Regulation of von Willebrand Factor - platelet binding by Force and Interdomain I

通过力和域间 I 调节血管性血友病因子 - 血小板结合

• 批准号: 8024519
• 负责人: Wendy E Thomas
• 金额: $ 44.55万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-12-01 至 2015-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8024519
• 关键词: Adhesions; Adhesives; Adverse effects; Antibodies; Behavior; Binding; Biological Assay; Blood Platelets; Blood Vessels; Cardiovascular Diseases; Cause of Death; Cells; Cloning; Disease; Elements; Event; Exposure to; Glycoprotein Ib; Hemorrhage; Hemostatic function; Knowledge; Lead; Measures; Mechanics; Mediating; Microfluidics; Mutation; Myocardial Infarction; Nucleic Acid Regulatory Sequences; Physiological; Physiology; Plasma Proteins; Platelet aggregation; Point Mutation; Process; Protein Engineering; Recombinants; Regulation; Research Proposals; Role; Scanning Probe Microscopes; Site; Spectrum Analysis; Stress; Stretching; Stroke; Surface; System; Testing; Therapeutic Agents; Thrombosis; Thrombus; Work; base; fluid flow; in vivo; novel; prevent; receptor; research study; shear stress; single molecule; theories; von Willebrand Disease; von Willebrand Factor

DESCRIPTION (provided by applicant): Arterial thrombosis occurs in conditions of high fluid flow and is mediated by the binding of platelets via their receptor Glycoprotein Ib (GPIb) to the plasma protein called von Willebrand Factor (VWF) via the A1 domain. This process is normally down-regulated to prevent spontaneous adhesion and thrombosis, but is activated by high shear stress. The proposed work will test the hypothesis that binding of GPIb to A1 is down-regulated by the neighboring domains or flanking regions within VWF, so that VWF displays interdomain auto-inhibition. It will also test the hypothesis that mechanical force associated with high shear stress activates platelet binding by pulling these regulatory regions away from the A1 domain to relieve the auto- inhibition. The final hypothesis to be tested is that conditions that increase the spontaneous binding of platelets to VWF, including type 2B von Willebrand disease, do so by damaging the normal interdomain auto-inhibition. Regulatory domains and elements within VWF will be identified by cloning different regions of VWF and determining the adhesive behavior of both soluble and surface-immoblized molecules. The effect of mechanical force and shear stress on this regulation will be determined using an atomic force microscope and microfluidics after immobilizing the molecules in an oriented fashion. PUBLIC HEALTH RELEVANCE: Thrombosis prevents bleeding at the site of damaged blood vessels but can also lead to heart attacks or strokes, the leading causes of death in cardiovascular disease. Current treatments for cardiovascular disease often cause excessive bleeding, while treatments for bleeding disorders can cause thrombotic disorders. To develop new therapies without these fundamental trade- offs, we need to better understand how the thrombotic process is regulated in the body.

描述（由申请人提供）：动脉血栓形成发生在高流体流动的条件下，并通过其受体糖蛋白IB（GPIB）与称为von Willebrand因子（VWF）的血浆蛋白与血浆蛋白的结合介导。该过程通常会下调以防止自发粘附和血栓形成，但被高剪切应力激活。拟议的工作将检验以下假设：GPIB与A1的结合被VWF内的相邻域或侧翼区域降低，因此VWF显示域间自动抑制。它还将检验以下假设：与高剪切应力相关的机械力通过将这些调节区域拉出A1结构域来减轻自身抑制，从而激活血小板结合。要测试的最终假设是，增加了血小板与VWF自发结合的条件，包括2B型Von Willebrand疾病，通过损害正常的域间自动抑制来做到这一点。通过克隆VWF的不同区域并确定可溶性和表面不大化分子的粘合性行为，可以确定VWF内的调节域和元素。机械力和剪切应力对这种调节的影响将使用原子力显微镜和微流体以定向方式固定分子后确定。 公共卫生相关性：血栓形成阻止受损的血管部位出血，但也可能导致心脏病发作或中风，这是心血管疾病中死亡的主要原因。目前对心血管疾病的治疗通常会导致过度出血，而出血疾病的治疗可能会引起血小板疾病。为了开发新的疗法，没有这些基本的贸易，我们需要更好地了解体内的血栓形成过程。