Thromboregulation in Occlusive Vascular Diseases

闭塞性血管疾病中的血栓调节

• 批准号: 7657802
• 负责人: Aaron Jacob Marcus
• 金额: $ 73.06万
• 依托单位: WEILL MEDICAL COLL OF CORNELL UNIV
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-05-01 至 2013-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7657802
• 关键词: ATP phosphohydrolase; Acute; Affect; Agonist; Alternative Splicing; Anabolism; Animal Model; Anus; Apyrase; Aspirin; Atherosclerosis; Autacoids; Biochemical; Biological; Blood; Blood Cells; Blood Platelets; Blood Vessels; Brain; Brain Diseases; Cardiovascular Diseases; Cardiovascular system; Carotid Arteries; Cell membrane; Cells; Cerebrovascular Disorders; Cessation of life; Cholesterol; Clinical; Clinical Trials; Coagulation Process; Comprehension; Coronary; Coronary Arteriosclerosis; Coronary Occlusions; Data; Diagnosis; Disease; Drug Formulations; Eicosanoids; Endothelial Cells; Enzymes; Exhibits; Family suidae; Fibrinolytic Agents; Genes; Goals; Hemostatic function; Human; In Vitro; Inflammation; Ischemic Stroke; Knockout Mice; Lead; Length; Leukocytes; Liquid substance; Mediator of activation protein; Membrane Microdomains; Metabolic; Modality; Modeling; Molecular Biology; Morbidity - disease rate; Mus; Nitric Oxide; Nucleotidases; Pathogenesis; Patients; Pharmaceutical Preparations; Phase; Platelet Activation; Prevention approach; Property; Prophylactic treatment; Prostaglandins I; Public Health; Publishing; RNA Splicing; Rattus; Recombinants; Recruitment Activity; Regulation; Research; Research Personnel; Safety; Signal Transduction; SolCD39; Stroke; Surface; System; Therapeutic; Thromboplastin; Thrombosis; Thrombus; Tissues; Variant; Vascular Diseases; atherothrombosis; base; cell type; ectoADPase; enzyme activity; fluidity; improved; in vivo; inhibitor/antagonist; monocyte; mortality; multidisciplinary; next generation; novel strategies; novel therapeutics; nucleotidase; prototype; public health relevance; response; tissue culture

DESCRIPTION (provided by applicant): Morbidity and mortality from stroke and coronary artery disease (CAD) are probably the most critical public health problems in the US. Both ischemic stroke and coronary occlusion are complications of atherosclerosis and inflammation which culminate in platelet activation, recruitment and vascular occlusion - a breach of the control of blood fluidity. Advances in therapeutic modalities for stroke and CAD have reached a plateau with an absence of novel approaches. The broad, long term objectives and specific aims of this proposal are to bring the ecto-enzyme CD39/NTPDase1 to clinical fruition via the in vitro and ex vivo studies described herein. Our recent studies of CD39 have demonstrated that it constitutes the main control system for blood fluidity (thromboregulation). CD39 has a unique mode of action whereby it inhibits platelet activation and recruitment, acting in the fluid phase. This occurs via metabolic deletion of prothrombotic ADP, the final common mediator in platelet-induced vascular occlusion. Thus the mechanism of action of CD39 is radically different from currently available therapeutic modalities for treatment of stroke and CAD. Our successful treatment of stroke with the soluble form of CD39 (solCD39) in 3 animal models, as previously published, suggests its safety and efficacy. Extensive preliminary molecular biology data indicate that expression of different CD39 alternative splice variants regulates CD39 expression, assembly into cholesterol-rich domains, and enzymatic and biological activity. This enhances our comprehension of the mechanisms of action of the enzyme, and its regulation. CD39 will be studied in cryptogenic and atherothrombotic stroke patients. Thus, we will evaluate the profile of CD39 variant expression in normals and patients with cryptogenic and atherothrombotic stroke. Platelet reactivity and markers of platelet activation will be evaluated in our cryptogenic and atherothrombotic patients and controls using a spectrum of platelet agonists and coagulation parameters, including circulating tissue factor, in the setting of their medication status. This also includes characterization of leukocyte-platelet aggregates (with emphasis on monocyte-platelet aggregates) and vascular cell-derived microparticles. We anticipate that our research will afford novel therapeutic opportunities for CD39 in prophylaxis and management of stroke and CAD. In this application we have developed a critical mass of patients and investigators in order to further our comprehension of the pathogenesis and treatment of platelet-driven cardiovascular diseases and to develop CD39 as the prototype of the next generation of antithrombotic agents. Thus, this revised submission is a broadly based multidisciplinary, molecularly and clinically focused application. PUBLIC HEALTH RELEVANCE: Vascular occlusion in the brain (stroke) is initiated by excessive blood platelet reactivity and recruitment, leading to extensive morbidity and mortality. This reactivity can be neutralized by the enzyme CD39. The relevance of this research is that it is provides a new and safe approach to the management of stroke for which there is no satisfactory treatment. We anticipate that the research proposed herein will lead to formulation of a clinical trial of a human soluble apyrase such as solCD39.

