Maintenance of Adhesion and Barrier Function during Epithelial Cell Shape Changes

上皮细胞形状变化期间粘附和屏障功能的维持

基本信息

批准号：
10470721
负责人：
Ann Louise Miller
金额：
$ 31.34万
依托单位：
UNIVERSITY OF MICHIGAN AT ANN ARBOR
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-08-01 至 2024-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10470721
关键词：
Actins Actomyosin Adherens Junction Adhesions African Apical Biological Biological Assay Biological Models Calcium Cell Adhesion Cell Maintenance Cell Shape Cell division Cell-Cell Adhesion Cells Cellular biology Cytokinesis Cytoskeletal Proteins Cytoskeleton Data Development Devices Disease Embryo Ensure Epithelial Epithelial Cells Event Flare Goals Heterotrimeric GTP-Binding Proteins Homeostasis Image Inflammatory Inflammatory Bowel Diseases Intercellular Junctions Intestines Knowledge Literature Maintenance Mechanics Mediating Molecular Morphogenesis Neoplasm Metastasis Organ Pathway interactions Permeability Physiological Piezo 1 ion channel Property Proteins Psychological reinforcement Public Health Regulation Research Shapes Signal Transduction Signaling Protein Site Stretching Testing Tight Junctions Tissues Vinculin Xenopus Xenopus laevis anillin cancer cell chemokine innovation insight macromolecule mechanical force mechanotransduction optogenetics recruit repaired response rho seal success tool transmission process

项目摘要

PROJECT SUMMARY / ABSTRACT Cell-cell junctions adhere epithelial cells to one another, transmit forces from cell to cell, and generate biological barriers that selectively regulate what can pass between cells in an epithelial tissue. Fundamental questions about how epithelial cell-cell junctions dynamically remodel in response to physiological forces that that challenge cell adhesion and barrier function remain unanswered. In addition to being absolutely essential for development and maintenance of organ homeostasis, disruption of adhesion and barrier function contributes to diseases including cancer cell metastasis and Inflammatory Bowel Disease. Therefore, it is critical to determine the mechanisms that control cell-cell junction remodeling as epithelial cells change shape. This proposal builds on recent discoveries from the lab showing that Rho flares locally reinforce tight junctions following leaks in barrier function, and that adherens junctions are reinforced by recruitment of Vinculin to the cleavage furrow of dividing epithelial cells. The overall objective of this application is to identify mechanisms that promote maintenance of adhesion and barrier function at sites of epithelial cell division and junction elongation. Our central hypothesis is that locally applied mechanical forces challenge adherens junctions and tight junctions and elicit actomyosin-mediated reinforcement required for maintenance of adhesion and barrier function at these sites. The central hypothesis will be tested by pursuing three specific aims: 1) Identify how mechanically-induced tight junction leaks trigger Rho flares; 2) Determine how Rho flare-mediated junction contractility repairs tight junctions; 3) Define mechanisms that mediate tension transmission and barrier maintenance at sites of locally increased tension. The proposed research is innovative because it applies powerful experimental tools including: a developing vertebrate model system (Xenopus laevis embryos), a live imaging barrier assay recently developed in the lab, proven approaches to locally or globally manipulate tension in the intact epithelium, probes for live imaging of active Rho dynamics as well as a host of cytoskeletal proteins, junction proteins, Rho regulators, and cytoplasmic calcium, and specialized analysis tools to quantitatively analyze live imaging data. The proposed research is significant because it will advance our knowledge about a fundamentally important problem in epithelial cell biology: how epithelial cells undergo dramatic cell shape changes like cytokinesis yet maintain tissue integrity and barrier function.

项目摘要 /摘要细胞 - 细胞连接将上皮细胞彼此粘附，将力从细胞传播到细胞，并产生生物屏障有选择地调节上皮组织中的细胞之间可以传递的东西。基本的关于上皮细胞 - 细胞连接如何对生理力进行动态重塑的问题这种挑战细胞粘附和屏障功能仍未得到解答。除了绝对必要为了开发和维护器官稳态，粘附和障碍功能的破坏导致包括癌细胞转移和炎症性肠病在内的疾病。因此，是对于确定在上皮细胞变化时控制细胞 - 细胞连接重塑的机制至关重要。该提议建立在实验室的最新发现的基础上，表明Rho在当地加强了紧密的连接在屏障功能泄漏之后，通过募集Vinculin招募到粘附器连接处分裂上皮细胞的裂解沟。该应用的总体目的是确定机制促进上皮细胞分裂部位的粘附和屏障功能的维持伸长。我们的中心假设是，本地应用机械力挑战了粘附连接和紧密连接和引起肌动蛋白介导的粘附和屏障所需的加固在这些站点的功能。中心假设将通过追求三个具体目标来检验：1）确定如何机械诱导的紧密连接泄漏触发了Rho Flares； 2）确定Rho Flare介导的连接收缩性维修紧密的连接； 3）定义介导张力传递和障碍的机制在局部张力增加的部位维护。拟议的研究具有创新性，因为它适用强大的实验工具，包括：开发的脊椎动物模型系统（Xenopus laevis胚胎），现场成像屏障测定最近在实验室中开发的，可验证的局部或全球操纵方法完整上皮的张力，活性Rho动力学的实时成像以及大量细胞骨架的探针蛋白质，连接蛋白，RHO调节剂和细胞质钙以及专门的分析工具定量分析实时成像数据。拟议的研究很重要，因为它将推动我们的关于上皮细胞生物学中根本重要问题的知识：上皮细胞如何经历戏剧性的细胞形状发生变化，例如细胞因子，但保持组织完整性和屏障功能。