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.

抽象的 血小板与受刺激的内皮细胞的粘附有助于与以下疾病相关的血栓形成事件： 动脉血栓形成和缺血性中风。尽管抗血小板药物已经减少了不利 但众所周知，出血或复发性血栓事件的风险仍然存在。 血管性血友病因子（VWF）蛋白在动脉血栓形成中的病理生理功能 中风已通过实验和临床研究得到验证。 VWF 很重要 介导流动血小板在表面结合的 VWF 上的束缚，从而导致血小板 粘附、激活和聚集——最终导致血管闭塞。最近，我们 描述了血小板细胞表面表达的波形蛋白与 VWF 结合并有助于 高剪切应力下的血小板粘附。我们还表明，重组 A2 结构域 具有波形蛋白结合活性的 VWF（“A2 蛋白”）可阻断波形蛋白与波形蛋白之间的相互作用。 VWF。使用内皮细胞 (EC) 进行其他实验，该细胞也表达波形蛋白 细胞表面，揭示 A2 蛋白阻止超大（长）VWF 的形成 正常 EC 上存在多聚体或 VWF 串，无法与波形蛋白缺陷的 EC 相互作用。 此外，在受刺激的内皮表面观察到大量 VWF 线。 野生型 (WT) 小鼠的脑动脉与动脉上明显减少的线形成对比 来自波形蛋白缺陷小鼠。这些数据首次证明了 脑血管系统中的超粘附性 VWF 线以及这些线在 脑血管病变。最后，损伤诱发远端中脑血栓形成 与 WT 小鼠相比，Vim(-/-) 小鼠的 MCA 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 弦的域，从而有助于将 VWF 弦锚定到受刺激的 内皮和循环血小板的附着。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和进一步损伤 缺血再灌注期间。提出了三个目标： 目标 1) 定义重要站点 波形蛋白和 VWF 参与血小板和内皮细胞上波形蛋白/VWF 相互作用 细胞；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；和 目标 3) 证明破坏波形蛋白-VWF 相互作用的治疗益处 中风。这些目标的完成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。