Force Transduction Mechanisms At Adherens Junctions

机械力是细胞和组织发育的主要决定因素。在组织发育过程中，细胞经历了许多机械力，包括来自周围组织形状变化的拉伸或压缩力以及来自管状结构中流体流动的剪切力。在肺的内壁，血管和肾细胞排列在管状结构上，通过连接纤维状的内部细胞骨架直接与它们的邻居相连。细胞间的连接在细胞间传递力。细胞间的连接是肺、肾和血管衬里细胞测量流体流动或空气压力的一部分。这项研究的目的是模拟流体流动的力量是如何耦合到细胞-细胞连接的，以及它们是如何引起细胞结构的变化，使细胞正常工作的。细胞不能正确地感知和传递机械信号会导致肾脏、血管系统和肺部疾病的进展。由于缺乏测量活细胞中特定蛋白质中的力的工具，我们对力传递途径的理解受到限制。本研究将采用一种将基于FRET的力传感器与微流体相结合的新方法，这将有机会建立流动诱导的细胞骨架力与负责细胞重塑的粘附连接（AJ）动力学之间的联系。细胞骨架力的分布及其对特定蛋白质的时间依赖性将在受到流量刺激的细胞中使用FRET探针进行绘制。粘附体的连接动力学将与荧光标记的连接蛋白同时测量。这项研究的结果将是对活细胞中剪切力转导动力学的新理解，也许最重要的是，对细胞中能量流动的因果关系的测量。FRET力探针还将为科学界研究细胞力学提供一套新的研究工具。更广泛的教育影响将通过一个独特的通过实验或E2E的教育项目来实现

Mechanical forces are major determinants of cell and tissue development. During tissue development, cells experience numerous mechanical forces including tensile or compressive forces from shape change of the surrounding tissues and shear force from fluid flow in tubular structures. In the lining of the lungs, blood vessels and the kidney cells that line tubular structures adhere directly to their neighbors by junctions that join the fibrous internal cellular cytoskeleton. The cell-to-cell connections transmit forces between the cells. The intercellular connections are part of how lung, kidney and vascular lining cells can measure the forces of fluid flow or air pressure. The objective of this research is to mimic how the forces of fluid flow are coupled to the cell-cell junctions and how they cause cell structural changes in the cells that make them function properly. Failure of cells properly to sense and transmit mechanical signals can cause progression of diseases of the kidney, vasculature and lung.Our understanding of force transmission pathways has been limited due to a lack of tools to measure forces in specific proteins in live cells. This research will use a novel approach of combining FRET based force sensors with microfluidics that presents an opportunity to establish the link between flow-induced cytoskeletal forces and adherens junction (AJ) dynamics responsible for cell remodeling. The distribution of cytoskeletal force and its time-dependence for specific proteins will be mapped using FRET probes in cells subjected to flow stimuli. Adherens junction dynamics will be measured simultaneously with fluorescently labeled junction proteins. The outcome of this research will be a new understanding of the dynamics of shear force transduction in living cells, and perhaps most importantly, a measure of the cause and effect relationships of energy flow in the cell. The FRET force probes would also provide a new research toolset to the scientific community to study cell mechanics. The educational broader impacts will be accomplished through a unique program on Education through Experimentation or E2E