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 血小板与刺激内皮细胞的黏附导致血栓事件 动脉血栓形成和缺血性中风。尽管抗血小板药物减少了对 考虑到患者的预后，出血或血栓复发的风险仍然存在。 血管性血友病因子在动脉血栓形成中的病理生理作用 中风已得到实验和临床研究的验证。VWF对于 调节流动的血小板在表面结合的VWF上的拴系，导致血小板 黏附、激活和聚集--最终导致血管闭塞。最近，我们 描述了表达在血小板细胞表面的波形蛋白与VWF结合并有助于 高切应力下的血小板粘附性。我们还证明了一个重组的A2结构域 具有波形蛋白结合活性的VWF(A2蛋白)阻断了波形蛋白与肌动蛋白的相互作用 VWF。使用内皮细胞(ECs)的其他实验，内皮细胞也表达波形蛋白 表明A2蛋白阻止了超大(长)VWF的形成 正常内皮细胞上的多聚体或VWF串，并且无法与波形蛋白缺陷的内皮细胞相互作用。 此外，在受刺激的血管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明显较少的线条形成对比 来自波形蛋白缺陷小鼠。这些数据首次证明了 脑血管系统中的超黏附VWF串以及这些串在脑血管中的潜力 脑血管病理学。最后，损伤诱导的大脑中脑远端血栓形成 与WT小鼠相比，Vim(-/-)小鼠的大脑中动脉(MCA)和缺血性中风的严重程度都有所降低。这个 这一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串的区域，从而有助于将VWF串锚定到受激者 血管内皮细胞与循环中的血小板的附着。此应用程序还建议测试 破坏Vimentin/VWF相互作用可减少血栓形成和进一步损害的假说 在缺血再灌流期间。提出了三个目标：目标1)定义 参与Vimentin/VWF与血小板和血管内皮细胞相互作用的Vimentin和VWF 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；以及 目的3)展示干扰Vimentin-VWF相互作用的治疗益处 卒中。这些目标的实现将通过展示一种新的受体来影响公共健康 VWF，并将为测试这种新的相互作用作为预防的治疗靶点奠定基础 动脉血栓形成和治疗缺血性中风。