EXTRACELLULAR PROTEINASES IN RETINAL NEOVASCULARIZATION

描述：（改编自申请人摘要）视网膜 新血管形成或从预先存在的新血管生长 在美国，视网膜血管是导致失明的主要原因 在工作年龄的成年人（糖尿病视网膜病变）和婴儿（视网膜病变 早产）。公认缺氧是最初的刺激 在这些条件下视网膜中的新血管形成或血管生成。 血管生成过程中的最后一个途径是细胞的侵入。 细胞外基质和微血管内皮细胞迁移。 这一阶段依赖于多种细胞外因子的表达。 蛋白酶，包括丝氨酸蛋白酶尿激酶和 基质金属蛋白酶（MMP）家族。这项提案将考验 以下假设：视网膜新生血管化是由 缺氧诱导视网膜细胞外蛋白酶表达 微血管内皮细胞这项研究的广泛、长期目标是 研究这些蛋白酶在视网膜血管生成中的作用， 为了测试这些蛋白酶的拮抗剂在抑制 新船编队。该提案的具体目标是：（1） 表征蛋白酶和抑制剂的表达， 血管生成的活跃阶段，通过使用生物化学，组织学， 分子生物学技术的时空表达 这些蛋白酶及其抑制剂将在井中进行研究， 视网膜新生血管形成的特征小鼠模型。的作用 在这个过程中，视网膜中的不同细胞类型将由 免疫组织化学和原位杂交技术。(2)以确定 分离的视网膜毛细血管内皮细胞是否能产生反应 与缺氧无关，其蛋白酶/抑制剂发生变化 profile.这将通过暴露培养的视网膜内皮细胞来研究。 细胞缺氧和/或血管内皮生长因子（VEGF）。还有， 将检测缺氧是否在蛋白质表达中起任何作用， 通过缺氧诱导因子（HIF-1）的产生和/或激活， 1 α）。(3)为了确定视网膜新生血管形成的过程 可以通过使用特定的蛋白酶抑制剂来抑制 小鼠模型

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.