title: Evaluation of transcriptomic signature as a valuable tool to study drug-induced cholestasis in primary human hepatocytes.
abstract: Primary human hepatocyte (PHH) sandwich cultures from five different donors were daily exposed to cyclosporine A (CsA), ibuprofen (IBU), chlorpromazine (CPZ), amiodarone (AMI) and paracetamol (APAP) at their respective Cmax(total) for short-term (1-3 days) and long-term treatment (14 days). Whole genome mRNA profiles (34,693 genes in total) were conducted using an Illumina microarray platform. The impact of compound treatments on gene signatures involved in liver differentiation, cholestasis and in bile acid homeostasis was evaluated. Notably, PHH from the five donors showed a highly comparable phenotype of terminally differentiated hepatocytes. As expected, PHH exposed to 100 µM APAP showed no signs of hepatotoxicity both after short- and long-term treatment. CsA at 0.7 µM, IBU at 100 µM, AMI at 2.5 µM and CPZ at 0.1-0.2 µM presented, in line with their cholestatic syndromes reported at therapeutic doses, transcriptomic signatures of cholestasis in PHH cultures; deregulation of genes involved in bile acid homeostasis further confirmed this finding. The strength of the cholestasis signature obtained after treatment with CsA, IBU and AMI could be directly related to the basal expression of the respective drug metabolizing enzymes in the various PHH cultures from different individuals. Our data show that the PHH model system combined with transcriptomics carries the future promise to identify individual gene expression profiles predictive of increased cholestasis risk. As the present work suggests possible correlation between mRNA levels of ADME relevant genes and a transcriptomic signature of cholestasis, particular focus on this research question could be the emphasis of additional data collection.
authors.affiliation: KaLy-Cell, 20A rue du Général Leclerc, 67115, Plobsheim, France.
authors.forename: Céline
authors.initials: C
authors.lastname: Parmentier
authors.affiliation: Novartis Institutes of Biomedical Research, Basel, Switzerland.
authors.forename: Philippe
authors.initials: P
authors.lastname: Couttet
authors.affiliation: Novartis Institutes of Biomedical Research, Basel, Switzerland.
authors.forename: Armin
authors.initials: A
authors.lastname: Wolf
authors.affiliation: Centre Hospitalier de Mulhouse, 20 Avenue du Docteur René Laennec, 68100, Mulhouse, France.
authors.forename: Thomas
authors.initials: T
authors.lastname: Zaccharias
authors.affiliation: Hôpital Jean Minjoz, 3 Boulevard Alexandre Fleming, 25000, Besançon, France.
authors.forename: Bruno
authors.initials: B
authors.lastname: Heyd
authors.affiliation: Hôpital de Hautepierre, Avenue Molière, 67100, Strasbourg, France.
authors.forename: Philippe
authors.initials: P
authors.lastname: Bachellier
authors.affiliation: Novartis Institutes of Biomedical Research, Basel, Switzerland.
authors.forename: Marianne
authors.initials: M
authors.lastname: Uteng
authors.affiliation: KaLy-Cell, 20A rue du Général Leclerc, 67115, Plobsheim, France. l.richert@kaly-cell.com.|Laboratoire de Toxicologie Cellulaire, Université de Bourgogne Franche-Comté, EA 4267, Besançon, France. l.richert@kaly-cell.com.
authors.forename: Lysiane
authors.initials: L
authors.lastname: Richert
mesh_terms: D000328:Adult; D000368:Aged; D001647:Bile Acids and Salts / Q000378:metabolism; D056486:Chemical and Drug Induced Liver Injury / Q000209:etiology* / Q000235:genetics; D002779:Cholestasis / Q000139:chemically induced* / Q000235:genetics; D005260:Female; D005786:Gene Expression Regulation / Q000187:drug effects; D022781:Hepatocytes / Q000187:drug effects*; D006801:Humans; D008297:Male; D008875:Middle Aged; D012333:RNA, Messenger / Q000378:metabolism; D013997:Time Factors; D059467:Transcriptome
chemical_list: D001647:Bile Acids and Salts; D012333:RNA, Messenger
keywords: Cholestasis; Donor-donor variability; Primary Human Hepatocytes; Transcriptomic