FIBROBLAST GROWTH FACTOR I IN VASCULAR PATHOBIOLOGY

血管生成是许多疾病的特征，包括糖尿病 视网膜病变，类风湿性关节炎，动脉粥样硬化，以及生长和 实体瘤的转移与成纤维细胞的原型膜 生长因子家族是血管生成生长因子家族中的一员。 直接作用于血管细胞，诱导内皮细胞生长和 迁移。虽然这个应用程序的长期目标是阐明 成纤维细胞生长因子-1启动的信号通路最终可能揭示新的 抗血管生成治疗的策略，当前的目标是 研究成纤维细胞生长因子受体-1(FGFR-10)与皮质蛋白的相互作用， 最近发现的成纤维细胞生长因子受体的底物，以及src家族和 成纤维细胞生长因子-1诱导的信号转导通路的潜在成分。 我们将研究这些信号调节器在成纤维细胞生长因子-1中的功能 介导的细胞在体外和体外的迁移和增殖 体内血管生成。具体来说，我们将确定酪氨酸 皮质蛋白中对成纤维细胞生长因子-1反应发生磷酸化的残基 磷酸肽图和磷酸氨基酸分析及其功能 这些位点中的一个将通过定点突变进行研究。它是 预计这些研究的结果将为 确定与酪氨酸有关的真正的激酶(S) 皮质蛋白的磷酸化。虽然我们的初步数据表明 C-src和FGFR-1都能与Cortactin结合，我们将 通过研究确定哪一种与Cortactin更相关的激酶 体内成纤维细胞生长因子-1依赖的酪氨酸磷酸化。一旦 定义了FGFR-1、Cortactin和c-src之间的相互关系，我们将 进一步启动删除、定点突变研究以解剖 它们与蛋白质结合所涉及的氨基酸序列 细胞骨架，并检查负责共同- 皮质蛋白和c-src在细胞骨架中的定位。基于 根据这些研究的结果，我们将尝试设计一系列 可能干扰相互作用的显性负性突变体 在野生型FGFR-1，c=-src和Cortactin之间。这些变种人会 最终被利用来研究它们在成纤维细胞生长因子-1中的生物学作用- 介导的DNA合成和趋化作用及其作为 体内抗血管生成药物。

Angiogenesis is a feature of many diseases, including diabetic retinopathy, rheumatoid arthritis, atherogenesis, and the growth and metastasis of solid tumors and the prototype membranes of fibroblast growth factor (FGF) family are among the angiogenic growth factors that act directly on vascular cells to induce endothelial cell growth and migration. While the long-term goal of this application is to elucidate the signaling pathway initiated by FGF-1 which may ultimately reveal new strategies for antiangiogenic therapy, the immediate goal is to characterize the interaction of FGF receptor-1 (FGFR-10) with cortactin, a recently identified substrate for the FGF receptor, and src family and potential components of the FGF-1 induced signal transduction pathway. We will examine the function of these signaling modulators in FGF-1 mediated cell migration and proliferation in vitro and during angiogenesis in vivo. Specifically, we will identify the tyrosine residues in cortactin that are phosphorylated in response to FGF-1 using phosphopeptide mapping and phosphoamino acid analysis and the function of these sites will be studied by site-directed mutagenesis. It is anticipated that the results from these studies will provide criteria to define the authentic kinase(s) responsible for the tyrosine phosphorylation of cortactin. While our preliminary data suggest that both c-src and FGFR-1 are able to associate with cortactin, we will ascertain which is the more relevant kinase for cortactin by studying FGF-1-dependent tyrosine phosphorylation in vivo. Once the interrelationship among FGFR-1, cortactin and c-src are defined, we will further initiate deletion, site-directed mutagenesis studies to dissect the amino acid sequences involved in their association with the cytoskeleton and examine the mechanism responsible for the co- localization of cortactin and c-src with the cytoskeleton. Based on the results from these studies, we will attempt to design a series of dominant negative mutants that may interfere with the interactions between wild-type FGFR-1, c=-src and cortactin. These mutants will ultimately be exploited to study their biological roles in the FGF-1- mediated DNA synthesis and chemotaxis, and their potential as antiangiogenics in vivo.