INHIBITION OF ANGIOGENESIS-- PROTEINS BINDING TO TSP2

细胞外基质蛋白血栓反应蛋白（TSP） 2，像TSP1一样，与许多细胞表面受体和细胞外基质蛋白相互作用。这项资助的直接目标是关注和分析与抑制血管生成功能相关的TSP结构特征和蛋白质伙伴。这些目标的追求是由刺激，设备齐全，智力丰富的研究环境提供的工作表现促进。这些发现将为理解血管生物学功能的分子基础奠定长期目标的框架。现在人们普遍认识到，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。血管生成抑制剂在血管异常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，血管生成的增强剂在血液供应不足的情况下是有用的，例如伤口愈合、冠状动脉疾病和中风。由于TSP1和TSP2抑制血管生成的能力，最近在医学科学中引起了特别的兴趣。最近的体内研究表明，TSP1和TSP2蛋白在血管生成调控中的重要性得到了强调，这些研究表明，聚二甲基硅氧烷植入物中，tsp依赖性诱导伴随的肿瘤抵抗、肿瘤生长抑制和血管化抑制。然而，其作用机制尚不清楚。我们知识上的这一差距反映在基于血栓反应蛋白的抗血管生成合成肽的低效力上，这比完整的TSP1低一个数量级。由于使用合成肽和敲除研究的抑制试验表明可能引发多种信号通路或启动基质反应，我们希望表征与TSP2相互作用的蛋白质。对TSP2知之甚少，因为大多数研究都是关于TSP2的。在酵母双杂交系统中使用TSP2 1型重复序列，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控的TSP结构决定因素，分析TSP1和TSP2与促血管生成的MMP2相互作用的意义，并鉴定新的TSP结合蛋白并对其进行功能表征。希望这些研究可以作为设计更有效的抗血管生成试剂的基础。

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.