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.

项目概要 将循环白细胞募集到感染部位在身体防御病毒方面发挥着重要作用 病原体。白细胞从血液动员到周围组织发生多步粘附 级联，由几个粘附分子家族介导。整合素淋巴细胞功能- 相关 1 (LFA1) 通过结合表达的细胞内粘附分子 (ICAM) 介导牢固附着 通过血管内皮细胞，最终导致通过血管壁的迁移。申请人，博士。 Travis Moore 在 Timothy Springer 博士的实验室中进行了该领域的重要研究。目标 该应用程序的目的是加深摩尔博士在这一重要研究领域的专业知识和发现，以便能够 发展成为一项重要的独立研究计划。摩尔博士最近领导的研究表明 整合素与肌动蛋白细胞骨架的连接以及沿逆行肌动蛋白方向产生的拉力 流动导致 LFA1 在细胞表面的定向。 LFA1 的这种排列依赖于底物 并与整合素在与逆行肌动蛋白流相同的方向上倾斜一致。这些发现强烈 支持肌动蛋白细胞骨架在细胞粘附性调节中的作用。摩尔博士将测试 假设肌动蛋白细胞骨架通过调节整合素构象平衡和亲和力 机械力施加到整合素胞质结构域并充当变构效应器以稳定 高亲和力、延伸开放的整合素构象。这项工作将利用创新方法来衡量 LFA1 的内在亲和力和构象平衡，并使用最先进的超分辨率 显微镜定量肌动蛋白细胞骨架与整合素细胞质尾部的连接并直接测量 白细胞细胞表面的整合素构象。这些实验将加深我们对 细胞粘附性和迁移的调节，为未来提供新颖和创新的方法 研究，并通过蛋白质化学和纯化方面的专业知识扩展摩尔博士的多元化背景， 超分辨率显微镜和计算图像分析、基因组修饰和定量 细胞表面受体热力学。摩尔博士将把他 100% 的时间投入到这个职业中 波士顿儿童医院的 Timothy Springer 博士作为他的导师获得了发展奖。