INHIBITION OF ANGIOGENESIS-- PROTEINS BINDING TO TSP2

抑制血管生成——与 TSP2 结合的蛋白质

• 批准号: 6388663
• 负责人: Kiflai Bein
• 金额: $ 13.61万
• 依托单位: DARTMOUTH COLLEGE
• 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/6388663
• 关键词: angiogenesis; angiogenesis inhibitors; binding proteins; cell growth regulation; cell migration; cell proliferation; chimeric proteins; clone cells; collagenase; complementary DNA; disulfide bond; enzyme activity; extracellular matrix proteins; gene deletion mutation; gene expression; metalloproteins; nucleic acid repetitive sequence; protein binding; protein protein interaction; protein structure function; recombinant proteins; thrombospondins; vascular endothelium; western blottings; yeast two hybrid system

The extracellular matrix protein thrombospondin (TSP) 2, like TSP1, interacts with a number of cell surface receptor and extracellular matrix proteins. The immediate goals of this grant are to focus on and analyze the TSP structural features and protein partners relevant to the function of inhibition of angiogenesis. The pursuit of these goals is facilitated by the stimulating, well-equipped and intellectually enriched research environment available for the performance of the work. The findings will lay the framework for long-term goals aimed at understanding the molecular basis of vascular biological functions in vivo. It is widely recognized nowadays that in many pathological conditions a common underlying process is a derangement in angiogenesis. Inhibitors of angiogenesis have potential clinical application in disease states where abnormal blood vessel formation is related to disease progression e.g. solid tumors, hemangiomas and inflammatory diseases. On the other hand, enhancers of angiogenesis would be useful in conditions resulting from insufficient blood supply e.g. wound healing, coronary artery disease and stroke. TSP1 and TSP2 have been of particular interest in the medical sciences recently because of their ability to inhibit angiogenesis. The importance of the TSP1 and TSP2 proteins in the regulation of angiogenesis has been underscored by recent in vivo studies that demonstrated TSP-dependent induction of concomitant tumor resistance, suppression of tumor growth, and inhibition of vascularization of polydimethylsiloxane implants. However, the mechanism(s) of action remains unknown. This gap in our knowledge is reflected in the low potency of thrombospondin-based anti-angiogenic synthetic peptides, which was an order of magnitude less than that of intact TSP1. Since inhibition assays using synthetic peptides and knockout studies suggested the possibility of multiple signal pathways elicited or matrix reactions initiated, we wanted to characterize proteins that interact with TSP2. There is very little known about TSP2 for most studies have been on TSP1. Use of the TSP2 type 1 repeats in a yeast two-hybrid system led to the isolation of 36 independent clones. The aims of the proposed investigation are to study the TSP structural determinants involved in regulation of angiogenesis, analyze the significance of the interaction of TSP1 and TSP2 with the proangiogenic MMP2, and identify and functionally characterize novel TSP-binding proteins. It is hoped that the studies will serve as the basis for designing more potent antiangiogenic reagents.

细胞外基质蛋白血小板反应蛋白（TSP）2，如TSP 1，与许多细胞表面受体和细胞外基质蛋白相互作用。 该基金的近期目标是关注和分析TSP的结构特征和与抑制血管生成功能相关的蛋白质伴侣。 这些目标的追求是由刺激，设备齐全，知识丰富的研究环境，可用于工作的性能提供便利。 这些发现将为旨在了解体内血管生物学功能的分子基础的长期目标奠定框架。 现在广泛认识到，在许多病理条件下，一个共同的潜在过程是血管生成的紊乱。 血管生成抑制剂在异常血管形成与疾病进展相关的疾病状态中具有潜在的临床应用，例如实体瘤、血管瘤和炎性疾病。 另一方面，血管生成的增强剂可用于由血液供应不足引起的病症，例如伤口愈合、冠状动脉疾病和中风。 TSP1和TSP2由于其抑制血管生成的能力，最近在医学科学中特别感兴趣。 最近的体内研究已经强调了TSP 1和TSP 2蛋白在调节血管生成中的重要性，这些研究证明了TSP依赖性诱导伴随的肿瘤抗性、抑制肿瘤生长和抑制聚二甲基硅氧烷植入物的血管形成。 然而，作用机制仍然未知。 我们知识中的这一差距反映在基于血小板反应蛋白的抗血管生成合成肽的效力较低，比完整的TSP1的效力低一个数量级。 由于使用合成肽的抑制试验和敲除研究表明可能引发多个信号通路或引发基质反应，因此我们希望表征与TSP2相互作用的蛋白质。关于TSP2的了解很少，因为大多数研究都是关于TSP1的。在酵母双杂交系统中使用TSP2 1型重复导致分离36个独立克隆。 拟议的调查的目的是研究TSP的结构决定因素参与调节血管生成，分析的意义的相互作用的TSP 1和TSP 2与促血管生成MMP 2，并确定和功能特性的新TSP结合蛋白。 希望这些研究能为设计更有效的抗血管生成药物奠定基础。