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：
7644952
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/7644952
关键词：
Activins Address Alleles Allergic Antigens Applications Grants Attenuated Automobile Driving Biochemical Bleomycin Breathing Cell Culture Techniques Cell Line Cicatrix Collagen Complex Data Dependence Deposition Development Dimerization Dominant-Negative Mutation E-Cadherin Enterobacteria phage P1 Cre recombinase Environment Epithelial Epithelial Cells Epithelium Etiology Evaluation Event 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 Outcome Ovalbumin Pathway interactions 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

Activins Address Alleles Allergic Antigens Applications Grants Attenuated Automobile Driving Biochemical Bleomycin Breathing Cell Culture Techniques Cell Line Cicatrix Collagen Complex Data Dependence Deposition Development Dimerization Dominant-Negative Mutation E-Cadherin Enterobacteria phage P1 Cre recombinase Environment Epithelial Epithelial Cells Epithelium Etiology Evaluation Event 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 Outcome Ovalbumin Pathway interactions 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

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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）：纤维化的发展代表了气道重塑的关键参数，并且可能是不同细胞类型中众多信号传导级联反应的结果。促纤维化细胞因子转化生长因子β-1（TGF-1）在纤维化发展中起着基本的作用，TGF-1诱导的信号传导的结果高度取决于细胞环境，以及与其他信号通路的合作。来自我们实验室的令人兴奋的新初步研究已经确定了C-Jun-N-终端激酶（JNK）在增强TGF-1的促纤维化作用方面的关键作用，这与气道上皮细胞（EMT）间充质过渡的病因有关。我们已经证明了这一点； 1）JNK主要在卵巢蛋白敏感和挑战的小鼠的气道上皮中激活； 2）缺乏JNK1（JNK1 - / - ）的小鼠无法诱导间充质基因或响应卵巢蛋白挑战或TGF-1或施用白血病而产生纤维化； 3）衍生自JNK1 - / - 小鼠的原发性气管上皮细胞对TGF-1诱导的间充质基因和蛋白质的表达难治性； 4）TGF-1诱导的上皮到间质转变需要JNK1。我们的数据表明，气道上皮衍生的JNK依赖性信号在编排气道纤维化中的关键作用。该提议的假设是，在气道上皮中的JNK1激活在气道重塑的鼠模型中，需要通过促进上皮上皮到间质转变（EMT）来发展上皮下纤维化。我们假设JNK1通过受体SMAD的协调接头磷酸化和C-Jun的磷酸化增强了TGF-1信号传导，从而增强了间充质基因的表达，对TGF-1诱导的纤维化至关重要。在特定的目标＃1中，我们将确定在TGF-1诱导的上皮过渡中JNK1在气管上皮细胞原代培养物中的要求。在特定的目标＃2中，我们将确定气道上皮中间充质基因表达的增加以及小鼠中上皮纤维化的发展是否需要JNK1-TGF信号轴。 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在OVA或TGF-1诱导的气道纤维化的编排中。我们的研究的结果基于细胞培养和遗传改变的小鼠模型，将更好地理解导致气道胶原蛋白沉积（疤痕）的分子信号，以及在此过程中上皮细胞的相关性。项目叙述：排队呼吸道（呼吸管）的上皮细胞在防御吸入材料中起着至关重要的作用，并且对于维持正常的肺功能很重要。但是，我们的实验室已经确定上皮细胞在促进肺组织僵硬，从而降低肺功能方面起负面作用。我们已经确定了一些关键的生化信号，这些信号促进了这一可能的负面事件。项目叙述：该赠款提案具有四个特定的目的，目的是测试这些生化信号在肺部僵化中的重要性，并将使用小鼠气道，人类上皮细胞系的原代细胞培养物以及遗传改变的小鼠的构建。该项目的完成将为肺部僵硬过程提供新的见解，并可能导致新的治疗剂的发展以限制肺部僵硬，并有可能扭转这一过程。