Mechanoregulation of the Leukocyte Specific Integrin LFA-1 by the Actin Cytoskeleton

肌动蛋白细胞骨架对白细胞特异性整合素 LFA-1 的机械调节

• 批准号: 9985152
• 负责人: Travis I Moore
• 金额: $ 16.96万
• 依托单位: UNIVERSITY OF TEXAS HLTH SCI CTR HOUSTON
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-08-15 至 2023-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9985152
• 关键词: Actins; Adhesions; Adhesiveness; Affect; Affinity; Area; Bacteria; Bacterial Infections; Binding; Binding Proteins; Blood Circulation; Blood Vessels; Boston; Cell Adhesion; Cell Adhesion Molecules; Cell membrane; Cell surface; Cells; Cellular Structures; Clustered Regularly Interspaced Short Palindromic Repeats; Complex; Coupling; Crystallization; Cutaneous; Cytoplasmic Tail; Cytoskeleton; Development; Disease; Family; Flow Cytometry; Fluorescence; Free Energy; Gene-Modified; Genome; Goals; HL60; ICAM1 gene; Image Analysis; Immune response; Infection; Integrins; K-Series Research Career Programs; Knock-out; Laboratories; Lead; Leukocytes; Libraries; Ligand Binding; Ligand Binding Domain; Ligands; Lighting; Link; Lymphocyte Function; Lymphocyte Function-Associated Antigen-1; Measurement; Measures; Mechanics; Mediating; Membrane; Mentors; Methodology; Microscopy; Modification; Molecular; Molecular Conformation; Multiple Sclerosis; Optics; Pathology; Pediatric Hospitals; Play; Point Mutation; Population; Protein Chemistry; Protein Conformation; Proteins; Regulation; Research; Resolution; Rheumatoid Arthritis; Role; Series; Signal Pathway; Signal Transduction; Signal Transduction Pathway; Site; Structural Models; Structure; Surface; T-Lymphocyte; Techniques; Testing; Thermodynamics; Time; Tissues; Virus; Virus Diseases; Work; cell growth regulation; cell motility; cell type; chemokine receptor; experimental study; extracellular; improved; innovation; macrophage; mechanical force; migration; mutant; novel; pathogen; peripheral blood; photoactivation; programs; protein purification; receptor; recruit; small molecule; tumor

PROJECT SUMMARY Recruitment of circulating leukocytes to sites of infection plays an important role in the body's defense against pathogens. Mobilization of leukocytes from the blood to the surrounding tissue occurs in a multistep adhesion cascade, which is mediated by several adhesion molecule families. The integrin lymphocyte function- associated 1 (LFA1) mediates firm attachment by binding Intracellular Adhesion Molecules (ICAMs) expressed by the vascular endothlilium and ultimately leads to transmigration through the vessel wall. The applicant, Dr. Travis Moore has preformed important studies in this area in the laboratory of Dr. Timothy Springer. The goal of this application is to deepen Dr. Moore's expertise and findings in this important research area so that it can be developed into a significant independent research program. Dr. Moore recently lead research to show that integrin linkage to the actin cytoskeleton and the tensile force generated along the direction of retrograde actin flow resulted in the orientation of LFA1 on the cell surface. This alignment of LFA1 was substrate dependent and consistent with integrins being tilted in the same direction as retrograde actin flow. These findings strongly support a role for the actin cytoskeleton in the regulation of cellular adhesiveness. Dr. Moore will test the hypothesis that the actin cytoskeleton regulates integrin conformational equilibria and affinity through mechanical force applied to the integrin cytoplasmic domain and acts as an allosteric effector to stabilize the high-affinity, extended-open integrin conformation. This work will utilize innovative methodologies to measure the intrinsic affinities and conformational equilibria of LFA1 and use state-of-the-art super-resolution microscopy to quantitate the actin cytoskeleton linkage to the integrin cytoplasmic tail and directly measure integrin conformation on the cell surface of leukocytes. These experiments will advance our understanding of the regulation of cell adhesiveness and migration, produce novel and innovative methodologies for future research, and expand Dr. Moore's diverse background with expertise in protein chemistry and purification, super-resolution microscopy and computational image analysis, genome modification, and quantitating receptor thermodynamics on the cell surface. Dr. Moore will devote 100 % of his time to this Career Development Award with Dr. Timothy Springer as his mentor at Boston Children's Hospital.

项目摘要 在感染部位招募流通的白细胞在人体防御中起着重要作用 病原体。从血液到周围组织的白细胞动员发生在多步粘附中 级联反应是由几个粘附分子家族介导的。整联蛋白淋巴细胞功能 - 相关的1（LFA1）通过表达的细胞内粘附分子（ICAMS）介导企业的附着 通过血管内胸，最终导致通过血管壁移民。申请人，博士 特拉维斯·摩尔（Travis Moore）在蒂莫西·斯普林格（Timothy Springer）博士实验室中已经在这一领域进行了重要研究。目标 此应用是为了加深摩尔博士在这个重要研究领域的专业知识和发现，以便可以 被发展为重要的独立研究计划。摩尔博士最近领导研究表明 整合素连接到肌动蛋白细胞骨架和沿逆行肌动蛋白方向产生的拉伸力 流动导致LFA1在细胞表面的取向。 LFA1的这种比对是底物依赖性的 并且与整联蛋白在与逆行肌动蛋白流相同的方向上倾斜。这些发现强烈 支持肌动蛋白细胞骨架在调节细胞粘附性中的作用。摩尔博士将测试 肌动蛋白细胞骨架调节整联蛋白构象平衡和亲和力的假设 机械力应用于整联蛋白细胞质结构域，并作为变构效应器稳定 高亲和力，扩展打开的整联蛋白构象。这项工作将利用创新的方法来衡量 LFA1的固有亲和力和构象平衡并使用最先进的超分辨率 显微镜量化肌动蛋白细胞骨架链接到整合素细胞质尾巴并直接测量 白细胞细胞表面的整合素构象。这些实验将提高我们对 细胞粘附性和迁移的调节，为未来产生新颖的创新方法 研究并扩大了Moore博士的不同背景，具有蛋白质化学和纯化方面的专业知识， 超分辨率显微镜和计算图像分析，基因组修饰和定量 细胞表面的受体热力学。摩尔博士将100％的时间投入这一职业 蒂莫西·斯普林格（Timothy Springer）博士的开发奖是波士顿儿童医院的导师。