CAREER: Force-activated Protein Dynamics in Mechanobiology

该学院早期职业发展（CAREER）计划拨款将开创一种新方法，用于了解机械力对活细胞中特定蛋白质动力学的影响。疾病通常是通过生化手段来理解和治疗的;当我们生病时，我们通常会得到一片药。然而，除了生化信号，细胞还受到各种各样的机械信号的影响。在活组织内，细胞存在于复杂的机械环境中，该环境既是施加力的来源，也是机械支持的手段。细胞周围机械环境的改变是许多基本过程的有效调节剂，包括细胞如何生长，迁移和发育成不同的组织类型。目前，由于无法在活细胞分子水平上研究机械力和生化信号通路之间的相互作用，因此了解细胞如何感知、解释和响应机械信号受到限制。更好地理解这种关系将有助于开发癌症和动脉粥样硬化等疾病的治疗方法，其中机械环境被改变，以及为再生医学设计替代组织的努力。对细胞用于检测机械信号的过程（称为机械转导）的不完全理解阻碍了不同领域的进步，从形态发生过程中细胞集体运动的基础研究到通过引入相关的机械刺激来改进组织工程方法的应用研究。已知将力产生细胞骨架机械连接到细胞外环境的亚细胞结构（称为粘着斑）对机械力天生敏感并且在机械力转导中至关重要。决定这些力激活动力学的关键分子过程是黏着斑蛋白的机械加载，黏着斑蛋白是在粘着斑中发现的衔接蛋白。这项研究将创建和利用最先进的分子工具和实验方法，能够阐明介导活细胞和组织中机械转导的分子尺度机械和生物化学过程的相互依赖性和动力学。新的工具包括基因编码的分子张力传感器，具有合理设计的生物物理特性和定义的生化功能。此外，一个简单的，但新颖的，直接测量力激活蛋白质动力学的实验程序将被开发。这些将被结合起来测试的假设，不同形式的机械刺激检测和整合的细胞通过改变力激活动力的黏着斑蛋白。

This Faculty Early Career Development (CAREER) Program grant will pioneer a new approach for understanding the effects of mechanical force on the dynamics of specific proteins in living cells. Disease is generally understood and treated through biochemical means; when we get sick, we typically get a pill. However, in addition to biochemical signals, cells are subject to a wide variety of mechanical cues. Inside living tissues, cells exist in a complex mechanical environment that is both a source of applied forces and a means of mechanical support. Alterations in the mechanical environment surrounding cells are potent regulators of many fundamental processes, including how cells grow, migrate, and develop into different tissue types. Understanding how cells sense, interpret, and respond to mechanical cues is currently limited by the inability to study the interplay between mechanical forces and biochemical signaling pathways at the molecular level in living cells. A greater understanding of this relationship will aid endeavors to develop therapies for diseases like cancer and atherosclerosis, where the mechanical environment is altered, as well as efforts to engineer replacement tissues for regenerative medicine.An incomplete understanding of the processes cells use to detect mechanical cues, referred to as mechanotransduction, is preventing advancements in diverse fields, ranging from fundamental studies of the collective movements of cells during morphogenesis to applied research geared toward improving tissue engineering approaches by incorporating relevant mechanical stimuli. The sub-cellular structures that mechanically link the force-generating cytoskeleton to the extracellular environment, termed focal adhesions, are known to be innately sensitive to mechanical force and critically important in mechanotransduction. A key molecular process dictating these force-activated dynamics is the mechanical loading of vinculin, an adaptor protein found in focal adhesions. This research will create and utilize state of the art molecular tools and experimental approaches capable of elucidating the interdependence and dynamics of the molecular-scale mechanical and biochemical processes mediating mechanotransduction in living cells and tissues. Novel tools include genetically-encoded molecular tension sensors with rationally-designed biophysical properties and defined biochemical functions. Additionally, a simple, but novel, experimental procedure for directly measuring force-activated protein dynamics will be developed. These will be combined to test the hypothesis that distinct forms of mechanical stimulation are detected and integrated by the cell through alterations in the force-activated dynamics of vinculin.