Predicting drug-induced liver injury in a preclinical setting remains challenging, as cultured primary human hepatocytes (PHHs), pluripotent stem cell-derived hepatocyte-like cells (HLCs), and hepatoma cells exhibit poor drug biotransformation capacity. We here demonstrate that hepatic functionality depends more on cellular metabolism and extracellular nutrients than on developmental regulators. Specifically, we demonstrate that increasing extracellular amino acids beyond the nutritional need of HLCs and HepG2 cells induces glucose independence, mitochondrial function, and the acquisition of a transcriptional profile that is closer to PHHs. Moreover, we show that these high levels of amino acids are sufficient to drive HLC and HepG2 drug biotransformation and liver-toxin sensitivity to levels similar to those in PHHs. In conclusion, we provide data indicating that extracellular nutrient levels represent a major determinant of cellular maturity and can be utilized to guide stem cell differentiation to the hepatic lineage.
Bibliographical noteFunding Information:
S.M.F. acknowledges funding support from Marie Curie–CIG and FWO–Odysseus II. Funding to C.M.V. was from KU Leuven (ETH-C1900-PFC32/17/053 and C14/17/111 -3D-MuSYC), FWO–G067314N and FWO G.0D99.17N, IWT-SBO-HEPSTEM, IWT-SBO-HILIM-3D, EC-SEURAT-1-HEMIBIO, and H2020-EuTOX-Risk.
We thank Dr. Stefaan Soenen and Dr. Bella Manshian of the KU Leuven Biomedical MRI unit for their help in setting up the Operetta based toxicology screens. We also acknowledge the work of Professor Arvind Patel from the university of Glasgow in generating the immortalized HHL5 cell line and allowing us to use it for this manuscript. We greatly acknowledge the sequencing performed by VIB Nucleomics Core (www. nucleomics.be). R.B. was funded by IWT fellowship SB-121393 and by H2020-EuTOX-Risk. M.K. received funding from the European Union’s Horizon 2020 research and innovation program under the Marie Sklodowska-Curie-IF grant agreement number 657701. T.T. was funded by FWO (1185918N). L.O. was funded by IWT/OZM/090838, IACS BPAMER3/08/04, and Government of Aragon FMI048/08. P.R. was supported by Postdoctoral fellowship from Research Foundation - Flanders (FWO). P.R. is supported by Clinical Mandate from Belgian Foundation against Cancer (Stichting tegen Kanker) and receives speaking and consultancy fees from MSD Belgium. On behalf of D.C., the University of Leuven and University Hospitals Leuven have received research grants, travel and conference bursaries, speaker fees, and advisory board compensations from Genzyme/Sanofi, Shire, Actelion, Bayer, Roche, BMS, Schering-Plough, Synageva, Chiesi, and Alexion. None of these create a conflict of interest for the work represented here.
© 2020, The Author(s).
- Amino Acids/metabolism
- Carcinoma, Hepatocellular/metabolism
- Cell Differentiation/physiology
- Cell Line, Tumor
- Cytochrome P-450 CYP3A
- Cytochrome P-450 Enzyme System/metabolism
- Gene Knockout Techniques
- Hep G2 Cells
- Hepatocyte Nuclear Factor 1-alpha
- Hepatocyte Nuclear Factor 3-gamma
- High-Throughput Screening Assays
- Homeodomain Proteins
- Liver Neoplasms/metabolism
- Metabolic Engineering
- Metabolic Networks and Pathways
- Middle Aged
- Pluripotent Stem Cells
- Stem Cells
- Tumor Suppressor Proteins
PubMed: MeSH publication types
- Research Support, Non-U.S. Gov't
- Journal Article