描述（由申请人提供）：中风和冠状动脉疾病（CAD）的发病率和死亡率可能是美国最严重的公共卫生问题。缺血性中风和冠状动脉闭塞都是动脉粥样硬化和炎症的并发症，最终导致血小板活化、募集和血管闭塞——破坏血液流动性的控制。中风和 CAD 治疗方式的进展已达到一个平台期，缺乏新的方法。该提案的广泛、长期目标和具体目标是通过本文所述的体外和离体研究将胞外酶 CD39/NTPDase1 推向临床。我们最近对CD39的研究表明，它构成了血液流动性（血栓调节）的主要控制系统。 CD39 具有独特的作用模式，可在流体相中抑制血小板活化和募集。这是通过促血栓 ADP 的代谢缺失而发生的，促血栓 ADP 是血小板诱导的血管闭塞的最终常见介质。因此，CD39的作用机制与目前治疗中风和CAD的治疗方式截然不同。正如之前发表的，我们在 3 个动物模型中使用可溶性 CD39 (solCD39) 成功治疗中风，这表明其安全性和有效性。大量的初步分子生物学数据表明，不同 CD39 选择性剪接变体的表达可调节 CD39 表达、组装成富含胆固醇的结构域以及酶活性和生物活性。这增强了我们对酶的作用机制及其调节的理解。 CD39 将在隐源性和动脉粥样硬化性中风患者中进行研究。因此，我们将评估正常人和隐源性中风和动脉粥样硬化血栓性中风患者中 CD39 变异表达的概况。我们将使用一系列血小板激动剂和凝血参数（包括循环组织因子）在我们的隐源性和动脉粥样硬化血栓形成患者和对照患者的药物状态下评估血小板反应性和血小板活化标志物。这还包括白细胞-血小板聚集体（重点是单核细胞-血小板聚集体）和血管细胞衍生微粒的表征。我们预计我们的研究将为 CD39 在中风和 CAD 的预防和治疗中提供新的治疗机会。在此应用中，我们培养了足够数量的患者和研究人员，以进一步了解血小板驱动的心血管疾病的发病机制和治疗，并开发 CD39 作为下一代抗血栓药物的原型。因此，这次修订后的提交是一个广泛基础的多学科、分子和临床重点应用。公众健康相关性：大脑血管闭塞（中风）是由血小板过度反应和募集引发的，导致广泛的发病率和死亡率。这种反应性可以被酶 CD39 中和。这项研究的意义在于，它为目前尚无令人满意的治疗方法的中风治疗提供了一种新的、安全的方法。我们预计本文提出的研究将导致制定人类可溶性腺苷三磷酸双磷酸酶（例如 solCD39）的临床试验。