Mechanisms of interactions between von Willebrand factor and its binding partners

冯维勒布兰德因子与其结合伙伴之间的相互作用机制

基本信息

批准号：
10629031
负责人：
Hongxia Fu
金额：
$ 64.54万
依托单位：
UNIVERSITY OF WASHINGTON
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-09-01 至 2027-08-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10629031
关键词：
ADAMTS Adhesions Atomic Force Microscopy Behavior Binding Biophysics Blood Blood Coagulation Disorders Blood Platelets Blood Vessels Coagulation Process Complex Data Disease Dissociation Event Factor VIII Fibrin Fluorescence Fluorescence Microscopy Goals Hematological Disease Hemorrhage Human Image Individual Infusion procedures Injury Intravenous infusion procedures Label Laboratories Left Link Location Magnetism Maps Measures Mediating Methods Microfluidics Microscopy Modeling Molecular Molecular Conformation Mutation Patients Plasma Polysaccharides Preparation Process Property Proteins Reporting Role Sampling Shapes Spectrum Analysis Structure System Thrombin Thrombosis Thrombus Visualization Work body sense disulfide bond high risk imaging modality immunogenicity improved insight interdisciplinary approach monomer nanometer preservation prevent single molecule structural determinants superresolution imaging therapeutic development von Willebrand Disease von Willebrand Factor

ADAMTS Adhesions Atomic Force Microscopy Behavior Binding Biophysics Blood Blood Coagulation Disorders Blood Platelets Blood Vessels Coagulation Process Complex Data Disease Dissociation Event Factor VIII Fibrin Fluorescence Fluorescence Microscopy Goals Hematological Disease Hemorrhage Human Image Individual Infusion procedures Injury Intravenous infusion procedures Label Laboratories Left Link Location Magnetism Maps Measures Mediating Methods Microfluidics Microscopy Modeling Molecular Molecular Conformation Mutation Patients Plasma Polysaccharides Preparation Process Property Proteins Reporting Role Sampling Shapes Spectrum Analysis Structure System Thrombin Thrombosis Thrombus Visualization Work body sense disulfide bond high risk imaging modality immunogenicity improved insight interdisciplinary approach monomer nanometer preservation prevent single molecule structural determinants superresolution imaging therapeutic development von Willebrand Disease von Willebrand Factor

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项目摘要

SUMMARY Von Willebrand factor (VWF) has two major roles in blood. One is to facilitate platelet adhesion and aggregation, in which a critical step is to activate the VWF A1 domain to bind with platelet protein GPIbα under flow. The other is to protect coagulation factor VIII (FVIII) from degradation, which is important for fibrin clot formation. Mutations in VWF interfering with these binding interactions can cause thrombosis or von Willebrand disease. The goal of this project is to determine molecular mechanisms governing the interactions between VWF and its binding partners GPIbα and FVIII. Several questions regarding VWF interactions with GPIbα and FVIII persist. There has not been a consistent model for how VWF A1 domain is activated to bind with GPIbα. It still remains unclear which and how conformational changes actually happen during A1 activation. To date, the interaction between VWF multimer and FVIII has been studied primarily using bulk VWF preparations. How FVIII binds to individual VWF multimers is unknown. In intravenous infusion treatment for bleeding disorders, more free FVIII in the infusion is linked to higher risk of FVIII immunogenicity, but the mechanisms remain unclear. In this project, we will combine single molecule biophysics and super resolution imaging methods to study the following aims. (1) Understand flow-induced activation of VWF A1 binding with GPIbα. (2) Dissect the VWF- FVIII binding mode and its role in protecting FVIII in blood. This study will provide direct evidence for the mechanisms of how VWF binds with GPIbα and FVIII, and provide new insights into therapeutic development to treat thrombotic or bleeding disorders.

概括冯·威尔布兰德因子（VWF）在血液中有两个主要角色。一种是促进血小板粘附和聚集，其中关键步骤是激活VWF A1结构域与血小板蛋白GPIBα在下面结合流动。另一个是保护凝血因子VIII（FVIII）免受降解，这对于纤维蛋白克隆很重要格式。 VWF中的突变会干扰这些结合相互作用会导致血栓形成或von Willebrand 疾病。该项目的目的是确定管理VWF之间相互作用的分子机制及其结合伙伴GPIBα和FVIII。有关VWF与GPIBα和FVIII相互作用的几个问题持续存在。没有 VWF A1结构域如何激活与GPIBα结合的一致模型。仍然不清楚哪个以及如何构象变化实际上发生在A1激活期间。迄今为止，VWF多二聚体之间的相互作用 FVIII主要是使用散装VWF制剂进行研究的。 FVIII如何与单个VWF多二聚体结合是未知的。在静脉输注治疗出血疾病中，输注中更多的自由FVIII与 FVIII免疫原性的风险更高，但这些机制仍不清楚。在这个项目中，我们将结合单分子生物物理学和超级分辨率成像方法来研究以下目标。（1）了解与GPIBα结合VWF A1的流动诱导的激活。（2）剖析vwf- FVIII结合模式及其在保护血液中FVIII中的作用。这项研究将为 VWF如何与GPIBα和FVIII结合的机制，并为治疗发展提供新的见解治疗血栓形成或出血疾病。