Mechanical Regulation of Cell Adhesion by Dynamic Cytoskeletal Assemblies

项目摘要 动态细胞骨架组装对细胞黏附的机械调节 上皮组织是由连接相邻细胞的动态粘连形成的，即细胞-细胞连接 在保持组织凝聚力和屏障功能的同时，还允许像创伤这样的动态过程 愈合和组织形态发生。肌动球蛋白内产生的收缩力 细胞骨架被传输到细胞-细胞连接，以控制局部细胞的形状和运动， 塑造组织形态发生，启动控制细胞命运的下游信号通路。 了解细胞-细胞连接的生物物理特性是如何调节的，这一点已经得到了广泛的理解 理解和治疗胚胎发育过程中的缺陷对组织的影响 工程学与转移性肿瘤的诊断和治疗。这项提议充分利用了 细胞生物物理学、分子细胞生物学、活细胞成像、数学 建模和光遗传学研究RhoA信号如何调节驱动 细胞-细胞连接长度的变化，控制细胞的形状，最终控制组织的形态发生。 我们建议用实验来阐明力依赖的过程如何调节肌动球蛋白 收缩、膜重塑和RhoA信号相互反馈控制连接 长度和长度会发生变化。我们通过整合分子细胞生物学来解决这个问题。 细胞骨架动力学和生物物理的先进定量成像方法 测量。通过获得蛋白质在不同条件下的运动学和运动学(运动)特征 张力水平，我们确定了局灶粘连内的力传递机制和 肌动蛋白细胞骨架。然后，我们与理论物理学家密切合作，以检验这些预测 分析理论和模拟与我们的定量生物物理测量。这部作品 建立了对细胞黏附、张力和形状的生物物理理解，最终将 为组织形态发生的理论和模型提供框架，这些理论和模型将具有预测性 在复杂的生理过程中理解的能力。更广泛地说，这些战略 可以更普遍地应用于理解力敏感 细胞骨架内的反馈共同促进了细胞的形态发生过程。这将是 能够开发改进的治疗方法来治疗涉及组织动态平衡的疾病 目前，仅通过治疗分子靶标仍难以实现这一点。

Project Summary Mechanical Regulation of Cell Adhesion by Dynamic Cytoskeletal Assemblies Epithelial tissue is built by dynamic adhesions, cell-cell junctions, that connect neighboring cells to maintain tissue cohesion and barrier function yet also allow dynamic processes like wound healing and tissue morphogenesis. Contractile forces generated within the actomyosin cytoskeleton are transmitted to cell-cell junctions to control the local cell shape and motions that sculpt tissue morphogenesis and initiate downstream signaling pathways that control cell fate. Understanding how the biophysical properties of cell-cell junctions are regulated has widespread implications for understanding and treating defects during embryonic development, for tissue engineering and the diagnosis and treatment of metastatic tumors. This proposal leverages innovative combination of cell biophysics, molecular cell biology, live cell imaging, mathematical modeling and optogenetics to investigate how RhoA signals regulate contractile forces to drive changes in cell-cell junction length that control cell shape and, ultimately, tissue morphogenesis. We propose experiments to elucidate how force-dependent process regulating actomyosin contractility, membrane remodeling and RhoA signaling feedback to each other to control junction length and length changes. 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 biophysical understanding of cell adhesion, tension and shape that, ultimately, will provide the framework for theories and models of tissue morphogenesis that will have predictive power in understanding in complex physiological processes. More generally, the strategies developed in this proposal can be applied more generally to understand how force-sensitive feedbacks within the cytoskeletal conspire to facilitate cell morphogenic processes. This will enable the development of improved therapies to treat diseases involved in tissue homeostasis that currently remain elusive by solely treating molecular targets.