Subcellular Platelet Forces and Adhesions

亚细胞血小板力和粘附

• 批准号: 7713020
• 负责人: Nathan John Sniadecki
• 金额: $ 20.97万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-01 至 2011-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7713020
• 关键词: Adhesions; Adhesives; Affect; Anticoagulants; Area; Biological; Biological Assay; Biology; Blood Platelets; Blood Transfusion; Blood Vessels; Blood flow; Cells; Coagulation Process; Cytoskeletal Proteins; Cytoskeleton; Development; Devices; Dimensions; Elasticity; Event; Extracellular Matrix; Extracellular Matrix Proteins; Fibrin; Focal Adhesions; Goals; Heart Diseases; Hemorrhage; Hemostatic function; Hyperactive behavior; Image Analysis; Immunofluorescence Immunologic; Individual; Injury; Integrins; Laboratories; Lead; Ligand Binding; Ligands; Measurement; Measures; Mechanics; Mediating; Mentors; Methods; Microscopy; Monitor; Obstruction; Optics; Outcome Study; Pennsylvania; Physiological; Play; Postdoctoral Fellow; Printing; Proteins; Regulation; Reporting; Research; Research Design; Research Personnel; Resolution; Resources; Role; Silicone Elastomers; Spatial Distribution; Stents; Stroke; Structure; Supporting Cell; Techniques; Thrombosis; Thrombus; Tissues; Training; Work; atherothrombosis; base; biomechanical engineering; cell type; cellular development; conventional therapy; flexibility; improved; injured; insight; nanofabrication; nanoscale; novel; novel strategies; prevent; public health relevance; receptor; response; sensor; tool

DESCRIPTION (provided by applicant): This project focuses on the role that cytoskeletal forces play in platelet adhesions to mediate the formation and stability of clots. Platelet adhesion significantly influences thrombosis since platelets need to aggregate together to stop blood loss, but also to detach and disassemble in order to not obstruct flow to other tissue. It is unclear how platelets direct this dual response, but previous work in this area has shown that integrin adhesions and cytoskeletal forces both are necessary for controlling thrombus stability. However, the interactions between integrins and forces are not completely understood, in part because there are limited tools and techniques available to simultaneously monitor the mechanical interactions that occur at the subcellular level. This project will combine our expertise in measuring cellular forces and quantitative image analysis in order to study the relationship between cytoskeleton-generated platelet forces and integrin adhesivity in platelets. The approach involves the use of flexible post force-sensors with nanoscale dimensions that can directly measure cytoskeletal forces at focal adhesions of platelets and immunofluorescence image analysis to assess their cytoskeletal protein organization. The long-term goal of this project will be to develop a new method that quantitatively measures the magnitude of platelet forces without the confounding influences of platelet-fibrin interactions. As a proof-of-concept, this approach will be used to identify and characterize cytoskeletal proteins that promote platelet adhesion and thrombus stability. Specific Aim 1 will be to measure platelet forces using flexible nanopost force-sensors. Specific Aim 2 will be to investigate the development of platelet forces of activated platelets. Specific Aim 3 will be to examine the relationship of platelet forces with focal adhesion development and lay the groundwork for mechanistic studies of force and integrin interactions that regulate platelet adhesion. The device to be developed will enable physiological studies of platelets at the nanoscale that have not been possible previously. The increased understanding that this new approach will bring will have the potential to provide important input into efforts to control thrombosis, which is a critical event in the development of heart disease and stroke. PUBLIC HEALTH RELEVANCE: The device to be developed will enable physiological studies of platelets at the nanoscale that have not been possible previously. The increased understanding that this new approach will bring will have the potential to provide important input into efforts to control thrombosis, which is a critical event in the development of heart disease and stroke.

描述（由申请人提供）：该项目重点研究细胞骨架力在血小板粘附中所发挥的作用，以介导凝块的形成和稳定性。血小板粘附显着影响血栓形成，因为血小板需要聚集在一起以阻止失血，但也需要分离和分解，以免阻碍流向其他组织。目前尚不清楚血小板如何引导这种双重反应，但该领域之前的工作表明，整合素粘附和细胞骨架力都是控制血栓稳定性所必需的。然而，整合素和力之间的相互作用尚不完全清楚，部分原因是可用于同时监测亚细胞水平上发生的机械相互作用的工具和技术有限。该项目将结合我们在测量细胞力和定量图像分析方面的专业知识，以研究细胞骨架产生的血小板力与血小板中整合素粘附力之间的关系。该方法涉及使用具有纳米级尺寸的灵活后力传感器，可以直接测量血小板粘着斑处的细胞骨架力，并通过免疫荧光图像分析来评估其细胞骨架蛋白组织。该项目的长期目标是开发一种新方法，定量测量血小板力的大小，而不受血小板-纤维蛋白相互作用的混杂影响。作为概念验证，该方法将用于识别和表征促进血小板粘附和血栓稳定性的细胞骨架蛋白。具体目标 1 是使用柔性纳米柱力传感器测量血小板力。具体目标 2 将是研究活化血小板的血小板力的发展。具体目标 3 将是检查血小板力与粘着斑发展的关系，并为调节血小板粘附的力和整合素相互作用的机制研究奠定基础。即将开发的设备将能够在纳米尺度上对血小板进行生理学研究，这在以前是不可能的。这种新方法带来的更多了解将有可能为控制血栓形成的努力提供重要的投入，血栓形成是心脏病和中风发展的关键事件。公共健康相关性：即将开发的设备将能够在纳米尺度上对血小板进行生理研究，这在以前是不可能的。这种新方法带来的更多了解将有可能为控制血栓形成的努力提供重要的投入，血栓形成是心脏病和中风发展的关键事件。