title: 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.
abstract: In the pharmaceutical industry, improving the early detection of drug-induced hepatotoxicity is essential as it is one of the most important reasons for attrition of candidate drugs during the later stages of drug development. The first objective of this study was to better characterize different cellular models (i.e., HepG2, HepaRG cells, and fresh primary human hepatocytes) at the gene expression level and analyze their metabolic cytochrome P450 capabilities. The cellular models were exposed to three different CYP450 inducers; beta-naphthoflavone (BNF), phenobarbital (PB), and rifampicin (RIF). HepG2 cells responded very weakly to the different inducers at the gene expression level, and this translated generally into low CYP450 activities in the induced cells compared with the control cells. On the contrary, HepaRG cells and the three human donors were inducible after exposure to BNF, PB, and RIF according to gene expression responses and CYP450 activities. Consequently, HepaRG cells could be used in screening as a substitute and/or in complement to primary hepatocytes for CYP induction studies. The second objective was to investigate the predictivity of the different cellular models to detect hepatotoxins (16 hepatotoxic and 5 nonhepatotoxic compounds). Specificity was 100% with the different cellular models tested. Cryopreserved human hepatocytes gave the highest sensitivity, ranging from 31% to 44% (depending on the donor), followed by lower sensitivity (13%) for HepaRG and HepG2 cells (6.3%). Overall, none of the models under study gave desirable sensitivities (80-100%). Consequently, a high metabolic capacity and CYP inducibility in cell lines does not necessarily correlate with a high sensitivity for the detection of hepatotoxic drugs. Further investigations are necessary to compare different cellular models and determine those that are best suited for the detection of hepatotoxic compounds.
authors.affiliation: UCB Pharma, Brussels, Belgium. helga.gerets@ucb.com
authors.forename: H H J
authors.initials: HH
authors.lastname: Gerets
authors.forename: K
authors.initials: K
authors.lastname: Tilmant
authors.forename: B
authors.initials: B
authors.lastname: Gerin
authors.forename: H
authors.initials: H
authors.lastname: Chanteux
authors.forename: B O
authors.initials: BO
authors.lastname: Depelchin
authors.forename: S
authors.initials: S
authors.lastname: Dhalluin
authors.forename: F A
authors.initials: FA
authors.lastname: Atienzar
mesh_terms: D000328:Adult; D000368:Aged; D056486:Chemical and Drug Induced Liver Injury / Q000201:enzymology / Q000209:etiology* / Q000235:genetics; D002648:Child; D003577:Cytochrome P-450 Enzyme System / Q000096:biosynthesis / Q000378:metabolism*; D064420:Drug-Related Side Effects and Adverse Reactions; D004790:Enzyme Induction; D005260:Female; D015870:Gene Expression / Q000187:drug effects; D020869:Gene Expression Profiling; D056945:Hep G2 Cells; D022781:Hepatocytes / Q000187:drug effects* / Q000201:enzymology / Q000378:metabolism; D006801:Humans; D020128:Inhibitory Concentration 50; D007928:Lethal Dose 50; D008297:Male; D008875:Middle Aged; D008954:Models, Biological; D011237:Predictive Value of Tests; D025341:Principal Component Analysis; D012333:RNA, Messenger / Q000235:genetics*; D018675:Toxicity Tests / Q000379:methods*; D043922:Toxicogenetics
chemical_list: D012333:RNA, Messenger; D003577:Cytochrome P-450 Enzyme System