title: Hepatic 3D spheroid models for the detection and study of compounds with cholestatic liability.
abstract: Drug-induced cholestasis (DIC) is poorly understood and its preclinical prediction is mainly limited to assessing the compound's potential to inhibit the bile salt export pump (BSEP). Here, we evaluated two 3D spheroid models, one from primary human hepatocytes (PHH) and one from HepaRG cells, for the detection of compounds with cholestatic liability. By repeatedly co-exposing both models to a set of compounds with different mechanisms of hepatotoxicity and a non-toxic concentrated bile acid (BA) mixture for 8 days we observed a selective synergistic toxicity of compounds known to cause cholestatic or mixed cholestatic/hepatocellular toxicity and the BA mixture compared to exposure to the compounds alone, a phenomenon that was more pronounced after extending the exposure time to 14 days. In contrast, no such synergism was observed after both 8 and 14 days of exposure to the BA mixture for compounds that cause non-cholestatic hepatotoxicity. Mechanisms behind the toxicity of the cholestatic compound chlorpromazine were accurately detected in both spheroid models, including intracellular BA accumulation, inhibition of ABCB11 expression and disruption of the F-actin cytoskeleton. Furthermore, the observed synergistic toxicity of chlorpromazine and BA was associated with increased oxidative stress and modulation of death receptor signalling. Combined, our results demonstrate that the hepatic spheroid models presented here can be used to detect and study compounds with cholestatic liability.
authors.affiliation: Department of Physiology and Pharmacology, Section of Pharmacogenetics, Karolinska Institutet, Stockholm, Sweden.
authors.forename: Delilah F G
authors.initials: DF
authors.lastname: Hendriks
authors.affiliation: Department of Physiology and Pharmacology, Section of Pharmacogenetics, Karolinska Institutet, Stockholm, Sweden.
authors.forename: Lisa
authors.initials: L
authors.lastname: Fredriksson Puigvert
authors.affiliation: InSphero AG, Schlieren, Canton of Zürich, Switzerland.
authors.forename: Simon
authors.initials: S
authors.lastname: Messner
authors.affiliation: InSphero AG, Schlieren, Canton of Zürich, Switzerland.
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Mortiz
authors.affiliation: Department of Physiology and Pharmacology, Section of Pharmacogenetics, Karolinska Institutet, Stockholm, Sweden.
authors.forename: Magnus
authors.initials: M
authors.lastname: Ingelman-Sundberg
mesh_terms: D001647:Bile Acids and Salts / Q000009:adverse effects / Q000378:metabolism; D001692:Biological Transport; D018929:Cell Culture Techniques; D002460:Cell Line; D056486:Chemical and Drug Induced Liver Injury / Q000378:metabolism / Q000473:pathology; D002746:Chlorpromazine / Q000009:adverse effects / Q000031:analogs & derivatives; D002779:Cholestasis / Q000209:etiology / Q000378:metabolism* / Q000473:pathology; D022781:Hepatocytes / Q000378:metabolism* / Q000473:pathology; D006801:Humans; D018384:Oxidative Stress / Q000187:drug effects; D053220:Receptors, TNF-Related Apoptosis-Inducing Ligand / Q000378:metabolism; D015398:Signal Transduction; D018874:Spheroids, Cellular
chemical_list: D001647:Bile Acids and Salts; D053220:Receptors, TNF-Related Apoptosis-Inducing Ligand; C054028:chlorproethazine; D002746:Chlorpromazine