Epithelial JNK-TGFb1 Signaling Axis in Airway Remodeling

气道重塑中的上皮 JNK-TGFb1 信号轴

基本信息

批准号：
7367482
负责人：
Yvonne M. W. Janssen-Heininger
金额：
$ 37.63万
依托单位：
UNIVERSITY OF VERMONT & ST AGRIC COLLEGE
依托单位国家：
美国
项目类别：
财政年份：
2008
资助国家：
美国
起止时间：
2008-07-01 至 2012-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7367482
关键词：
Activins Address Alleles Allergic Antigens Applications Grants Attenuated Automobile Driving Biochemical Bleomycin Breathing Causations Cell Line Cicatrix Collagen Complex Cultured Cells Data Dependence Deposition Development Dimerization Dominant-Negative Mutation E-Cadherin End Point Enterobacteria phage P1 Cre recombinase Environment Epithelial Epithelial Cells Epithelium Evaluation Event Facility Construction Funding Category Fibroblasts Fibronectins Fibrosis Gene Expression Gene Proteins Gene Targeting Genes Human Human Cell Line In Vitro Inflammation JUN gene Knock-out Laboratories Lead Link Lung MADH2 gene MADH3 gene MADH4 gene MAPK8 gene Measures Mediating Mediator of activation protein Mesenchymal Modeling Molecular Molecular Target Mus Other Finding Outcome Ovalbumin Pathway interactions Personal Satisfaction Phosphorylation Phosphotransferases Play Primary Cell Cultures Process Production Proline Proteins Publishing Refractory Relative (related person)Resistance Respiratory physiology Role Serine Signal Pathway Signal Transduction Site Smooth Muscle Actin Staining Method Source Structure of parenchyma of lung TGF-beta type I receptor Testing Transcription Factor AP-1 Transcriptional Activation Transgenic Mice Tube airway epithelium airway hyperresponsiveness airway remodeling allergic airway disease attenuation base cell type cytokine epithelial to mesenchymal transition fibrogenesis human TGFB1 protein improved in vivo insight mouse model mutant novel novel therapeutics protein expression receptor response stress-activated protein kinase 1

项目摘要

DESCRIPTION (provided by applicant): The development of fibrosis represents a critical parameter of airway remodeling, and likely is the outcome of numerous signaling cascades operative in different cell types. The pro-fibrotic cytokine, transforming growth factor beta-1 (TGF-1) plays a cardinal role in the development of fibrosis, and the outcome of TGF-1 induced signaling is highly dependent upon the cellular environment, and the cooperation with other signaling pathways. Exciting new preliminary studies from our laboratory have identified a critical role for c-Jun-N- terminal-kinase (JNK) in augmenting the pro-fibrotic effects of TGF-1, in association with the causation of a mesenchymal transition of airway epithelial cells (EMT). We have demonstrated that; 1) JNK is predominantly activated in airway epithelium from ovalbumin sensitized and challenged mice; 2) Mice lacking JNK1 (JNK1-/-) fail to induce mesenchymal genes or develop fibrosis in response to ovalbumin challenge or TGF-1, or bleomycin administration; 3) Primary tracheal epithelial cells derived from JNK1-/- mice are refractory to TGF-1-induced expression of mesenchymal genes and proteins; and 4) TGF-1 induced epithelial-to mesenchymal transition requires JNK1. Our data, suggest a critical role of airway epithelium-derived JNK- dependent signals in orchestrating airways fibrosis. The hypothesis of this proposal is that activation of JNK1 in the airway epithelium is required for the development of subepithelial fibrosis in murine models of airway remodeling by promoting epithelial to mesenchymal transition (EMT). We hypothesize that JNK1 enhances TGF-1 signaling, via coordinated linker phosphorylation of receptor SMADs, and phosphorylation of c-Jun, resulting in enhanced expression of mesenchymal genes, critical to TGF-1-induced fibrosis. In Specific Aim #1, we will determine the requirement for JNK1 in TGF-1-induced epithelial to mesenchymal transition in primary cultures of tracheal epithelial cells. In Specific Aim #2, we will determine whether increases in mesenchymal gene expression in airway epithelium, and development of subepithelial fibrosis in mice require a JNK1-TGF- signaling axis. Specific Aim #3 proposes to elucidate the relative importance of JNK1- dependent phosphorylation of c-Jun or rSMADs linker regions in augmenting TGF-1 induced signaling, whereas Specific Aim #4 will unravel whether JNK1 activation, c-Jun and SMAD3 linker phosphorylation, and subsequent c-Jun and SMAD4 dependent transcriptional responses within the airway epithelium are critical in the orchestration of OVA or TGF-1-induced airways fibrosis. The outcome of our study, which is based on cell culture and genetically altered mouse models, will provide a better understanding of the molecular signals that lead to collagen deposition (scarring) of airways, and the relevance of epithelial cells in this process. Project Narrative: Epithelial cells that line the airways (breathing tubes) are now well known to play a critical role in the defense against inhaled materials, and are important in maintaining normal lung function. However, our laboratories have identified that epithelial cells play a negative role in promoting stiffening of the lung tissue, thereby decreasing lung function. We have identified some critical biochemical signals that promote this possibly negative event. Project Narrative: This grant proposal has four specific aims to test the importance of these biochemical signals in lung stiffening, and will use primary cell cultures from mouse airways, a human epithelial cell line, and the construction of genetically altered mice. Completion of this project will provide new insights into the process of lung stiffening, and may lead to the development of new therapeutics to limit lung stiffening, and also potentially reverse this process.

