EXTRACELLULAR PROTEINASES IN RETINAL NEOVASCULARIZATION

描述：(改编自申请人的摘要)视网膜 原有血管的新生血管或新血管的生长 视网膜血管是美国致盲的主要原因 在工作年龄成人(糖尿病视网膜病变)和婴儿(视网膜病变)之间 早产)。众所周知，低氧是最初的刺激因素。 在这些情况下视网膜中的新生血管或血管生成。 血管生成过程中的最后一条途径是血管的侵入。 细胞外基质与微血管内皮细胞迁移。 这一阶段依赖于多种胞外因子的表达 蛋白水解酶，包括丝氨酸蛋白水解酶、尿激活酶和 基质金属蛋白酶(MMPs)家族。这项提案将考验 以下假设：视网膜新生血管是由 缺氧诱导视网膜细胞外蛋白水解酶表达 微血管内皮细胞。这项研究的广泛、长期目标 研究这些蛋白在视网膜血管生成中的作用，以及 目的：检测这些蛋白水解酶拮抗剂的抑制作用。 新的舰船编队。该建议的具体目标是：(1) 描述蛋白水解酶和抑制物在体内的表达。 血管生成的活跃期通过生化、组织学和 分子生物学技术。时间和空间上的表达 这些蛋白酶及其抑制物将在油井中进行研究- 描述了视网膜新生血管的小鼠模型。的作用 在这一过程中，视网膜中不同的细胞类型将由 免疫组织化学和原位杂交技术。(二)确定 分离的视网膜毛细血管内皮细胞是否有反应 蛋白水解酶/抑制物的变化独立于低氧 侧写。这将通过暴露培养的视网膜内皮细胞来研究。 细胞对低氧和/或血管内皮生长因子(VEGF)的作用。另外， 我们将研究低氧对蛋白质表达是否有影响。 通过产生和/或激活缺氧诱导因子(HIF- 1阿尔法)。(3)确定视网膜新生血管的形成过程 可以通过使用特定的蛋白酶抑制剂来抑制 老鼠模型。

DESCRIPTION: (Adapted from applicant's abstract) Retinal neovascularization or growth of new blood vessels from pre-existing retinal vessels is the leading cause of blindness in the United States among working-age adults (diabetic retinopathy) and infants (retinopathy of prematurity). It is well accepted that hypoxia is the initial stimulus for neovascularization or angiogenesis in the retina in these conditions. The final, pathway in the angiogenic process is the invasion of extracellular matrix and migration of microvascular endothelial cells. This phase is dependent upon the expression of a variety of extracellular proteinases including the serine proteinase urokinase and members of the matrix metalloproteinase (MMP) family. This proposal will test the following hypothesis: retinal neovascularization is facilitated by the hypoxia-induced expression of extracellular proteinases by retinal microvascular endothelial cells. The broad, long-term goals of this study are to examine the role of these proteinases in retinal angiogenesis, and to test the efficacy of antagonists to these proteinases in inhibition of new vessel formation. Specific aims of the proposal are: (1) To characterize the expression of proteinases and inhibitors during the active stages of angiogenesis by using biochemical, histological, and molecular biological techniques. The spatial and temporal expression of these proteinases and their inhibitors will be studied in the well- characterized mouse model of retinal neovascularization. The role of different cell types in the retina in this process will be determined by immunohistochemical and in situ hybridization techniques. (2) To determine whether isolated retinal capillary endothelial cells can respond independently to hypoxia with a change in their proteinase/inhibitor profile. This will be studied by exposing cultured retina endothelial cells to hypoxia and/or vascular endothelial growth factor (VEGF). Also, it will be examined whether hypoxia plays any role in protein expression through the production and/or activation of hypoxia inducible factor (HIF- 1 alpha). (3) To determine if the process of retinal neovascularization can be inhibited through the use of specific proteinase inhibitors in the mouse model.