title: Determination of liver specific toxicities in rat hepatocytes by high content imaging during 2-week multiple treatment.
abstract: DILI is a major safety issue during drug development and one of the leading causes for market withdrawal. Despite many efforts made in the past, the prediction of DILI using in vitro models remains very unreliable. In the present study, the well-established hepatocyte Collagen I-Matrigel™ sandwich culture was used, mimicking chronic drug treatment after multiple incubations for 14 days. Ten drugs associated with different types of specific preclinical and clinical liver injury were evaluated at non-cytotoxic concentrations. Mrp2-mediated transport, intracellular accumulation of neutral lipids and phospholipids were selected as functional endpoints by using Cellomics™ Arrayscan® technology and assessed at five timepoints (day 1, 3, 7, 10, 14). Liver specific functional impairments after drug treatment were enhanced over time and could be monitored by HCI already after few days and before cytotoxicity. Phospholipidosis-inducing drugs Chlorpromazine and Amiodarone displayed the same response as in vivo. Cyclosporin A, Chlorpromazine, and Troglitazone inhibited Mrp2-mediated biliary transport, correlating with in vivo findings. Steatosis remained difficult to be reproduced under the current in vitro testing conditions, resulting into false negative and positive responses. The present results suggest that the repeated long-term treatment of rat hepatocytes in the Collagen I-Matrigel™ sandwich configuration might be a suitable tool for safety profiling of the potential to induce phospholipidosis and impair Mrp2-mediated transport processes, but not to predict steatosis.
authors.affiliation: Discovery and Investigative Safety, Preclinical Safety, Novartis Institutes for Biomedical Research, CH-4057 Basel, Switzerland.
authors.forename: Davide
authors.initials: D
authors.lastname: Germano
authors.affiliation: Discovery and Investigative Safety, Preclinical Safety, Novartis Institutes for Biomedical Research, CH-4057 Basel, Switzerland.
authors.forename: Marianne
authors.initials: M
authors.lastname: Uteng
authors.affiliation: Discovery and Investigative Safety, Preclinical Safety, Novartis Institutes for Biomedical Research, CH-4057 Basel, Switzerland.
authors.forename: Francois
authors.initials: F
authors.lastname: Pognan
authors.affiliation: Discovery and Investigative Safety, Preclinical Safety, Novartis Institutes for Biomedical Research, CH-4057 Basel, Switzerland.
authors.forename: Salah-Dine
authors.initials: SD
authors.lastname: Chibout
authors.affiliation: Discovery and Investigative Safety, Preclinical Safety, Novartis Institutes for Biomedical Research, CH-4057 Basel, Switzerland. Electronic address: armin.wolf@novartis.com.
authors.forename: Armin
authors.initials: A
authors.lastname: Wolf
mesh_terms: D000638:Amiodarone / Q000008:administration & dosage / Q000633:toxicity; D000818:Animals; D002478:Cells, Cultured; D002746:Chlorpromazine / Q000008:administration & dosage / Q000633:toxicity; D002839:Chromans / Q000008:administration & dosage / Q000633:toxicity; D046508:Culture Techniques; D016572:Cyclosporine / Q000008:administration & dosage / Q000633:toxicity; D018492:Dopamine Antagonists / Q000008:administration & dosage / Q000633:toxicity; D005786:Gene Expression Regulation / Q000187:drug effects; D022781:Hepatocytes / Q000187:drug effects*; D007004:Hypoglycemic Agents / Q000008:administration & dosage / Q000633:toxicity; D007166:Immunosuppressive Agents / Q000008:administration & dosage / Q000633:toxicity; D008297:Male; D026902:Potassium Channel Blockers / Q000008:administration & dosage / Q000633:toxicity; D051381:Rats; D017208:Rats, Wistar; D045162:Thiazolidinediones / Q000008:administration & dosage / Q000633:toxicity; D000077288:Troglitazone
chemical_list: D002839:Chromans; D018492:Dopamine Antagonists; D007004:Hypoglycemic Agents; D007166:Immunosuppressive Agents; D026902:Potassium Channel Blockers; D045162:Thiazolidinediones; D016572:Cyclosporine; D000077288:Troglitazone; D000638:Amiodarone; D002746:Chlorpromazine
keywords: Hepatotoxicity; High content imaging; In vitro model; Long-term culture; Rat hepatocytes; Safety profiling