Orchestration of adhesion signalling by the mechanosensors talin and vinculin.

细胞持续感知并产生其周围环境，该环境由细胞可以附着的纤维状材料组成，称为细胞外基质（ECM）。细胞-ECM通信在需要对变化的环境做出特定细胞响应的发育或再生过程中特别重要。细胞反应包括运动行为（例如伤口闭合）、收缩性（例如心血管系统的功能）的变化，但也包括为了形成新的功能组织而对其ECM的主动重塑。细胞如何感知外界环境是目前研究的热点，但对于细胞如何感知外界环境信号，以及这些信号如何在细胞内转化为信号，从而促进特定的细胞反应的机制，我们还远未了解。细胞在发育、正常衰老、损伤和某些疾病中所处的环境会发生巨大的变化。例如，ECM的机械性质被认为影响肿瘤进展，并且增加的乳腺基质硬度与较差的存活率相关。ECM硬化也会导致心血管功能障碍。有趣的是，细胞一方面有助于特定基质的产生，另一方面也对这种产生的环境做出反应。因此，了解细胞如何感知和产生ECM环境是至关重要的，如果我们想更接近治疗疾病的根源并促进再生，细胞可以通过对其施加力并探测其变形来感受或感知其环境。为了传递力，它们使用称为整合素的表面蛋白质“抓住”邻近的结构。这些整合素不仅与细胞的环境结合，而且还连接到细胞内的骨架。这种联系不是直接的，而是由将两者耦合或解耦的组件调节的。我们发表了大量的手稿，表明其中两种耦合蛋白，称为talin和vinculin，是感知环境变化的核心。它们对于测量其环境的硬度特别重要，它们控制细胞迁移以及细胞生长和分化。在这项提案中，我们还提出了重要的试点数据表明，黏着斑蛋白是ECM重塑的关键。然而，他们如何做到这一点仍然不清楚。为了研究这些蛋白质如何调节对环境的反应，以及它们在多大程度上参与告诉细胞如何行为，曼彻斯特大学着名的细胞基质中心的两个实验室将合作并将其长期的专业知识与整合素信号传导和细胞基质相互作用领域相结合。这项研究的目的是（i）了解机械信号在talin和vinculin激活中的作用，（ii）这种激活如何帮助黏着斑蛋白和talin与大量其他蛋白质结合，以发挥特定的信号（例如细胞迁移或细胞生长）（iii）黏着斑蛋白与新发现的另一种称为张力蛋白的蛋白质的结合如何有助于ECM环境的形成和重塑。为了实现我们的目标，我们不仅将使用尖端的显微镜，生物化学和分子生物学技术，而且还将使用新生成的细胞内系统，使我们能够将蛋白质靶向细胞中的特定隔室（线粒体），从而使我们能够在规定的条件下可视化和探测分子相互作用和行为。我们的研究结果将被结合到一个模型中，该模型概述并可能预测细胞如何解释和重塑其环境。最终，所获得的知识可能会导致我们目前如何看待环境变化及其对疾病的影响的重要变化。这也可能导致患者治疗的变化，因此，例如，它可能有助于改善疾病预防和再生过程。

Cells continuously sense and produce their surrounding environment, which consists of fibrillar material the cells can attach to and is called extracellular matrix (ECM). Cell-ECM communication is particularly important during development or regeneration processes that require specific cellular responses to changing environments. Cellular responses comprise changes in motile behaviour (e.g. closing of wounds), contractility (e.g. functioning of the cardiovascular system) but also active remodelling of their ECM for the purpose of formation of new functional tissue. Many studies have focused on how cells sense their environment, but we are still far from understanding the mechanisms how cells perceive environmental signals and how they are translated into signals within cells that promote specific cellular responses.The environment of cells alters enormously during development, normal ageing, injury and certain diseases. For example, the mechanical properties of the ECM is thought to influence tumour progression and increased breast matrix stiffness is associated with poor survival. Stiffening of ECM also causes cardiovascular malfunctioning. Intriguingly cells contribute to the production of specific matrix on one hand but also respond to this produced environment on the other hand. Therefore, understanding how cells sense and produce their ECM environment is critically important if we want to get a step closer to treating the roots of diseases and promote regeneration.Cells can feel or sense their environment by exerting forces on it and probing its deformation. To transmit forces, they 'grab' neighbouring structures using surface proteins, which are called integrins. These integrins not only bind to the environment of the cells but also connect to a skeleton inside the cells. This link is not direct but is regulated by components that couple or uncouple the two. We published a number of manuscripts showing that two of these coupling proteins, called talin and vinculin, are central to sensing of environmental changes. They are particularly important to measure the stiffness of their environment, they control cell migration, as well as cell growth and differentiation. In this proposal we also present important pilot data demonstrating that vinculin is critical for ECM remodelling. However how they do this is still unclear. In order to investigate how these proteins regulate the response to their environment, and to what extent they are involved in telling cells how to behave, the two laboratories in the prestigious Cell-Matrix Centre at the University of Manchester will team up and combine their long-standing expertise with the field of integrins signalling and cell-matrix interactions. The proposed research aims to to (i) understand the role of mechanical signals in the activation of talin and vinculin, (ii) how this activation helps vinculin and talin to associate with a large number other proteins that serve to exert specific signals (e.g. cell migration or cell growth) (iii) how vinculin with the newly found association of another protein called tensin is contributing to the formation and remodelling of ECM environment. To reach our goals, we will not only use cutting edge microscopy, biochemisty and molecular biology techniques but also a newly generated intracellular system whereby we can target proteins to specific compartments (mitochondria) in the cells which enable us to visualise and probe molecular interactions and behaviour under defined conditions. Our results will be combined into a model that outlines and potentially predicts how cells interpret and remodel their environment. Ultimately, the knowledge gained may lead to important changes in how we currently envisage environmental changes and their contribution to diseases. This may also lead to changes in treatment of patients, and it might thus, for example, contribute to improvements in disease prevention and in regeneration processes.