| Autor: |
Okuda K; Marsico Lung Institute/Cystic Fibrosis Research Center and., Dang H; Marsico Lung Institute/Cystic Fibrosis Research Center and., Kobayashi Y; Department of Cell Biology, School of Medicine, Duke University, Durham, North Carolina., Carraro G; Department of Medicine, Cedars-Sinai Medical Center, Los Angeles, California., Nakano S; Marsico Lung Institute/Cystic Fibrosis Research Center and., Chen G; Marsico Lung Institute/Cystic Fibrosis Research Center and., Kato T; Marsico Lung Institute/Cystic Fibrosis Research Center and., Asakura T; Marsico Lung Institute/Cystic Fibrosis Research Center and., Gilmore RC; Marsico Lung Institute/Cystic Fibrosis Research Center and., Morton LC; Marsico Lung Institute/Cystic Fibrosis Research Center and., Lee RE; Marsico Lung Institute/Cystic Fibrosis Research Center and., Mascenik T; Marsico Lung Institute/Cystic Fibrosis Research Center and., Yin WN; Marsico Lung Institute/Cystic Fibrosis Research Center and., Barbosa Cardenas SM; Marsico Lung Institute/Cystic Fibrosis Research Center and., O'Neal YK; Marsico Lung Institute/Cystic Fibrosis Research Center and., Minnick CE; Marsico Lung Institute/Cystic Fibrosis Research Center and., Chua M; Marsico Lung Institute/Cystic Fibrosis Research Center and., Quinney NL; Marsico Lung Institute/Cystic Fibrosis Research Center and., Gentzsch M; Marsico Lung Institute/Cystic Fibrosis Research Center and., Anderson CW; Center for Gastrointestinal Biology and Disease, School of Medicine, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina., Ghio A; Clinical Research Branch, National Health and Environmental Effects Research Laboratory, U.S. Environmental Protection Agency, Chapel Hill, North Carolina., Matsui H; Center for Respiratory Disease, National Hospital Organization Tokyo Hospital, Kiyose, Tokyo, Japan; and., Nagase T; Department of Respiratory Medicine, The University of Tokyo, Tokyo, Japan., Ostrowski LE; Marsico Lung Institute/Cystic Fibrosis Research Center and., Grubb BR; Marsico Lung Institute/Cystic Fibrosis Research Center and., Olsen JC; Marsico Lung Institute/Cystic Fibrosis Research Center and., Randell SH; Marsico Lung Institute/Cystic Fibrosis Research Center and., Stripp BR; Department of Medicine, Cedars-Sinai Medical Center, Los Angeles, California., Tata PR; Department of Cell Biology, School of Medicine, Duke University, Durham, North Carolina., O'Neal WK; Marsico Lung Institute/Cystic Fibrosis Research Center and., Boucher RC; Marsico Lung Institute/Cystic Fibrosis Research Center and. |
| Abstrakt: |
Rationale: Identification of the specific cell types expressing CFTR (cystic fibrosis [CF] transmembrane conductance regulator) is required for precision medicine therapies for CF. However, a full characterization of CFTR expression in normal human airway epithelia is missing. Objectives: To identify the cell types that contribute to CFTR expression and function within the proximal-distal axis of the normal human lung. Methods: Single-cell RNA (scRNA) sequencing (scRNA-seq) was performed on freshly isolated human large and small airway epithelial cells. scRNA in situ hybridization (ISH) and single-cell qRT-PCR were performed for validation. In vitro culture systems correlated CFTR function with cell types. Lentiviruses were used for cell type-specific transduction of wild-type CFTR in CF cells. Measurements and Main Results: scRNA-seq identified secretory cells as dominating CFTR expression in normal human large and, particularly, small airway superficial epithelia, followed by basal cells. Ionocytes expressed the highest CFTR levels but were rare, whereas the expression in ciliated cells was infrequent and low. scRNA ISH and single-cell qRT-PCR confirmed the scRNA-seq findings. CF lungs exhibited distributions of CFTR and ionocytes similar to those of normal control subjects. CFTR mediated Cl - secretion in cultures tracked secretory cell, but not ionocyte, densities. Furthermore, the nucleotide-purinergic regulatory system that controls CFTR-mediated hydration was associated with secretory cells and not with ionocytes. Lentiviral transduction of wild-type CFTR produced CFTR-mediated Cl - secretion in CF airway secretory cells but not in ciliated cells. Conclusions: Secretory cells dominate CFTR expression and function in human airway superficial epithelia. CFTR therapies may need to restore CFTR function to multiple cell types, with a focus on secretory cells. |
