Proteolytic Pathways in Venous Thrombus Resolution

静脉血栓溶解中的蛋白水解途径

• 批准号: 10369353
• 负责人: Toni M Antalis
• 金额: --
• 依托单位: BALTIMORE VA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-07-01 至 2025-12-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10369353
• 关键词: 3-Dimensional; Acceleration; Acute; Adjuvant; Adoptive Transfer; Affect; Age; Agonist; Air; American; Anti-Inflammatory Agents; Anticoagulant therapy; Anticoagulants; Anticoagulation; Award; Biological Assay; Blood; Blood coagulation; Cardiovascular Diseases; Cell Survival; Cells; Cessation of life; Chronic; Clinical; Clinical Management; Clinical Research; Coagulation Process; Competence; Complication; Constitution; Data; Deep Vein Thrombosis; Dehydration; Disease; Embolism; Environment; Event; Experimental Models; Fibrinolysis; Fibrosis; Foundations; Gene Expression; Genetic; Goals; Health; Health Care Costs; Hospitalization; Human; Immune; Immune response; Inflammation; Inflammatory; Knowledge; Leg; Leukocytes; Longitudinal Studies; Malignant Neoplasms; Measurement; Mediating; Medical center; Methods; Military Personnel; Molecular; Morbidity - disease rate; Mus; Neutrophil Activation; Obstruction; Operative Surgical Procedures; Pain; Paralysed; Pathogenesis; Pathway interactions; Patients; Pattern; Peptide Hydrolases; Peptides; Phenotype; Plasminogen Activator Inhibitor 2; Play; Postphlebitic Syndrome; Pre-Clinical Model; Process; Prognosis; Property; Pulmonary Embolism; RNA Interference; Recurrence; Regulation; Regulator Genes; Research; Resolution; Risk; Risk Factors; Serine Proteinase Inhibitors; Serpins; Serum; Signal Pathway; Skin; Source; Swelling; Testing; Therapeutic; Thrombus; Time; Tobacco use; Trauma; Treatment Efficacy; United States Department of Veterans Affairs; Urokinase; Vascular remodeling; Venous; Venous Thrombosis; Veterans; antagonist; arm; base; candidate marker; chemokine; clinically relevant; cytokine; deep vein; disability; effective therapy; experimental study; gene network; genetic signature; immunoregulation; improved; inhibitor; insight; macrophage; military veteran; molecular marker; mortality; mouse model; neutrophil; novel; novel therapeutics; peripheral blood; potential biomarker; premature; prevent; productivity loss; prognostic value; programs; severe injury; skin ulcer; therapeutic evaluation; thrombolysis; thrombotic complications; transcriptome; translational potential; urokinase inhibitor; venous thromboembolism

Background / Rationale: VTE (Venous Thromboembolism) is a common and serious cardiovascular disease with significant mortality and morbidity. Prompt anticoagulation of patients with deep venous thrombosis (DVT) reduces fatal pulmonary embolism (PE) but does not prevent long-term morbidity of VTE. Post-thrombotic syndrome occurs in 25-50% of patients with clots in the deep veins of the arms and legs that consists of pain, swelling, and recurrent skin ulceration. Common risk factors for DVT include cancer, major trauma, surgery, paralysis, prolonged periods of immobility, and older age. DVT and its complications have increased in the Veteran population over the last decade. Deployed military personnel are at an increased risk due to prolonged air and ground transport, dehydration, tobacco use, and extended immobility during hospitalizations for severe injuries. Thrombus resolution is a critical factor in the pathogenesis of post-thrombotic syndrome since incomplete thrombus resolution can result in obstruction of flow and loss of venous valve function. Using novel 3D serial measurements of thrombus volume; we demonstrated that DVT patients with similar initial DVTs have widely varying rates of thrombus resolution over time despite adequate anticoagulation. Longitudinal studies show that patients with more rapid thrombus resolution have a better prognosis than those patients whose thrombus resolves much slower. Despite its clinical importance, the cellular and molecular mechanisms involved in DVT are poorly understood, and there currently is no therapy to accelerate this process. Objectives: Using clinically relevant experimental models of DVT, a comprehensive picture of interconnected cell-mediated molecular processes that orchestrate a precise inflammatory program is starting to emerge and forms the foundation for this proposal. Our objectives are to build on our previous VA Merit findings to: 1) define mechanisms by which plasminogen activator inhibitor-2 (PAI-2) deficiency modulates inflammatory leukocytes to accelerate venous thrombus resolution; 2) test the therapeutic efficacy of blocking the PAI-2 pathway to accelerate venous thrombus resolution; and 3) determine specific gene signatures for the temporal inflammatory vascular remodeling events that occur during venous thrombus resolution and evaluate the prognostic value of candidate biomarkers in patients with evolving and maladaptive thrombus resolution after VTE. Methods: Studies will utilize genetically deficient mice in experimental models of DVT that accurately mimic many of the clinical and pathophysiological features observed in human DVT. We will define mechanisms by which PAI-2 deficiency calibrates immune regulation to accelerate venous thrombus resolution using ex vivo thrombolysis assays, transmigration assays, and neutrophil adoptive transfer experiments. The translational potential of suppressing PAI-2 expression or activity to accelerate venous thrombus resolution will be tested in human cells and in preclinical models. Finally, we will use our well-established clinically relevant mouse models of DVT resolution to identify gene regulatory signatures/potential biomarkers and then evaluate changes in gene networks over time in blood from patients with evolving thrombus resolution after VTE. Findings/Results: Molecular mechanisms that modulate inflammation during venous thrombus resolution in experimental models and in human patients will be identified and therapies based on these mechanisms tested in preclinical models. Status: This is a new project arising from substantial supportive preliminary data from a previous VA Merit Award. Impact: New knowledge from these studies regarding the inflammatory signatures in DVT patients may form the basis for novel therapies for accelerating this process, and in combination with anticoagulants, control excessive fibrosis, and prevent this disease.

