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Review Articles

Through a glass, darkly? HepaRG and HepG2 cells as models of human phase I drug metabolism

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Pages 46-62 | Received 19 Jan 2022, Accepted 03 Feb 2022, Published online: 23 Feb 2022

References

  • Aninat C, Piton A, Glaise D, Le Charpentier T, Langouët S, Morel F, Guguen-Guillouzo C, Guillouzo A. 2006. Expression of cytochromes P450, conjugating enzymes and nuclear receptors in human hepatoma HepaRG cells. Drug Metab Dispos. 34(1):75–83.
  • Anthérieu S, Chesné C, Li R, Camus S, Lahoz A, Picazo L, Turpeinen M, Tolonen A, Uusitalo J, Guguen-Guillouzo C, et al. 2010. Stable expression, activity, and inducibility of cytochromes P450 in differentiated HepaRG cells. Drug Metab Dispos. 38(3):516–525.
  • Berger B, Donzelli M, Maseneni S, Boess F, Roth A, Krähenbühl S, Haschke M. 2016. Comparison of liver cell models using the basel phenotyping cocktail. Front Pharmacol. 7:443.
  • Berger E, Vega N, Weiss-Gayet M, Géloën A. 2015. Gene network analysis of glucose linked signaling pathways and their role in human hepatocellular carcinoma cell growth and survival in HuH7 and HepG2 cell lines. Biomed Res Int. 2015:821761.
  • Bernasconi C, Pelkonen O, Andersson TB, Strickland J, Wilk-Zasadna I, Asturiol D, Cole T, Liska R, Worth A, Müller-Vieira U, et al. 2019. Validation of in vitro methods for human cytochrome P450 enzyme induction: outcome of a multi-laboratory study. Toxicol In Vitro. 60:212–228.
  • Bonn B, Svanberg P, Janefeldt A, Hultman I, Grime K. 2016. Determination of human hepatocyte intrinsic clearance for slowly metabolized compounds: comparison of a primary hepatocyte/stromal cell co-culture with plated primary hepatocytes and HepaRG. Drug Metab Dispos. 44(4):527–533.
  • Buccitelli C, Selbach M. 2020. mRNAs, proteins and the emerging principles of gene expression control. Nat Rev Genet. 21(10):630–644.
  • Chen L, Bao Y, Piekos SC, Zhu K, Zhang L, Zhong XB. 2018. A transcriptional regulatory network containing nuclear receptors and long noncoding RNAs controls basal and drug-induced expression of cytochrome P450s in HepaRG cells. Mol Pharmacol. 94(1):749–759.
  • Choi JM, Oh SJ, Lee JY, Jeon JS, Ryu CS, Kim YM, Lee K, Kim SK. 2015. Prediction of drug-induced liver injury in HepG2 cells cultured with human liver microsomes. Chem Res Toxicol. 28(5):872–885.
  • Chu V, Einolf HJ, Evers R, Kumar G, Moore D, Ripp S, Silva J, Sinha V, Sinz M, Skerjanec A. 2009. In vitro and in vivo induction of cytochrome P450: a survey of the current practices and recommendations: a pharmaceutical research and manufacturers of America perspective. Drug Metab Dispos. 37(7):1339–1354.
  • Dawson JR, Adams DJ, Wolf CR. 1985. Induction of drug metabolizing enzymes in human liver cell line HepG2. FEBS Lett. 183(2):219–222.
  • Dearfield KL, Jacobson-Kram D, Brown NA, Williams JR. 1983. Evaluation of a human hepatoma cell line as a target cell in genetic toxicology. Mutat Res. 108(1–3):437–449.
  • Easterbrook J, Lu C, Sakai Y, Li AP. 2001. Effects of organic solvents on the activities of cytochrome P450 isoforms, UDP-dependent glucuronyl transferase, and phenol sulfotransferase in human hepatocytes. Drug Metab Dispos. 29(2):141–144.
