title: Advantageous use of HepaRG cells for the screening and mechanistic study of drug-induced steatosis.
abstract: Only a few in vitro assays have been proposed to evaluate the steatotic potential of new drugs. The present study examines the utility of HepaRG cells as a cell-based assay system for screening drug-induced liver steatosis. A high-content screening assay was run to evaluate multiple toxicity-related cell parameters in HepaRG cells exposed to 28 compounds, including drugs reported to cause steatosis through different mechanisms and non-steatotic compounds. Lipid content was the most sensitive parameter for all the steatotic drugs, whereas no effects on lipid levels were produced by non-steatotic compounds. Apart from fat accumulation, increased ROS production and altered mitochondrial membrane potential were also found in the cells exposed to steatotic drugs, which indicates that all these cellular events contributed to drug-induced hepatotoxicity. These findings are of clinical relevance as most effects were observed at drug concentrations under 100-fold of the therapeutic peak plasmatic concentration. HepaRG cells showed increased lipid overaccumulation vs. HepG2 cells, which suggests greater sensitivity to drug-induced steatosis. An altered expression profile of transcription factors and the genes that code key proteins in lipid metabolism was also found in the cells exposed to drugs capable of inducing liver steatosis. Our results generally indicate the value of HepaRG cells for assessing the risk of liver damage associated with steatogenic compounds and for investigating the molecular mechanisms involved in drug-induced steatosis.
authors.affiliation: Unidad de Hepatología Experimental, Instituto de Investigación Sanitaria La Fe, Valencia 46026, Spain.
authors.forename: Laia
authors.initials: L
authors.lastname: Tolosa
authors.affiliation: Unidad de Hepatología Experimental, Instituto de Investigación Sanitaria La Fe, Valencia 46026, Spain; CIBERehd, FIS, Barcelona 08036, Spain.
authors.forename: M José
authors.initials: MJ
authors.lastname: Gómez-Lechón
authors.affiliation: Unidad de Hepatología Experimental, Instituto de Investigación Sanitaria La Fe, Valencia 46026, Spain.
authors.forename: Nuria
authors.initials: N
authors.lastname: Jiménez
authors.affiliation: Biostatistics Unit, Instituto de Investigación Sanitaria La Fe, Valencia 46026, Spain.
authors.forename: David
authors.initials: D
authors.lastname: Hervás
authors.affiliation: Unidad de Hepatología Experimental, Instituto de Investigación Sanitaria La Fe, Valencia 46026, Spain; CIBERehd, FIS, Barcelona 08036, Spain; Departamento de Bioquímica y Biología Molecular, Facultad de Medicina, Universidad de Valencia, Valencia 46010, Spain.
authors.forename: Ramiro
authors.initials: R
authors.lastname: Jover
authors.affiliation: Unidad de Hepatología Experimental, Instituto de Investigación Sanitaria La Fe, Valencia 46026, Spain; CIBERehd, FIS, Barcelona 08036, Spain; Departamento de Bioquímica y Biología Molecular, Facultad de Medicina, Universidad de Valencia, Valencia 46010, Spain. Electronic address: donato_mte@gva.es.
authors.forename: M Teresa
authors.initials: MT
authors.lastname: Donato
mesh_terms: D045744:Cell Line, Tumor; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D064420:Drug-Related Side Effects and Adverse Reactions; D005234:Fatty Liver / Q000139:chemically induced*; D006801:Humans; D050356:Lipid Metabolism / Q000235:genetics
keywords: Drug; HepaRG; Screening; Steatosis