Cytochrome P450 expression, induction and activity in human induced pluripotent stem cell-derived intestinal organoids and comparison with primary human intestinal epithelial cells and Caco-2 cells

• Published in: Archives of toxicology 2021-03, Vol.95 (3), p.907-922
• Main Authors: Janssen, Aafke W. F., Duivenvoorde, Loes P. M., Rijkers, Deborah, Nijssen, Rosalie, Peijnenburg, Ad A. C. M., van der Zande, Meike, Louisse, Jochem
• Format: Article
• Language: English
• Subjects: Adenocarcinoma; Biomedical and Life Sciences; Biomedicine; Biotransformation; Computer architecture; Cytochrome; Cytochrome P450; Cytochromes P450; Enterocytes; Environmental Health; Enzymes; Epithelial cells; Epithelium; Gene expression; Goblet cells; Intestine; Occupational Medicine/Industrial Medicine; Organoids; Paneth cells; Pharmacology/Toxicology; Phenobarbital; Pluripotency; Rifampin; Statistical analysis; Stem cells; Tight junctions; Toxicokinetics and Metabolism
• ISSN: 0340-5761; 1432-0738
• DOI: 10.1007/s00204-020-02953-6

Human intestinal organoids (HIOs) are a promising in vitro model consisting of different intestinal cell types with a 3D microarchitecture resembling native tissue. In the current study, we aimed to assess the expression of the most common intestinal CYP enzymes in a human induced pluripotent stem cell (hiPSC)-derived HIO model, and the suitability of that model to study chemical-induced changes in CYP expression and activity. We compared this model with the commonly used human colonic adenocarcinoma cell line Caco-2 and with a human primary intestinal epithelial cell (IEC)-based model, closely resembling in vivo tissue. We optimized an existing protocol to differentiate hiPSCs into HIOs and demonstrated that obtained HIOs contain a polarized epithelium with tight junctions consisting of enterocytes, goblet cells, enteroendocrine cells and Paneth cells. We extensively characterized the gene expression of CYPs and activity of CYP3A4/5, indicating relatively high gene expression levels of the most important intestinal CYP enzymes in HIOs compared to the other models. Furthermore, we showed that CYP1A1 and CYP1B1 were induced by β-naphtoflavone in all three models, whereas CYP3A4 was induced by phenobarbital and rifampicin in HIOs, in the IEC-based model (although not statistically significant), but not in Caco-2 cells. Interestingly, CYP2B6 expression was not induced in any of the models by the well-known liver CYP2B6 inducer phenobarbital. In conclusion, our study indicates that hiPSC-based HIOs are a useful in vitro intestinal model to study biotransformation of chemicals in the intestine.