Role of vimentin in thrombosis and stroke

波形蛋白在血栓形成和中风中的作用

基本信息

批准号：
9927689
负责人：
Miguel Angel Cruz
金额：
$ 39.11万
依托单位：
BAYLOR COLLEGE OF MEDICINE
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-09-01 至 2021-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9927689
关键词：
Acute Adhesions Antiplatelet Drugs Arteries Attenuated Binding Biological Blood Platelets Cell surface Cerebrovascular system Clinical Research Data Disease Distal Effectiveness Endothelial Cells Endothelium Event Foundations Hemorrhage Histamine Immobilization Injury Ischemic Stroke Knock-out Laser-Doppler Flowmetry Ligands Mediating Modeling Molecular Mus Myocardial Infarction Outcome Pathogenesis Pathologic Pathology Pharmacology Plasma Platelet Activation Platelet aggregation Proteins Public Health Publishing Recombinants Recovery of Function Recurrence Reperfusion Therapy Risk Role Severities Site Stroke Surface Testing Therapeutic Thrombosis Thrombus Time Vimentin Wild Type Mouse cerebral artery cerebrovascular pathology experimental study improved in vivo intravital microscopy ischemic injury middle cerebral artery novel novel therapeutic intervention novel therapeutics prevent protein function shear stress stroke model stroke outcome therapeutic target von Willebrand Factor von Willebrand factor receptor

项目摘要

ABSTRACT Adhesion of platelets to stimulated endothelium contributes to thrombotic events associated with arterial thrombosis and ischemic stroke. Although anti-platelet agents have reduced unfavorable outcomes, it is well appreciated that the risk for bleeding or recurrent thrombotic events persists. The pathophysiological function of the protein von Willebrand factor (VWF) in arterial thrombosis and stroke has been validated by experimental and clinical studies. VWF is important for mediating the tethering of the flowing platelets over the surface-bound VWF that result in platelet adhesion, activation, and aggregation – and ultimately occlusion of the vessel. Recently, we described that vimentin expressed on the cell surface of platelets binds to VWF and contributes to platelet adhesion at high shear stress. We also have shown that a recombinant A2 domain of VWF (“A2 protein”) with binding activity for vimentin blocked the interaction between vimentin and VWF. Additional experiments employing endothelial cells (ECs), which also express vimentin on the cell surface, revealed that that A2 protein prevented the formation of ultra large (long) VWF multimers or VWF strings on normal ECs and failed to interact with vimentin-deficient ECs. Moreover, the large number of VWF strings observed at the endothelial surface of stimulated cerebral arteries from wild type (WT) mice contrast to the significantly less strings seen on arteries from vimentin deficient mice. These data demonstrate for the first time the presence of hyperadhesive VWF strings in the cerebrovasculature and the potential for these strings in cerebrovascular pathologies. Finally, injury-induced thrombus formation in distal middle cerebral artery (MCA) and severity of ischemic stroke was reduced in Vim(-/-) compared to WT mice. The central hypothesis of this application is that cell-surface expressed vimentin interacts with the A2 domain of VWF strings, thus contributing to anchoring of VWF strings to the stimulated endothelium and attachment to circulating platelets. This application also proposes to test the hypothesis that disrupting the vimentin/VWF interaction reduces thrombosis and further damage during ischemic reperfusion. Three aims are proposed: Aim 1) Define the essential sites in vimentin and VWF that are involved in the vimentin/VWF interaction on platelets and endothelial cells; Aim 2) Determine the role of platelet and endothelial vimentin in thrombosis in vivo; and Aim 3) Demonstrate therapeutic benefit of disrupting the vimentin-VWF interaction following stroke. Completion of these aims will impact public health by demonstrating a novel receptor for VWF, and will lay the foundation to test this new interaction as a therapeutic target to prevent arterial thrombosis and treat ischemic stroke.

抽象的血小板对刺激内皮的粘附有助于与虽然反板剂剂减少了不利结果，人们非常感谢，出血或复发性血栓事件的风险仍然存在。蛋白质von Willebrand因子（VWF）的病理生理功能并通过实验和临床研究证实了中风。 vwf对介导流动血小板在表面结合的VWF上的束缚，导致血小板粘附，激活和聚集 - 并最终阻塞血管。最近，我们描述的是，在血小板的细胞表面表达的波形蛋白与VWF结合，并有助于高剪切应力下的血小板粘附。我们还表明，重组A2域 VWF（“ A2蛋白”）具有与波形蛋白的结合活性，阻止了波形蛋白与 vwf。采用内皮细胞（EC）的其他实验，也表达波形蛋白在细胞表面表明A2蛋白阻止了超大（长）VWF的形成正常EC上的多聚体或VWF字符串，未能与缺乏波形蛋白的EC相互作用。此外，在刺激的内皮表面上观察到的大量VWF串来自野生型（WT）小鼠的脑动脉与动脉上看到的明显较少的弦来自波形蛋白缺乏小鼠。这些数据首次证明了脑血管管中的高粘性VWF弦，以及这些字符串的潜力脑血管病理。最后，损伤引起的远端中部大脑的血栓形成与WT小鼠相比，VIM（ - / - ）的动脉（MCA）和缺血性中风的严重程度降低。这该应用的中心假设是细胞表面表达的波形蛋白与A2相互作用 VWF字符串的域，从而有助于将VWF字符串锚定在刺激上内皮和附着在循环血小板上。该应用程序还建议测试破坏波形蛋白/VWF相互作用的假设可减少血栓形成并进一步损害在缺血性再灌注期间。提出了三个目标：目标1）定义中的基本站点在血小板和内皮上涉及的波形蛋白/VWF相互作用中涉及的波形蛋白和VWF 细胞；目标2）确定血小板和内皮波形蛋白在体内血栓形成中的作用；和目的3）在以下中风。这些目标的完成将通过展示一种新颖的受体来影响公共卫生 vwf，并将奠定基础，以测试这种新互动作为一种治疗目标，以防止动脉血栓形成和治疗缺血性中风。