In vitro platform to model the function of ionocytes in the human airway epithelium
Issued Date
2024-12-01
Resource Type
ISSN
14659921
eISSN
1465993X
Scopus ID
2-s2.0-85191261579
Journal Title
Respiratory Research
Volume
25
Issue
1
Rights Holder(s)
SCOPUS
Bibliographic Citation
Respiratory Research Vol.25 No.1 (2024)
Suggested Citation
Vilà-González M., Pinte L., Fradique R., Causa E., Kool H., Rodrat M., Morell C.M., Al-Thani M., Porter L., Guo W., Maeshima R., Hart S.L., McCaughan F., Granata A., Sheppard D.N., Floto R.A., Rawlins E.L., Cicuta P., Vallier L. In vitro platform to model the function of ionocytes in the human airway epithelium. Respiratory Research Vol.25 No.1 (2024). doi:10.1186/s12931-024-02800-7 Retrieved from: https://repository.li.mahidol.ac.th/handle/123456789/98196
Title
In vitro platform to model the function of ionocytes in the human airway epithelium
Author's Affiliation
Berliner Institut für Gesundheitsforschung
Department of Physics
Department of Medicine
Department of Clinical Neurosciences
Fundació Institut d'Investigació Sanitaria Illes Balears
Royal Papworth Hospital NHS Foundation Trust
University of Cambridge
Max Planck Institute for Molecular Genetics
Wellcome Trust/ Cancer Research UK Gurdon Institute
Humanitas Research Hospital
University of Bristol
UCL Great Ormond Street Institute of Child Health
Universitat de les Illes Balears
Institute of Molecular Biosciences, Mahidol University
Department of Physics
Department of Medicine
Department of Clinical Neurosciences
Fundació Institut d'Investigació Sanitaria Illes Balears
Royal Papworth Hospital NHS Foundation Trust
University of Cambridge
Max Planck Institute for Molecular Genetics
Wellcome Trust/ Cancer Research UK Gurdon Institute
Humanitas Research Hospital
University of Bristol
UCL Great Ormond Street Institute of Child Health
Universitat de les Illes Balears
Institute of Molecular Biosciences, Mahidol University
Corresponding Author(s)
Other Contributor(s)
Abstract
Background: Pulmonary ionocytes have been identified in the airway epithelium as a small population of ion transporting cells expressing high levels of CFTR (cystic fibrosis transmembrane conductance regulator), the gene mutated in cystic fibrosis. By providing an infinite source of airway epithelial cells (AECs), the use of human induced pluripotent stem cells (hiPSCs) could overcome some challenges of studying ionocytes. However, the production of AEC epithelia containing ionocytes from hiPSCs has proven difficult. Here, we present a platform to produce hiPSC-derived AECs (hiPSC-AECs) including ionocytes and investigate their role in the airway epithelium. Methods: hiPSCs were differentiated into lung progenitors, which were expanded as 3D organoids and matured by air-liquid interface culture as polarised hiPSC-AEC epithelia. Using CRISPR/Cas9 technology, we generated a hiPSCs knockout (KO) for FOXI1, a transcription factor that is essential for ionocyte specification. Differences between FOXI1 KO hiPSC-AECs and their wild-type (WT) isogenic controls were investigated by assessing gene and protein expression, epithelial composition, cilia coverage and motility, pH and transepithelial barrier properties. Results: Mature hiPSC-AEC epithelia contained basal cells, secretory cells, ciliated cells with motile cilia, pulmonary neuroendocrine cells (PNECs) and ionocytes. There was no difference between FOXI1 WT and KO hiPSCs in terms of their capacity to differentiate into airway progenitors. However, FOXI1 KO led to mature hiPSC-AEC epithelia without ionocytes with reduced capacity to produce ciliated cells. Conclusion: Our results suggest that ionocytes could have role beyond transepithelial ion transport by regulating epithelial properties and homeostasis in the airway epithelium.