  • El-Sankary W, Plant NJ, Gibson GG, Moore DJ. 2000. Regulation of the CYP3A4 gene by hydrocortisone and xenobiotics: role of the glucocorticoid and pregnane X receptors. Drug Metab Dispos. 28(5):493–496.
  • Gerets HH, Tilmant K, Gerin B, Chanteux H, Depelchin BO, Dhalluin S, Atienzar FA. 2012. Characterization of primary human hepatocytes, HepG2 cells, and HepaRG cells at the mRNA level and CYP activity in response to inducers and their predictivity for the detection of human hepatotoxins. Cell Biol Toxicol. 28(2):69–87.
  • Gripon P, Rumin S, Urban S, Le Seyec J, Glaise D, Cannie I, Guyomard C, Lucas J, Trepo C, Guguen-Guillouzo C. 2002. Infection of a human hepatoma cell line by hepatitis B virus. Proc Natl Acad Sci USA. 99(24):15655–15660.
  • Hammer H, Schmidt F, Marx-Stoelting P, Pötz O, Braeuning A. 2021. Cross-species analysis of hepatic cytochrome P450 and transport protein expression. Arch Toxicol. 95(1):117–133.
  • Harmsen S, Koster AS, Beijnen JH, Schellens JH, Meijerman I. 2008. Comparison of two immortalized human cell lines to study nuclear receptor-mediated CYP3A4 induction. Drug Metab Dispos. 36(6):1166–1171.
  • Henderson CJ, Kapelyukh Y, Scheer N, Rode A, McLaren AW, MacLeod AK, Lin D, Wright J, Stanley LA, Wolf CR. 2019. An extensively humanized mouse model to predict pathways of drug disposition and drug/drug interactions, and to facilitate design of clinical trials. Drug Metab Dispos. 47(6):601–615.
  • Hurrell T, Segeritz CP, Vallier L, Lilley KS, Cromarty AD. 2018. Proteomic comparison of various hepatic cell cultures for preclinical safety pharmacology. Toxicol Sci. 164(1):229–239.
  • Jackson JP, Li L, Chamberlain ED, Wang H, Ferguson SS. 2016. Contextualizing hepatocyte functionality of cryopreserved HepaRG cell cultures. Drug Metab Dispos. 44(9):1463–1479.
  • Jossé R, Aninat C, Glaise D, Dumont J, Fessard V, Morel F, Poul JM, Guguen-Guillouzo C, Guillouzo A. 2008. Long-term functional stability of human HepaRG hepatocytes and use for chronic toxicity and genotoxicity studies. Drug Metab Dispos. 36(6):1111–1118.
  • Kanebratt KP, Andersson TB. 2008a. Evaluation of HepaRG cells as an in vitro model for human drug metabolism studies. Drug Metab Dispos. 36(7):1444–1452.
  • Kanebratt KP, Andersson TB. 2008b. HepaRG cells as an in vitro model for evaluation of cytochrome P450 induction in humans. Drug Metab Dispos. 36(1):137–145.
  • Kaneko A, Kato M, Sekiguchi N, Mitsui T, Takeda K, Aso Y. 2009. In vitro model for the prediction of clinical CYP3A4 induction using HepaRG cells. Xenobiotica. 39(11):803–810.
  • Kelly JH, Sussman NL. 2000. A fluorescent cell-based assay for cytochrome P-450 isozyme 1A2 induction and inhibition. J Biomol Screen. 5(4):249–254.
  • Khoo KC, Mendels J, Rothbart M, Garland WA, Colburn WA, Min BH, Lucek R, Carbone JJ, Boxenbaum HG, Kaplan SA. 1980. Influence of phenytoin and phenobarbital on the disposition of a single oral dose of clonazepam. Clin Pharmacol Ther. 28(3):368–375.
  • Knowles BB, Howe CC, Aden DP. 1980. Human hepatocellular carcinoma cell lines secrete the major plasma proteins and hepatitis B surface antigen. Science. 209(4455):497–499.
  • Koutsounas I, Theocharis S, Patsouris E, Giaginis C. 2013. Pregnane X receptor (PXR) at the crossroads of human metabolism and disease. Curr Drug Metab. 14(3):341–350.
