Traditional toxicity risk assessment approaches have until recently focussed mainly on histochemical readouts for cell death. Modern toxicology methods attempt to deduce a mechanistic understanding of pathways involved in the development of toxicity, by using transcriptomics and other big data-driven methods such as high-content screening. Here, we used a recently described optimised method to differentiate human induced pluripotent stem cells (hiPSCs) to hepatocyte-like cells (HLCs), to assess their potential to classify hepatotoxic and non-hepatotoxic chemicals and their use in mechanistic toxicity studies. The iPSC-HLCs could accurately classify chemicals causing acute hepatocellular injury, and the transcriptomics data on treated HLCs obtained by TempO-Seq technology linked the cytotoxicity to cellular stress pathways, including oxidative stress and unfolded protein response (UPR). Induction of these stress pathways in response to amiodarone, diclofenac, and ibuprofen, was demonstrated to be concentration and time dependent. The transcriptomics data on diclofenac-treated HLCs were found to be more sensitive in detecting differentially expressed genes in response to treatment, as compared to existing datasets of other diclofenac-treated in vitro hepatocyte models. Hence iPSC-HLCs generated by transcription factor overexpression and in metabolically optimised medium appear suitable for chemical toxicity detection as well as mechanistic toxicity studies.
Bibliographical noteFunding Information:
Funding: This work was funded by in3 Marie Skłodowska-Curie Action-Innovative Training Network under grant no. 721975 (fellowship to S.G.) and Fonds Wetenschappelijk Onderzoek (FWO) 1S33916N (J.D.S.). T.T. was funded by FWO (1185918N), and F.N. by FWO (1151318N). C.M.V. received funding from KU Leuven C14/17/111-3D-MuSYC, IWT-HILIM-3D, FWO-SBO-QPG-359638-iPSC-LIMIC, FWO G0D9917N; and funding from the European Union’s Horizon 2020 research and innovation programme under grant agreement No 681002 (EU-ToxRisk).
© 2021 by the authors. Licensee MDPI, Basel, Switzerland.
- ER stress
- In vitro toxicology
- Mechanistic toxicity
- Stem cell derived