Study the role of integrin tension in cell migration, platelet contraction and invadopodium dynamics

项目概要/摘要 PI 的实验室在分子张力水平上研究细胞力学生物学。力量对于生命来说是必不可少的 细胞。细胞-基质界面处整合素传递的细胞力机械地驱动短期细胞 细胞粘附、收缩和迁移等功能。从长远来看，细胞力也会转化为 调节细胞增殖、分化、癌细胞转移等的生化信号。 至关重要的是，整合素传递的细胞力已被广泛研究在整体水平上 发达的细胞牵引力显微镜。然而，整合素张力，即个体传递的力 整合素分子尽管在整合素信号传导中发挥着重要作用，但在大多数细胞中对其了解甚少。 功能。 PI 的实验室专门使用创新的分子张力工具研究细胞力学生物学， 测量、绘制和操纵活细胞中的整合素张力。一种名为ITS的张力传感器是由 PI 将整合素张力转换为荧光信号，并通过荧光直接进行整合素张力图谱 高空间分辨率成像。开发了一种名为 TGT 的张力调节器来定量限制 设计水平下全细胞的整合素张力，用于研究整合素张力在特定细胞中的调节作用 细胞功能。借助这些工具，PI 的实验室开始了迁移细胞、血小板中整合素张力的研究 和癌细胞的微型侵袭伪足。初步实验证明了可行性和 这些整合素张力工具在分子张力水平的细胞力学生物学研究中具有多功能性。 借助这些创新的张力工具，现在可以可视化、量化和控制难以捉摸的整合素张力 以及活细胞中的细胞结构和生化因子，具有相当的分辨率、灵敏度和精度。 未来五年，PI实验室将研究整合素张力、细胞因子之间的相关性、共定位和因果关系。 活细胞的结构和局部生化活性。 MIRA 基金为 PI 实验室提供了灵活性 继续研究三个不同细胞过程中的整合素张力。 PI将结合分子张力 工具、遗传方法和荧光成像来研究其范围、来源、分布和功能 迁移细胞中的整合素张力，并揭示细胞如何局部发挥和控制整合素张力以协调 细胞突出和缩回。对于血小板研究，整合素张力的力量来源和生物学功能 将在 2D 和 3D 背景下研究血小板粘附、收缩和聚集。整合素 张力图也将作为诊断测定进行测试，以评估止血中的血小板活性。 对于入侵足研究，整合素张力在入侵足形成和基质降解中的作用 癌细胞的侵袭将得到全面研究。拟议的研究将揭示有价值的见解 整合素张力在这些不同的细胞过程中的基本作用。该项目还将提供 领域具有创新的分子张力传感器和张力调节器，预计将广泛用于 细胞力研究具有前所未有的分辨率和灵敏度。

Project Summary/Abstract The PI’s laboratory studies cell mechanobiology at the molecular tension level. Force is essential for the life of cells. Integrin-transmitted cellular force at the cell-matrix interface mechanically drives short-term cellular functions such as cell adhesion, contraction and migration. In long term, cellular force is also transduced to biochemical signals to regulate cell proliferation, differentiation, cancer cell metastasis, etc. Because of its fundamental importance, integrin-transmitted cellular force has been extensively studied at the bulk level by the well-developed cell traction force microscopies. However, integrin tension, the force transmitted by individual integrin molecules, despite its fundamental role in integrin signaling, is much less understood in most cellular functions. The PI’s lab specializes in studying cell mechanobiology using innovative molecular tension tools that measure, map and manipulate integrin tension in live cells. A tension sensor named ITS was developed by the PI to convert integrin tension to fluorescent signal and enable integrin tension mapping directly by fluorescence imaging with high spatial resolution. A tension modulator named TGT was developed to quantitatively restrict integrin tension in whole cells under a designed level for the study of regulative role of integrin tension in specific cellular functions. With these tools, the PI’s lab initiated the study of integrin tension in migrating cells, platelets and micro-sized invadopodia of cancer cells. The preliminary experiments have demonstrated the feasibility and versatility of these integrin tension tools in the research of cell mechanobiology at the molecular tension level. With these innovative tension tools, the elusive integrin tension can now be visualized, quantified and controlled along with cell structure and biochemical factors in live cells with comparable resolution, sensitivity and precision. In next five years, the PI’s lab will study the correlation, co-localization and causality among integrin tension, cell structure and local biochemical activities in live cells. The MIRA fund provides the PI’s lab with the flexibility to continue the study of integrin tension in three distinct cellular processes. The PI will combine molecular tension tools, genetic methods and fluorescence imaging to investigate the range, source, distribution and function of integrin tension in migrating cells, and reveal how cells exert and control integrin tension locally to coordinate cell protrusion and retraction. For platelet study, the force source and biological function of integrin tension during platelet adhesion, contraction and aggregation will be investigated in both 2D and 3D contexts. The integrin tension map will also be tested as a diagnostic assay for the assessment of platelet activity in stemming bleeding. For invadopodium study, the role of integrin tension in invadopodium formation and matrix degradation during cancer cell invasion will be comprehensively investigated. The proposed research will reveal valuable insights to the fundamental role of integrin tension in these various cellular processes. This project will also provide the field with innovative molecular tension sensor and tension modulator that are expected to be widely useful for cellular force study with unprecedented resolution and sensitivity.