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（申请人提供）：视网膜色素上皮细胞（RPE）和绒毛膜毛细血管（CC）在眼睛中有着密切的关系。作为最近的邻居，它们仅被布鲁赫膜隔开，它们相互依赖，相互帮助，以完成维持健康的光感受器细胞的关键作用。本研究探讨了这两种细胞类型从绒毛膜的发育或出生（妊娠5-23周）到两种不同形式的年龄相关性黄斑变性（AMD）、地理萎缩（GA）和渗出性AMD中任一细胞类型的功能障碍或死亡之间的关系。使用酶和免疫组织化学（IH）以及TUNEL标记来评估CC的代谢状态和活力。RPE细胞的活力和功能将通过IH和TUNEL标记进行研究。生长因子（FGF， VEGF， PIGF，血管生成素，红细胞生成素，干细胞因子，基质衍生因子-1），两种细胞类型产生并用于自身（自分泌）或邻近（旁分泌），及其同源受体将通过IH和组织漂白进行研究。在胎儿RPE存在或不存在的情况下，将使用中和抗体和siRNA评估生长因子的旁分泌和自分泌作用。IH将研究白细胞和补体对AMD中CC死亡和功能障碍以及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（TNFa、IL6、IL8、MCP-1）和血管生成（FGF、VEGF、PIGF、血管生成素）细胞因子。补体因子H、攻击复合体、白细胞和CC活力之间的空间关系也将被研究。利用我们的碱性磷酸酶平面包埋技术，可以量化CC血管密度和RPE覆盖的布鲁赫膜百分比，我们已经建立并将继续研究CC和RPE之间的互惠共生关系。这一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。这项建议的目标是更好地理解这两种相邻细胞类型之间的关系，并阐明可能有针对性地维持或改善这种健康关系的重要因素。