Deciphering mechanisms that drive collective cell migration

破译驱动集体细胞迁移的机制

基本信息

批准号：
10917532
负责人：
Pamela K Kreeger
金额：
$ 32.87万
依托单位：
UNIVERSITY OF WISCONSIN-MADISON
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-09-01 至 2026-08-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10917532
关键词：
3-Dimensional Actins Biochemical Biocompatible Materials Biological Assay Biophysical Process Biophysics Breast Cancer Cell Cell model Cell physiology Cells Cellular Structures Chemicals Chronic Collaborations Coupling Cues Cytoskeleton Decision Making EGF gene Elements Environment Epidermal Growth Factor Receptor Epithelium Event Fluorescence Resonance Energy Transfer Foundations Genetic Goals Growth Factor Health Human Human Development Impairment Individual Intercellular Junctions Intervention Knowledge Link Malignant Neoplasms Methods Molecular Monitor Motion Movement PLC gamma1 Pathologic Pathology Pathway interactions Pattern Process Reporter Signal Transduction Stimulus Tissues Traction Force Microscopy Translating Travel Work biophysical tools cell behavior cell motility experimental study healing human tissue innovation insight keratinocyte migration novel optogenetics repaired spatiotemporal tissue repair tool transmission process

项目摘要

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).

项目概要集体细胞迁移是体内最普遍的细胞迁移形式；它参与了建设以及几乎所有人体组织的修复。 PI 之前的工作提供了关于层次结构的新颖见解驱动集体迁移的个体细胞行为和细胞内信号。这项工作确定了激活细胞内信号 PLCg1 与集体迁移和细胞迁移的增加密切相关坚持。拟议的研究旨在阐明 PLCg1 驱动集体迁移的机制并将产生可用于控制这一过程的新颖工具和知识。我们将实现这一目标目标通过： 1) 确定 PLCg1 激活产生细胞骨架重塑的机制。我们将开发基于 FRET 的 PLCg1 报告基因，并识别驱动细胞骨架的 PLCg1 下游效应器重排，细胞运动的关键组成部分。 2）阐明PLCg1激活转化为定向集体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 PLCg1 的细胞模型并应用牵引力显微镜分析细胞骨架力如何传递以产生集体细胞移动。 3) 确定稳健集体迁移所需的 PLCg1 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 PLCg1 激活的不同时空动态在定向角质形成细胞迁移过程中。通过结合创新的分子和生物物理工具，我们的目标是揭示机制使我们能够控制集体上皮迁移的知识。确定途径 PLCg1 激活与下游信号及其在集体表，提供了规范健康和病态集体移民的机会范围的组织和病理。阐明这些途径和线索为针对特定因素，在其受损的情况下引发集体运动（例如，慢性病愈合）或在病理情况下（例如癌症）停止集体运动。