Role of vimentin in thrombosis and stroke

波形蛋白在血栓形成和中风中的作用

• 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
• 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

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.

摘要 血小板与刺激的内皮细胞的粘附有助于血栓形成事件， 动脉血栓形成和缺血性中风。虽然抗血小板药物已经减少了不利的 结果，充分认识到出血或复发性血栓形成事件的风险持续存在。 血管性血友病因子在动脉血栓形成中的病理生理作用 并且中风已经通过实验和临床研究得到验证。VWF对于 介导流动的血小板在表面结合的VWF上的束缚， 粘附、活化和聚集-并最终阻塞血管。最近我们 描述了血小板细胞表面表达的波形蛋白与VWF结合， 高剪切应力下的血小板粘附。我们还表明，重组的A2结构域， 对波形蛋白具有结合活性的VWF（“A2蛋白”）阻断波形蛋白与VWF之间的相互作用。 VWF。使用内皮细胞（EC）的另外的实验，所述内皮细胞也在血管内皮细胞上表达波形蛋白。 细胞表面显示A2蛋白阻止了超大（长）VWF形成 多聚体或VWF串在正常EC上，并且未能与波形蛋白缺陷的EC相互作用。 此外，在刺激的内皮细胞表面观察到大量的VWF串， 野生型（WT）小鼠的脑动脉与动脉上观察到的明显较少的弦形成对比 波形蛋白缺陷小鼠的基因这些数据首次表明， 高粘附性的VWF字符串在血管和潜在的这些字符串， 脑血管病最后，损伤诱导的大脑中动脉远端血栓形成 与WT小鼠相比，在Vim（-/-）中，缺血性中风的大脑中动脉（MCA）和严重程度降低。的 本申请的中心假设是细胞表面表达的波形蛋白与A2相互作用 结构域的VWF字符串，从而有助于锚定的VWF字符串的刺激 内皮和附着于循环血小板。本申请还提出测试 破坏波形蛋白/VWF相互作用减少血栓形成和进一步损伤的假说 在缺血再灌注期间。提出了三个目标：目标1）确定 波形蛋白和VWF参与血小板和内皮细胞上的波形蛋白/VWF相互作用， 目的2）确定血小板和内皮波形蛋白在体内血栓形成中的作用; 目的3）证明在VWF诱导后破坏波形蛋白-VWF相互作用的治疗益处。 中风这些目标的实现将通过展示一种新的受体来影响公共卫生， VWF，并将奠定基础，以测试这种新的相互作用作为一个治疗目标，以防止 动脉血栓形成和治疗缺血性中风。