title: Mouse precision-cut liver slices as an ex vivo model to study idiosyncratic drug-induced liver injury.
abstract: Idiosyncratic drug-induced liver injury (IDILI) has been the top reason for withdrawing drugs from the market or for black box warnings. IDILI may arise from the interaction of a drug's reactive metabolite with a mild inflammation that renders the liver more sensitive to injury resulting in increased toxicity (inflammatory stress hypothesis). Aiming to develop a robust ex vivo screening method to study inflammatory stress-related IDILI mechanisms and to find biomarkers that can detect or predict IDILI, mouse precision-cut liver slices (mPCLS) were coincubated for 24 h with IDILI-related drugs and lipopolysaccharide. Lipopolysaccharide exacerbated ketoconazole (15 μM) and clozapine (45 μM) toxicity but not their non-IDILI-related comparators, voriconazole (1500 μM) and olanzapine (45 μM). However, the other IDILI-related drugs tested [diclofenac (200 μM), carbamazepine (400 μM), and troglitazone (30 μM)] did not cause synergistic toxicity with lipopolysaccharide after 24 h of incubation. Lipopolysaccharide further decreased the reduced glutathione levels caused by ketoconazole or clozapine in mPCLS after 24 h of incubation, which was not the case for the other drugs. Lipopolysaccharide significantly increased nitric oxide (NO), cytokine, and chemokine release into the mPCLS media, while the treatment with the drugs alone did not cause any substantial change. All seven drugs drastically reduced lipopolysaccharide-induced NO production. Interestingly, only ketoconazole and clozapine increased the lipopolysaccharide-induced granulocyte colony-stimulating factor (G-CSF) and granulocyte-macrophage colony-stimulating factor (GM-CSF) release. Pilot experiments showed that diclofenac and troglitazone, but not carbamazepine, demonstrated synergistic toxicity with lipopolysaccharide after a longer incubation of 48 h in mPCLS. In conclusion, we have developed an ex vivo model to detect inflammatory stress-related liver toxicity and identified ketoconazole, clozapine, troglitazone, and diclofenac as drugs that showed synergistic toxicity with lipopolysaccharide. Reduced glutathione, G-CSF, and GM-CSF were identified to be potential biomarkers for IDILI-inducing drugs mediated by inflammatory stress, and mPCLS appear to be a promising screening tool to further unravel the mechanism of IDILI.
authors.affiliation: Division of Pharmacokinetics, Toxicology and Targeting, Department of Pharmacy, University of Groningen, Antonius Deusinglaan 1, 9713 AV Groningen, The Netherlands.
authors.forename: Mackenzie
authors.initials: M
authors.lastname: Hadi
authors.forename: Yixi
authors.initials: Y
authors.lastname: Chen
authors.forename: Viktoriia
authors.initials: V
authors.lastname: Starokozhko
authors.forename: Marjolijn T
authors.initials: MT
authors.lastname: Merema
authors.forename: Geny M M
authors.initials: GM
authors.lastname: Groothuis
mesh_terms: D000818:Animals; D001569:Benzodiazepines / Q000737:chemistry / Q000633:toxicity; D015415:Biomarkers / Q000378:metabolism; D002220:Carbamazepine / Q000737:chemistry / Q000633:toxicity; D002470:Cell Survival / Q000187:drug effects; D056486:Chemical and Drug Induced Liver Injury / Q000378:metabolism / Q000473:pathology*; D018925:Chemokines / Q000378:metabolism; D002839:Chromans / Q000737:chemistry / Q000633:toxicity; D003024:Clozapine / Q000737:chemistry / Q000633:toxicity; D016207:Cytokines / Q000378:metabolism; D004008:Diclofenac / Q000737:chemistry / Q000633:toxicity; D004357:Drug Synergism; D005260:Female; D005978:Glutathione / Q000378:metabolism; D016179:Granulocyte Colony-Stimulating Factor / Q000378:metabolism; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000378:metabolism; D066298:In Vitro Techniques; D007654:Ketoconazole / Q000737:chemistry / Q000633:toxicity; D008070:Lipopolysaccharides / Q000633:toxicity; D051379:Mice; D008810:Mice, Inbred C57BL; D009589:Nitrogen Oxides / Q000378:metabolism; D000077152:Olanzapine; D011743:Pyrimidines / Q000737:chemistry / Q000633:toxicity; D045162:Thiazolidinediones / Q000737:chemistry / Q000633:toxicity; D014230:Triazoles / Q000737:chemistry / Q000633:toxicity; D000077288:Troglitazone; D065819:Voriconazole
chemical_list: D015415:Biomarkers; D018925:Chemokines; D002839:Chromans; D016207:Cytokines; D008070:Lipopolysaccharides; D009589:Nitrogen Oxides; D011743:Pyrimidines; D045162:Thiazolidinediones; D014230:Triazoles; D001569:Benzodiazepines; D016179:Granulocyte Colony-Stimulating Factor; D004008:Diclofenac; D002220:Carbamazepine; D016178:Granulocyte-Macrophage Colony-Stimulating Factor; D005978:Glutathione; D000077288:Troglitazone; D003024:Clozapine; D065819:Voriconazole; D000077152:Olanzapine; D007654:Ketoconazole