Ectonucleotidases in Atherothrombosis and Stroke

外切核苷酸酶在动脉粥样硬化血栓形成和中风中的作用

• 批准号: 7341621
• 负责人: David J. Pinsky
• 金额: $ 36.41万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-01-19 至 2011-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7341621
• 关键词: 5' -Nucleotidase; Abbreviations; Accounting; Adam11 gene; Adenosine; Adenosine Diphosphate; Adenosine Triphosphate; Adhesions; Alteplase; American; Anti-Inflammatory Agents; Anti-inflammatory; Apolipoprotein E; Apyrase; Atherosclerosis; Automobile Driving; Blood; Blood Platelets; Blood Vessels; Brain; Breeding; CCL17 gene; CCL22 gene; Catalytic Domain; Cell Adhesion Molecules; Cell Communication; Cell Line; Cells; Cerebral Ischemia; Cerebral hemisphere hemorrhage; Cerebrovascular Circulation; Cerebrum; Chemokine, Other; Chloride Ion; Chlorides; Coagulants; Coagulation Process; Core-Binding Factor; Data; Deposition; Dose; Endothelial Cells; Endothelium; Engineering; Environment; Event; Exhibits; Factor VIII-Related Antigen; Fibrinolytic Agents; Genes; Genetic; Glossary; Goals; Hour; In Vitro; Infarction; Inflammation; Inflammatory; Injury; Integrins; Intercellular adhesion molecule 1; Ischemic Stroke; Knockout Mice; Lead; Leukocytes; Lipids; Mediating; Metabolism; Middle Cerebral Artery Occlusion; Mus; Nucleotidases; Nucleotides; Outcome; Pathogenesis; Pathologic; Peptides; Phenotype; Platelet Activation; Platelet Factor 4; Platelet Glycoproteins; Property; Proteins; Public Health; RANTES; Reaction; Recombinants; Recruitment Activity; Relative (related person); Research Personnel; Risk; Role; Site; Small Inducible Cytokine A18; SolCD39; Stroke; Stroke prevention; Stromal Cell-Derived Factor 1; Supplementation; Symptoms; Testing; Therapeutic; Thrombin; Thrombolytic Therapy; Thrombosis; Treatment Efficacy; VWF gene; Vascular Cell Adhesion Molecule-1; adenosine monophosphatase; adenylyl(3' -5' )cytidine-3' -phosphate; atherogenesis; atheroprotective; atherothrombosis; carbene; cerebrovascular; chemokine; chlorambucil/dactinomycin/methotrexate protocol; extracellular; human CCL17 protein; improved; inhibitor/antagonist; injured; macrophage-derived chemokine; middle cerebral artery; migration; monocyte; neutrophil; novel therapeutics; nucleotidase; prevent; programs; research study; therapeutic target; thrombolysis; trafficking; triphenyltetrazolium; von Willebrand Factor

ADP released from activated platelets recruits nearby platelets, resulting in explosive accretion of a thrombotic nidus. ATP, released from activated platelets or injured cells, promotes inflammation. The coordinated phosphohydrolysis of extracellular nucleotides, by the sequential actions of the ectonucleotidases CD39 (nucleotide diphosphohydrolase 1, converts ATP-> ADP->AMP) and CD73 (51 nucleotidase, converts AMP->adenosine), is an important endothelial homeostatic mechanism which limits thrombosis and inflammation at the blood-vessel interface. CD39 gene null mice exhibit a latent prothrombotic phenotype and worse outcomes than controls in the setting of focal cerebral ischemia. These mice can be rescued by recombinant soluble CD39, which retains apyrase activity, without increasing intracerebral hemorrhage. Furthermore, hypercholesterolemic ApoE/CD39 double knockout mice exhibit exaggerated atherogenesis, consistent with a role for platelet and inflammatory cell recruitment into the developing plaque. These data suggest that ectonucleotidases protect against atherothrombotic events and are relevant to the pathoaenesis of stroke. Experiments will elucidate mechanisms by which CD39 and CD73 are atheroprotective, focusing on their ability to suppress platelet activation and inflammatory cascades, using wild-type, cd39- or cd73-gene null mice in control or hypercholesterolemic backgrounds. Experiments will test whether ectonucleotidases improve ischemic stroke outcomes in an atherosclerosis-prone cerebrovascular milieu, using solCD39 (and/or purified CD73) as monotherapy or as an adjunct to low dose thrombolytic therapy. The overarching goal here is to delineate an endogenous cascade which protects vessels against atherothrombosis, and determine whether it may be therapeutically harnessed to ameliorate ischemic stroke outcomes. Relevance to Public Health: This project will explore how two proteins, which exist on cells lining blood vessels, degrade circulating substances which would otherwise promote clotting, inflammation, and atherosclerosis. Harnessing the activity of these proteins might lead to a new way of preventing clot formation and atherosclerosis, and thereby lead to a completely new and perhaps safer treatment for stroke.

活化的血小板释放的 ADP 会招募附近的血小板，从而导致爆炸 血栓病灶的增生。 ATP，从活化的血小板或受伤的细胞中释放出来，促进 炎。细胞外核苷酸的协调磷酸水解，通过顺序 核酸外切酶 CD39（核苷酸二磷酸水解酶 1，将 ATP 转化为 ADP->AMP）和CD73（51核苷酸酶，转化AMP->腺苷），是重要的内皮细胞 限制血管界面血栓形成和炎症的稳态机制。 CD39 基因缺失小鼠表现出潜在的血栓形成表型，并且结果比对照组更差 局灶性脑缺血的情况。这些小鼠可以被重组可溶性CD39拯救， 它保留腺苷三磷酸双磷酸酶活性，而不增加脑出血。此外， 高胆固醇血症 ApoE/CD39 双基因敲除小鼠表现出过度的动脉粥样硬化形成， 与血小板和炎症细胞募集到正在形成的斑块中的作用一致。 这些数据表明核酸外切酶可预防动脉粥样硬化血栓事件，并且 与中风的发病机制有关。实验将阐明 CD39 和 CD73 具有动脉粥样硬化保护作用，重点是抑制血小板活化和炎症的能力 级联反应，使用野生型、cd39 或 cd73 基因缺失小鼠作为对照或高胆固醇血症 背景。实验将测试核酸外切酶是否可以改善缺血性中风的结果 在易发生动脉粥样硬化的脑血管环境中，使用 solCD39（和/或纯化的 CD73）作为 单一疗法或作为低剂量溶栓疗法的辅助疗法。这里的总体目标是 描绘出保护血管免受动脉粥样硬化血栓形成的内源性级联，并确定 是否可以在治疗上利用它来改善缺血性中风的结果。 与公共健康的相关性：该项目将探讨细胞内壁上存在的两种蛋白质如何 血管，降解原本会促进凝血的循环物质， 炎症、动脉粥样硬化。利用这些蛋白质的活性可能会产生一种新的 预防血栓形成和动脉粥样硬化的方法，从而导致一种全新的、 也许治疗中风更安全。