Endothelial Cell Migration and Retinal Angiogenesis

内皮细胞迁移和视网膜血管生成

• 批准号: 7057341
• 负责人: ROBERT S FISCHER
• 金额: $ 18.33万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-05-01 至 2007-09-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7057341
• 关键词: RNA interference; actin binding protein; actins; angiogenesis; cell migration; cytoskeleton; fluorescence microscopy; laboratory mouse; method development; protein structure function; retina; stem cells; time resolved data; vascular endothelial growth factors; vascular endothelium

DESCRIPTION (provided by applicant): Endothelial cell migration is critical to normal neovascularization, which is in turn essential for retinal development and function. Conversely, abnormal angiogenesis is the primary cause in the vast majority of diseases that result in vision loss, including age-related macular degeneration and diabetic retinopathy. While there has been some progress in understanding the factors that promote retinal angiogenesis, the mechanisms that drive the underlying endothelial cell migration process are not well understood. The research outlined here will develop a novel direct correlation approach to mechanistically dissect the cell locomotion functions required for endothelial migration both in vitro and in vivo. As a test case, this proposal will investigate the role of cofilin, an actin binding protein known to be important for motility in other cell types. In Aim 1, the function of cofilin in endothelial cell migration in vitro will be examined by expression of cofilin mutants and RNA interference. Quantitative fluorescence image analysis and live-cell, time-lapse microscopy will be used to determine how the actin dynamics are altered in cells with perturbed cofilin function, and how this affects cell migration behavior. In Aim 2, the functions of cofilin will be similarly perturbed in endothelial-precursor stem cells, which will then be intravitreally injected into neonatal mice. The ability of these stem cells to migrate during retinal neovascularization, and to integrate into the retinal vasculature will be monitored by quantitative fluorescence microscopy analyses of the retinal vasculature at various time intervals. This novel integrated approach will lead to a fundamental understanding of the endothelial cell migration processes that are specifically required for retinal neovascularization, and will lay the groundwork for future comprehensive studies of regulatory networks controlling retinal angiogenesis. Furthermore, this research will initiate development of in vitro assays with greater predictive power for identification of novel targets and compounds for anti-angiogenesis treatments.

描述（由申请人提供）：内皮细胞迁移对于正常新生血管形成至关重要，而正常新生血管形成又对于视网膜发育和功能至关重要。相反，血管生成异常是绝大多数导致视力丧失的疾病的主要原因，包括年龄相关性黄斑变性和糖尿病视网膜病变。虽然在了解促进视网膜血管生成的因素方面取得了一些进展，但驱动潜在内皮细胞迁移过程的机制尚不清楚。这里概述的研究将开发一种新颖的直接相关方法，以机械方式剖析体外和体内内皮迁移所需的细胞运动功能。作为一个测试案例，该提案将研究丝切蛋白的作用，丝切蛋白是一种肌动蛋白结合蛋白，已知对其他细胞类型的运动很重要。在目标1中，将通过肌动蛋白丝切蛋白突变体的表达和RNA干扰来检查肌动蛋白丝切蛋白在体外内皮细胞迁移中的功能。定量荧光图像分析和活细胞延时显微镜将用于确定丝动蛋白功能受到干扰的细胞中肌动蛋白动力学如何改变，以及这如何影响细胞迁移行为。在目标 2 中，内皮前体干细胞中的丝切蛋白功能将受到类似的干扰，然后将其玻璃体内注射到新生小鼠体内。这些干细胞在视网膜新生血管形成过程中迁移以及整合到视网膜脉管系统中的能力将通过视网膜的定量荧光显微镜分析来监测 不同时间间隔的脉管系统。这种新颖的综合方法将带来根本性的 了解视网膜特别需要的内皮细胞迁移过程 新生血管形成，将为未来的监管综合研究奠定基础 控制视网膜血管生成的网络。此外，这项研究将启动 具有更强预测能力的体外测定，可用于鉴定新靶点和化合物 抗血管生成治疗。