Investigating E-cadherin Mechanotransduction

项目总结/摘要 所有的细胞都受到力的作用。这些力被细胞表面粘附受体感知，并触发鲁棒性的粘附。 肌动蛋白细胞骨架重排和相关粘附复合物的生长以对抗所施加的力。 这个过程被称为细胞硬化或增强。硬化所需肌动蛋白重新排列是 能量成本高，表明存在耦合力传递和能量产生的机制。 以前我们的实验室确定了耦合力传递和能量利用的机制。我们 证明，响应于力，AMPK在E-钙粘蛋白粘附复合物处被募集和激活， 从而刺激肌动球蛋白收缩性、葡萄糖摄取和ATP产生。葡萄糖的增加 摄取和ATP提供了生长粘附复合物和加强肌动蛋白 细胞骨架尽管取得了这一进展，机械力如何调节葡萄糖摄取和葡萄糖 代谢还没有完全了解。本研究旨在确定葡萄糖转运蛋白-1（GLUT 1）如何影响 力诱导的代谢变化和细胞硬化。在这里，我们认为GLUT 1是力敏葡萄糖 负责粘附复合物生长所必需的葡萄糖摄取的转运蛋白， 加强肌动蛋白细胞骨架。为了进一步支持这一观点，我们表明GLUT 1被招募到 细胞-细胞连接，并与E-钙粘蛋白形成复合物，以响应力。此外，我们提出证据， 抑制GLUT 1可以阻断力诱导的细胞硬化。本研究所提出的工作的第二个目标是 评估葡萄糖代谢如何与E-钙粘蛋白介导的细胞骨架重排偶联。多 糖酵解酶与丝状肌动蛋白（F-肌动蛋白）结合，如醛缩酶和磷酸果糖激酶-1。 先前的研究表明，F-肌动蛋白结合的醛缩酶在胰岛素刺激肌动蛋白时释放， 重塑我们认为，对E-钙粘蛋白施加力会导致F-肌动蛋白结合糖酵解的释放， 酶，如醛缩酶和PFK。此外，我们怀疑这些酶的胞浆释放介导了 糖酵解的增加和定位，这是力诱导能量产生所必需的。本研究提出 葡萄糖代谢与力诱导细胞能量密集过程之间的新联系 硬化

Project Summary/Abstract All cells experience force. These forces are sensed by cell surface adhesion receptors and trigger robust actin cytoskeletal rearrangements and growth of the associated adhesion complex to counter the applied forces. This process is known as cell stiffening or reinforcement. The actin re-arrangements necessary for stiffening are energetically costly suggesting that mechanisms coupling force transduction and energy production exist. Previously our laboratory identified a mechanism for coupling force transmission and energy utilization. We demonstrated that, in response to force, AMPK is recruited and activated at the E-cadherin adhesion complexes, thereby stimulating actomyosin contractility, glucose uptake, and ATP production. This increase in glucose uptake and ATP provides the energy necessary to grow the adhesion complexes and reinforce the actin cytoskeleton. Despite this advancement, how mechanical force modulates glucose uptake and glucose metabolism is not fully understood. This study aims to determine how glucose transporter-1 (GLUT1) affects force-induced metabolic changes and cell stiffening. Here we suggest that GLUT1 is the force-sensitive glucose transporter responsible for the glucose uptake necessary for the growth of adhesion complexes and reinforcement of the actin cytoskeleton. In further support of this notion, we show that GLUT1 is recruited to the cell-cell junctions and forms a complex with E-cadherin in response to force. Furthermore, we present evidence that inhibition of GLUT1 blocks force-induced cell stiffening. A second goal of the proposed work in this study is to assess how glucose metabolism is coupled to E-cadherin mediated cytoskeleton rearrangements. Multiple glycolytic enzymes are bound to filamentous actin (F-actin), such as aldolase and phosphofructokinase-1. Previous studies have demonstrated that F-actin bound aldolase is released upon insulin stimulated actin remodeling. We propose that the application of force to E-cadherin causes the release of F-actin-bound glycolytic enzymes, such as aldolase and PFK. Additionally, we suspect that cytosolic release of these enzymes mediates the increase and localization of glycolysis, necessary for force-induced energy production. This study proposes a novel connection between glucose metabolism and the energy-intensive process of force-induced cell stiffening.