Fibrotic effects and regulation of MMP proteins in thrombus resolution

MMP 蛋白在血栓溶解中的纤维化作用和调节

• 批准号: 7071000
• 负责人: RAJABRATA SARKAR
• 金额: $ 41.25万
• 依托单位: NORTHERN CALIFORNIA INSTITUTE/RES/EDU
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-05-12 至 2010-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7071000
• 关键词: chromatin immunoprecipitation; clinical research; fibrosis; gene expression; genetic mapping; genetically modified animals; human tissue; isozymes; laboratory mouse; medical complication; metalloendopeptidases; monocyte; pathologic process; protein localization; transcription factor; vascular endothelium; vascular smooth muscle; venous thrombosis

Description (provided by Applicant): Deep venous thrombosis (DVT) affects more than 2 million Americans per year. Post-phlebitic syndrome can affect 25-75% of patients following DVT and includes leg swelling, pain, skin changes and ulceration of the skin. This develops only in a subset of DVT patients, suggesting that thrombus resolution is critical in determining whether chronic venous obstruction and fibrosis will develop and lead to post-phlebitic syndrome. Matrix metalloproteinase (MMP) genes, critical to cell migration and tissue remodeling, are expressed and activated during thrombus resolution suggesting a critical role in this process. Their role in the pathogenesis of post-phlebitic syndrome will be defined with three Specific Aims: 1) To define the regions of the MMP-2 gene and cognate transcription factors critical to thrombus-induced MMP-2 expression, 2) To determine the role of MMP-2, MMP-9 and MMP-14 (MT-MMP-1) in DVT resolution and thrombus-induced vein wall fibrosis, 3) To determine if overexpression of MMP proteins (MMP-2, MMP-9 and MMP-14) accelerates DVT resolution and alters thrombus-induced vein wall fibrosis. A unique series of transgenic MMP-2 reporter mice will be used to determine which regions of the MMP-2 gene are essential for thrombus-induced MMP-2 transcription and chromatin immunoprecipitation will be used to identity the cognate transcription factors. Mice with targeted deletion of various MMP genes will be used to determine the role of these enzymes in thrombus recanalization, vein wall fibrosis and loss of vein wall compliance and elasticity using novel assays of mouse vein biomechanics. Tissue-specific transgenic overexpression of MMP-2, -14 and -9 as well as adenoviral vectors encoding these isoforms will be utilized to overexpress these proteins during thrombus resolution to determine resolution of DVT is accelerated and the effects on fibrosis and vein wall biomechanics. These studies will define the role of MMP proteins in the beneficial and detrimental aspects of thrombus resolution, and characterize potential molecular therapy to prevent post- phlebitic syndrome. RELEVANCE: This proposal will determine how matrix metalloproteinase proteins cause scarring and thickening of veins after blood clots, which can cause later leg pain, swelling and ulcers. These studies will increase knowledge of how veins are damaged by clots and test new treatments to prevent this damage.

描述（由申请人提供）： 深静脉血栓 (DVT) 每年影响超过 200 万美国人。静脉炎后综合征会影响 25-75% 的 DVT 患者，包括腿部肿胀、疼痛、皮肤变化和皮肤溃疡。这种情况仅发生在部分 DVT 患者中，这表明血栓消退对于确定慢性静脉阻塞和纤维化是否会发展并导致静脉炎后综合征至关重要。基质金属蛋白酶 (MMP) 基因对细胞迁移和组织重塑至关重要，在血栓溶解过程中表达和激活，表明在此过程中发挥着关键作用。它们在静脉炎后综合征发病机制中的作用将通过三个具体目标来定义：1) 确定 MMP-2 基因的区域和对血栓诱导的 MMP-2 表达至关重要的同源转录因子，2) 确定其作用MMP-2、MMP-9 和 MMP-14 (MT-MMP-1) 在 DVT 消退和血栓诱导的静脉壁纤维化中的作用，3) 确定是否过度表达MMP 蛋白（MMP-2、MMP-9 和 MMP-14）可加速 DVT 消退并改变血栓引起的静脉壁纤维化。一系列独特的转基因 MMP-2 报告小鼠将用于确定 MMP-2 基因的哪些区域对于血栓诱导的 MMP-2 转录至关重要，染色质免疫沉淀将用于鉴定同源转录因子。靶向删除各种 MMP 基因的小鼠将用于通过小鼠静脉生物力学的新测定来确定这些酶在血栓再通、静脉壁纤维化以及静脉壁顺应性和弹性丧失中的作用。 MMP-2、-14 和 -9 的组织特异性转基因过表达以及编码这些亚型的腺病毒载体将用于在血栓消退过程中过表达这些蛋白质，以确定 DVT 的消退是否加速以及对纤维化和静脉壁生物力学的影响。这些研究将明确 MMP 蛋白在血栓消退的有益和有害方面的作用，并表征预防静脉炎后综合征的潜在分子疗法。相关性：该提案将确定基质金属蛋白酶蛋白如何在血栓后导致静脉疤痕和增厚，从而导致随后的腿部疼痛、肿胀和溃疡。这些研究将增加人们对静脉如何被血栓损害的认识，并测试新的治疗方法来预防这种损害。