Proteolytic Pathways in Venous Thrombus Resolution

静脉血栓溶解中的蛋白水解途径

• 批准号: 10549748
• 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/10549748
• 关键词: 3-Dimensional; Acceleration; Acute; Adjuvant Therapy; Adoptive Transfer; Affect; Agonist; Air; American; Anti-Inflammatory Agents; Anticoagulant therapy; Anticoagulants; Anticoagulation; Award; Biological Assay; Blood; Blood coagulation; Calibration; Cardiovascular Diseases; Cell Survival; Cells; Cessation of life; Chronic; Clinical; Clinical Management; Clinical Research; Coagulation Process; Competence; Complication; Constitution; Constitutional; Data; Deep Vein Thrombosis; Dehydration; Disease; Embolism; Environment; Event; Experimental Models; Fibrinolysis; Fibrosis; Foundations; Gene Expression; Genetic; Goals; Health; Health Care Costs; Hospitalization; Human; Immobilization; Immune; Immune response; Inflammation; Inflammatory; Knowledge; Leg; Leukocytes; Longitudinal Studies; Macrophage; Malignant Neoplasms; Measurement; Mediating; Medical center; Methods; Migration Assay; 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; candidate marker; chemokine; clinically relevant; cytokine; deep vein; disability; effective therapy; experimental study; gene network; genetic signature; human old age (65+); immunoregulation; improved; inflammatory modulation; inhibitor; insight; 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 /理由：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。促使深静脉血栓形成（DVT）患者的抗凝治疗 减少致命的肺栓塞（PE），但不能防止VTE的长期发病率。施加后 综合征发生在25-50％的凝块患者的胳膊和腿的深静脉中，这些患者由疼痛组成， 肿胀和复发性皮肤溃疡。 DVT的常见危险因素包括癌症，重大创伤，手术， 瘫痪，长时间的固定时期和年龄。 DVT及其并发症在 过去十年中的退伍军人人口。部署的军事人员由于延长而面临增加的风险 在住院期间，空气和地面运输，脱水，烟草使用以及延长的固定性 受伤。血栓分辨率是施加后综合征发病机理的关键因素，因为 不完整的血栓分辨率会导致流量阻塞和静脉瓣功能的丧失。使用小说 血栓体积的3D串行测量；我们证明了具有相似初始DVT的DVT患者 尽管有足够的抗凝治疗，但随着时间的流逝，血栓分辨率的变化速率很大。纵向研究 表明血栓分辨率更快的患者比那些患者的预后更好 血栓解决得慢得多。尽管其临床重要性，但涉及细胞和分子机制 在DVT中，人们对DVT的了解很少，目前尚无疗法来加速这一过程。 目标：使用DVT的临床相关实验模型，互连的全面图片 精确炎症计划的细胞介导的分子过程开始出现，并且 构成了该提议的基础。我们的目标是建立在我们以前的VA优点发现的基础上：1）定义 纤溶酶原激活剂2（PAI-2）缺乏调节炎症白细胞的机制 加速静脉血栓分辨率； 2）测试阻断PAI-2途径的治疗功效 加速静脉血栓分辨率； 3）确定时间炎症的特定基因标志 静脉血栓分辨率期间发生的血管重塑事件，并评估 VTE后，患有不断发展和适应不良血栓的患者的候选生物标志物。 方法：研究将在DVT的实验模型中利用遗传缺陷的小鼠，以准确模仿 在人DVT中观察到的许多临床和病理生理特征。我们将通过 PAI-2缺乏可以校准免疫调节以使用离体加速静脉血栓分辨率 溶栓测定，移民测定和中性粒细胞的转移实验。翻译 抑制PAI-2表达或活性加速静脉血栓分辨率的潜力将在 人类细胞和临床前模型。最后，我们将使用良好的临床相关鼠标模型 DVT分辨率以识别基因调节特征/潜在生物标志物，然后评估基因的变化 VTE后血栓分辨率进化的患者的血液中的网络随着时间的流逝。 发现/结果：在静脉血栓分辨率调节炎症的分子机制 将鉴定出实验模型和人类患者，并根据这些机制进行疗法 在临床前模型中。 状态：这是一个新项目，由先前VA优点的大量支持初步数据引起 奖。 影响：这些研究中有关DVT患者炎症特征的新知识可能形成 用于加速这一过程并与抗凝剂结合使用的新型疗法的基础，控制过多 纤维化并预防这种疾病。