title: Cytotoxicity evaluation using cryopreserved primary human hepatocytes in various culture formats.
abstract: Sixteen training compounds selected in the IMI MIP-DILI consortium, 12 drug-induced liver injury (DILI) positive compounds and 4 non-DILI compounds, were assessed in cryopreserved primary human hepatocytes. When a ten-fold safety margin threshold was applied, the non-DILI-compounds were correctly identified 2h following a single exposure to pooled human hepatocytes (n=13 donors) in suspension and 14-days following repeat dose exposure (3 treatments) to an established 3D-microtissue co-culture (3D-MT co-culture, n=1 donor) consisting of human hepatocytes co-cultured with non-parenchymal cells (NPC). In contrast, only 5/12 DILI-compounds were correctly identified 2h following a single exposure to pooled human hepatocytes in suspension. Exposure of the 2D-sandwich culture human hepatocyte monocultures (2D-sw) for 3days resulted in the correct identification of 11/12 DILI-positive compounds, whereas exposure of the human 3D-MT co-cultures for 14days resulted in identification of 9/12 DILI-compounds; in addition to ximelagatran (also not identified by 2D-sw monocultures, Sison-Young et al., 2016), the 3D-MT co-cultures failed to detect amiodarone and bosentan. The sensitivity of the 2D human hepatocytes co-cultured with NPC to ximelagatran was increased in the presence of lipopolysaccharide (LPS), but only at high concentrations, therefore preventing its classification as a DILI positive compound. In conclusion (1) despite suspension human hepatocytes having the greatest metabolic capacity in the short term, they are the least predictive of clinical DILI across the MIP-DILI test compounds, (2) longer exposure periods than 72h of human hepatocytes do not allow to increase DILI-prediction rate, (3) co-cultures of human hepatocytes with NPC, in the presence of LPS during the 72h exposure period allow the assessment of innate immune system involvement of a given drug.
authors.affiliation: KaLy-Cell, 20A rue du Général Leclerc, 67115 Plobsheim, France; Université de Franche-Comté, EA 4267 Besançon, France. Electronic address: l.richert@kaly-cell.com.
authors.forename: Lysiane
authors.initials: L
authors.lastname: Richert
authors.affiliation: KaLy-Cell, 20A rue du Général Leclerc, 67115 Plobsheim, France. Electronic address: a.baze@kaly-cell.com.
authors.forename: Audrey
authors.initials: A
authors.lastname: Baze
authors.affiliation: KaLy-Cell, 20A rue du Général Leclerc, 67115 Plobsheim, France. Electronic address: c.parmentier@kaly-cell.com.
authors.forename: Céline
authors.initials: C
authors.lastname: Parmentier
authors.affiliation: UCB BioPharma SPRL, Non-Clinical Development, Chemin du Foriest, 1420 Braine-l'Alleud, Belgium. Electronic address: helga.gerets@ucb.com.
authors.forename: Helga H J
authors.initials: HHJ
authors.lastname: Gerets
authors.affiliation: MRC Centre for Drug Safety Science, Department of Molecular and Clinical Pharmacology, Sherrington Building, Ashton Street, University of Liverpool, Liverpool L69 3GE, UK. Electronic address: Rowena.Sison-Young@liverpool.ac.uk.
authors.forename: Rowena
authors.initials: R
authors.lastname: Sison-Young
authors.affiliation: Sanofi-Aventis Deutschland GmbH, R&D DSAR Preclinical Safety, Industriepark Hoechst, D-65926 Frankfurt, Germany. Electronic address: Martina.Dorau@sanofi.com.
authors.forename: Martina
authors.initials: M
authors.lastname: Dorau
authors.affiliation: GlaxoSmithKline, Safety Assessment, Stevenage, Hertfordshire, UK. Electronic address: cerys.a.lovatt@gsk.com.
authors.forename: Cerys
authors.initials: C
authors.lastname: Lovatt
authors.affiliation: Sanofi-Aventis Deutschland GmbH, R&D DSAR Preclinical Safety, Industriepark Hoechst, D-65926 Frankfurt, Germany. Electronic address: Andreas.Czich@sanofi.com.
authors.forename: Andreas
authors.initials: A
authors.lastname: Czich
authors.affiliation: MRC Centre for Drug Safety Science, Department of Molecular and Clinical Pharmacology, Sherrington Building, Ashton Street, University of Liverpool, Liverpool L69 3GE, UK. Electronic address: C.E.P.Goldring@liverpool.ac.uk.
authors.forename: Christopher
authors.initials: C
authors.lastname: Goldring
authors.affiliation: MRC Centre for Drug Safety Science, Department of Molecular and Clinical Pharmacology, Sherrington Building, Ashton Street, University of Liverpool, Liverpool L69 3GE, UK. Electronic address: B.K.Park@liv.ac.uk.
authors.forename: B Kevin
authors.initials: BK
authors.lastname: Park
authors.affiliation: Orion Corporation, R&D, In Vitro Biology, Orionintie 1A, P.O. Box 65, FI-02101 Espoo, Finland. Electronic address: satu.juhila@orionpharma.com.
authors.forename: Satu
authors.initials: S
authors.lastname: Juhila
authors.affiliation: Translational Safety, Drug Safety & Metabolism, AstraZeneca, Cambridge Science Park, Cambridge, UK. Electronic address: Alison.Foster2@astrazeneca.com.
authors.forename: Alison J
authors.initials: AJ
authors.lastname: Foster
authors.affiliation: Translational Safety, Drug Safety & Metabolism, AstraZeneca, Cambridge Science Park, Cambridge, UK. Electronic address: Dominic.P.Williams@astrazeneca.com.
authors.forename: Dominic P
authors.initials: DP
authors.lastname: Williams
mesh_terms: D016475:3T3 Cells; D000818:Animals; D061249:Batch Cell Culture Techniques; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D060833:Cellular Microenvironment / Q000187:drug effects*; D056486:Chemical and Drug Induced Liver Injury / Q000209:etiology; D018920:Coculture Techniques; D015925:Cryopreservation; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D015507:Drugs, Investigational / Q000009:adverse effects* / Q000378:metabolism; D022781:Hepatocytes / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D006801:Humans; D007113:Immunity, Innate / Q000187:drug effects; D007700:Kinetics; D007728:Kupffer Cells / Q000166:cytology / Q000187:drug effects / Q000276:immunology; D008070:Lipopolysaccharides / Q000819:agonists / Q000037:antagonists & inhibitors / Q000633:toxicity; D051379:Mice; D008958:Models, Molecular; D017154:Stromal Cells / Q000166:cytology / Q000187:drug effects / Q000502:physiology
chemical_list: D015507:Drugs, Investigational; D008070:Lipopolysaccharides
keywords: Cryopreserved human hepatocytes; DILI; In vitro models; Monoculture and co-culture; Short term and long term exposure