CEBPD-Mediated Mechanisms of Glucocorticoid Insensitivity in Severe Asthma

CEBPD 介导的严重哮喘糖皮质激素不敏感机制

• 批准号: 9914313
• 负责人: Blanca E Himes
• 金额: $ 63.59万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-04-01 至 2022-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9914313
• 关键词: Affect; Agonist; Airway Disease; American; Asthma; Binding; Binding Sites; Biological Assay; Biological Markers; Biology; Bronchodilator Agents; Budesonide; CCAAT-enhancer-binding protein-delta; Cell Differentiation process; ChIP-seq; Characteristics; Clinical; Complex; Cyclic AMP; DNA Binding; Data; Disease; Enzyme-Linked Immunosorbent Assay; Epigenetic Process; Event; Exposure to; Gene Expression; Gene Targeting; Genes; Genetic Transcription; Glucocorticoid Receptor; Glucocorticoids; Health Expenditures; Histones; Immune; Individual; Inflammatory; Inflammatory Response; Interleukin-1 beta; Life; Luciferases; Lung diseases; Measures; Mediating; Messenger RNA; Modification; Morbidity - disease rate; Patients; Pattern; Pharmaceutical Preparations; Physiological; Physiological Processes; Play; Post-Translational Protein Processing; Proteins; Proteomics; RNA analysis; Relaxation; Reporter; Research Personnel; Resources; Role; Signal Transduction; Small Interfering RNA; Stimulus; Structure; Testing; Therapeutic; Tissues; Transcript; Transcription Alteration; Validation; Work; airway hyperresponsiveness; asthmatic patient; base; biomarker development; cytokine; differential expression; health care service utilization; insight; knock-down; mRNA Expression; novel therapeutics; overexpression; respiratory smooth muscle; response; time use; transcription factor; transcriptome; transcriptome sequencing; transcriptomics

PROJECT SUMMARY Asthma is an episodic inflammatory disease that affects over 25 million Americans and manifests as airway hyperresponsiveness to specific environmental stimuli. Despite effective medications that control asthma in most individuals, 15% respond inadequately and suffer life-threatening exacerbations. A feature shared by most patients with severe disease is glucocorticoid insensitivity, a poorly understood physiological process. The airway smooth muscle (ASM), which plays an important role in asthma, is a target of glucocorticoids that act in part via modulation of gene transcription, alteration of histone post-translational modifications, and inhibition of transcription factors. CCAAT/Enhancer Binding Protein D (CEBPD) is a pleiotropic glucocorticoid- responsive transcription factor that regulates inflammatory responses, cell differentiation and tissue remodeling. Based on our data showing that CEBPD expression is (1) lower in fatal asthma vs. non-asthma ASM, (2) induced with glucocorticoid treatment in non-asthma ASM but unchanged in fatal asthma ASM, and that (3) decreasing CEBPD via knockdown resulted in increased IL1β-induced NFkB-luciferase expression with glucocorticoid stimulation, our central hypothesis states that low CEBPD levels in ASM elicits transcriptomic and epigenetic modifications that decrease glucocorticoid sensitivity. We will test this hypothesis by using the following unbiased and complementary `omic approaches to study the effect of CEBPD on glucocorticoid response in ASM from fatal asthma and non-asthma donors: (1) RNA-Seq to identify transcriptomic effects, (2) proteomics to determine global histone post-translational modification effects, and (3) ChIP-Seq to measure transcription factor binding of NFkB and the glucocorticoid receptor. An integrated analysis of RNA-Seq, proteomics and ChIP-Seq results will identify major targets of CEBPD that influence glucocorticoid response, whose role will be confirmed via NFkB-luciferase assays. Our project will offer insights into transcriptional and epigenetic signatures that are characteristic of severe asthma, enable glucocorticoid sensitivity biomarker development, and offer therapeutic insights that benefit the most vulnerable individuals with asthma.

项目摘要 哮喘是一种发作性炎症性疾病，影响超过2500万美国人，并表现为气道炎症。 对特定环境刺激的高反应性。尽管有效的药物控制哮喘， 大多数人，15%的反应不充分，并遭受危及生命的恶化。一个共同的特点， 大多数严重疾病的患者是糖皮质激素不敏感，这是一种了解甚少的生理过程。 在哮喘中起重要作用的气道平滑肌（ASM）是糖皮质激素的靶点， 部分通过调节基因转录、改变组蛋白翻译后修饰发挥作用， 抑制转录因子。CCAAT/增强子结合蛋白D（CEBPD）是一种多效性糖皮质激素- 调节炎症反应、细胞分化和组织的应答性转录因子 重塑基于我们的数据显示CEBPD表达（1）致命性哮喘比非哮喘低， ASM，（2）在非哮喘ASM中用糖皮质激素治疗诱导，但在致死性哮喘ASM中无变化，和 （3）通过敲低降低CEBPD导致IL 1 β诱导的NF κ B-荧光素酶表达增加， 糖皮质激素刺激，我们的中心假设指出，低CEBPD水平的ASM eliminating转录组， 和降低糖皮质激素敏感性的表观遗传修饰。我们将通过使用 采用无偏倚和互补的“组学”方法研究CEBPD对糖皮质激素的影响 来自致死性哮喘和非哮喘供体的ASM中的应答：（1）RNA-Seq以鉴定转录组学效应，（2） 蛋白质组学以确定全局组蛋白翻译后修饰效应，以及（3）ChIP-Seq以测量 NF κ B和糖皮质激素受体的转录因子结合。RNA-Seq的综合分析， 蛋白质组学和ChIP-Seq结果将鉴定影响糖皮质激素反应的CEBPD的主要靶标， 其作用将通过NF κ B-荧光素酶测定来证实。我们的项目将提供对转录和 表观遗传特征是严重哮喘的特征，使糖皮质激素敏感性生物标志物 发展，并提供有益于最脆弱的哮喘患者的治疗见解。