Controllable Rigidity Surfaces for T Cell Mechanobiology

T 细胞力学生物学的可控刚性表面

• 批准号: 9243969
• 负责人: Lance C Kam
• 金额: $ 19.66万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-03-15 至 2021-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9243969
• 关键词: Actins; Adaptive Immune System; Address; Adverse effects; Alpha Cell; Antibodies; Area; Atomic Force Microscopy; Biological Products; CD28 gene; CD3 Antigens; CD4 Positive T Lymphocytes; Cadherins; Cell Communication; Cell physiology; Cells; Chemical Agents; Chemicals; Communication; Development; Elastomers; Engineering; Event; Exhibits; Extracellular Matrix; Generations; Goals; Hour; Human; Immune response; Immunotherapy; Integrins; Knowledge; Language; Ligands; Magnetism; Measures; Mechanics; Modeling; Molecular; Molecular Immunology; Mus; Optics; Pathway interactions; Phase; Play; Process; Property; Role; Signal Transduction; Site; Substrate Interaction; Surface; System; T-Cell Activation; T-Lymphocyte; TCR Activation; Technology; Testing; Time; Traction; Translations; Tumor stage; Validation; Work; ZAP-70 Gene; adaptive immune response; base; biological systems; cell behavior; cellular targeting; density; human disease; immunological synapse; improved; insight; knowledge base; magnetic dipole; magnetic field; mechanical force; mechanotransduction; nanoscale; nanowire; public health relevance; tool

 DESCRIPTION: Mechanical forces play increasingly recognized roles in directing T cell activation and subsequent function. We recently discovered that T cells are sensitive to the rigidity of an underlying substrate presenting activating ligands to CD3 and CD28. Specifically, primary mouse CD4+ T cells exhibited increased activation on surfaces presenting antibodies to CD3 and CD28 as material rigidity increased. While established in other cell systems, predominantly in the context of integrin-ECM and cadherin-cadherin interactions, mechanosensing by T cells through the CD3/TCR and CD28 is not well understood. Addressing this gap in knowledge would provide a new system upon which the mechanosensing concepts developed in other systems could be tested, as well as provide new insight into T cell physiology and advanced tools for immunotherapy. The proposed study seeks to address the issue that T cell-substrate interaction, like other cells, occurs in multiple stages. The ability t control the rigidity of the substrate during each stage would reveal the role each one has in mechanosensing, focusing subsequent studies into the molecular pathways responsible for this ability. Such knowledge could provide new targets for replicating or improving upon the benefits of soft substrates, but using chemical or biological agents. Towards this goal, we propose a magnetically actuated system that provides on-demand, reversible, and repeatable control over the mechanical stiffness presented to an adherent cell. We first focus on development and validation of this new system, which is an adaptation of the elastomer pillar array technology used for traction force microscopy. This system is then used to independently assess T cell mechanosensing during early cell spreading and sustained contraction. In a complementary direction, we will also assess dynamics of CD3/TCR signaling as a function of substrate rigidity, a core concept in a developing model of T cell mechanosensing. Successful completion of the proposed study will be a key advance in the emerging interdisciplinary field of T cell mechanobiology. In addition, it is expected that the proposed system will be immediately and directly applicable towards other cellular systems.

 描述：机械力在指导T细胞激活和随后的功能中起越来越多的作用。我们最近发现，T细胞对呈现给CD3和CD28的激活配体的底物的刚度很敏感。具体而言，随着材料刚度的增加，原代小鼠CD4+ T细胞暴露于表面上的激活增加。虽然在其他细胞系统中建立，但主要是在整联蛋白ECM和钙粘蛋白 - 钙粘蛋白相互作用的背景下建立的，但T细胞通过CD3/TCR和CD28的机制尚不清楚。解决这一知识的差距将提供一个新的系统，可以测试其他系统中开发的机制抑制概念，并提供有关T细胞生理学和免疫疗法的高级工具的新见解。拟议的研究旨在解决与其他细胞一样在多个阶段发生的T细胞基底相互作用的问题。控制每个阶段底物的刚性的能力将揭示每个人在机械机制中的作用，从而将随后的研究集中在负责这种能力的分子途径上。这种知识可以为软化学或生物学剂提供复制或改善的新目标，以复制或改善软化底物的好处。为了实现这一目标，我们提出了一个磁性激活的系统，该系统可对呈现给粘附电池的机械刚度进行按需，可逆和可重复的控制。我们首先关注该新系统的开发和验证，这是用于用于牵引力显微镜的弹性体支柱阵列技术的适应。然后，该系统用于在早期细胞扩散和持续收缩期间独立评估T细胞机制。在一个完整的方向上，我们还将评估CD3/TCR信号传导的动力学，这是底物刚度的函数，这是T细胞机制开发模型的核心概念。成功完成拟议的研究将是T细胞机制的新兴跨学科领域的重要进步。此外，预计所提出的系统将立即并直接适用于其他蜂窝系统。