Evaluation of a human in vitro hepatocyte-NPC co-culture model for the prediction of idiosyncratic drug-induced liver injury: A pilot study

Anne Granitzny; Jan Knebel; Meike Müller; Armin Braun; Pablo Steinberg; Clemens Dasenbrock; Tanja Hansen

doi:10.1016/j.toxrep.2017.02.001

Evaluation of a human in vitro hepatocyte-NPC co-culture model for the prediction of idiosyncratic drug-induced liver injury: A pilot study

Toxicol Rep. 2017 Feb 12:4:89-103. doi: 10.1016/j.toxrep.2017.02.001. eCollection 2017.

Authors

Anne Granitzny¹, Jan Knebel¹, Meike Müller¹, Armin Braun², Pablo Steinberg³, Clemens Dasenbrock¹, Tanja Hansen¹

Affiliations

¹ Fraunhofer Institute for Toxicology and Experimental Medicine (ITEM), Nikolai-Fuchs-Straße 1, 30625 Hannover, Germany.
² Fraunhofer Institute for Toxicology and Experimental Medicine (ITEM), Member of the German Center for Lung Research (DZL), Biomedical Research in End stage and Obstructive Lung Disease (BREATH) research network, Member of the Cluster of Excellence Regenerative Biology to Reconstructive Therapy (REBIRTH), Nikolai-Fuchs-Straße 1, 30625 Hannover, Germany.
³ University of Veterinary Medicine Hannover (TiHo), Institute for Food Toxicology and Analytical Chemistry, Bischofsholer Damm 15, 30173 Hannover, Germany.

Abstract

Interactions between hepatocytes and immune cells as well as inflammatory episodes are frequently discussed to play a critical role in the alteration of the individual susceptibility to idiosyncratic drug-induced liver injury (iDILI). To evaluate this hypothesis and to face the urgent need for predictive in vitro models, we established two co-culture systems based on two human cell lines in presence or absence of pro-inflammatory factors (LPS, TNF), i.e. hepatoma HepG2 cells co-cultured with monocytic or macrophage-like THP-1 cells. HepG2 monocultures served as control scenario. Mono- or co-cultures were treated with iDILI reference substances (Troglitazone [TGZ], Trovafloxacin [TVX], Diclofenac [DcL], Ketoconazole [KC]) or their non-iDILI partner compounds (Rosiglitazone, Levofloxacin, Acetylsalicylic Acid, Fluconazole). The liver cell viability was subsequently determined via WST-Assay. An enhanced cytotoxicity (synergy) or a hormetic response compared to the drug effect in the HepG2 monoculture was considered as iDILI positive. TGZ synergized in co-cultures with monocytes without an additional pro-inflammatory stimulus, while DcL and KC showed a hormetic response. All iDILI drugs synergized with TNF in the simple HepG2 monoculture, indicating its relevance as an initiator of iDILI. KC showed a synergy when co-exposed to both, monocytes and LPS, while TVX and DcL showed a synergy under the same conditions with macrophages. All described iDILI responses were not observed with the corresponding non-iDILI partner compounds. Our first results confirm that an inflammatory environment increases the sensitivity of liver cells towards iDILI compounds and point to an involvement of pro-inflammatory factors, especially TNF, in the development of iDILI.

Keywords: CD, cluster of differentiation; Co-culture model; DAMP, damage-associated molecular pattern; Drug-induced liver injury; EC, effective concentration; EpCAM, epithelial cellular adhesion molecule; HSP, heat shock protein; Idiosyncratic; Inflammation; JNK, c-Jun N-terminal kinase; LPS, bacterial lipopolysaccharide; NF-κB, nuclear factor kappa B; NPC, non-parenchymal cell; NSAID, nonsteriodal anti-inflammatory drug; PAMP, pathogen-associated molecular pattern; Preclinical research; SD, standard deviation; TNF, tumor necrosis factor; iDILI, idiosyncratic drug-induced liver injury.