Mechanical Regulation of Cell Adhesion by Dynamic Cytoskeletal Assemblies - Resubmission 01

 描述（由申请人提供）：细胞粘附和形态受动态细胞骨架组装体调节，动态细胞骨架组装体介导力穿过细胞并传递到周围环境。细胞力产生和粘附的时空调节驱动不同生理过程中的形态发生变化，包括细胞迁移、组织形态发生和ECM重塑。虽然已经取得了显着的进展，了解细胞粘附和力产生的分子机制，我们缺乏一个框架来了解如何从动态的细胞骨架蛋白合奏的粘附斑块和肌动蛋白细胞骨架的复杂的生物物理行为。我们假设，了解力传递内的粘附斑和肌动蛋白细胞骨架将提供必要的见解，翻译分子机制，复杂的物理行为的细胞。我们提出的实验，将阐明力传递机制，通过局灶性粘连，细胞-细胞粘连和肌动蛋白细胞骨架，以及如何协调这些调节力在多细胞组织中的传输。我们通过将分子细胞生物学方法与细胞骨架动力学和生物物理测量的先进定量成像相结合来解决这个问题。通过获得动力学和运动学（运动）签名的蛋白质在不同水平的张力，我们确定的机制内的焦点粘连和肌动蛋白细胞骨架的力传递。然后，我们与理论物理学家密切合作，用我们的定量生物物理测量来测试分析理论和模拟的预测。这项工作建立了对细胞粘附，张力和形状的物理学的定量理解，最终将为细胞迁移和组织形态发生的理论和模型提供框架，这些理论和模型将在理解复杂的生理过程中具有预测能力。通过这些目标中获得的知识，我们将确定细胞-ECM和细胞-细胞粘附之间的机械耦合在控制多细胞组织中的形态学重排中的作用。这将使得能够开发改进的疗法来治疗涉及组织稳态的疾病，这些疾病目前仍然难以通过单独治疗分子靶点来实现。

 DESCRIPTION (provided by applicant): Cell adhesion and morphology are regulated by dynamic cytoskeletal assemblies that mediate force transmission across the cell and to the surrounding environment. Spatiotemporal regulation of cellular force generation and adhesion drive morphogenic changes in diverse physiological processes including cell migration, tissue morphogenesis and ECM remodeling. While significant progress has been made to understand the molecular mechanisms of cell adhesion and force generation, we lack a framework to understand how the complex biophysical behaviors of adhesion plaques and the actin cytoskeleton emerge from dynamic ensembles of cytoskeletal proteins. We hypothesize that understanding force transmission within adhesion plaques and the actin cytoskeleton will provide the necessary insight to translate molecular mechanisms to complex physical behaviors of cells. We propose experiments that will elucidate mechanisms of force transmission through focal adhesions, cell-cell adhesions and the actin cytoskeleton and how these are coordinated to regulate force transmission in multicellular tissue. We approach this problem by integrating molecular cell biology approaches with advanced quantitative imaging of cytoskeletal dynamics and biophysical measurements. By obtaining kinetic and kinematic (motion) signatures of proteins at varying levels of tension, we identify mechanisms of force transmission within focal adhesions and the actin cytoskeleton. We then collaborate closely with theoretical physicists to test the predictions of analytical theory and simulations with our quantitative biophysical measurements. This work builds a quantitative understanding of the physics of cell adhesion, tension and shape that, ultimately, will provide the framework for theories and models of cell migration and tissue morphogenesis that will have predictive power in understanding complex physiological processes. Through knowledge gained in these aims, we will identify the role of mechanical coupling between cell-ECM and cell-cell adhesions in controlling morphological rearrangements in multi-cellular tissue. This will enable the development of improved therapies to treat diseases involved in tissue homeostasis that currently remain elusive by solely treating molecular targets.