Kinase Control of Synergistic Cell Migration Mechanics

项目摘要 细胞迁移是发育所必需的基本细胞过程， 癌症转移我们的长期目标是阐明控制迁移和癌症侵袭的信号， 减少病理性迁移和癌症转移的治疗策略可以得到改进。波动 在细胞迁移力控制前沿升降周期，但我们不知道是什么控制 力的波动。这里的总体目标是了解控制和整合的信号机制 细胞迁移的波动分子力信号通路可以通过引导空间定位的和 肌动蛋白，粘附和膜张力参数的协调波动（指导性）。可选择地， 信号传导途径可以指示某些过程，而在其他过程中没有时空精确性（允许的）。 我们将阐明细胞迁移的控制机制，通过解剖的时间和空间的调节， 蛋白激酶ERK及其在未转化细胞和癌细胞中的信号输出。ERK作用于多个步骤， 伸缩循环疾病相关的癌细胞模拟高活性状态，其中ERK 活性由于致癌突变而上调。我们的中心假设是ERK在空间上指导- 肌动蛋白装配、粘附寿命和边缘运动的膜张力的局部协同波动 和细胞迁移。对于第一个目标，我们将测量ERK活性在边缘过程中的时间波动， 使用修饰的ERK生物传感器进行突出和缩回。我们将结合实验观察到的 活动波动到计算模型和实验测试，以确定哪些模式决定 突出速度和持久性。对于第二个目标，我们将确定是否空间组织ERK活性 控制边运动。我们将测试膜和粘附激活的ERK诱导突出的能力。 实验上和计算上。我们还将测试ERK在细胞膜中的保留模式， 粘附域有助于突出能力和宽度。对于第三个目标，我们将测试ERK是否为对照 膜张力和粘附寿命对突出速度的影响。我们将通过Zyxin和Ezrin测试信号 肌动蛋白作为ERK控制这些额外的分子力的可能机制。拟议 这项研究在概念上是创新的，因为它测试了波动的ERK信号在细胞调节中的作用。 迁移它在新的光遗传学工具的开发和使用方面具有技术创新性， 模型这项研究意义重大，因为它有可能揭示一个新的原理， 力被整合以产生运动。它还将识别控制局部ERK活性的支架和信号 这些波动可以适用于新的治疗策略，以控制细胞粘附和迁移。

Project Abstract Cell migration is a fundamental cellular process necessary for development and coopted in diseases like cancer metastasis. Our long-term goal is to elucidate the signals that control migration and cancer invasion, so that treatment strategies to reduce pathological migration and cancer metastasis can be improved. Fluctuations in cell migration forces control leading edge protrusion-retraction cycles, but we do not know what controls the force fluctuations. The overall objective here is to understand the signaling mechanisms that control and integrate the fluctuating molecular forces of cell migration. Signaling pathways can act by directing spatially-localized and coordinated fluctuations in actin, adhesion, and membrane tension paramters (instructive). Alternatively, signaling pathways may instruct some processes and act without spatiotemporal precision (permissive) in others. We will elucidate the cell migration control mechanisms by dissecting the temporal and spatial regulation of the protein kinase ERK and its signaling outputs in untransformed and cancer cells. ERK acts on multiple steps in the protrusion-retraction cycle. The disease-relevant cancer cells model a high-activity state, in which ERK activity is upregulated due to onocogenic mutations. Our central hypothesis is that ERK instructs spatially- localized synergistic fluctuations in actin assembly, adhesion lifetime, and membrane tension for edge motion and cell migration. For the first aim, we will measure the temporal fluctuations in ERK activity during edge protrusion and retraction using modified ERK biosensors. We will incorporate the experimentally-observed activity fluctuations into a computational model and experimental tests to determine which patterns dictate protrusion velocity and persistence. For the second aim, we will determine if spatiatially-organized ERK activity controls edge motion. We will test membrane and adhesion-activated ERK for the ability to induce protrusion experimentally and computationally. We will also test how the pattern of ERK retention in the membrane and adhesion domains contributes to protrusion power and width. For the third aim, we will test if ERK is controls membrane tension and adhesion lifetime for protrusion velocity. We will test signaling through Zyxin and Ezrin to actin as possible mechanisms by which ERK controls these additional for molecular forces. The proposed research is conceptually innovative because it tests the role of fluctuating ERK signals in the regulation of cell migration. It is technically innovative in the development and use of new optogenetics tools and computational models. The research is significant because it has the potential to reveal a new principle about how molecular forces are integrated to bring about motion. It will also identify scaffolds and signals that control local ERK activity fluctuations that could be adapted for new therapeutic strategies to control cell adhesion and migration.