  • Kratochwil NA, Meille C, Fowler S, Klammers F, Ekiciler A, Molitor B, Simon S, Walter I, McGinnis C, Walther J, et al. 2017. Metabolic profiling of human long-term liver models and hepatic clearance predictions from in vitro data using nonlinear mixed-effects modeling. Aaps J. 19(2):534–550.
  • Kvist AJ, Kanebratt KP, Walentinsson A, Palmgren H, O'Hara M, Björkbom A, Andersson LC, Ahlqvist M, Andersson TB. 2018. Critical differences in drug metabolic properties of human hepatic cellular models, including primary human hepatocytes, stem cell derived hepatocytes, and hepatoma cell lines. Biochem Pharmacol. 155:124–140.
  • Lübberstedt M, Müller-Vieira U, Mayer M, Biemel KM, Knöspel F, Knobeloch D, Nüssler AK, Gerlach JC, Zeilinger K. 2011. HepaRG human hepatic cell line utility as a surrogate for primary human hepatocytes in drug metabolism assessment in vitro. J Pharmacol Toxicol Methods. 63(1):59–68.
  • McGinnity DF, Zhang G, Kenny JR, Hamilton GA, Otmani S, Stams KR, Haney S, Brassil P, Stresser DM, Riley RJ. 2009. Evaluation of multiple in vitro systems for assessment of CYP3A4 induction in drug discovery: human hepatocytes, pregnane X receptor reporter gene, and Fa2N-4 and HepaRG cells. Drug Metab Dispos. 37(6):1259–1268.
  • McMahon M, Ding S, Jimenez LA, Terranova R, Gerard MA, Vitobello A, Moggs J, Henderson CJ, Wolf CR. 2019. Constitutive androstane receptor 1 is constitutively bound to chromatin and ‘primed’ for transactivation in hepatocytes. Mol Pharmacol. 95(1):97–105.
  • Negishi M, Kobayashi K, Sakuma T, Sueyoshi T. 2020. Nuclear receptor phosphorylation in xenobiotic signal transduction. J Biol Chem. 295(45):15210–15225.
  • Nelson LJ, Morgan K, Treskes P, Samuel K, Henderson CJ, LeBled C, Homer N, Grant MH, Hayes PC, Plevris JN. 2017. Human hepatic HepaRG cells maintain an organotypic phenotype with high intrinsic CYP450 activity/metabolism and significantly outperform standard HepG2/C3A cells for pharmaceutical and therapeutic applications. Basic Clin Pharmacol Toxicol. 120(1):30–37.
  • Nofziger C, Turner AJ, Sangkuhl K, Whirl-Carrillo M, Agúndez JAG, Black JL, Dunnenberger HM, Ruano G, Kennedy MA, Phillips MS, et al. 2020. PharmVar GeneFocus: CYP2D6. Clin Pharmacol Ther. 107(1):154–170.
  • OECD. 2019. Determination of Cytochrome P450 (CYP) enzyme activity induction using differentiated human hepatic cells. [accessed 2021 Aug 2]. https://www.oecd.org/chemicalsafety/testing/Draft_TG_CYP_induction_for_2nd_WNT_review.pdf
  • Parent R, Marion MJ, Furio L, Trépo C, Petit MA. 2004. Origin and characterization of a human bipotent liver progenitor cell line. Gastroenterology. 126(4):1147–1156.
  • Preiss LC, Liu R, Hewitt P, Thompson D, Georgi K, Badolo L, Lauschke VM, Petersson C. 2021. Deconvolution of cytochrome P450 induction mechanisms in HepaRG nuclear hormone receptor knockout cells. Drug Metab Dispos. 49(8):668–678.
  • Ramaiahgari SC, Waidyanatha S, Dixon D, DeVito MJ, Paules RS, Ferguson SS. 2017. From the cover: three-dimensional (3D) HepaRG spheroid model with physiologically relevant xenobiotic metabolism competence and hepatocyte functionality for liver toxicity screening. Toxicol Sci. 159(1):124–136.
