A Single-Cell Atlas of Large and Small Airways at Birth in a Porcine Model of Cystic Fibrosis

• Published in: American journal of respiratory cell and molecular biology Vol. 66; no. 6; pp. 612 - 622
• Main Authors: Thurman, Andrew L, Li, Xiaopeng, Villacreses, Raul, Yu, Wenjie, Gong, Huiyu, Mather, Steven E, Romano-Ibarra, Guillermo S, Meyerholz, David K, Stoltz, David A, Welsh, Michael J, Thornell, Ian M, Zabner, Joseph, Pezzulo, Alejandro A
• Format: Journal Article
• Language: English
• Published: United States American Thoracic Society 01.06.2022
• Subjects: Airway management; Animal models; Animals; Asthma; Basal cells; Birth; Cystic fibrosis; Cystic Fibrosis - metabolism; Cystic Fibrosis Transmembrane Conductance Regulator - metabolism; Epithelial cells; Epithelial Cells - metabolism; Humans; Immunohistochemistry; Inflammation; Inflammation - metabolism; Ion Transport; Neonates; Original Research; Pulmonary fibrosis; Respiratory System - metabolism; Respiratory tract; Swine; Transcriptomes; Transcriptomics; cystic fibrosis transmembrane conductance regulator; single-cell RNA-seq; airway epithelia; goblet cells; basal cells; small airway
• ISSN: 1044-1549; 1535-4989
• DOI: 10.1165/rcmb.2021-0499OC

Lack of CFTR (cystic fibrosis transmembrane conductance regulator) affects the transcriptome, composition, and function of large and small airway epithelia in people with advanced cystic fibrosis (CF); however, whether lack of CFTR causes cell-intrinsic abnormalities present at birth versus inflammation-dependent abnormalities is unclear. We performed a single-cell RNA-sequencing census of microdissected small airways from newborn CF pigs, which recapitulate CF host defense defects and pathology over time. Lack of minimally affected the transcriptome of large and small airways at birth, suggesting that infection and inflammation drive transcriptomic abnormalities in advanced CF. Importantly, common small airway epithelial cell types expressed a markedly different transcriptome than corresponding large airway cell types. Quantitative immunohistochemistry and electrophysiology of small airway epithelia demonstrated basal cells that reach the apical surface and a water and ion transport advantage. This single cell atlas highlights the archetypal nature of airway epithelial cells with location-dependent gene expression and function.