描述（由申请人提供）：纤维化的发展代表气道重塑的关键参数，并且可能是在不同细胞类型中起作用的众多信号级联的结果。促纤维化细胞因子转化生长因子β-1 (TGF-1) 在纤维化的发展中起着重要作用，TGF-1 诱导的信号传导的结果高度依赖于细胞环境以及与其他信号传导的配合途径。我们实验室令人兴奋的新初步研究发现，c-Jun-N-末端激酶 (JNK) 在增强 TGF-1 的促纤维化作用中发挥着关键作用，与气道上皮细胞的间质转化有关（急救医疗队）。我们已经证明； 1) JNK 主要在卵清蛋白致敏和攻击小鼠的气道上皮中被激活； 2) 缺乏 JNK1 (JNK1-/-) 的小鼠无法诱导间充质基因或在卵清蛋白攻击或 TGF-1 或博来霉素给药后发生纤维化； 3) JNK1-/-小鼠的原代气管上皮细胞对TGF-1诱导的间质基因和蛋白的表达具有抵抗力； 4) TGF-1 诱导的上皮间质转化需要 JNK1。我们的数据表明，气道上皮衍生的 JNK 依赖性信号在协调气道纤维化中发挥着关键作用。该提议的假设是，在气道重塑的小鼠模型中，通过促进上皮间质转化（EMT），气道上皮中 JNK1 的激活是上皮下纤维化发展所必需的。我们假设 JNK1 通过协调受体 SMAD 接头磷酸化和 c-Jun 磷酸化来增强 TGF-1 信号传导，从而增强间充质基因的表达，这对 TGF-1 诱导的纤维化至关重要。在具体目标#1中，我们将确定气管上皮细胞原代培养物中 TGF-1 诱导的上皮向间质转化对 JNK1 的需求。在具体目标#2中，我们将确定小鼠气道上皮间充质基因表达的增加以及上皮下纤维化的发展是否需要JNK1-TGF-信号轴。具体目标 #3 旨在阐明 c-Jun 或 rSMAD 接头区域的 JNK1 依赖性磷酸化在增强 TGF-1 诱导信号传导中的相对重要性，而具体目标 #4 将阐明 JNK1 激活、c-Jun 和 SMAD3 接头磷酸化是否气道上皮内随后的 c-Jun 和 SMAD4 依赖性转录反应对于 OVA 或 TGF-1 诱导的气道的协调至关重要纤维化。我们的研究结果基于细胞培养和基因改造的小鼠模型，将有助于更好地理解导致气道胶原沉积（疤痕）的分子信号，以及上皮细胞在此过程中的相关性。项目叙述：众所周知，气道（呼吸管）内衬的上皮细胞在防御吸入物质方面发挥着关键作用，并且对于维持正常的肺功能也很重要。然而，我们的实验室已发现上皮细胞在促进肺组织硬化方面发挥负面作用，从而降低肺功能。我们已经确定了一些促进这一可能的负面事件的关键生化信号。项目叙述：这项拨款提案有四个具体目标，以测试这些生化信号在肺硬化中的重要性，并将使用来自小鼠气道的原代细胞培养物、人类上皮细胞系以及基因改造小鼠的构建。该项目的完成将为肺部硬化过程提供新的见解，并可能导致开发新的疗法来限制肺部硬化，并有可能逆转这一过程。