Nuclear Receptor REV-ERB alpha Role in the Pathophysiology of Allergic Asthma

摘要：过敏性哮喘是一种慢性炎症性疾病，表现出时间依赖性的变化。 在临床症状和严重程度上。尽管吸入的(或全身的)皮质类固醇和交感神经抑制剂 通常用于缓解哮喘对呼吸道功能的直接影响，这些药物大多是 效果不佳。有趣的是，哮喘患者表现出肺功能和粘液产生的异常昼夜节律。 与肺部炎症增加和病情加重有关。REV-ERBα是一种核受体 转录因子在昼夜节律系统中起关键作用的转录因子，其作用是维持昼夜节律 基因表达与免疫、炎症和新陈代谢有关。我们的初步数据显示， 家尘螨暴露对小鼠肺上皮细胞REV-ERBα表达的影响 (HDM)变应原与哮喘肺表型增加(呼吸道炎症增加， 呼吸道高反应性、Th2细胞因子、血浆IgE和粘液化生)。这一数据与 研究表明，REV-ERBα在哮喘小鼠模型和人类模型中表达下调 哮喘患者的呼吸道细胞已恢复。综上所述，这些数据表明REV-ERBα可能有助于 过敏性哮喘的病理生理机制及减轻免疫炎症的新靶点 回应。然而，没有数据描述REV-ERBα可能起作用的分子机制 过敏性哮喘的病理生物学。我们假设变应原诱导的REV-ERBα的破坏 表达导致生物钟功能紊乱和免疫炎症反应增强 肺部。目的1：确定REV-ERBα在免疫细胞募集到肺组织中的作用 过敏性哮喘。我们将随时间和全球昼夜节律测量肺免疫-炎症反应 急性HDM暴露后一天两个不同时间的RNA测序转录组(RNA-SEQ) (AM与PM)在REV-ERBα基因敲除小鼠和野生型小鼠中。目标2：确定小分子是否激活了 REV-ERBα可通过以下途径预防和/或减轻呼吸道炎症和哮喘肺表型 体内抑制NFIL3-STAT3轴。我们将采用遗传和药理学方法来 确定激活REV-ERBα是否可以保护和/或减轻过敏性哮喘。靶标特异性和细胞- 将使用RNA-seq分析来确定类型相关性。目标3：如何确定机制 HDM/Th2型细胞因子抑制上皮细胞REV-ERBα表达导致上皮屏障 功能障碍和体外培养的杯状细胞增生。我们将治疗原代人支气管上皮细胞和 EpiAirway组织(正常和哮喘患者)经或不经选择性REV-1治疗后对HDM/Th2细胞因子的影响 ERBα激动剂/拮抗剂或STAT3抑制剂。这将决定NFIL3-STAT3轴如何抑制REV- ERBα在Th2型细胞因子诱导的屏障功能障碍和杯状细胞增生中的作用总体而言，这项研究将 描述REV-ERBα在过敏性哮喘的时钟依赖的病理生理反应中的新作用。

SUMMARY: Allergic asthma is a chronic inflammatory disease that displays time-of-day dependent variations in clinical symptoms and severity. Though inhaled (or systemic) corticosteroids and sympathetic inhibitors are commonly used to alleviate the immediate impact of asthma on airway function, these drugs are mostly ineffective. Intriguingly, asthmatics show abnormal circadian rhythms of lung function, and mucus production associated with increased lung inflammation and exacerbations. REV-ERBα is a nuclear receptor and transcription factor that plays a critical role in the circadian timing system, acting to maintain daily rhythms of gene expression linked to immunity, inflammation and metabolism. Our preliminary data show that reduced expression of REV-ERBα in the airway epithelium of mouse lungs following exposure to the House Dust Mite (HDM) allergen is associated with augmented asthmatic lung phenotypes (increased airway inflammation, airway hyperresponsiveness, Th2 cytokines, plasma IgE and mucous metaplasia). This data agrees with studies showing that REV-ERBα expression is downregulated in mouse models of asthma and in human airway cells recovered from asthmatics. Together, these data suggests that REV-ERBα may contribute to the pathophysiology of allergic asthma and represent a novel target for alleviating the immune-inflammatory response. However, there is no data describing the molecular mechanism, whereby REV-ERBα may contribute to the pathobiology of allergic asthma. We hypothesize that allergen-induced disruption of REV-ERBα expression leads to irregular clock function and enhanced immune-inflammatory response in the lungs. Aim 1: Determine the role of REV-ERBα in recruitment of immune cells into the lung during allergic asthma. We will measure lung immune-inflammatory response over time and global circadian transcriptome by RNA-sequencing (RNA-seq) following acute HDM exposure at two different times of the day (AM vs. PM) in Rev-erbα knockout and wild-type mice. Aim 2: Determine if small molecule activation of REV-ERBα can prevent and/or attenuate airway inflammation and asthmatic lung phenotypes by repressing NFIL3-STAT3 axis in vivo. We will employ both genetic and pharmacological approaches to determine if activation of REV-ERBα can protect and/or attenuate allergic asthma. Target specificity and cell- type dependency will be determined using RNA-seq analysis. Aim 3: Determine the mechanism how HDM/Th2 cytokines suppress REV-ERBα expression in the epithelium leading to epithelial barrier dysfunction and goblet cell hyperplasia in vitro. We will treat primary human bronchial epithelial cells and EpiAirway tissues (normal and asthmatics) to HDM/Th2 cytokines with or without treatment with selective REV- ERBα agonists/antagonists or a STAT3 inhibitor. This will determine how NFIL3-STAT3 axis represses REV- ERBα function in Th2 cytokine-induced barrier dysfunction and goblet cell hyperplasia. Overall, this study will delineate the novel role of REV-ERBα in the clock-dependent pathophysiological response to allergic asthma.