Influences of nanomechanical forces on T cells

纳米机械力对 T 细胞的影响

• 批准号: 8932016
• 负责人: MANISH J BUTTE
• 金额: $ 32.23万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-09-25 至 2019-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8932016
• 关键词: Actins; Antigen-Presenting Cells; Antigens; Atomic Force Microscopy; Autoimmune Diabetes; Autoimmune Diseases; Autoimmune Process; Autoimmunity; Automobile Driving; Biochemical; Biological; Biological Models; Biomechanics; Biomimetics; Blood Vessels; Blood flow; Cell Surface Receptors; Cells; Cellular biology; Confocal Microscopy; Cues; Cytoskeletal Filaments; Cytoskeletal Proteins; Cytoskeleton; Data; Diabetes Mellitus; Disease; Endothelial Cells; Environment; Esthesia; Extracellular Matrix; Generations; Glaucoma; Goals; Graft Rejection; Health; Hydrogels; Hypersensitivity; Immune; Immune system; Immunology; Infection Control; Inflammation; Inflammatory; Insulin-Dependent Diabetes Mellitus; Kidney Diseases; Knowledge; Language; Life; Ligation; Link; Lymphoid Tissue; Malignant Neoplasms; Measures; Mechanics; Mediating; Methods; Molecular; Motor; Muscular Dystrophies; Nanotechnology; Osteoblasts; Outcome; Play; Premature aging syndrome; Process; Property; Receptor Signaling; Research Methodology; Role; Scanning Probe Microscopes; Signal Pathway; Signal Transduction; Skeleton; Structure; Surface; T cell regulation; T cell response; T-Cell Activation; T-Cell Receptor; T-Lymphocyte; Techniques; Testing; Therapeutic; Therapeutic Intervention; Time; Tissues; Work; bone; cantilever; cell type; cellular imaging; in vivo; innovation; insight; islet; mouse model; nanofabrication; nanomechanical; nanoscale; novel; prevent; receptor; response; single molecule; tool

DESCRIPTION (provided by applicant): Virtually every cell in the body is exposed to mechanical forces that are transduced through its cytoskeleton and influence its activity. Recent studies using T cells, the master organizing cells of the adaptive immune system, have shown that mechanical forces acting upon the T cell receptor (TCR) and through the cytoskeleton can trigger cellular responses. It is unclear how the cytoskeleton plays this role, or how T cells interpret the interplay of forces and signals of receptor ligation. We have discovered that T cells are more sensitive to antigen (i.e., have a lower threshold of activation) when their actin cytoskeleton is untethered and mechanically "soft." Furthermore, we have discovered that in autoimmune diabetes, mechanical forces from inflamed extracellular matrix (ECM) can provoke T cell autoimmunity. The gaps in our knowledge are due to a lack of tools that can measure and deliver nanoscale forces to live cells. That T cells encode their threshold of activation in cytoskeletal structures provides impetus to study how cells communicate in a language that combines mechanical forces with receptor signals. Here, we propose to 1) Determine the mechanosensitivity of the T cell receptor at the molecular level, using an advanced atomic force microscope (AFM) to convey mechanical forces and antigens to single-molecule TCRs; 2) Identify the cytoskeletal networks required for mechanosensing, using a new generation of AFM cantilevers that can measure cytoskeletal changes in live cells; and 3) Determine influence of mechanical forces due to inflammatory ECM, using a mouse model of autoimmune diabetes and a 3D biomimetic matrix to emulate the inflamed ECM and its mechanical effects on T cell activation. Our lab, working with our collaborators at Stanford, has pioneered breakthroughs in biological atomic force microscopy (AFM) and nanofabrication that make these innovative studies possible by allowing us to precisely ligate receptors and exert minute forces while measuring mechanical responses in live T cells, all while imaging cells and their cytoskeletal changes using live confocal microscopy. These studies will help us decipher the "language" of mechanical forces in cell signaling, and will have a major impact on our understanding of the mechanical effects of inflammatory ECM in autoimmune diseases. We expect our findings will spur novel immune therapeutics. Our new methods of using AFM in cell biology have the potential to revolutionize studies of mechanobiology and receptor signaling pathways implicated in many diseases.

描述（由申请人提供）：几乎体内的每个细胞都暴露于通过其细胞骨架转导并影响其活性的机械力。最近使用T细胞（适应性免疫系统的主要组织细胞）的研究表明，作用于T细胞受体（TCR）并通过细胞骨架的机械力可以触发细胞反应。目前尚不清楚细胞骨架如何发挥这一作用，或者T细胞如何解释受体连接的力和信号的相互作用。我们发现T细胞 对抗原更敏感（即，具有较低的激活阈值）。“此外，我们发现，在自身免疫性糖尿病中，炎症细胞外基质（ECM）的机械力可以引起T细胞自身免疫。我们知识的差距是由于缺乏可以测量和向活细胞传递纳米级力量的工具。T细胞在细胞骨架结构中编码其激活阈值，这为研究细胞如何以结合机械力和受体信号的语言进行交流提供了动力。在这里，我们提出1）在分子水平上确定T细胞受体的机械敏感性，使用先进的原子力显微镜（AFM）将机械力和抗原传递到单分子TCR; 2）识别机械敏感所需的细胞骨架网络，使用新一代的AFM杠杆，可以测量活细胞中细胞骨架的变化;和3）使用自身免疫性糖尿病的小鼠模型和3D仿生基质来模拟发炎的ECM及其对T细胞活化的机械作用，确定由于发炎的ECM引起的机械力的影响。我们的实验室与我们在斯坦福大学的合作者合作，开创了生物原子力显微镜（AFM）和纳米纤维的突破，使这些创新研究成为可能，使我们能够精确地连接受体并施加微小的力，同时测量活T细胞中的机械反应，同时使用活共聚焦显微镜成像细胞及其细胞骨架变化。这些研究将帮助我们破译细胞信号传导中机械力的“语言”，并将对我们理解炎症性ECM在自身免疫性疾病中的机械作用产生重大影响。我们希望我们的发现将刺激新的免疫疗法。我们在细胞生物学中使用AFM的新方法有可能彻底改变与许多疾病有关的机械生物学和受体信号通路的研究。