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: 7392800
• 负责人: RAJABRATA SARKAR
• 金额: $ 40.05万
• 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/7392800
• 关键词: Adenovirus Vector; Affect; American; Biological Assay; Biomechanics; Blood Clot; Blood Vessels; Blood coagulation; Chronic; Cicatrix; Coagulation Process; Collagen; Deep Vein Thrombosis; Development; Disease regression; Elasticity; End Point; Endopeptidases; Endothelial Cells; Enzymes; Essential Genes; Event; Fibrosis; Functional disorder; Gelatinase A; Gelatinase B; Genes; Genetic Transcription; Human; Interstitial Collagenase; Knowledge; Lead; Leg; Localized; Matrix Metalloproteinases; Metalloproteinase Gene; Molecular; Mus; Obstruction; Pain; Pain in lower limb; Pathogenesis; Patients; Pattern; Peptide Hydrolases; Play; Postphlebitic Syndrome; Process; Protein Isoforms; Protein Overexpression; Proteins; Regulation; Reporter; Research Personnel; Research Proposals; Resolution; Role; Series; Skin; Smooth Muscle Myocytes; Swelling; Testing; Thrombophlebitis; Thrombus; Tissues; Transgenic Organisms; Ulcer; Veins; Venous; activating transcription factor; cell motility; cell type; chromatin immunoprecipitation; clinically significant; human MMP14 protein; monocyte; mouse model; novel; prevent; programs; research study; transcription factor

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-f) 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 proposalwill 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.

深静脉血栓 (DVT) 每年影响超过 200 万美国人。静脉炎后综合征 25-75% 的 DVT 患者会受到影响，包括腿部肿胀、疼痛、皮肤变化和溃疡 皮肤。这种情况仅发生在部分 DVT 患者中，表明血栓溶解对于 DVT 患者至关重要。 确定慢性静脉阻塞和纤维化是否会发展并导致静脉炎后 综合症。基质金属蛋白酶 (MMP) 基因对于细胞迁移和组织重塑至关重要， 在血栓溶解过程中表达并激活，表明在此过程中发挥关键作用。他们的角色 静脉炎后综合征的发病机制将通过三个具体目标来定义： 1) 定义 MMP-2 基因区域和对血栓诱导的 MMP-2 至关重要的同源转录因子 表达，2) 确定 MMP-2、MMP-9 和 MMP-14 (MT-MMP-f) 在 DVT 缓解和治疗中的作用 血栓诱发的静脉壁纤维化，3) 确定 MMP 蛋白（MMP-2、MMP-9）是否过度表达 和 MMP-14）可加速 DVT 消退并改变血栓引起的静脉壁纤维化。一个独特的系列 转基因 MMP-2 报告小鼠将用于确定 MMP-2 基因的哪些区域是必需的 对于血栓诱导的 MMP-2 转录和染色质免疫沉淀将用于鉴定 同源转录因子。将使用靶向删除各种 MMP 基因的小鼠来确定 这些酶在血栓再通、静脉壁纤维化和静脉壁顺应性丧失中的作用和 使用小鼠静脉生物力学的新测定法测定弹性。组织特异性转基因过表达 MMP-2、-14 和 -9 以及编码这些亚型的腺病毒载体将用于过表达 这些蛋白质在血栓溶解过程中加速了 DVT 的溶解，并且对 纤维化和静脉壁生物力学。这些研究将确定 MMP 蛋白在有益和静脉壁生物力学中的作用。 血栓溶解的不利方面，并描述潜在的分子治疗以预防血栓溶解后 静脉炎综合征。 相关性：该提案将确定基质金属蛋白酶蛋白如何导致疤痕和 血栓后静脉增厚，可能导致随后的腿部疼痛、肿胀和溃疡。这些研究将 增加对血栓如何损害静脉的了解，并测试新的治疗方法以防止这种损害。