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/原理：静脉血栓栓塞（VTE）是一种常见且严重的心血管疾病 死亡率和发病率都很高。深静脉血栓形成（DVT）患者的及时抗凝治疗 减少致命性肺栓塞（PE），但不能预防VTE的长期发病率。血栓后 综合征发生在25-50%的手臂和腿部深静脉血栓的患者中， 肿胀和复发性皮肤溃疡。DVT的常见风险因素包括癌症、严重创伤、手术， 瘫痪、长时间不动和年老。DVT及其并发症的增加， 在过去的十年里。部署的军事人员面临的风险增加， 空中和地面运输，脱水，烟草使用，以及严重急性呼吸道感染住院期间的长期不动 受伤血栓消退是血栓后综合征发病机制中的关键因素， 不完全血栓溶解可导致血流阻塞和静脉瓣膜功能丧失。使用新颖 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 尽管抗凝充分，但随着时间的推移，血栓消退率差异很大。纵向研究 显示血栓消退更快的患者比那些 血栓消退得更慢。尽管其临床重要性，涉及的细胞和分子机制 在DVT中，人们对这一过程知之甚少，目前还没有加速这一过程的治疗方法。 目的：使用DVT的临床相关实验模型，全面了解DVT的相互联系， 细胞介导的分子过程，精心策划一个精确的炎症程序开始出现， 这是这项提议的基础。我们的目标是建立在我们以前的VA Merit调查结果的基础上：1）定义 纤溶酶原激活物抑制剂-2（派-2）缺乏调节炎性白细胞的机制 加速静脉血栓消退; 2）测试阻断派-2通路的治疗效果， 加速静脉血栓消退;以及3）确定颞叶炎性血管炎的特异性基因特征。 静脉血栓消退期间发生的血管重塑事件，并评价 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：研究将利用遗传缺陷小鼠在DVT的实验模型，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： 派-2缺陷校准免疫调节以使用离体方法加速静脉血栓消退， 血栓溶解试验、移行试验和中性粒细胞过继转移试验。平移 抑制派-2表达或活性以加速静脉血栓消退的潜力将在 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分辨率，以识别基因调控特征/潜在生物标志物，然后评价基因 静脉血栓栓塞（VTE）后血栓消退患者血液中随时间推移的血栓网络。 发现/结果：在静脉血栓消退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，来自之前VA Merit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，并与抗凝剂联合使用，控制过度 纤维化，预防这种疾病。