title: Transcriptional, Functional, and Mechanistic Comparisons of Stem Cell-Derived Hepatocytes, HepaRG Cells, and Three-Dimensional Human Hepatocyte Spheroids as Predictive In Vitro Systems for Drug-Induced Liver Injury.
abstract: Reliable and versatile hepatic in vitro systems for the prediction of drug pharmacokinetics and toxicity are essential constituents of preclinical safety assessment pipelines for new medicines. Here, we compared three emerging cell systems-hepatocytes derived from induced pluripotent stem cells, HepaRG cells, and three-dimensional primary human hepatocyte (PHH) spheroids-at transcriptional and functional levels in a multicenter study to evaluate their potential as predictive models for drug-induced hepatotoxicity. Transcriptomic analyses revealed widespread gene expression differences between the three cell models, with 8148 of 17,462 analyzed genes (47%) being differentially expressed. Expression levels of genes involved in the metabolism of endogenous as well as xenobiotic compounds were significantly elevated in PHH spheroids, whereas genes involved in cell division and endocytosis were significantly upregulated in HepaRG cells and hepatocytes derived from induced pluripotent stem cells, respectively. Consequently, PHH spheroids were more sensitive to a panel of drugs with distinctly different toxicity mechanisms, an effect that was amplified by long-term exposure using repeated treatments. Importantly, toxicogenomic analyses revealed that transcriptomic changes in PHH spheroids were in compliance with cholestatic, carcinogenic, or steatogenic in vivo toxicity mechanisms at clinically relevant drug concentrations. Combined, the data reveal important phenotypic differences between the three cell systems and suggest that PHH spheroids can be used for functional investigations of drug-induced liver injury in vivo in humans.
authors.affiliation: Section of Pharmacogenetics, Department of Physiology and Pharmacology, Karolinska Institutet, Stockholm, Sweden (C.C.B., V.M.L., S.U.V., T.B.A., M.I.-S.); Cardiovascular and Metabolic Diseases, Innovative Medicines and Early Development Biotech Unit, AstraZeneca, Mölndal, Sweden (H.P., T.B.A.); and CXR Biosciences Ltd., Dundee, United Kingdom (R.D.).
authors.forename: Catherine C
authors.initials: CC
authors.lastname: Bell
authors.affiliation: Section of Pharmacogenetics, Department of Physiology and Pharmacology, Karolinska Institutet, Stockholm, Sweden (C.C.B., V.M.L., S.U.V., T.B.A., M.I.-S.); Cardiovascular and Metabolic Diseases, Innovative Medicines and Early Development Biotech Unit, AstraZeneca, Mölndal, Sweden (H.P., T.B.A.); and CXR Biosciences Ltd., Dundee, United Kingdom (R.D.) volker.lauschke@ki.se.
authors.forename: Volker M
authors.initials: VM
authors.lastname: Lauschke
authors.affiliation: Section of Pharmacogenetics, Department of Physiology and Pharmacology, Karolinska Institutet, Stockholm, Sweden (C.C.B., V.M.L., S.U.V., T.B.A., M.I.-S.); Cardiovascular and Metabolic Diseases, Innovative Medicines and Early Development Biotech Unit, AstraZeneca, Mölndal, Sweden (H.P., T.B.A.); and CXR Biosciences Ltd., Dundee, United Kingdom (R.D.).
authors.forename: Sabine U
authors.initials: SU
authors.lastname: Vorrink
authors.affiliation: Section of Pharmacogenetics, Department of Physiology and Pharmacology, Karolinska Institutet, Stockholm, Sweden (C.C.B., V.M.L., S.U.V., T.B.A., M.I.-S.); Cardiovascular and Metabolic Diseases, Innovative Medicines and Early Development Biotech Unit, AstraZeneca, Mölndal, Sweden (H.P., T.B.A.); and CXR Biosciences Ltd., Dundee, United Kingdom (R.D.).
authors.forename: Henrik
authors.initials: H
authors.lastname: Palmgren
authors.affiliation: Section of Pharmacogenetics, Department of Physiology and Pharmacology, Karolinska Institutet, Stockholm, Sweden (C.C.B., V.M.L., S.U.V., T.B.A., M.I.-S.); Cardiovascular and Metabolic Diseases, Innovative Medicines and Early Development Biotech Unit, AstraZeneca, Mölndal, Sweden (H.P., T.B.A.); and CXR Biosciences Ltd., Dundee, United Kingdom (R.D.).
authors.forename: Rodger
authors.initials: R
authors.lastname: Duffin
authors.affiliation: Section of Pharmacogenetics, Department of Physiology and Pharmacology, Karolinska Institutet, Stockholm, Sweden (C.C.B., V.M.L., S.U.V., T.B.A., M.I.-S.); Cardiovascular and Metabolic Diseases, Innovative Medicines and Early Development Biotech Unit, AstraZeneca, Mölndal, Sweden (H.P., T.B.A.); and CXR Biosciences Ltd., Dundee, United Kingdom (R.D.).
authors.forename: Tommy B
authors.initials: TB
authors.lastname: Andersson
authors.affiliation: Section of Pharmacogenetics, Department of Physiology and Pharmacology, Karolinska Institutet, Stockholm, Sweden (C.C.B., V.M.L., S.U.V., T.B.A., M.I.-S.); Cardiovascular and Metabolic Diseases, Innovative Medicines and Early Development Biotech Unit, AstraZeneca, Mölndal, Sweden (H.P., T.B.A.); and CXR Biosciences Ltd., Dundee, United Kingdom (R.D.).
authors.forename: Magnus
authors.initials: M
authors.lastname: Ingelman-Sundberg
mesh_terms: D018929:Cell Culture Techniques / Q000379:methods*; D056486:Chemical and Drug Induced Liver Injury / Q000191:economics / Q000453:epidemiology / Q000209:etiology; D020869:Gene Expression Profiling; D022781:Hepatocytes / Q000187:drug effects / Q000378:metabolism*; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000187:drug effects / Q000378:metabolism*; D020128:Inhibitory Concentration 50; D008954:Models, Biological; D018874:Spheroids, Cellular / Q000187:drug effects / Q000378:metabolism*; D015262:Xenobiotics / Q000378:metabolism / Q000633:toxicity*
chemical_list: D015262:Xenobiotics