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

项目总结/摘要 PI的实验室在分子张力水平上研究细胞机械生物学。力是生命的基本要素 细胞细胞-基质界面处的整合素传递的细胞力机械地驱动短期细胞内的细胞粘附。 细胞粘附、收缩和迁移等功能。从长远来看，细胞力也被转换为 由于其具有生物化学信号以调节细胞增殖、分化、癌细胞转移等， 最重要的是，整合素传递的细胞力已被广泛研究，在散装水平的 发育良好的细胞牵引力显微镜。然而，整合素张力，由个体传递的力， 整合素分子，尽管其在整合素信号传导中的基本作用，在大多数细胞中了解得少得多， 功能协调发展的PI的实验室专门研究细胞机械生物学，使用创新的分子张力工具， 测量、绘制和操纵活细胞中的整合素张力。ITS张力传感器是一种新型的张力传感器， PI将整合素张力转化为荧光信号，并直接通过荧光进行整合素张力映射 高空间分辨率的图像。开发了一种张力调节器TGT， 整联蛋白张力在整个细胞在一个设计的水平下，用于研究整联蛋白张力的调节作用， 细胞功能。有了这些工具，PI的实验室开始研究迁移细胞，血小板， 和癌细胞的微型侵入伪足。初步实验证明了该方法的可行性， 这些整合素张力工具在分子张力水平的细胞机械生物学研究中的通用性。 有了这些创新的张力工具，难以捉摸的整合素张力现在可以可视化，量化和控制 沿着活细胞中的细胞结构和生化因子，具有可比的分辨率、灵敏度和精确度。 在未来的五年里，PI的实验室将研究整合素张力，细胞凋亡和细胞凋亡之间的相关性，共定位和因果关系。 结构和局部生化活性。MIRA基金为PI的实验室提供了灵活性， 继续研究三种不同细胞过程中的整合素张力。PI将联合收割机分子张力 工具，遗传学方法和荧光成像，以调查范围，来源，分布和功能， 整合素张力迁移细胞，并揭示细胞如何发挥和控制整合素张力局部协调 细胞突起和收缩。对于血小板的研究，整合素张力的力源和生物学功能， 将在2D和3D环境中研究血小板粘附、收缩和聚集。整联蛋白 还将测试张力图作为诊断测定，用于评估血小板活性以阻止出血。 对于侵袭性伪足的研究，整合素张力在侵袭性伪足形成和基质降解中的作用， 将全面研究癌细胞的侵袭。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 创新的分子张力传感器和张力调制器，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.