INHIBITION OF ANGIOGENESIS-- PROTEINS BINDING TO TSP2

抑制血管生成——与 TSP2 结合的蛋白质

• 批准号: 6750772
• 负责人: Kiflai Bein
• 金额: $ 13.72万
• 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/6750772
• 关键词: angiogenesis; angiogenesis inhibitors; binding proteins; cell growth regulation; cell migration; cell proliferation; cell surface receptors; 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 与 TSP1 一样，与许多细胞表面受体和细胞外基质蛋白相互作用。 这笔资助的直接目标是关注和分析与血管生成抑制功能相关的 TSP 结构特征和蛋白质伴侣。 可用于开展工作的刺激性、设备齐全且智力丰富的研究环境有助于实现这些目标。 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。 如今人们广泛认识到，在许多病理状况下，共同的潜在过程是血管生成紊乱。 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、血管瘤和炎症性疾病。 另一方面，血管生成增强剂对于血液供应不足引起的病症（例如，高血压）很有用。伤口愈合、冠状动脉疾病和中风。 TSP1 和 TSP2 最近在医学科学中受到特别关注，因为它们具有抑制血管生成的能力。 最近的体内研究强调了 TSP1 和 TSP2 蛋白在血管生成调节中的重要性，这些研究证明 TSP 依赖性诱导伴随的肿瘤抵抗、抑制肿瘤生长和抑制聚二甲基硅氧烷植入物的血管化。 然而，作用机制仍然未知。 我们知识上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低效力上，其比完整的 TSP1 低一个数量级。 由于使用合成肽的抑制测定和敲除研究表明可能引发多种信号通路或引发基质反应，因此我们想要表征与 TSP2 相互作用的蛋白质。人们对 TSP2 知之甚少，因为大多数研究都是针对 TSP1 的。在酵母双杂交系统中使用 TSP2 1 型重复序列导致分离出 36 个独立克隆。 本研究的目的是研究参与血管生成调节的 TSP 结构决定因素，分析 TSP1 和 TSP2 与促血管生成 MMP2 相互作用的重要性，并鉴定和功能表征新型 TSP 结合蛋白。 希望这些研究能够成为设计更有效的抗血管生成试剂的基础。