  • Rowe C, Gerrard DT, Jenkins R, Berry A, Durkin K, Sundstrom L, Goldring CE, Park BK, Kitteringham NR, Hanley KP, et al. 2013. Proteome-wide analyses of human hepatocytes during differentiation and dedifferentiation. Hepatology. 58(2):799–809.
  • Sangkuhl K, Claudio-Campos K, Cavallari LH, Agundez JAG, Whirl-Carrillo M, Duconge J, Del Tredici AL, Wadelius M, Rodrigues Botton M, Woodahl EL, et al. 2021. PharmVar GeneFocus: CYP2C9. Clin Pharmacol Ther. 110(3):662–676.
  • Scheer N, Ross J, Rode A, Zevnik B, Niehaves S, Faust N, Wolf CR. 2008. A novel panel of mouse models to evaluate the role of human pregnane X receptor and constitutive androstane receptor in drug response. J Clin Invest. 118(9):3228–3239.
  • Seo JE, Tryndyak V, Wu Q, Dreval K, Pogribny I, Bryant M, Zhou T, Robison TW, Mei N, Guo X. 2019. Quantitative comparison of in vitro genotoxicity between metabolically competent HepaRG cells and HepG2 cells using the high-throughput high-content CometChip assay. Arch Toxicol. 93(5):1433–1448.
  • Seo JE, Wu Q, Bryant M, Ren L, Shi Q, Robison TW, Mei N, Manjanatha MG, Guo X. 2020. Performance of high-throughput CometChip assay using primary human hepatocytes: a comparison of DNA damage responses with in vitro human hepatoma cell lines. Arch Toxicol. 94(6):2207–2224.
  • Shi J, Wang X, Lyu L, Jiang H, Zhu HJ. 2018. Comparison of protein expression between human livers and the hepatic cell lines HepG2, Hep3B, and Huh7 using SWATH and MRM-HR proteomics: Focusing on drug-metabolizing enzymes. Drug Metab Pharmacokinet. 33(2):133–140.
  • Sison-Young RL, Mitsa D, Jenkins RE, Mottram D, Alexandre E, Richert L, Aerts H, Weaver RJ, Jones RP, Johann E, et al. 2015. Comparative proteomic characterization of 4 human liver-derived single cell culture models reveals significant variation in the capacity for drug disposition, bioactivation, and detoxication. Toxicol Sci. 147(2):412–424.
  • Stanley LA, Horsburgh BC, Ross J, Scheer N, Wolf CR. 2006. PXR and CAR: nuclear receptors which play a pivotal role in drug disposition and chemical toxicity. Drug Metab Rev. 38(3):515–597.
  • Tascher G, Burban A, Camus S, Plumel M, Chanon S, Le Guevel R, Shevchenko V, Van Dorsselaer A, Lefai E, Guguen-Guillouzo C, et al. 2019. In-depth proteome analysis highlights HepaRG cells as a versatile cell system surrogate for primary human hepatocytes. Cells. 8(2):192.
  • Thomas M, Bayha C, Vetter S, Hofmann U, Schwarz M, Zanger UM, Braeuning A. 2015. Activating and inhibitory functions of WNT/β-catenin in the induction of cytochromes P450 by nuclear receptors in HepaRG cells. Mol Pharmacol. 87(6):1013–1020.
  • Tolosa L, Gómez-Lechón MJ, López S, Guzmán C, Castell JV, Donato MT, Jover R. 2016. Human upcyte hepatocytes: characterization of the hepatic phenotype and evaluation for acute and long-term hepatotoxicity routine testing. Toxicol Sci. 152(1):214–229.
  • Turpeinen M, Tolonen A, Chesne C, Guillouzo A, Uusitalo J, Pelkonen O. 2009. Functional expression, inhibition and induction of CYP enzymes in HepaRG cells. Toxicol In Vitro. 23(4):748–753.
  • Ulvestad M, Nordell P, Asplund A, Rehnström M, Jacobsson S, Holmgren G, Davidson L, Brolén G, Edsbagge J, Björquist P, et al. 2013. Drug metabolizing enzyme and transporter protein profiles of hepatocytes derived from human embryonic and induced pluripotent stem cells. Biochem Pharmacol. 86(5):691–702.
