title: Comparative Proteomic Characterization of 4 Human Liver-Derived Single Cell Culture Models Reveals Significant Variation in the Capacity for Drug Disposition, Bioactivation, and Detoxication.
abstract: In vitro preclinical models for the assessment of drug-induced liver injury (DILI) are usually based on cryopreserved primary human hepatocytes (cPHH) or human hepatic tumor-derived cell lines; however, it is unclear how well such cell models reflect the normal function of liver cells. The physiological, pharmacological, and toxicological phenotyping of available cell-based systems is necessary in order to decide the testing purpose for which they are fit. We have therefore undertaken a global proteomic analysis of 3 human-derived hepatic cell lines (HepG2, Upcyte, and HepaRG) in comparison with cPHH with a focus on drug metabolizing enzymes and transport proteins (DMETs), as well as Nrf2-regulated proteins. In total, 4946 proteins were identified, of which 2722 proteins were common across all cell models, including 128 DMETs. Approximately 90% reduction in expression of cytochromes P450 was observed in HepG2 and Upcyte cells, and approximately 60% in HepaRG cells relative to cPHH. Drug transporter expression was also lower compared with cPHH with the exception of MRP3 and P-gp (MDR1) which appeared to be significantly expressed in HepaRG cells. In contrast, a high proportion of Nrf2-regulated proteins were more highly expressed in the cell lines compared with cPHH. The proteomic database derived here will provide a rational basis for the context-specific selection of the most appropriate 'hepatocyte-like' cell for the evaluation of particular cellular functions associated with DILI and, at the same time, assist in the construction of a testing paradigm which takes into account the in vivo disposition of a new drug.
authors.affiliation: *Department of Molecular and Clinical Pharmacology, MRC Centre for Drug Safety Science, Liverpool L69 3GE, UK;
authors.forename: Rowena L C
authors.initials: RL
authors.lastname: Sison-Young
authors.affiliation: *Department of Molecular and Clinical Pharmacology, MRC Centre for Drug Safety Science, Liverpool L69 3GE, UK;
authors.forename: Dimitra
authors.initials: D
authors.lastname: Mitsa
authors.affiliation: *Department of Molecular and Clinical Pharmacology, MRC Centre for Drug Safety Science, Liverpool L69 3GE, UK;
authors.forename: Rosalind E
authors.initials: RE
authors.lastname: Jenkins
authors.affiliation: *Department of Molecular and Clinical Pharmacology, MRC Centre for Drug Safety Science, Liverpool L69 3GE, UK;
authors.forename: David
authors.initials: D
authors.lastname: Mottram
authors.affiliation: KaLy-Cell, Research & Development, 67115 Plobsheim, France;
authors.forename: Eliane
authors.initials: E
authors.lastname: Alexandre
authors.affiliation: KaLy-Cell, Research & Development, 67115 Plobsheim, France;
authors.forename: Lysiane
authors.initials: L
authors.lastname: Richert
authors.affiliation: Biologie Servier, 905 Route de Saran, 45520, Gidy, France;
authors.forename: Hélène
authors.initials: H
authors.lastname: Aerts
authors.affiliation: Biologie Servier, 905 Route de Saran, 45520, Gidy, France;
authors.forename: Richard J
authors.initials: RJ
authors.lastname: Weaver
authors.affiliation: *Department of Molecular and Clinical Pharmacology, MRC Centre for Drug Safety Science, Liverpool L69 3GE, UK;
authors.forename: Robert P
authors.initials: RP
authors.lastname: Jones
authors.affiliation: North Western Hepatobiliary Unit, Aintree University Hospital NHS Foundation Trust, Longmoor Lane, Liverpool L9 7AL, UK;
authors.forename: Esther
authors.initials: E
authors.lastname: Johann
authors.affiliation: Merck KGaA, Merck Serono, Non-Clinical Safety, 64293 Darmstadt, Germany; and.
authors.forename: Philip G
authors.initials: PG
authors.lastname: Hewitt
authors.affiliation: Section of Pharmacogenetics, Department of Physiology and Pharmacology, Karolinska Institutet, SE-17177 Stockholm, Sweden.
authors.forename: Magnus
authors.initials: M
authors.lastname: Ingelman-Sundberg
authors.affiliation: *Department of Molecular and Clinical Pharmacology, MRC Centre for Drug Safety Science, Liverpool L69 3GE, UK;
authors.forename: Christopher E P
authors.initials: CE
authors.lastname: Goldring
authors.affiliation: *Department of Molecular and Clinical Pharmacology, MRC Centre for Drug Safety Science, Liverpool L69 3GE, UK; neilk@liverpool.ac.uk.
authors.forename: Neil R
authors.initials: NR
authors.lastname: Kitteringham
authors.affiliation: *Department of Molecular and Clinical Pharmacology, MRC Centre for Drug Safety Science, Liverpool L69 3GE, UK;
authors.forename: B Kevin
authors.initials: BK
authors.lastname: Park
mesh_terms: D015153:Blotting, Western; D002478:Cells, Cultured; D056945:Hep G2 Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D022781:Hepatocytes / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D008099:Liver / Q000187:drug effects* / Q000378:metabolism; D008954:Models, Biological; D040901:Proteomics / Q000379:methods*
keywords: DILI; DMETS; hepatocytes; metabolism