title: High-content screening of drug-induced mitochondrial impairment in hepatic cells: effects of statins.
abstract: A frequent mechanism for drug-induced liver injury (DILI) is mitochondrial impairment, and early evaluation of new drugs for their potential to cause mitochondrial dysfunction is becoming an important task for drug development. To this end, we designed a high-content screening assay to study mitochondrial-induced hepatotoxicity in HepG2 cells in detail. Simultaneous assessment of mitochondrial mass and cell viability in cells exposed for 24 h to compounds provides preliminary information on the mitochondrial- or nonmitochondrial-related hepatotoxic potential of compounds. To fully address the mechanisms implicated in mitochondrial impairment, prelethal changes in mitochondrial superoxide production, mitochondrial membrane potential, mitochondrial permeability transition, intracellular calcium concentration and apoptotic cell death were studied in cells incubated for 1 h with compounds. The assay correctly classified a set of well-known mitochondrial toxicants and negative controls and revealed high sensitivity for the detection of mitochondrial DILI and the establishment of different mitochondrial toxicity risks (low to high). This procedure was used for analysing the potential mitochondrial impairment of six statins to determine their clinical risk. All the tested statins produced mitochondrial impairment, although they showed different levels of toxicity (low-medium toxicity risk). The results suggest that this cell-based assay is a promising in vitro approach to predict the potential of drug candidates to induce mitochondrial-associated hepatotoxicity.
authors.affiliation: Unidad de Hepatología Experimental, Torre A, Instituto Investigación Sanitaria La Fe (IIS La Fe), Bulevard Sur s/n, 46026, Valencia, Spain. laiatolosa@hotmail.com.
authors.forename: Laia
authors.initials: L
authors.lastname: Tolosa
authors.affiliation: Unidad de Hepatología Experimental, Torre A, Instituto Investigación Sanitaria La Fe (IIS La Fe), Bulevard Sur s/n, 46026, Valencia, Spain.
authors.forename: Antonio
authors.initials: A
authors.lastname: Carmona
authors.affiliation: Unidad de Hepatología Experimental, Torre A, Instituto Investigación Sanitaria La Fe (IIS La Fe), Bulevard Sur s/n, 46026, Valencia, Spain.|Departamento de Bioquímica y Biología Molecular, Facultad de Medicina, Universidad de Valencia, 46010, Valencia, Spain.|CIBERehd, FIS, 08036, Barcelona, Spain.
authors.forename: José V
authors.initials: JV
authors.lastname: Castell
authors.affiliation: Unidad de Hepatología Experimental, Torre A, Instituto Investigación Sanitaria La Fe (IIS La Fe), Bulevard Sur s/n, 46026, Valencia, Spain. gomez_mjo@gva.es.|CIBERehd, FIS, 08036, Barcelona, Spain. gomez_mjo@gva.es.
authors.forename: M José
authors.initials: MJ
authors.lastname: Gómez-Lechón
authors.affiliation: Unidad de Hepatología Experimental, Torre A, Instituto Investigación Sanitaria La Fe (IIS La Fe), Bulevard Sur s/n, 46026, Valencia, Spain.|Departamento de Bioquímica y Biología Molecular, Facultad de Medicina, Universidad de Valencia, 46010, Valencia, Spain.|CIBERehd, FIS, 08036, Barcelona, Spain.
authors.forename: M Teresa
authors.initials: MT
authors.lastname: Donato
mesh_terms: D017209:Apoptosis / Q000187:drug effects; D002118:Calcium / Q000378:metabolism; D002470:Cell Survival / Q000187:drug effects; D056486:Chemical and Drug Induced Liver Injury / Q000209:etiology* / Q000473:pathology; D056945:Hep G2 Cells; D022781:Hepatocytes / Q000187:drug effects / Q000473:pathology; D006801:Humans; D019161:Hydroxymethylglutaryl-CoA Reductase Inhibitors / Q000633:toxicity*; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D008930:Mitochondria, Liver / Q000187:drug effects* / Q000473:pathology
chemical_list: D019161:Hydroxymethylglutaryl-CoA Reductase Inhibitors; D002118:Calcium
keywords: Hepatotoxicity; High-content screening; Mitochondrial injury; Statins