Characterization of an In Vitro Assay for the Detection of Drug‐Induced Cholestasis Using the HepaRG Cell Line

Abstract

Drug‐induced cholestasis (DIC) is a relevant concern in the pharmaceutical industry, and reliable early detection of DIC is of great value in the discovery and development of safe drugs. Traditional animal models are poorly predictive of cholestasis risk in humans, partly because of biological differences. Moreover, limited robust in vitro methods to predict DIC are available. Recently, an in vitro model for the detection of DIC was developed using primary hepatocytes in sandwich configuration. Following incubation of cells with test compounds in the presence or absence of a concentrated bile acid mixture, a DIC Index (DICI) is calculated as a measure of the relative reduction in cellular function. This model was evaluated as part of the European Federation of Pharmaceutical Industries and Associations (EFPIA) Innovative Medicine Initiative project MIP‐DILI. Here, we assessed several test compounds in a modified version of this assay using the HepaRG cell line. Several tool compounds with or without known cholestasis risk were tested at relevant concentrations: five compounds with clinical cholestasis reports (troglitazone, cyclosporin A, chlorpromazine, bosentan, and nefazodone), one toxicant inducing biliary injury (ANIT), two non‐cholestatic compounds known to induce hepatocyte injury (APAP, perhexiline), and one compound associated with rare instances of liver injury in humans (pioglitazone). Albumin and ATP production were assessed as parameters to evaluate the overall functionality and integrity of the cells and DICIs were calculated. Based on change in these parameters relative to the vehicle, the assay accurately predicted 6/6 cholestasis positive compounds and 2/3 cholestasis negative compounds. Results were repeatable. From these data, the HepaRG model for the detection of DIC may be beneficial for hazard identification (i.e., biliary injury) during lead optimization.Support or Funding InformationAuthor Disclosures: All authors are employees of AbbVie. The design, study conduct, and financial support for this research was provided by AbbVie. AbbVie participated in the interpretation of data, review, and approval of the publication.