PDF(Pubmed)
Abstract：
UNASSIGNED: The airway epithelium plays a central role in the pathogenesis of chronic respiratory diseases such as asthma and chronic rhinosinusitis with nasal polyps (CRSwNP), but the mechanisms by which airway epithelial cells (EpCs) maintain inflammation are poorly understood.
UNASSIGNED: We hypothesized that transcriptomic assessment of sorted airway EpCs across the spectrum of differentiation would allow us to define mechanisms by which EpCs perpetuate airway inflammation.
UNASSIGNED: Ethmoid sinus EpCs from adult patients with CRS were sorted into 3 subsets, bulk RNA sequenced, and analyzed for differentially expressed genes and pathways. Single cell RNA-seq (scRNA-seq) datasets from eosinophilic and non-eosinophilic CRSwNP and bulk RNA-seq of EpCs from mild/moderate and severe asthma were assessed. Immunofluorescent staining and ex vivo functional analysis of sinus EpCs were used to validate our findings.
UNASSIGNED: Analysis within and across purified EpC subsets revealed an enrichment in glycolytic programming in CRSwNP vs CRSsNP. Correlation analysis identified mammalian target of rapamycin complex 1 (mTORC1) as a potential regulator of the glycolytic program and identified EpC expression of cytokines and wound healing genes as potential sequelae. mTORC1 activity was upregulated in CRSwNP, and ex vivo inhibition demonstrated that mTOR is critical for EpC generation of CXCL8, IL-33, and CXCL2. Across patient samples, the degree of glycolytic activity was associated with T2 inflammation in CRSwNP, and with both T2 and non-T2 inflammation in severe asthma.
UNASSIGNED: Together, these findings highlight a metabolic axis required to support epithelial generation of cytokines critical to both chronic T2 and non-T2 inflammation in CRSwNP and asthma.
UNASSIGNED: We hypothesized that transcriptomic assessment of sorted airway EpCs across the spectrum of differentiation would allow us to define mechanisms by which EpCs perpetuate airway inflammation.
UNASSIGNED: Ethmoid sinus EpCs from adult patients with CRS were sorted into 3 subsets, bulk RNA sequenced, and analyzed for differentially expressed genes and pathways. Single cell RNA-seq (scRNA-seq) datasets from eosinophilic and non-eosinophilic CRSwNP and bulk RNA-seq of EpCs from mild/moderate and severe asthma were assessed. Immunofluorescent staining and ex vivo functional analysis of sinus EpCs were used to validate our findings.
UNASSIGNED: Analysis within and across purified EpC subsets revealed an enrichment in glycolytic programming in CRSwNP vs CRSsNP. Correlation analysis identified mammalian target of rapamycin complex 1 (mTORC1) as a potential regulator of the glycolytic program and identified EpC expression of cytokines and wound healing genes as potential sequelae. mTORC1 activity was upregulated in CRSwNP, and ex vivo inhibition demonstrated that mTOR is critical for EpC generation of CXCL8, IL-33, and CXCL2. Across patient samples, the degree of glycolytic activity was associated with T2 inflammation in CRSwNP, and with both T2 and non-T2 inflammation in severe asthma.
UNASSIGNED: Together, these findings highlight a metabolic axis required to support epithelial generation of cytokines critical to both chronic T2 and non-T2 inflammation in CRSwNP and asthma.
摘要：
■气道上皮在慢性呼吸道疾病如哮喘和慢性鼻窦炎伴鼻息肉（CRSwNP）的发病机理中起着核心作用，但是对气道上皮细胞（EpCs）维持炎症的机制知之甚少。
■我们假设分类的气道EpCs在分化谱中的转录组学评估将允许我们定义EpCs延续气道炎症的机制。
■来自患有CRS的成年患者的筛窦EpCs分为3个亚群，批量RNA测序，并分析了差异表达的基因和途径。评估了来自嗜酸性粒细胞和非嗜酸性粒细胞CRSwNP的单细胞RNA-seq（scRNA-seq）数据集以及来自轻度/中度和重度哮喘的EpCs的大量RNA-seq。免疫荧光染色和离体功能分析的窦EpCs被用来验证我们的发现。
■纯化的EpC亚群内和跨纯化的EpC亚群的分析揭示了CRSwNP与CRSsNP中糖酵解编程的富集。相关分析确定哺乳动物雷帕霉素复合物1（mTORC1）是糖酵解程序的潜在调节剂，并确定细胞因子和伤口愈合基因的EpC表达是潜在的后遗症。mTORC1活性在CRSwNP中上调，和离体抑制表明mTOR对于CXCL8、IL-33和CXCL2的EpC生成至关重要。在患者样本中，糖酵解活性的程度与CRSwNP的T2炎症有关，严重哮喘患者同时伴有T2和非T2炎症。
■一起，这些发现强调了在CRSwNP和哮喘中支持上皮生成对慢性T2和非T2炎症都至关重要的细胞因子所需的代谢轴.
结论：CRSwNP中上皮mTORC1活性上调。mTOR调节EpC细胞因子的产生。CRSwNP中上皮代谢重编程与T2炎症相关，在哮喘中伴有T2和非T2炎症。
结论：mTORC1在CRSwNP中介导EpC细胞因子的产生。
■我们假设分类的气道EpCs在分化谱中的转录组学评估将允许我们定义EpCs延续气道炎症的机制。
■来自患有CRS的成年患者的筛窦EpCs分为3个亚群，批量RNA测序，并分析了差异表达的基因和途径。评估了来自嗜酸性粒细胞和非嗜酸性粒细胞CRSwNP的单细胞RNA-seq（scRNA-seq）数据集以及来自轻度/中度和重度哮喘的EpCs的大量RNA-seq。免疫荧光染色和离体功能分析的窦EpCs被用来验证我们的发现。
■纯化的EpC亚群内和跨纯化的EpC亚群的分析揭示了CRSwNP与CRSsNP中糖酵解编程的富集。相关分析确定哺乳动物雷帕霉素复合物1（mTORC1）是糖酵解程序的潜在调节剂，并确定细胞因子和伤口愈合基因的EpC表达是潜在的后遗症。mTORC1活性在CRSwNP中上调，和离体抑制表明mTOR对于CXCL8、IL-33和CXCL2的EpC生成至关重要。在患者样本中，糖酵解活性的程度与CRSwNP的T2炎症有关，严重哮喘患者同时伴有T2和非T2炎症。
■一起，这些发现强调了在CRSwNP和哮喘中支持上皮生成对慢性T2和非T2炎症都至关重要的细胞因子所需的代谢轴.
结论：CRSwNP中上皮mTORC1活性上调。mTOR调节EpC细胞因子的产生。CRSwNP中上皮代谢重编程与T2炎症相关，在哮喘中伴有T2和非T2炎症。
结论：mTORC1在CRSwNP中介导EpC细胞因子的产生。
