Regulating retinal microvascular morphogenesis

调节视网膜微血管形态发生

• 批准号: 7101741
• 负责人: IRA M HERMAN
• 金额: $ 35.92万
• 依托单位: TUFTS UNIVERSITY BOSTON
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-08-01 至 2008-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7101741
• 关键词: angiogenesis; biological signal transduction; cell cell interaction; cell migration; cell proliferation; cytoskeleton; epithelium; eye; genetic regulation; genetically modified animals; laboratory mouse; macrophage; pathologic process; retina disorder; tissue /cell culture

DESCRIPTION (provided by applicant): Retinal microvascular morphogenesis is a complex and highly coordinated process, which occurs during embryonic development, post-natally and in association with several visually-impairing diseases, including retinopathy of prematurity (ROP), age-related macular degeneration and diabetic retinopathy. Recent work carried out in the principal investigator's laboratory has revealed that isoactin-based cytoskeletal remodeling is integral to the microvascular migration and proliferation observed during developmental and pathologic angiogenesis, including the pericyte-based remodeling seen during retinal microvascular maturation. In a focused, inter-disciplinary research plan that will take advantage of in vitro and in vivo models using a spectrum of well-established molecular, cell biology-based and molecular genetic approaches, we aim to reveal the molecular mechanisms and the isoactin-based signaling cascades regulating (i) retinal endothelial migration driving normal and pathologic angiogenesis, and (ii) pericyte-based control of endothelial proliferation and capillary contractility. Quantitative analyses of retinal microvascular endothelial cell cultures will be performed in conjunction with experiments aimed at over-expressing the novel beta-actin specific binding and filament capping protein, betacap73, discovered in the lab. To reveal the molecular mechanisms driving isoactin-based control of developmental and pathologic angiogenesis, we will take advantage of a 'two-mouse' transgenic approach, where we will specifically induce betacap73 over-expression within the post-natal vascular endothelium. These combined results, revealing alterations in endothelial motility and impaired angiogenesis, will serve to guide cDNA expression array analyses aimed at identifying key signaling effectors controlling these pivotal microvascular events. Further, to reveal the molecular signaling mechanisms regulating pericyte contractility and retinal endothelial growth, we will characterize the role that Rho GTPase family members play in signaling vascular cytoskeletal remodeling and isoactin dynamics during microvascular morphogenesis. Based on the preliminary data recently obtained and the experimental approaches proposed, we anticipate that our research plan will provide important new insights into the molecular mechanisms regulating retinal microvascular morphogenesis during normal development and in association with the pathologic angiogenesis accompanying diabetic retinopathy.

描述（申请人提供）：视网膜微血管形态发生是一个复杂且高度协调的过程，发生在胚胎发育、出生后，并与几种视力损害疾病有关，包括早产儿视网膜病变（ROP）、年龄相关性黄斑变性和糖尿病性视网膜病变。最近在首席研究员实验室开展的工作表明，在发育和病理性血管生成过程中，以异肌动蛋白为基础的细胞骨架重塑是微血管迁移和增殖的组成部分，包括在视网膜微血管成熟过程中以周细胞为基础的重塑。在一个专注的跨学科研究计划中，我们将利用体外和体内模型，利用一系列成熟的分子、细胞生物学和分子遗传学方法，揭示分子机制和基于等肌动蛋白的信号级联调节(i)视网膜内皮细胞迁移驱动正常和病理血管生成，以及（ii）基于周细胞的内皮细胞增殖和毛细血管收缩性控制。视网膜微血管内皮细胞培养的定量分析将与实验室中发现的新型β -肌动蛋白特异性结合和丝盖蛋白betacap73的过度表达实验一起进行。为了揭示驱动基于异肌动蛋白的发育和病理性血管生成控制的分子机制，我们将利用“双小鼠”转基因方法，在那里我们将在出生后血管内皮中特异性诱导betacap73过表达。这些综合结果揭示了内皮运动的改变和血管生成的受损，将有助于指导cDNA表达阵列分析，旨在识别控制这些关键微血管事件的关键信号效应物。此外，为了揭示调节周细胞收缩和视网膜内皮生长的分子信号机制，我们将描述Rho GTPase家族成员在微血管形态发生过程中血管细胞骨架重塑和等肌动蛋白动力学信号传导中的作用。基于最近获得的初步数据和提出的实验方法，我们预计我们的研究计划将为正常发育过程中调节视网膜微血管形态发生的分子机制以及与糖尿病视网膜病变相关的病理性血管生成提供重要的新见解。