Deciphering mechanisms that drive collective cell migration

项目摘要 集体细胞迁移是体内最普遍的细胞迁移形式;它参与了细胞的构建。 和修复几乎所有人体组织。PI以前的工作提供了关于以下层次结构的新颖见解： 个体细胞行为和细胞内信号驱动集体迁移。这项工作确定了激活 细胞内信号PLCg 1与增加的集体迁移和细胞迁移强烈相关 坚持不懈该研究旨在阐明PLCg 1驱动集体迁移的机制 并将产生可用于控制该过程的新工具和知识。我们将实现这一目标 目标： 1)确定PLCg 1激活产生细胞骨架重塑的机制。我们将开发 基于FRET的PLCg 1报告基因，并鉴定驱动细胞骨架的PLCg 1下游效应子 重排是细胞运动的重要组成部分。 2)阐明PLCg 1激活转化为定向集体表达的生物物理机制 迁移我们将开发具有不同水平的组成性活性PLCg 1的细胞模型，并应用于 牵引力显微镜分析细胞骨架力是如何传递的， 运动 3)确定强大的集体迁移所需的PLCg 1激活的最佳模式和程度。我们 将开发一种光遗传学方法来刺激PLCg 1激活的不同时空动力学 在定向角质形成细胞迁移期间。 通过采用创新的分子和生物物理工具的组合，我们的目标是揭示机制 这些知识使我们能够控制集体上皮迁移。确定以下途径： PLCg 1激活连接到下游信号，以及其在细胞内的空间和时间动态。 集体表，提供了机会，以调节健康和病理性集体迁移， 一系列组织和病理。这些途径和线索的阐明提供了重要的基础， 以特定元素为目标以在其受损的条件下诱导集体运动（例如，慢性会 愈合）或在病态的情况下停止集体运动（例如，癌症）。

PROJECT SUMMARY Collective cell migration is the most prevalent form of cell migration in the body; it is involved in the construction and repair of almost all human tissues. Previous work by the PIs has provided novel insight on the hierarchy of individual cell behaviors and intracellular signals that drive collective migration. This work identified activation of the intracellular signal PLCg1 as strongly correlated with increased collective migration and cell migration persistence. The proposed study aims to elucidate the mechanisms by which PLCg1 drives collective migration and will yield both novel tools and knowledge that may be applied to control this process. We will achieve this goal by: 1) Determining the mechanism(s) by which PLCg1 activation yields cytoskeletal remodeling. We will develop a FRET-based PLCg1 reporter and identify the downstream effectors of PLCg1 that drive cytoskeletal rearrangement, a critical component of cell movement. 2) Elucidating the biophysical mechanisms by which PLCg1 activation translates into directed collective migration. We will develop cellular models with varying levels of constitutively active PLCg1 and apply traction force microscopy to analyze how cytoskeletal forces are transmitted to yield collective cell movement. 3) Determining optimal patterns and extent of PLCg1 activation needed for robust collective migration. We will develop an optogenetic approach to stimulate different spatio-temporal dynamics of PLCg1 activation during directed keratinocyte migration. By employing a combination of innovative molecular and biophysical tools, we aim to uncover mechanistic knowledge that allows us to gain control over collective epithelial migration. Identifying the pathway(s) by which PLCg1 activation connects to downstream signals, as well as its spatial and temporal dynamics within the collective sheet, provides opportunities to regulate both healthy and pathological collective migration across a range of tissues and pathologies. Elucidation of these pathways and cues provides an important foundation for targeting specific elements to induce collective movement in conditions where it is impaired (e.g., chronic would healing) or to halt collective movement in conditions where it is pathological (e.g., cancer).