Circadian Signatures in Chronic Lung Disease

项目摘要/摘要 昼夜节律是慢性下呼吸道疾病的一个重要但知之甚少的特征。在呼吸道中 如哮喘和慢性阻塞性肺疾病(COPD)等疾病，夜间加重是一种 疾病定义的特征，即呼吸道阻力的正常昼夜波动是病理性的 放大了。昼夜节律是由一个细胞自主的分子时钟产生的，它协调组织- 基因表达的特定节律，导致包括肺在内的生理振荡。研究 提示分子钟在呼吸道上皮细胞中的特异性作用调节先天抗菌药物 回应。这种反应在慢性肺部疾病中至关重要，因为它们可能决定 遇到病毒或其他病原体会引发临床恶化。目前，还没有关于 人体肺内的昼夜节律程序以及慢性疾病可能如何重写这一规则以 驱使病机。在这里，我们展示了慢性肺部疾病改变了生物钟基因表达的节律。 在患者的人支气管上皮细胞(HBECs)中。我们证明HBECs表达昼夜节律 转录本包含数百个基因，并显示出可以进一步改变的疾病特异性模式 由急性病毒感染引起。因此，我们假设慢性呼吸道疾病改变了昼夜节律基因。 在呼吸道细胞中的表达，从而影响病毒的敏感性。这个项目的首要目标是 通过以下方式将昼夜生物学转化为慢性呼吸道疾病患者的疾病修正治疗 减轻或预防病毒引发的临床恶化。目标1：确定特定疾病的昼夜节律 调节哮喘和慢性阻塞性肺病患者抗病毒反应的基因表达程序。使用唯一的 我们小组开发的从人肺外植体中分离的HBECs的库，我们将确定 哮喘和慢性阻塞性肺疾病患者HBECs的昼夜转录本及其与供者的比较 没有慢性呼吸道疾病。同时，我们将根据HBECs对急性胰腺炎的易感性进行表型分析。 甲型流感病毒(IAV)感染时存在或不存在生物钟干扰。HBECs中的观测 将得到昼夜节律的单细胞成像和对活体人类呼吸道的首个此类分析的支持 器官移植供者术后连续呼吸道刷检标本的昼夜节律基因表达 他们在等待器官购买时脑死亡。目标2：发现进化保守的昼夜节律 慢性呼吸道疾病的重新编程。我们将使用一种具有良好特性的哮喘/慢性阻塞性肺病小鼠模型- 就像已知的抗病毒反应的时钟调节的呼吸道重塑。利用这个模型，我们将检查肺部 慢性呼吸道疾病基因表达和白细胞转运的昼夜节律重编程 这一程序对呼吸道上皮细胞外周时钟的依赖性。这些研究将得到以下支持 使用生物发光报告器对小鼠呼吸道时钟基因表达的活体成像。

PROJECT SUMMARY/ABSTRACT Circadian rhythms are an important yet poorly understood feature of chronic lower respiratory disease. In airway diseases such as asthma and chronic obstructive pulmonary disease (COPD), nocturnal exacerbations are a disease-defining characteristic in which normal circadian swings in airway resistance are pathologically amplified. Circadian rhythms are generated by a cell-autonomous molecular clock that orchestrates tissue- specific rhythms in gene expression, leading to oscillations in physiology including in the lung. Research suggests that molecular clock function specifically within airway epithelial cells regulates innate antimicrobial responses. Such responses are critical in chronic lung disease because they may determine whether an encounter with a virus or other pathogen triggers a clinical exacerbation. Currently, there is no information on the circadian regulatory program within the human lung and how chronic disease might rewrite this regulation to drive pathogenesis. Here we show that chronic lung disease alters rhythms in circadian clock gene expression in human bronchial epithelial cells (HBECs) from patients. We demonstrate that HBECs express circadian transcriptomes encompassing hundreds of genes and exhibit disease-specific patterns that can be further altered by acute viral infection. As such, we hypothesize that chronic airway disease reprograms circadian gene expression in airway cells thereby influencing virus susceptibility. The overarching goal of this project is to translate circadian biology into disease-modifying treatments for patients with chronic airway disease by mitigating or preventing virally triggered clinical exacerbations. Aim 1: Identify disease-specific circadian gene expression programs regulating antiviral responses in asthma and COPD patients. Using a unique repository of HBECs developed by our group that were isolated from human lung explants, we will determine the circadian transcriptomes of HBECs obtained from asthma and COPD patients and compare these to donors without chronic airway disease. In parallel, we will phenotype HBECs in terms of their susceptibility to acute influenza A virus (IAV) infection in the presence or absence of circadian clock disruption. Observations in HBECs will be backed by single cell imaging of circadian rhythms and a first-of-its-kind analysis of in vivo human airway circadian gene expression using serial airway brushing samples obtained from organ transplant donors after brain death while they await organ procurement. Aim 2: Discover evolutionarily conserved circadian reprogramming in chronic airway disease. We will use a well characterized mouse model of asthma/COPD- like airway remodeling with known clock regulation of antiviral responses. Using this model, we will examine lung circadian reprogramming of gene expression and leukocyte trafficking by chronic airway disease and determine the dependency of this program on the airway epithelial cell peripheral clock. These studies will be backed by in vivo imaging of airway clock gene expression in mice using bioluminescent reporters.