ERO1 alpha in platelet activity and thrombosis

ERO1 α 在血小板活性和血栓形成中的作用

基本信息

批准号：
10285785
负责人：
Jaehyung Cho
金额：
$ 40.41万
依托单位：
WASHINGTON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-02-10 至 2024-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10285785
关键词：
Adhesions Adhesives Affect Animals Antiplatelet Drugs Binding Binding Proteins Biochemical Biological Bleeding time procedure Blocking Antibodies Blood Platelets Cell surface Cells Cessation of life Complex Coronary Data Development Disease Endoplasmic Reticulum Endothelial Cells Environment Extracellular Protein Family member Fibrin Fibrinolytic Agents Generations Glycoproteins Goals Hemorrhage Hemostatic function Impairment In Vitro Integrins Isomerase Knockout Mice Lead Ligand Binding Lung Mediating Modeling Molecular Mus Oxidases Oxidation-Reduction Oxidative Stress Oxides Oxidoreductase Pathologic Pathology Peptides Peripheral arterial disease Permeability Pharmacology Platelet aggregation Protein Disulfide Isomerase Reporting Reticulum Risk Role Series Signal Transduction Site Sulfhydryl Compounds Surface Tail Testing Thrombosis Thrombus base conditional knockout disulfide bond effective therapy extracellular in vivo inhibitor/antagonist insight intravital microscopy new therapeutic target novel novel therapeutic intervention platelet function protein function small molecule inhibitor targeted agent targeted treatment thromboinflammation thrombotic vascular injury

项目摘要

Project Summary Despite advances in our understanding of the mechanisms mediating platelet thrombus formation, current antiplatelet drugs increase the risk of major bleeding. In an effort to identify novel therapeutic targets, we and others showed that intravascular protein disulfide isomerase (PDI) is crucial for full activation of αIIbβ3 integrin and platelet accumulation in arterial thrombosis, providing insights into a new antithrombotic agent. However, blocking the oxidoreductase activity of intravascular PDI prolongs tail bleeding times in mice. Our preliminary data have demonstrated that endoplasmic reticulum (ER) oxidoreductin 1α (Ero1α), a key oxidase of PDI in the ER, is released from activated platelets and that inhibition or deletion of platelet Ero1α alters the activity of platelet surface-bound PDI and impairs platelet aggregatory function. Furthermore, we have found that global deletion of Ero1α reduces the size of platelet thrombus formation without affecting initial platelet adhesion and tail bleeding times following vascular injury. Using biochemical, cellular and in vivo animal studies with novel pharmacological inhibitors and Ero1α conditional knockout and global knockout mice developed by our lab, we will test the hypothesis that platelet-released Ero1α regulates the function of PDI and αIIbβ3 integrin on the cell surface and contributes to the propagation step of platelet thrombus formation without affecting hemostasis following vascular injury. In Aim 1, we will identify the molecular mechanism by which extracellular Ero1α promotes platelet aggregation. In Aim 2, we will test whether arterial thrombotic conditions alter the function of extracellular Ero1α. In Aim 3, we will determine the pathophysiological role of intravascular and platelet-derived Ero1α in platelet adhesion and accumulation and vessel occlusion under thrombotic conditions. The proposed studies will identify a central regulatory mechanism of platelet thrombus formation and lead to the discovery of novel therapeutic strategies for the safe and effective treatment of thrombotic disease.

项目摘要尽管我们了解了介导血小板血栓形成的机制，但抗血小板药物会增加大量出血的风险。为了确定新型的治疗目标，我们和其他人则表明，血管内蛋白二硫化物异构酶（PDI）对于完全激活αIIBβ3整合素至关重要和血小板在动脉血栓形成中的积累，为新的抗血栓形成剂提供了见解。然而，阻塞血管内PDI的氧化还原酶活性延长了小鼠的尾巴出血时间。我们的初步数据表明，内质网（ER）氧化还原素1α（ERO1α），PDI的关键氧化酶 ER从活化的血小板中释放出来，抑制或缺失血小板ERO1α改变了活性血小板结合的PDI并损害血小板聚集功能。此外，我们发现全球 ERO1α的缺失减少了血小板形成的大小，而不会影响初始血小板粘合剂和血管损伤后的尾部出血时间。使用新型的生化，细胞和体内动物研究药理学抑制剂和ERO1α条件敲除和我们实验室开发的全球敲除小鼠，我们将检验以下假设，即血小板发行的ERO1α调节PDI和αIIBβ3整合素对细胞的功能表面并有助于血小板形成的传播步骤，而不会影响止血血管损伤。在AIM 1中，我们将确定细胞外ERO1α的分子机制促进血小板聚集。在AIM 2中，我们将测试伪影是否会改变细胞外ERO1α。在AIM 3中，我们将确定血管内和血小板衍生的病理生理作用血小板条件下的血小板粘附，积累和血管阻塞的ERO1α。提议研究将确定血小板形成的中心调节机制，并导致发现用于安全有效治疗血栓性疾病的新型治疗策略。