Ero1α in platelet activity and thrombosis

Ero1α 在血小板活性和血栓形成中的作用

基本信息

批准号：
9884277
负责人：
Jaehyung Cho
金额：
$ 45.91万
依托单位：
UNIVERSITY OF ILLINOIS AT CHICAGO
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：
2020-02-01 至 2021-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9884277
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 therapeutics platelet function protein function small molecule inhibitor targeted agent targeted treatment thromboinflammation 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.

项目概要尽管我们对介导血小板血栓形成的机制的理解取得了进展，但目前抗血小板药物会增加大出血的风险，为了寻找新的治疗靶点，我们和其他人表明血管内蛋白二硫键异构酶 (PDI) 对于 αIIbβ3 整合素的完全激活至关重要和动脉血栓形成中的血小板积聚，为新的抗血栓药物提供了见解。阻断血管内 PDI 的氧化还原酶活性可延长小鼠尾部出血时间。数据表明，内质网 (ER) 氧化还原素 1α (Ero1α) 是 PDI 的关键氧化酶，在 ER 由活化的血小板释放，抑制或删除血小板 Ero1α 会改变 ER 的活性此外，我们还发现，血小板表面结合的 PDI 会损害血小板的聚集功能。删除 Ero1α 可减少血小板血栓形成的大小，而不影响初始血小板粘附和使用新颖的生化、细胞和体内动物研究来确定血管损伤后的尾部出血时间。我们实验室开发的药理抑制剂和Ero1α条件敲除和全局敲除小鼠，我们将检验血小板释放的 Ero1α 调节细胞上 PDI 和 αIIbβ3 整合素功能的假设表面并有助于血小板血栓形成的传播步骤而不影响止血在目标 1 中，我们将确定细胞外 Ero1α 的分子机制。在目标 2 中，我们将测试动脉血栓状况是否会改变血小板的功能。在目标 3 中，我们将确定血管内和血小板衍生的细胞外 Ero1α 的病理生理作用。 Ero1α 在血栓条件下血小板粘附和积聚以及血管闭塞中的作用。研究将确定血小板血栓形成的中央调节机制，并导致发现安全有效治疗血栓性疾病的新治疗策略。