Inhibiting biomechanical platelet aggregation to prevent arterial thrombosis without compromising hemostasis

抑制生物力学血小板聚集以预防动脉血栓形成而不影响止血

• 批准号: 10039481
• 负责人: Yunfeng Chen
• 金额: $ 10.12万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-21 至 2022-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10039481
• 关键词: Adhesions; Affect; American; Antibodies; Arteries; Award; Binding; Biochemical; Biological Assay; Biology; Biomechanics; Biomedical Engineering; Blood Platelets; Blood Vessels; Blood coagulation; Blood coagulation tests; Blood flow; Clinical Treatment; Data; Diabetes Mellitus; Disease; Effectiveness; Evaluation; Fibrinolytic Agents; Foundations; Glycoproteins; Goals; Hematology; Hemorrhage; Hemostatic function; Hot Spot; In Vitro; Incidence; Integrins; Mediating; Mentors; Microfluidics; Modeling; Molecular; Monoclonal Antibodies; Morbidity - disease rate; Mus; Mutation; Myocardial Infarction; Names; Pathologic; Performance; Phase; Plasma Proteins; Platelet Aggregation Inhibition; Platelet aggregation; Program Development; Property; Research; Research Personnel; Risk; Safety; Screening procedure; Stenosis; Stroke; Supervision; System; Testing; Therapeutic; Thrombosis; Work; base; career; career development; crosslink; design; diabetic patient; in vivo; inhibitor/antagonist; mechanical force; mechanotransduction; mortality; mutant; pre-clinical; prevent; prototype; receptor; screening; shear stress; therapeutic development; thrombogenesis; translational approach; von Willebrand Factor

Project Summary/Abstract Arterial thrombosis, which describes the formation of abnormal blood clots in the artery, is a highly fatal disease that claims ~500,000 American lives per year. A symbolic feature of arterial thrombosis is the elevated shear rate and shear force generated by stenosis, which facilitates platelet aggregation towards vessel occlusion. Unfortunately, current therapeutic strategies are ineffective in inhibiting the prothrombotic effects of the pathological blood flow, and have a major risk of excessive bleeding. The overall objective of this proposal is to establish a mechanism-driven strategy to better treat arterial thrombosis. Our lab's previous works and my research identified that the prothrombotic effects of pathological blood flow result from a phenomenon called “biomechanical platelet aggregation”, in which mechanical force drives platelets to crosslink. Von Willebrand factor (VWF) is a plasma protein that mediates platelet crosslinking by binding to platelet receptor GPIbα. In my preliminary study, I worked on an artificially designed triple-residue VWF mutation named `M13', which inhibited VWF in mediating biomechanical platelet aggregation and shear-induced thrombogenesis, but did not affect platelet adhesion or hemostasis. Based on these findings, I hypothesize that: biomechanical platelet aggregation is a key factor to arterial thrombosis, and its inhibition can suppress arterial thrombosis without compromising hemostasis. I propose 4 aims to step-by-step establish anti-thrombotic approaches targeting the biomechanical platelet aggregation. Aim 1 will combine biomechanics and hematology assays to study the mechanism underlying M13's function in inhibiting biomechanical platelet aggregation but not adhesion. Aim 2 will establish a microfluidic-based assay that concurrently assesses platelet adhesion and shear-induced thrombogenesis, which allows the screening of anti-thrombotics targeting biomechanical platelet aggregation. In Aim 3, a monoclonal antibody against VWF, NMC4, was identified to be functionally aligned with my anti-thrombotic strategy. I will collaborate with my postdoctoral lab and use NMC4 as a prototype to design and produce anti-thrombotic agents. A 3-step screening procedure will be established to select candidate agents with the best functional performance. Lastly in Aim 4, I will expand my anti-thrombotic strategy to another platelet receptor also important to biomechanical platelet aggregation: integrin αIIbβ3. I will explore the efficacy and safety of inhibiting both VWF- and αIIbβ3-mediated biomechanical platelet aggregation in treating arterial thrombosis exacerbated by diabetes. Overall, this research will be accomplished in the setting of a comprehensive career development program designed to help me achieve my career goal as an independent researcher in the interdisciplinary field of vascular biology, mechanobiology and bioengineering. During the K99 phase, I will continue to gain expertise in biochemical, preclinical and translational approaches. My mentor, collaborators and consultants will together guide me in the steps towards successful transition to independence over the course of the award period.

项目摘要/摘要 动脉血栓形成描述了动脉异常血凝块的形成，是一种高度致命的 每年约有500,000次生活的疾病。动脉血栓形成的符号特征是升高 狭窄产生的剪切速率和剪切力，该设施血小板聚集到血管 阻塞。不幸的是，当前的治疗策略无效地抑制 病理血液流动，并具有过量出血的主要风险。该提议的总体目标 是建立一种机制驱动的策略，以更好地治疗伪影血栓形成。我们实验室以前的作品以及我的 研究表明，病理血流的促血栓性作用是由一种称为的现象引起的 “生物力学血小板聚集”，其中机械力驱动血小板交叉链接。冯·威勒布兰德 因子（VWF）是一种血浆蛋白，通过与血小板受体GPIBα结合来介导血小板交联。在 我的初步研究，我研究了一个人为设计的三重剩余vwf突变，名为“ M13”，该突变 抑制VWF在介导生物力学血小板聚集和剪切诱导的血栓形成时，但没有 影响血小板粘合剂或止血。基于这些发现，我假设：生物力学血小板 聚集是动脉血栓形成的关键因素，其抑制作用可以抑制动脉血栓形成 没有损害止血。我提出4个目标，旨在逐步建立反栓性方法 靶向生物力学血小板聚集。 AIM 1将结合生物力学和血液学测定法 研究M13在抑制生物力学血小板聚集中的功能的机制，但不 粘附。 AIM 2将建立基于微流体的评估，同时评估血小板粘合剂和 剪切诱导的血栓形成，允许筛查靶向生物力学血小板 聚合。在AIM 3中，针对VWF，NMC4的单克隆抗体被确定为功能对齐 有了我的反栓性策略。我将与博士后实验室合作，并将NMC4用作原型 设计并生产抗栓性剂。将建立三步筛选程序以选择 具有最佳功能性能的候选代理商。最后，在AIM 4中，我将扩大我的反栓性 另一个血小板受体的策略对生物力学血小板聚集也很重要：整联蛋白αIIBβ3。我会 探索抑制VWF-和αIIBβ3介导的生物力学血小板聚集的效率和安全性 在治疗动脉血栓形成时，糖尿病加剧了。总体而言，这项研究将在 设定一项全面的职业发展计划，旨在帮助我实现自己的职业目标 血管生物学，机械生物学和生物工程跨学科领域的独立研究人员。 在K99阶段，我将继续获得生化，临床前和翻译方法的专业知识。 我的心理，合作者和顾问将共同指导我成功过渡到 在整个奖项期间的独立性。