过IH和组织漂白研究两种细胞类型产生并用于自身（自分泌）或其邻近细胞（旁分泌）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。将使用中和抗体和siRNA评估在存在或不存在胎儿RPE的情况下由胎儿人内皮细胞体外形成管的生长因子的旁分泌和自分泌效应。将通过IH研究白细胞和补体对AMD中CC死亡和功能障碍以及对CNV的贡献，以鉴定细胞类型、白细胞数量（非特异性酯酶、CD 45、CD 68）、白细胞粘附分子（ICAM-1、VCAM-1、P-选择素）和促炎（TNF α、IL 6、IL 8、MCP-1）和血管生成（FGF、VEGF、PIGF、血管生成素）细胞因子。还将研究补体因子H、攻击复合物、白细胞和CC活力之间的空间关系。使用我们的碱性磷酸酶平面包埋技术，其中CC血管密度和百分比的Bruch的膜覆盖的RPE可以量化，我们已经建立并将继续调查CC和RPE之间的互惠共生关系。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，并阐明可以靶向维持或改善这种健康关系的重要因素。