Cell-based assay using glutathione-depleted HepaRG and HepG2 human liver cells for predicting drug-induced liver injury

Toxicol In Vitro. 2018 Apr;48:286-301. doi: 10.1016/j.tiv.2018.01.019. Epub 2018 Jan 31.

Abstract

Immortalized liver cells have been used for evaluating the toxicity of compounds; however, excessive glutathione is considered to lessen cytotoxicity. In this study, we compared the effects of glutathione depletion on cytotoxicities of drugs using HepaRG and HepG2 cells, which express and lack drug-metabolizing enzymes, respectively, for predicting drug-induced liver injury (DILI) risks. These cells were pre-incubated with L-buthionine-S,R-sulfoximine (BSO) and then exposed to 34 test compounds with various DILI risks for 24 h. ATP level exhibited the highest predictability of DILI among tested parameters. BSO treatment rendered cells susceptible to drug-induced cytotoxicity when evaluated by cell viability and caspase 3/7 activity with the sensitivity of cell viability from 50% in non-treated HepaRG cells to 71% in BSO-treated HepaRG cells. These results indicate that cytotoxicity assays using GSH-depleted HepaRG cells improve the predictability of DILI risks. However, HepaRG cells were not always superior to HepG2 cells when assessed by ATP level. The combination of HepG2 and HepaRG cells index produced the best prediction in the cases of caspase 3/7 acitivity and ATP level. In conclusions, the developed highly sensitive cell-based assay using GSH-reduced cells would be useful for predicting potential DILI risks at an early stage of drug development.

Keywords: Cell-based assay; Drug-induced liver injury; Glutathione; HepaRG; Reactive metabolites.

MeSH terms

  • Adenosine Triphosphate / metabolism
  • Caspases / metabolism
  • Cell Line
  • Cell Survival / drug effects
  • Chemical and Drug Induced Liver Injury / pathology*
  • Cytochrome P-450 CYP3A / metabolism
  • Drug-Related Side Effects and Adverse Reactions / pathology*
  • Glutathione / metabolism*
  • Hep G2 Cells
  • Hepatocytes / drug effects
  • Humans
  • Liver / enzymology
  • Liver / pathology*
  • Predictive Value of Tests

Substances

  • Adenosine Triphosphate
  • Cytochrome P-450 CYP3A
  • CYP3A4 protein, human
  • Caspases
  • Glutathione