背景/基本原理：VTE（静脉血栓栓塞）是一种常见且严重的心血管疾病 具有显着的死亡率和发病率。深静脉血栓（DVT）患者及时抗凝 减少致命性肺栓塞 (PE)，但不能预防 VTE 的长期发病率。血栓后 25-50% 的手臂和腿部深静脉有血栓的患者会出现这种综合征，包括疼痛、 肿胀和反复出现的皮肤溃疡。 DVT 的常见危险因素包括癌症、重大创伤、手术、 瘫痪、长时间不动和年老。 DVT 及其并发症在 过去十年的退伍军人人口。由于长期部署，部署的军事人员面临更大的风险 空中和地面交通、脱水、吸烟以及重症患者住院期间长时间不动 受伤。血栓消退是血栓后综合征发病机制的关键因素，因为 血栓溶解不完全可能导致血流受阻和静脉瓣膜功能丧失。使用小说 血栓体积的 3D 连续测量；我们证明，具有相似初始 DVT 的 DVT 患者具有 尽管进行了充分的抗凝治疗，但随着时间的推移，血栓溶解率差异很大。纵向研究 表明血栓消退较快的患者比血栓消退较快的患者预后更好 血栓的溶解速度要慢得多。尽管其临床重要性，但所涉及的细胞和分子机制 人们对 DVT 的认识知之甚少，目前还没有治疗方法可以加速这一过程。 目标：使用临床相关的 DVT 实验模型，全面了解相互关联的情况 协调精确炎症程序的细胞介导的分子过程开始出现，并且 构成本提案的基础。我们的目标是在之前 VA Merit 调查结果的基础上：1) 定义 纤溶酶原激活物抑制剂-2 (PAI-2) 缺乏调节炎症白细胞的机制 加速静脉血栓溶解； 2）测试阻断PAI-2途径的治疗效果 加速静脉血栓溶解； 3) 确定颞部炎症的特定基因特征 静脉血栓溶解过程中发生的血管重塑事件并评估预后价值 VTE 后血栓消退进展和适应不良患者的候选生物标志物。 方法：研究将利用基因缺陷小鼠建立 DVT 实验模型，准确模拟 在人类 DVT 中观察到的许多临床和病理生理学特征。我们将通过以下方式定义机制 PAI-2 缺陷可通过体外校准免疫调节来加速静脉血栓溶解 溶栓测定、迁移测定和中性粒细胞过继转移实验。翻译的 抑制 PAI-2 表达或活性以加速静脉血栓溶解的潜力将在 人类细胞和临床前模型。最后，我们将使用我们完善的临床相关小鼠模型 DVT 解析以确定基因调控特征/潜在生物标志物，然后评估基因变化 随着时间的推移，静脉血栓栓塞（VTE）后血栓消退患者的血液中存在网络。 研究结果/结果：在静脉血栓溶解过程中调节炎症的分子机制 将确定实验模型和人类患者，并测试基于这些机制的疗法 在临床前模型中。 状态：这是一个新项目，源自之前 VA Merit 提供的大量支持性初步数据 奖。 影响：这些研究中关于 DVT 患者炎症特征的新知识可能会形成 加速这一过程的新疗法的基础，并与抗凝剂结合使用，控制过度 纤维化，并预防这种疾病。