  • van der Mark VA, Rudi de Waart D, Shevchenko V, Elferink RP, Chamuleau RA, Hoekstra R. 2017. Stable overexpression of the constitutive androstane receptor reduces the requirement for culture with dimethyl sulfoxide for high drug metabolism in HepaRG cells. Drug Metab Dispos. 45(1):56–67.
  • Vermet H, Raoust N, Ngo R, Esserméant L, Klieber S, Fabre G, Boulenc X. 2016. Evaluation of normalization methods to predict CYP3A4 induction in six fully characterized cryopreserved human hepatocyte preparations and HepaRG cells. Drug Metab Dispos. 44(1):50–60.
  • Wang Z, Luo X, Anene-Nzelu C, Yu Y, Hong X, Singh NH, Xia L, Liu S, Yu H. 2015. HepaRG culture in tethered spheroids as an in vitro three-dimensional model for drug safety screening. J Appl Toxicol. 35(8):909–917.
  • Wang ZY, Li WJ, Li QG, Jing HS, Yuan TJ, Fu GB, Tang D, Zhang HD, Yan HX, Zhai B. 2019. A DMSO-free hepatocyte maturation medium accelerates hepatic differentiation of HepaRG cells in vitro. Biomed Pharmacother. 116:109010.
  • Weaver RJ, Betts C, Blomme EAG, Gerets HHJ, Gjervig Jensen K, Hewitt PG, Juhila S, Labbe G, Liguori MJ, Mesens N, et al. 2017. Test systems in drug discovery for hazard identification and risk assessment of human drug-induced liver injury. Expert Opin Drug Metab Toxicol. 13(7):767–782.
  • Weaver RJ, Blomme EA, Chadwick AE, Copple IM, Gerets HHJ, Goldring CE, Guillouzo A, Hewitt PG, Ingelman-Sundberg M, Jensen KG, et al. 2020. Managing the challenge of drug-induced liver injury: a roadmap for the development and deployment of preclinical predictive models. Nat Rev Drug Discov. 19(2):131–148.
  • Williamson B, Lorbeer M, Mitchell MD, Brayman TG, Riley RJ. 2016. Evaluation of a novel PXR-knockout in HepaRG™ cells. Pharmacol Res Perspect. 4(5):e00264.
  • Wiśniewski JR, Vildhede A, Norén A, Artursson P. 2016. In-depth quantitative analysis and comparison of the human hepatocyte and hepatoma cell line HepG2 proteomes. J Proteomics. 136:234–247.
  • Wong N, Lai P, Pang E, Leung TW, Lau JW, Johnson PJ. 2000. A comprehensive karyotypic study on human hepatocellular carcinoma by spectral karyotyping. Hepatology. 32(5):1060–1068.
  • Xie Y, Ke S, Chen J, Ouyang N, Tian Y. 2020. Epigenetic sensitization of pregnane X receptor-regulated gene expression by dimethyl sulfoxide. Toxicol Lett. 321:131–137.
  • Yokoyama Y, Sasaki Y, Terasaki N, Kawataki T, Takekawa K, Iwase Y, Shimizu T, Sanoh S, Ohta S. 2018. Comparison of drug metabolism and its related hepatotoxic effects in HepaRG, cryopreserved human hepatocytes, and HepG2 cell cultures. Biol Pharm Bull. 41(5):722–732.
  • Zanger UM, Schwab M. 2013. Cytochrome P450 enzymes in drug metabolism: regulation of gene expression, enzyme activities, and impact of genetic variation. Pharmacol Ther. 138(1):103–141.
  • Zhou B, Ho SS, Greer SU, Spies N, Bell JM, Zhang X, Zhu X, Arthur JG, Byeon S, Pattni R, et al. 2019. Haplotype-resolved and integrated genome analysis of the cancer cell line HepG2. Nucleic Acids Res. 47(8):3846–3861.