Circadian Signatures in Chronic Lung Disease

项目总结/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。气道 对于哮喘和慢性阻塞性肺疾病（COPD）等疾病， 疾病定义特征，其中气道阻力的正常昼夜节律波动在病理上是 放大了昼夜节律是由细胞自主分子钟产生的，它协调组织- 基因表达的特定节律，导致生理学的振荡，包括在肺中。研究 提示气道上皮细胞内的分子钟功能特异性调节先天性抗微生物 应答这种反应在慢性肺病中是至关重要的，因为它们可能决定了是否有一种 遇到病毒或其他病原体会引发临床恶化。目前，没有关于 人体肺部的昼夜节律调节程序以及慢性疾病如何改写这种调节， 驱动发病机制。在这里，我们表明，慢性肺部疾病改变了生物钟基因表达的节奏 在来自患者的人支气管上皮细胞（HBEC）中。我们证明HBEC表达昼夜节律， 转录组包含数百个基因，并表现出可以进一步改变的疾病特异性模式 急性病毒感染。因此，我们假设慢性气道疾病重新编程昼夜节律基因， 在气道细胞中表达，从而影响病毒易感性。该项目的总体目标是 将昼夜节律生物学转化为慢性气道疾病患者的疾病改善治疗， 减轻或预防病毒触发的临床恶化。目的1：确定疾病特异性昼夜节律 调节哮喘和COPD患者抗病毒反应的基因表达程序。使用唯一 由我们小组开发的从人肺外植体分离的HBEC库，我们将确定 从哮喘和COPD患者中获得的HBEC的昼夜节律转录组，并将其与供体进行比较 没有慢性呼吸道疾病与此同时，我们将根据HBEC对急性胰腺炎的易感性对HBEC进行表型分析。 在存在或不存在生物钟中断的情况下，甲型流感病毒（IAV）感染。HBEC中的观察结果 将由昼夜节律的单细胞成像和体内人体气道的首次分析支持 使用从器官移植供体获得的连续气道刷拭样品， 在等待器官移植的过程中脑死亡目标2：发现进化保守的昼夜节律 慢性气道疾病中的重编程我们将使用一种表征良好的哮喘/COPD小鼠模型- 如已知的抗病毒反应的时钟调节的气道重塑。使用这个模型，我们将检查肺 慢性气道疾病基因表达和白细胞运输的昼夜节律重编程， 该程序对气道上皮细胞外周时钟的依赖性。这些研究将得到 使用生物发光报告物对小鼠气道时钟基因表达进行体内成像。

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.