IL 13-responsive genes in goblet cell metaplasia in asthma

哮喘杯状细胞化生中的 IL 13 反应基因

• 批准号: 7105256
• 负责人: Mary C Rose
• 金额: $ 20.75万
• 依托单位: CHILDREN'S RESEARCH INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-03-01 至 2008-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7105256
• 关键词: asthma; cell differentiation; gene expression; genes; goblet cells; metaplasia

DESCRIPTION (provided by applicant): An increased number of goblet cells (goblet cell hyperplasia, GCH) in the airway epithelium is largely responsible for the deleterious overproduction of mucin glycoproteins in response to allergens and environmental toxins in patients with asthma. The mechanisms that result in GCH have not been elucidated. This project focuses on identifying genes and pathways that direct the appearance of goblet cells (goblet cell metaplasia, GCM) in the interleukin (IL)13-induced murine model of allergic asthma, which is mediated by activation of the Stat6 signal transduction pathway. The overall hypotheses to be tested in this Exploratory/Developmental Research project is that the IMS-induced activation of Stat6 activates master genes, e.g. transcription factors with Stat6 cis-sequences, and that master gene products subsequently regulate switch genes that mediate goblet cell differentiation. We have recently performed temporal expression array profiling of murine trachea following exposure to IL13 or PBS and identified four candidate master genes upregulated early (1 h) following IL13 exposure and several candidate switch genes, which exhibit altered expression at an intermediate time (3.5 h) and return to base line levels at a later time (6 h). In Aim 1, four candidate master genes will be evaluated to determine whether one or more are primary targets of Stat6. Candidate master genes will be evaluated for co-localization with IL4R and for binding to Stat6 by chromatin immunoprecipitation assay (ChIP) and functionally evaluated in vitro in murine tracheal epithelial (MTE) cells. In Aim 2, candidate switch gene products will be evaluated for cellular and sub- cellular co-localization with master genes using confocal microscopy. Binding of master gene products to cis-elements in switch gene promoters will be evaluated. The ability of candidate switch genes to functionally induce GCM will be monitored following siRNA or transduction of switch genes into MTE cells. This study should define for the first time a pathway for goblet cell differentiation in a mammalian airway system. This will contribute to the development of pharmacological agents to control GCH and mucus overproduction in patients with asthma and possibly other chronic airway diseases

描述（由申请人提供）：气道上皮中的杯状细胞（Goblet细胞增生，GCH）的数量增加，这在很大程度上是造成肌蛋白的有害过量生产，响应于哮喘患者的过敏原和环境毒素。导致GCH的机制尚未阐明。该项目的重点是识别指导白介素（IL）13诱导的过敏性哮喘的鼠模型中的杯状细胞（Goblet细胞化生，GCM）出现的基因和途径，该模型是由STAT6信号转导途径的激活介导的。在此探索性/发育研究项目中需要检验的总体假设是IMS诱导的STAT6激活激活了主基因，例如具有STAT6顺式序列的转录因子，并且主要基因产物随后调节介导杯状细胞分化的开关基因。暴露于IL13或PBS后，我们最近进行了鼠气管的时间表达阵列分析，并确定了IL13暴露后早期（1 h）上调的四个候选主基因和几个候选开关基因，这些基因在中间时间（3.5 h）表现出改变的表达（3.5 h），并在以后的时间（6 h）返回基准水平（6 h）。在AIM 1中，将评估四个候选主基因，以确定一个或多个是STAT6的主要目标。将评估候选主基因与IL4R共定位，并通过染色质免疫沉淀测定法（CHIP）与STAT6结合，并在鼠类气管上皮（MTE）细胞中在功能上进行了功能评估。在AIM 2中，将使用共聚焦显微镜评估候选开关基因产物与主基因的细胞和亚细胞共定位。将评估主基因产物与开关基因启动子中的顺式元素的结合。在siRNA或将开关基因转导向MTE细胞之后，将监测候选开关基因在功能上诱导GCM的能力。这项研究应首次定义哺乳动物气道系统中的杯状细胞分化途径。这将有助于开发药理学剂，以控制哮喘患者以及其他慢性气道疾病患者的GCH和粘液过量生产