Mechanisms and Consequences of Gene Induction by Glucocorticoids in Airway Smooth Muscle

糖皮质激素在气道平滑肌中基因诱导的机制和后果

基本信息

批准号：
10641892
负责人：
ANTHONY N GERBER
金额：
$ 76.79万
依托单位：
NATIONAL JEWISH HEALTH
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-04-15 至 2026-02-28
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10641892
关键词：
Address Advanced Development American Anti-Inflammatory Agents Applications Grants Asthma Binding Biochemical Bioinformatics Biological Assay Cells ChIP-seq Chromatin Chromatin Loop Clinical Complex DNA DNA Sequence Data Data Set Disease Enhancers Exhibits Family Functional disorder Gene Expression Gene Expression Regulation Genes Genetic Genetic Transcription Genomic approach Genomics Glucocorticoid Receptor Glucocorticoids Goals Health Care Costs Human Immune Immunomodulators Infection Inflammation Inflammatory Inhalation Knowledge Ligand Binding Ligands Link Lung Maps Mediating Methods Modeling Molecular Molecular Conformation Muscle function NF-kappa B Odds Ratio Pathogenesis Pathway interactions Patients Persons Pharmaceutical Preparations Pharmacogenomics RNA Receptor Signaling Regulation Reporter Repression Research Response Elements Risk Signal Pathway Signal Transduction Single Nucleotide Polymorphism Site Slice Small Interfering RNA Steroids Symptoms Systemic Therapy Testing Therapeutic Therapeutic Effect Tissues Transcript Transcriptional Regulation Viral analysis pipeline asthma exacerbation asthmatic patient cell type chromosome conformation capture cytokine gene induction genetic association genetic signature genome-wide genomic signature glycosyltransferase improved knock-down member novel novel therapeutics pharmacologic promoter receptor receptor binding respiratory smooth muscle response side effect therapeutic evaluation transcription factor translational approach translational model virtual

项目摘要

Glucocorticoids (GCs, generically known as “steroids”) are exceptionally effective drugs in the control of a wide variety of immune-mediated diseases, including asthma. They are also used to treat viral-induced inflammation in the context of asthma exacerbations. Through reducing inflammation in tissues such as airway smooth muscle (ASM), inhaled GCs are very effective in the majority of asthma patients, whereas systemic therapy, which causes side effects, is used to treat asthma exacerbations. Despite using best therapies, over one million Americans continue to be afflicted with difficult-to-control asthma and frequent exacerbations. Thus, new pharmacologic approaches aimed at steroid signaling mechanisms in the lung, including ASM, are urgently needed to improve treatment for asthma. GCs primary biochemical function is to bind to the GC receptor (GR), leading to regulation of transcription directly by GR, which is a transcription factor. ASM is a key target tissue of GCs in asthma, and ASM exhibits hyper-contractility, remodeling and exuberant cytokine expression in asthma. Previous efforts to exploit steroid- controlled pathways as a method to improve asthma therapies directed at ASM have been predicated on the so called “transrepression” model of GR activity, in which GR is thought to reduce inflammation by antagonistic tethering to pro-inflammatory transcription factors such as NFkB. Entirely contrary to dogma, this grant application is focused instead on transcriptional induction by GR and cooperation between GR and NFkB. Our preliminary data implicate GR-NFkB cooperation as central to controlling inflammation in ASM, thus likely contributing significantly to efficacy of steroids. Mechanistically, our data suggest this pathway involves transcription of enhancer RNAs and formation of chromatin loops, and our data also suggest this mechanism is druggable. Based on our promising findings on GR-induced pathways, we have also created a novel method to interrogate GR-induced enhancer RNA signatures for genetic links to asthma. Our overarching hypothesis is that GR-mediated transcriptional induction and GR-NFkB cooperation in ASM is crucial for steroid efficacy in asthma and these pathways are linked to risk of developing asthma. We will test this hypothesis with three specific aims, which will also address crucial molecular knowledge gaps. In Aim 1, we will determine the comprehensive set of enhancer RNAs that are induced cooperatively by GR/NFkB and determine if these enhancer RNAs loop to the promoters of anti-inflammatory genes. In Aim 2, we will determine the DNA sequence requirements and additional factors involved in cooperative regulation by GR and NFkB, and we will test the therapeutic relevance of this pathway in translational models. In Aim 3, we will identify novel genetic associations between SNPs, GR-induced enhancers and asthma, and we will determine the relevance of these SNPs to GR signaling and ASM function.

糖皮质激素（GC，通称“类固醇”）是控制多种疾病的极其有效的药物。各种免疫介导的疾病，包括哮喘，它们也用于治疗病毒引起的炎症。在哮喘恶化的情况下，通过减少气道平滑肌等组织的炎症。 (ASM)，吸入GC对大多数哮喘患者非常有效，而全身治疗则需要引起副作用，尽管使用了最好的疗法，但仍用于治疗哮喘发作，但仍有超过一百万。美国人继续受到难以控制的哮喘和频繁恶化的影响。迫切需要针对肺部类固醇信号传导机制（包括 ASM）的药理学方法需要改善哮喘的治疗。 GC 的主要生化功能是与 GC 受体 (GR) 结合，从而调节转录 GR 是一种转录因子，ASM 是哮喘中 GC 的关键靶组织，ASM 表现出这种作用。哮喘中的过度收缩、重塑和细胞因子表达旺盛。控制途径作为改善针对 ASM 的哮喘治疗的方法已被预测为 GR 活性被称为“反式抑制”模型，其中 GR 被认为通过拮抗作用减少炎症与促炎转录因子（如 NFkB）结合，这与教条完全相反。应用程序的重点是 GR 的转录诱导以及 GR 和 NFkB 之间的合作。初步数据表明 GR-NFkB 合作是控制 ASM 炎症的核心，因此可能从机制上讲，我们的数据表明该途径涉及类固醇的功效。增强子 RNA 的转录和染色质环的形成，我们的数据也表明这种机制是基于我们对 GR 诱导途径的有希望的发现，我们还创建了一种新方法探究 GR 诱导的增强子 RNA 特征与哮喘的遗传联系。 ASM 中 GR 介导的转录诱导和 GR-NFkB 合作对于类固醇功效至关重要哮喘以及这些途径与患哮喘的风险有关，我们将用三个方面来检验这一假设。具体目标，这也将解决关键的分子知识差距在目标 1 中，我们将确定 GR/NFkB 协同诱导的综合增强子 RNA 集，并确定这些是否在目标 2 中，我们将确定 DNA 序列。 GR 和 NFkB 合作监管涉及的要求和其他因素，我们将测试在目标 3 中，我们将确定该通路在转化模型中的治疗相关性。 SNP、GR 诱导的增强子和哮喘之间的关系，我们将确定这些 SNP 与 GR 的相关性信令和ASM功能。