PDF(Pubmed)
Abstract：
Rationale: CFTR (cystic fibrosis transmembrane conductance regulator) modulator drugs restore function to mutant channels in patients with cystic fibrosis (CF) and lead to improvements in body mass index and lung function. Although it is anticipated that early childhood treatment with CFTR modulators will significantly delay or even prevent the onset of advanced lung disease, lung neutrophils and inflammatory cytokines remain high in patients with CF with established lung disease despite modulator therapy, underscoring the need to identify and ultimately target the sources of this inflammation in CF lungs. Objectives: To determine whether CF lungs, like chronic obstructive pulmonary disease (COPD) lungs, harbor potentially pathogenic stem cell \"variants\" distinct from the normal p63/Krt5 lung stem cells devoted to alveolar fates, to identify specific variants that might contribute to the inflammatory state of CF lungs, and to assess the impact of CFTR genetic complementation or CFTR modulators on the inflammatory variants identified herein. Methods: Stem cell cloning technology developed to resolve pathogenic stem cell heterogeneity in COPD and idiopathic pulmonary fibrosis lungs was applied to end-stage lungs of patients with CF (three homozygous CFTR:F508D, one CFTR F508D/L1254X; FEV1, 14-30%) undergoing therapeutic lung transplantation. Single-cell-derived clones corresponding to the six stem cell clusters resolved by single-cell RNA sequencing of these libraries were assessed by RNA sequencing and xenografting to monitor inflammation, fibrosis, and mucin secretion. The impact of CFTR activity on these variants after CFTR gene complementation or exposure to CFTR modulators was assessed by molecular and functional studies. Measurements and Main Results: End-stage CF lungs display a stem cell heterogeneity marked by five predominant variants in addition to the normal lung stem cell, of which three are proinflammatory both at the level of gene expression and their ability to drive neutrophilic inflammation in xenografts in immunodeficient mice. The proinflammatory functions of these three variants were unallayed by genetic or pharmacological restoration of CFTR activity. Conclusions: The emergence of three proinflammatory stem cell variants in CF lungs may contribute to the persistence of lung inflammation in patients with CF with advanced disease undergoing CFTR modulator therapy.
摘要：
背景：“CFTR调节剂”药物可恢复囊性纤维化（CF）患者的突变通道功能，并改善体重指数和肺功能，尤其是在年轻患者中。虽然预期早期使用CFTR调节剂治疗将显著延迟晚期肺病的发作,肺中性粒细胞和炎性细胞因子在已确诊肺病的调质治疗的CF患者中仍然很高，强调需要确定并最终靶向CF肺部这种炎症的来源。
目的:检查CF肺的干细胞异质性以鉴定可能是CF中慢性肺部炎症的基础的干细胞变体以及CFTR遗传互补或CFTR调节剂对本文鉴定的干细胞变体的炎症特性的影响。
方法：将干细胞克隆技术应用于CF肺。通过RNA测序和异种移植评估单细胞来源的克隆，以监测炎症。纤维化，和粘蛋白分泌。通过分子和功能研究评估了在基因互补或暴露于CFTR调节剂后CFTR活性对这些变体的影响。
结果：CF肺表现出干细胞异质性，其特征为6种主要变体，其中3种在基因表达水平和在异种移植物中驱动中性粒细胞炎症的能力方面都是促炎的。这些变体的促炎功能未通过CFTR活性的遗传或药理学恢复而减轻。
结论：CF肺中促炎干细胞变异体的出现可能解释了接受CFTR调节剂治疗的CF患者肺部炎症的持续存在。
目的:检查CF肺的干细胞异质性以鉴定可能是CF中慢性肺部炎症的基础的干细胞变体以及CFTR遗传互补或CFTR调节剂对本文鉴定的干细胞变体的炎症特性的影响。
方法：将干细胞克隆技术应用于CF肺。通过RNA测序和异种移植评估单细胞来源的克隆，以监测炎症。纤维化，和粘蛋白分泌。通过分子和功能研究评估了在基因互补或暴露于CFTR调节剂后CFTR活性对这些变体的影响。
结果：CF肺表现出干细胞异质性，其特征为6种主要变体，其中3种在基因表达水平和在异种移植物中驱动中性粒细胞炎症的能力方面都是促炎的。这些变体的促炎功能未通过CFTR活性的遗传或药理学恢复而减轻。
结论：CF肺中促炎干细胞变异体的出现可能解释了接受CFTR调节剂治疗的CF患者肺部炎症的持续存在。
