Physical Signaling Mechanisms That Regulate Intestinal Architecture

调节肠道结构的物理信号机制

基本信息

批准号：
9885833
负责人：
MICHAEL A PACK
金额：
$ 41.69万
依托单位：
UNIVERSITY OF PENNSYLVANIA
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-05-19 至 2023-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9885833
关键词：
Animal Model Animals Architecture Biological Birth Cell membrane Cells Colorectal Cancer Digestive System Disorders Disease Engineering Enteric Nervous System Epithelial Epithelial Cells Epithelium Esophageal Atresia Esophagus Esophagus motility Familial colorectal cancer Family Gastrointestinal Motility Gastrointestinal Neoplasms Gastrointestinal tract structure Genes Germ-Line Mutation Goals Heritability Heterozygote Homozygote Human In Vitro Injury Intestines Knock-in Laboratories Lesion Life Link MYH11 gene Malignant Neoplasms Mammalian Cell Mediating Membrane Modeling Mus Muscle Contraction Mutation Myosin ATPase Myosin Heavy Chains Oncogenic Organ Oxidation-Reduction Oxidative Stress Pathologic Pathway interactions Phenotype Physiology Play Process Proteins Regulation Risk Factors Role Signal Pathway Signal Transduction Small Intestines Smooth Muscle Smooth Muscle Myosins Stomach Stress Syndrome Tissues Translations Variant WNT Signaling Pathway Work Zebrafish cancer cell cell behavior cell motility clinically relevant design experimental study in vivo intestinal epithelium motility disorder mouse model mutant nervous system development novel pup repaired response stressor tissue regeneration tumor progression

项目摘要

PROJECT SUMMARY Physical signals are increasingly recognized as playing an important role in modulating cell behavior. The goal of this proposal is to characterize the cellular response to force-mediated signaling in the intestine and esophagus using zebrafish and mouse models. We have shown that an activating mutation in smooth muscle myosin heavy chain gene myh11 disrupts intestinal architecture in zebrafish meltdown mutants. Physical signals arising from the mutant myosin activate a conserved redox signaling pathway in the intestinal epithelium that drives the formation of plasma membrane protrusions known as invadopodia that degrade matrix proteins. The invadopodia drive the invasive transformation and cystic expansion of the epithelium. Animals that are heterozygous for the meltdown mutation develop normally but are sensitized to form the homozygous cell invasion phenotype when oncogenic signaling pathways activated. The proposal consists of three aims designed to understand how physical signals from unregulated myosin are processed by digestive epithelia and how they may be risk factors for digestive disease. The goal of the first aim are to understand how the mutant smooth muscle myosin initiates invadopodia in the epithelium of meltdown mutants and to compare this to mechanisms that regulate invadopodia formation in mammalian cells. Invadopodia have rarely if ever been observed in vivo, thus this aim offers the opportunity to understand their regulation in a live animal model. The goal of the second aim is to understand how co- activation of KRas and Wnt signaling sensitize heterozygous mutants to invasive triggers. The experiments proposed for this aim have both basic and clinical relevance, as both pathways are activated in human digestive cancers. The goal of the third aim is to characterize a recently engineered mouse model of the meltdown mutation. This includes characterization of esophageal and intestinal phenotypes in homozygous mutants, and comparison with mice newly engineered to carry knock-in mutations that are identical to human MYH11 mutations associated with heritable motility syndromes. Collectively, the proposed experiments will define novel factors and signaling mechanisms that establish and maintain digestive organ architecture and function.

项目摘要物理信号越来越被认为是调节细胞行为的重要作用。这该建议的目标是表征肠中强制介导的信号传导的细胞反应，食管使用斑马鱼和小鼠模型。我们已经表明，平滑的激活突变肌肉肌球蛋白重链基因MYH11破坏斑马鱼熔化突变体中的肠结构。突变肌球蛋白引起的物理信号激活了保守的氧化还原信号通路肠上皮驱动质膜突起的形成，称为Invadopodia 降解基质蛋白。 Invadopia驱动了侵入性转化和囊性扩张上皮。对于熔融突变而杂合的动物正常发展，但对激活致癌信号通路时形成纯合细胞侵袭表型。提案由三个目的组成，旨在了解如何处理不受管制的肌球蛋白的物理信号通过消化性上皮以及它们如何成为消化疾病的危险因素。第一个目的的目的是了解突变平滑肌肌球蛋白如何启动Invadopodia 熔化突变体的上皮，并将其与调节Invadodia形成的机制进行比较在哺乳动物细胞中。如果在体内观察到Invadopia，很少有人观察到，因此此目的提供了有机会在现场动物模型中了解他们的调节。第二个目标的目标是了解KRAS和WNT信号的共同激活如何使杂合突变体敏感到侵入性触发器。为此目标提出的实验既具有基本和临床相关性，因为这两种途径在人类消化癌中被激活。第三个目标的目的是表征最近设计的熔化突变的小鼠模型。这包括食管和肠道的表征纯合突变体的表型，以及与新设计的小鼠进行比较以携带敲门与与遗传运动综合征相关的人类MYH11突变相同的突变。总的来说，提出的实验将定义建立和保持消化器官架构和功能。