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

• 批准号: 10446915
• 负责人: ANTHONY N GERBER
• 金额: $ 75.96万
• 依托单位: NATIONAL JEWISH HEALTH
• 依托单位国家: 美国
• 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/10446915
• 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; Infection; Inflammation; Inflammatory; Inhalation; Knowledge; Ligand Binding; Ligands; Link; Lung; Mediating; Methods; Modeling; Molecular; Molecular Conformation; Muscle function; Odds Ratio; Pathogenesis; Pathway interactions; Patients; Persons; Pharmaceutical Preparations; Pharmacogenomics; Pharmacology; 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; base; cell type; chromosome conformation capture; cytokine; gene induction; genetic association; genetic signature; genome-wide; genomic signature; glycosyltransferase; improved; knock-down; member; novel; novel therapeutics; promoter; receptor; receptor binding; respiratory smooth muscle; response; side effect; therapeutic evaluation; transcription factor; translational approach; translational model; virtual

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（GCS，通常称为“类固醇”）是在控制范围的极有效药物 包括哮喘在内的各种免疫介导的疾病。它们还用于治疗病毒诱发的炎症 在哮喘恶化的背景下。通过减少诸如气道平滑肌等组织的炎症 （ASM），吸入的GC在大多数哮喘患者中非常有效，而全身疗法则是 引起副作用，用于治疗哮喘加重。尽管使用了最佳疗法，但超过一百万 美国人继续遭受难以控制的哮喘和经常恶化的折磨。那是新的 涉及肺中的立体信号传导机制（包括ASM）的药理学方法是迫切的 需要改善哮喘治疗。 GCS主要生化功能是与GC受体（GR）结合，导致转录的调节 直接通过GR，这是转录因子。 ASM是哮喘中GC的关键目标组织，ASM表现出 哮喘中的超收缩，重塑和旺盛的细胞因子表达。以前利用类固醇的努力 在SO上预测了受控途径作为改善针对ASM的哮喘疗法的方法 GR活性的“反抑制”模型，其中GR被认为可以减少拮抗作用的炎症 将促炎性转录因子（例如NFKB）束缚。这笔赠款完全与教条形成鲜明对比 而是将应用集中在GR和GR和NFKB之间的合作上的转录诱导上。我们的 初步数据暗示GR​​-NFKB的合作是控制ASM注入的核心，因此很可能 对类固醇的有效性做出了重大贡献。从机械上讲，我们的数据表明此途径涉及 增强子RNA的转录和染色质环的形成，我们的数据还表明这种机制是 可吸毒。根据我们对GR诱导的途径的承诺结果，我们还创建了一种新颖的方法 询问GR诱导的增强子RNA特征以与哮喘的遗传联系。我们的总体假设是 ASM中GR介导的转录诱导和GR-NFKB合作对于类固醇的效率至关重要 哮喘和这些途径与患哮喘的风险有关。我们将用三个 具体目的，这也将解决关键的分子知识差距。在AIM 1中，我们将确定 全面由GR/NFKB合作诱导的增强剂RNA并确定这些是否是否 增强子RNA环向抗炎基因的启动子。在AIM 2中，我们将确定DNA序列 GR和NFKB合作监管中涉及的其他因素以及我们将测试 这种途径在翻译模型中的治疗相关性。在AIM 3中，我们将确定新颖的遗传关联 在SNP，GR诱导的增强子和哮喘之间，我们将确定这些SNP与GR的相关性 信号和ASM功能。