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（GCS，通常称为“类固醇”）是在控制范围方面具有异常有效的药物各种免疫介导的诱导，涉及哮喘。在哮喘的情况下，通过减少组织平滑肌的炎症（ASM），吸入的GC在大多数哮喘患者中都是有效的，而全身疗法则有效引起副作用，用于治疗哮喘加重。美国人继续遭受Difficalt-to-Control哮喘的折磨和自由发育针对肺部类固醇机制的药理方法，包括ASM，迫切地是需要改善哮喘治疗。 GCS主要生化功能是与GC受体（GR）结合，导致转录的调节直接是GR，这是ASM的转录因子。哮喘中的超合同，重塑和旺盛的细胞因子表达。受控途径是改善定向ASM哮喘疗法的方法 GR活性的“变性”模型，其中GRR虽然是通过拮抗作用的将促炎性转录因子（例如NFKB）束缚。相反，应用于GR和NFKB之间的转录诱导初步数据暗示GR-NFKB的合作是控制ASM炎症的核心，因此很可能对类固醇的功效产生了显着贡献。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根据我们对GR诱导的途径的有前途的发现，我们还创建了一种新方法质疑GR诱导的增强子RNA签名与我们的哮喘的遗传联系 ASM中GR介导的转录指示和GR-NFKB合作对于类固醇的功效至关重要哮喘和这些途径与患哮喘的风险有关。特定的目的也将解决目标1的关键分子知识差距。 gr/nfkb在不固定上是综合的增强剂RNA，并确定这些是否是否增强子RNA循环到AIM 2中的抗炎基因的启动子。 GRU和NFKB合作监管中涉及的要求和其他因素，我们将测试在AIM 3中，该途径的治疗相关性。在SNP，GR诱导的增强子和哮喘之间，我们将确定这些SNPSE SNP与GR的相关性信号和ASM功能。