Multiparametric assay using HepaRG cells for predicting drug-induced liver injury.
The utility of HepaRG cells as an in vitro cell-based assay system for assessing drug-induced liver injury (DILI) risk was investigated. Seventeen DILI-positive and 15 DILI-negative drugs were selected for the assay. HepaRG cells were treated with each drug for 24h at concentrations that were 1.6-, 6.3-, 25-, and 100-fold the therapeutic maximum plasma concentration (Cmax). After treatment, the cell viability, glutathione content, caspase 3/7 activity, lipid accumulation, leakage of lactate dehydrogenase, and albumin secretion were measured. The sensitivity and specificity were calculated to assess the ability of the assay to predict DILI. Our multiparametric assay using HepaRG cells exhibited a 67% sensitivity and 73% specificity at a 100-fold concentration of Cmax and a 41% sensitivity and 87% specificity at a 25-fold concentration of Cmax. When a 25-fold Cmax cut-off was applied, approximately 70% of drugs exhibiting positive responses were classified into the high DILI risk category. HepaRG cells distinguished relatively safe drugs from their high-risk analogs. Our study indicates that HepaRG cells may be of use to (1) prioritize drug analogs, (2) analyze the mechanism of DILI, and (3) assess the risk for DILI in the early drug discovery stage.
Pharmacokinetics and Safety Department, Drug Research Center, Kyoto Research Center, Kaken Pharmaceutical Co., LTD., Kyoto 607-8042, Japan. Electronic address: tomida_takafumi@kaken.co.jp.
Takafumi
T
Tomida
Pharmacokinetics and Safety Department, Drug Research Center, Kyoto Research Center, Kaken Pharmaceutical Co., LTD., Kyoto 607-8042, Japan.
Hayao
H
Okamura
Pharmacokinetics and Safety Department, Drug Research Center, Shizuoka Research Center, Kaken Pharmaceutical Co., LTD., Shizuoka 426-8464, Japan.
Masahiro
M
Satsukawa
Department of Drug Safety Sciences, Nagoya University Graduate School of Medicine, Nagoya 466-8550, Japan.
Tsuyoshi
T
Yokoi
Pharmacokinetics and Safety Department, Drug Research Center, Kyoto Research Center, Kaken Pharmaceutical Co., LTD., Kyoto 607-8042, Japan.
Yoshihiro
Y
Konno
D053148:Caspase 3 / Q000378:metabolism; D053179:Caspase 7 / Q000378:metabolism; D045744:Cell Line, Tumor; D002463:Cell Membrane Permeability / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D056486:Chemical and Drug Induced Liver Injury / Q000201:enzymology / Q000378:metabolism*; D055808:Drug Discovery / Q000379:methods*; D004353:Drug Evaluation, Preclinical; D015507:Drugs, Investigational / Q000145:classification / Q000494:pharmacology* / Q000633:toxicity; D005978:Glutathione / Q000378:metabolism; D022781:Hepatocytes / Q000187:drug effects* / Q000378:metabolism; D006801:Humans; D050356:Lipid Metabolism / Q000187:drug effects; D008954:Models, Biological; D009994:Osmolar Concentration; D012372:ROC Curve; D015203:Reproducibility of Results; D018570:Risk Assessment; D012709:Serum Albumin / Q000378:metabolism; D000075462:Serum Albumin, Human
C422455:ALB protein, human; D015507:Drugs, Investigational; D012709:Serum Albumin; C505407:CASP3 protein, human; C505520:CASP7 protein, human; D053148:Caspase 3; D053179:Caspase 7; D005978:Glutathione; D000075462:Serum Albumin, Human
Drug-induced liver injury; HepaRG cells; Multiparametric assay