Differentiation of Human Induced Pluripotent Stem Cells from Patients with Severe COPD into Functional Airway Epithelium

• Published in: Cells (Basel, Switzerland) Switzerland), 2022-08, Vol.11 (15), p.2422
• Main Authors: Ahmed, Engi, Fieldes, Mathieu, Bourguignon, Chloé, Mianné, Joffrey, Petit, Aurélie, Jory, Myriam, Cazevieille, Chantal, Boukhaddaoui, Hassan, Garnett, James P, Hirtz, Christophe, Massiera, Gladys, Vachier, Isabelle, Assou, Said, Bourdin, Arnaud, De Vos, John
• Format: Article
• Language: English
• Subjects: airway epithelium; Analysis; Cell differentiation; Chemokine receptors; Chronic obstructive pulmonary disease; Cloning; CXCR4 protein; Development and progression; disease modeling; Endoderm; Epithelium; Epithelium - metabolism; Flow cytometry; Flow velocity; Foregut; Growth factors; Health aspects; Homeobox; human induced pluripotent stem cells; Humans; Induced Pluripotent Stem Cells; Kinases; Leukocytes (mononuclear); Leukocytes, Mononuclear - pathology; Life Sciences; Lung diseases; Lung diseases, Obstructive; Obstructive lung disease; Peripheral blood mononuclear cells; Phosphatase; Pluripotency; Proteins; Pulmonary Disease, Chronic Obstructive - metabolism; Respiratory Mucosa - pathology; Respiratory tract; Stem cells; Thyroid transcription factor 1; Transcription factors
• ISSN: 2073-4409; 2073-4409
• DOI: 10.3390/cells11152422

Background: Chronic Obstructive Pulmonary Disease (COPD), a major cause of mortality and disability, is a complex disease with heterogeneous and ill-understood biological mechanisms. Human induced pluripotent stem cells (hiPSCs) are a promising tool to model human disease, including the impact of genetic susceptibility. Methods: We developed a simple and reliable method for reprogramming peripheral blood mononuclear cells into hiPSCs and to differentiate them into air−liquid interface bronchial epithelium within 45 days. Importantly, this method does not involve any cell sorting step. We reprogrammed blood cells from one healthy control and three patients with very severe COPD. Results: The mean cell purity at the definitive endoderm and ventral anterior foregut endoderm (vAFE) stages was >80%, assessed by quantifying C-X-C Motif Chemokine Receptor 4/SRY-Box Transcription Factor 17 (CXCR4/SOX17) and NK2 Homeobox 1 (NKX2.1) expression, respectively. vAFE cells from all four hiPSC lines differentiated into bronchial epithelium in air−liquid interface conditions, with large zones covered by beating ciliated, basal, goblets, club cells and neuroendocrine cells, as found in vivo. The hiPSC-derived airway epithelium (iALI) from patients with very severe COPD and from the healthy control were undistinguishable. Conclusions: iALI bronchial epithelium is ready for better understanding lung disease pathogenesis and accelerating drug discovery.