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 细胞激活和后续功能方面发挥着越来越重要的作用。我们最近发现 T 细胞对呈现 CD3 和 CD28 激活配体的底层基质的刚性敏感。具体来说，随着材料刚性的增加，原代小鼠 CD4+ T 细胞在呈递 CD3 和 CD28 抗体的表面上表现出活化增加。虽然在其他细胞系统中建立，主要是在整合素-ECM 和钙粘蛋白-钙粘蛋白相互作用的背景下，但 T 细胞通过 CD3/TCR 和 CD28 进行的机械传感尚不清楚。解决这一知识空白将提供一个新的系统，可以在该系统上测试其他系统中开发的机械传感概念，并为 T 细胞生理学和免疫治疗的先进工具提供新的见解。 拟议的研究旨在解决 T 细胞与底物相互作用与其他细胞一样发生在多个阶段的问题。在每个阶段控制基底刚性的能力将揭示每个阶段在机械传感中的作用，将后续研究的重点放在负责这种能力的分子途径上。这些知识可以为复制或改进软基质的好处提供新的目标，但使用化学或生物制剂。为了实现这一目标，我们提出了一种磁驱动系统，该系统可以对贴壁细胞的机械刚度提供按需、可逆和可重复的控制。我们首先关注这个新系统的开发和验证，该系统是对用于牵引力显微镜的弹性体柱阵列技术的改编。然后，该系统用于独立评估早期细胞扩散和持续收缩过程中的 T 细胞机械传感。在互补的方向上，我们还将评估 CD3/TCR 信号传导作为基质刚性函数的动态，这是 T 细胞机械传感发展模型的核心概念。 拟议研究的成功完成将是 T 细胞力学生物学这一新兴跨学科领域的关键进展。此外，预计所提出的系统将立即直接应用于其他蜂窝系统。