Mechanoregulatory mechanisms of von Willebrand disease and thrombosis

血管性血友病和血栓形成的机械调节机制

• 批准号: 10164845
• 负责人: Hongxia Fu
• 金额: $ 17.81万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-01 至 2023-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10164845
• 关键词: Adhesions; Affect; Affinity; Air Pressure; Area; Binding; Binding Sites; Biochemical; Biological Assay; Biomedical Engineering; Blood; Blood Circulation; Blood Coagulation Disorders; Blood Platelets; Blood Vessels; Blood coagulation; Blood flow; Cell model; Cells; Cellular biology; Complement; Complex; Diagnostic; Disease; Disease model; Elongation Factor; Endothelial Cells; Fluorescence; Fluorescence Microscopy; Foundations; Glycoproteins; Hemostatic function; Human; Human Cell Line; Individual; Inherited; Ionic Strengths; Laboratories; Length; Ligand Binding; Link; Membrane; Mentors; Mentorship; Methods; Microfluidics; Modeling; Molecular Conformation; Monitor; Mutation; Patients; Physiological; Platelet Glycoproteins; Play; Polymers; Population; Positioning Attribute; Process; Property; Regulation; Reporter; Research; Research Personnel; Resolution; Role; Science; Shapes; Structure; Surface; System; Tertiary Protein Structure; Testing; Therapeutic; Thrombosis; Time; Training; Urea; Variant; Weibel-Palade Bodies; Work; base; body sense; career development; conformational conversion; extracellular; force sensor; gain of function; genome editing; hydrodynamic flow; improved; in vivo; innovation; insight; interdisciplinary approach; molecular imaging; mutant; novel; novel strategies; physiologic model; prevent; programs; receptor; recruit; single molecule; solute; stem cells; von Willebrand Disease; von Willebrand Factor

PROJECT SUMMARY A critical initiating step in hemostasis and thrombosis is adhesion of platelet membrane receptor glycoprotein Ibα (GPIbα) to von Willebrand factor (VWF), a large, multidomain, polymeric blood glycoprotein. The precise mechanisms whereby VWF promotes platelet GPIbα adhesion to its A1 domain only during hemostasis or thrombosis, but not in normal circulation, are not yet clear. Understanding these mechanisms is essential for developing more effective diagnostics and therapeutics for vascular thrombosis and the most common hereditary bleeding disorder, von Willebrand disease (VWD). The applicant, Dr. Hongxia Fu, will develop innovative single-molecule approaches to study full-length VWF concatemers hemostatic function in the laboratory of the mentor, Dr. Timothy Springer. This system can be utilized to monitor both intramolecular VWF conformational transitions and GPIbα binding simultaneously by combining rapid air pressure-actuated shear flow with total internal fluorescence microscopy (TIRF). Utilizing this system, Dr. Fu will test the hypothesis that hydrodynamic flow directly induces a conformational transition in VWF concatemers from a compact to an elongated form, thereby exposing high-affinity, force- dependent binding sites to recruit both GPIbα (platelet adhesion) and additional VWF molecules (VWF self- association). To expand this system to include complex features of physiological and pathophysiological blood flow, she will furthermore develop a new fluorescence-based assay for VWF function in bulk solutions and its expression in Weibel-Palade bodies inside endothelial cells with or without VWD-relevant mutations. This work will provide direct insight into the regulatory mechanisms governing primary hemostasis, thrombosis, and bleeding disorder, establishing a paradigm for mechanosensory control of receptor-ligand binding affinity. It also will provide Dr. Fu with additional training in cell biology, genome editing, stem cells, and biomedicine, complementing her expertise in quantitative sciences. Dr. Fu will devote 100 % of her time to research under the direct mentorship of Dr. Springer. Dr. Fu's research program will establish new quantitative assays for VWF function and VWF-related diseases, from the single molecule to the cellular scale, providing a firm foundation for continued research in this area and career development to the independent investigator stage in biomedicine.

项目概要 止血和血栓形成的关键起始步骤是血小板膜受体的粘附 糖蛋白 Ibα (GPIbα) 到冯维勒布兰德因子 (VWF)，一种大的、多结构域的聚合血液 VWF 促进血小板 GPIbα 与其 A1 结构域粘附的精确机制。 仅在止血或血栓形成期间，但不在正常循环中，目前尚不清楚。 机制对于开发更有效的血管血栓形成诊断和治疗方法至关重要 以及最常见的遗传性出血性疾病，冯维勒布兰德病（VWD）。 傅红霞将开发创新的单分子方法来研究全长 VWF 串联体 导师 Timothy Springer 博士的实验室的止血功能可以利用该系统。 同时监测分子内 VWF 构象转变和 GPIbα 结合 将快速气压驱动剪切流与全内荧光显微镜 (TIRF) 相结合。 利用该系统，傅博士将测试流体动力流动直接诱导构象的假设 VWF 多联体从紧凑形式转变为细长形式，从而暴露出高亲和力、力- 依赖结合位点来募集 GPIbα（血小板粘附）和其他 VWF 分子（VWF 自 关联）扩展该系统以包括生理学和病理生理学的复杂特征。 血流量，她将进一步开发一种新的基于荧光的检测方法，用于检测散装溶液中的 VWF 功能 及其在有或没有 VWD 相关突变的内皮细胞内 Weibel-Palade 小体中的表达。 这项工作将提供对初级止血调节机制的直接见解， 血栓形成和出血性疾病，建立受体-配体机械感觉控制的范例 它还将为傅博士提供细胞生物学、基因组编辑、干细胞、 傅博士将投入 100% 的时间来补充她在定量科学方面的专业知识。 在施普林格博士的直接指导下进行研究将建立新的研究计划。 从单分子到细胞的 VWF 功能和 VWF 相关疾病的定量分析 规模，为该领域的继续研究和职业发展提供坚实的基础 生物医学的独立研究者阶段。

项目成果

Illustrated State-of-the-Art Capsules of the ISTH 2022 Congress.

ISTH 2022 大会最先进的胶囊插图。

• 发表时间: 2022-07
• 作者: Ariëns, Robert A;Hunt, Beverley J;Agbani, Ejaife O;Ahnström, Josefin;Ahrends, Robert;Alikhan, Raza;Assinger, Alice;Bagoly, Zsuzsa;Balduini, Alessandra;Barbon, Elena;Barrett, Christopher D;Batty, Paul;Carneiro, Jorge David Aivazoglou;Chan, Wee
• 通讯作者: Chan, Wee