Role of TWEAK and Fn14 in Vascular Biology

TWEAK 和 Fn14 在血管生物学中的作用

• 批准号: 6370325
• 负责人: JEFFREY A WINKLES
• 金额: $ 26.99万
• 依托单位: AMERICAN NATIONAL RED CROSS
• 依托单位国家: 美国
• 财政年份: 1988
• 资助国家: 美国
• 起止时间: 1988-03-01 至 2006-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6370325
• 关键词: angiogenesis; autocrine; biological signal transduction; cell growth regulation; cell population study; cell proliferation; chemoattractants; cytokine; cytokine receptors; embryo /fetus cell /tissue; gene expression; gene targeting; genetically modified animals; human tissue; immunoprecipitation; intermolecular interaction; laboratory mouse; mammalian embryology; membrane proteins; molecular site; protein structure function; site directed mutagenesis; tissue /cell culture; vascular endothelium; western blottings; yeast two hybrid system

The long-term goal of the research in our laboratory is to understand the molecular mechanisms of vascular endothelial cell (EC) and smooth muscle cell (SMC) growth control in vivo. These two cell types are generally in a quiescent growth state; however, the onset of EC proliferation is associated with the pathogenesis of numerous "angiogenic diseases" (e.g., solid tumor growth) while SMC accumulation occurs during the development of atherosclerosis and restenosis after vessel wall injury. We have used a gene discovery approach to study polypeptide growth factor-stimulated cell proliferation. Several novel growth factor-inducible genes have been identified, and one of these genes, named Fn14, is the subject of this competitive renewal application. Recent studies have indicated that the Fn14 plasma membrane protein is a receptor for the tumor necrosis factor (TNF)-related cytokine named TWEAK. This cytokine is a type II transmembrane protein that occurs in both cell-associated and soluble forms. Although TWEAK has been reported to be a weak inducer of cellular apoptosis, it is thought to primarily function as a survival and growth factor. Indeed, it was reported in 1999 that TWEAK is an EC mitogen and an angiogenic factor in the rat cornea angiogenesis assay. Additional studies are required to further evaluate the biological functions of TWEAK and Fn14 using both cell culture- and animal-based experimental systems. Accordingly, the Specific Aims of this proposal are: 1) to confirm and extend previous results indicating that TWEAK can stimulate EC proliferation in vitro and promote angiogenesis in vivo. In addition, we will determine whether TWEAK is a chemotactic factor for EC and if TWEAK biological activity on EC is mediated via binding to the Fn14 receptor; 2) To determine whether endogenously- expressed TWEAK can function in an autocrine manner via the Fn14 receptor to regulate EC survival/proliferation in vitro; 3) To begin investigating the TWEAK signal transduction pathway by identifying proteins that bind the Fn14 cytoplasmic tail and by determining whether TWEAK addition to quiescent EC or Fn14 overexpression in transfected HEK293 cells activates signaling pathways that promote cell survival/proliferation; and 4) To determine the phenotypic consequences of Fn14 deficiency in vivo. It is anticipated that these studies will provide important information on the functions of the TWEAK and Fn14 proteins and their role in vascular cell biology.

该研究中该研究的长期目标是了解体内血管内皮细胞（EC）和平滑肌细胞（SMC）生长控制的分子机制。 这两种细胞类型通常处于静止的生长状态。但是，EC增殖的发作与许多“血管生成疾病”（例如实体瘤生长）的发病机理有关，而SMC的积累发生在血管壁损伤后动脉粥样硬化和再狭窄的过程中发生。 我们已经使用了一种基因发现方法来研究多肽生长因子刺激的细胞增殖。 已经鉴定出了几种新型生长因子诱导因子基因，其中一个名为FN14的基因之一是这种竞争性更新应用的主题。 最近的研究表明，FN14质膜蛋白是肿瘤坏死因子（TNF）相关的细胞因子的受体。 该细胞因子是一种II型跨膜蛋白，以细胞相关和可溶性形式出现。 尽管据报道调整是细胞凋亡的较弱诱导剂，但被认为主要是生存和生长因子。 实际上，据报道，1999年据报道，调整是大鼠角膜血管生成测定法中的EC有丝分裂原和血管生成因子。需要进行其他研究来进一步评估使用基于细胞和动物的实验系统的Tweak和FN14的生物学功能。因此，该提案的具体目的是：1）确认并扩展先前的结果，表明调整可以在体外刺激EC增殖并促进体内血管生成。 此外，我们将确定调整是否是EC的趋化因子，以及通过与FN14受体结合介导的EC的TWEAK生物学活性。 2）确定内源性表达的调整是否可以通过FN14受体以自分泌方式起作用，以调节体外的EC生存/增殖； 3）通过鉴定结合FN14细胞质尾巴的蛋白质以及确定在转染的HEK293细胞中激活促进细胞存活/增殖的信号途径中，通过鉴定结合FN14细胞质尾巴的蛋白质并确定是否在静脉EC或FN14过表达中添加调整来开始研究调整信号转导途径； 4）确定体内FN14缺乏的表型后果。 可以预料，这些研究将提供有关调整和FN14蛋白功能及其在血管细胞生物学中的作用的重要信息。