TAP20, NOVEL INTEGRIN MODULATING PROTEIN

TAP20，新型整合素调节蛋白

• 批准号: 6130914
• 负责人: J Anthony Ware
• 金额: $ 27.87万
• 依托单位: ALBERT EINSTEIN COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-06-01 至 2005-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6130914
• 关键词: angiogenesis; biological signal transduction; cell migration; disease /disorder model; enzyme activity; focal adhesion kinase; guanosinetriphosphatases; human tissue; immunocytochemistry; integrins; laboratory mouse; laboratory rat; membrane proteins; myocardial ischemia /hypoxia; protein kinase; receptor expression; tissue /cell culture; transfection /expression vector; vascular endothelial growth factors; vascular endothelium; vitronectin

DESCRIPTION: (Applicant's Description) Integrin adhesion receptors mediate the interactions between endothelial cells and the extracellular matrix and thus are key determinants of angiogenesis. Modification of certain integrins, in particular, the endothelial cell victronectin receptions alpha-v-beta-3 and alpha-v-beta-5, has been shown to be successful in preventing angiogenesis. These integrin receptors can convey signals from the extracellular matrix to the cell interior; conversely, intracellular signal transduction can influence the affinity of integrins for their ligands. The two integrin receptors for victronectin appear to mediate separate pathways to angiogenesis, which may be regulate separately. Our laboratory has recently identified a new 20 kilodalton intracellular mediator, which we have designated as TAP20 this mediator is regulated by PKCO and modulates the function of the alpha-v-beta-5 integrin. Preliminary data presented in this proposal shows that overexpression of TAP20 in endothelial cells enhances their ability to migrate and form tubules in three dimensional cell culture. The primary focus of this proposal is to investigate whether TAP20 is an important mediator of pathological angiogenesis and is useful in therapeutic angiogenesis in ischemic cardiovascular disease. The molecular mechanism by which TAP20 mediate signal transduction and how that may affect endothelial cell migration and tube formation will be determined. The specific aims for this proposal are first, to test the hypothesis the TAP20 enhances endothelial cell migration and tubule formation by altering release of known autologous agonist or antagonist and/or by affecting intercellular mediators resulting from "outside in" integrin signaling. Whether endothelial cells that overexpress TAP20 release increased amounts of FGF2, VEGF or IL8 and whether the release of antiangiogenic substances such as thrombospondin 2 or TGF-beta is suppressed will be determined. We also will determine whether TAP20 affects cellular migration and tube formation by signaling events associated with alpha-v-beta-5 integrin function, such as activation of ERK, GTPase in the Rho family, Src kinase activity, and focal adhesion formation and kinase (FAK) activity. Antisense for TAP20 to prevent those events will be used whether TAP20 enhances migration specifically in endothelial cells by overexpressing it in non endothelial cells that express alpha-v-beta-5 and in non alpha-v-beta-5 containing cells will be determined. An antibody raised against TAP20 will be used in immunohistochemistry to determine whether the expression of TAP20 in a variety of angiogenic tissues is increased over that in similar tissues in which angiogenesis is not prominent. These tissues include various cancers and ischemic myocardium. Finally whether TAP20 enhances angiogenesis in vivo will be determined by utilizing a corneal model of angiogenesis with endothelial cell that overexpress TAP20 inserted and to inject the TAP20 adenovirus into ischemic rat myocardium to determine whether it enhances angiogenesis following ischemia. We anticipate that these studies should provide data that will be useful in designing new strategic for pharmacologic intervention in ischemic vascular disease and cancer.

描述：（申请人的描述） 整联蛋白粘附受体介导内皮细胞之间的相互作用 细胞外基质，因此是血管生成的关键决定因素。 尤其是某些整合素的修饰内皮细胞 维克特列内染蛋白接收alpha-v-beta-3和alpha-v-beta-5已显示为 成功预防血管生成。 这些整合素受体可以传达 从细胞外基质到细胞内部的信号；反过来， 细胞内信号转导可以影响整联蛋白的亲和力 他们的配体。 victronectin的两个整合素受体似乎介导 单独的血管生成途径，可以分别调节。 我们的 实验室最近确定了一个新的20公斤细胞内介体， 我们已将其指定为TAP20，该调解人由PKCO和 调节α-V-Beta-5整合素的功能。 初步数据 在此提案中提出的表明，内皮中TAP20的过表达 细胞增强其在三维中迁移和形成小管的能力 细胞培养。 该提案的主要重点是调查是否 TAP20是病理血管生成的重要介体，在 缺血性心血管疾病中的治疗性血管生成。 分子 TAP20介导信号转导的机制及其可能如何影响 将确定内皮细胞迁移和管形成。 这 该提案的具体目的是首先测试TAP20的假设 通过更改释放来增强内皮细胞迁移和小管形成 已知的自体激动剂或拮抗剂和/或通过影响细胞间 介体由“外部”整合素信号传导产生。 是否内皮 过表达TAP20释放的细胞增加了FGF2，VEGF或IL8的量 以及抗血管生成物质（例如血小板传播2）是否释放 或将抑制TGF-beta。 我们还将确定是否 TAP20通过信号事件影响细胞迁移和管形成 与alpha-v-beta-5整合素功能相关，例如ERK的激活， RHO家族，SRC激酶活性和局灶性粘附形成的GTPase 和激酶（FAK）活性。 TAP20的反义以防止这些事件 使用TAP20是否通过在内皮细胞中专门增强迁移 在表达alpha-v-beta-5的非内皮细胞中过表达它，在 将确定非α-V-beta-5的非α-V-beta-5。 提出了一种抗体 反对TAP20将用于免疫组织化学来确定是否是否 TAP20在多种血管生成组织中的表达增加了 在类似的组织中，血管生成并不突出。 这些组织 包括各种癌症和缺血性心肌。 最后是否Tap20 增强体内血管生成将通过使用角膜模型来确定 用过表达TAP20插入的内皮细胞的血管生成的作用 将TAP20腺病毒注入缺血大鼠心肌，以确定是否是否 它在缺血后增强了血管生成。 我们预计这些研究 应该提供对设计新战略有用的数据 缺血性血管疾病和癌症的药理干预。