Cytochrome P450 expression, induction and activity in human induced pluripotent stem cell-derived intestinal organoids a
- PDF / 7,001,219 Bytes
- 16 Pages / 595.276 x 790.866 pts Page_size
- 105 Downloads / 214 Views
TOXICOKINETICS AND METABOLISM
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 Aafke W. F. Janssen1 · Loes P. M. Duivenvoorde1 · Deborah Rijkers1 · Rosalie Nijssen1 · Ad A. C. M. Peijnenburg1 · Meike van der Zande1 · Jochem Louisse1 Received: 3 July 2020 / Accepted: 12 November 2020 © The Author(s) 2020
Abstract 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. Keywords Cytochrome P450 (CYP) · Gastrointestinal tract · Organotypic models · Stem cells · Toxicology Abbreviations AhR Aryl hydrocarbon receptor CAR Constitutive androstane receptor CYP Cytochrome P450 DE Definitive endoderm HG Hindgut endoderm IEC Intestinal epithelial cells Meike van der Zande and Jochem Louisse contributed equally to this work. Electronic supplementary material The online version of this article (https://doi.org/10.1007/s00204-020-02953-6) contains supplementary material, which is available to authorized users. * Aafke W. F. Janssen [email protected] 1
Wageningen Food Safety Research (WFSR), Wageningen University and Research, Akkermaalsbos 2, 6708 WB Wageningen, The Netherlands
hiPSCs Human induced pluripotent stem cells HIOs Human intestinal organoids PXR Pregnane X receptor
Introduction Cytochrome P450 (CYP) enzymes play crucial roles in detoxification or bioactivation of chemicals. Of the 18 CYP P450 families, members of t
Data Loading...
