title: Exploring Chronic Drug Effects on Microengineered Human Liver Cultures Using Global Gene Expression Profiling.
abstract: Global gene expression profiling is useful for elucidating a drug's mechanism of action on the liver; however, such profiling in rats is not very sensitive for predicting human drug-induced liver injury, while dedifferentiated monolayers of primary human hepatocytes (PHHs) do not permit chronic drug treatment. In contrast, micropatterned cocultures (MPCCs) containing PHH colonies and 3T3-J2 fibroblasts maintain a stable liver phenotype for 4-6 weeks. Here, we used MPCCs to test the hypothesis that global gene expression patterns in stable PHHs can be used to distinguish clinical hepatotoxic drugs from their non-liver-toxic analogs and understand the mechanism of action prior to the onset of overt hepatotoxicity. We found that MPCCs treated with the clinical hepatotoxic/non-liver-toxic pair, troglitazone/rosiglitazone, at each drug's reported and non-toxic Cmax (maximum concentration in human plasma) for 1, 7, and 14 days displayed a total of 12, 269, and 628 differentially expressed genes, respectively, relative to the vehicle-treated control. Troglitazone modulated >75% of transcripts across pathways such as fatty acid and drug metabolism, oxidative stress, inflammatory response, and complement/coagulation cascades. Escalating rosiglitazone's dose to that of troglitazone's Cmax increased modulated transcripts relative to the lower dose; however, over half the identified transcripts were still exclusively modulated by troglitazone. Last, other hepatotoxins (nefazodone, ibufenac, and tolcapone) also induced a greater number of differentially expressed genes in MPCCs than their non-liver-toxic analogs (buspirone, ibuprofen, and entacapone) following 7 days of treatment. In conclusion, MPCCs allow evaluation of time- and dose-dependent gene expression patterns in PHHs treated chronically with analog drugs.
authors.affiliation: School of Biomedical Engineering, Colorado State University, Fort Collins, Colorado, USA.|Department of Bioengineering, University of Illinois at Chicago, Chicago, Illinois, USA.
authors.forename: Brenton R
authors.initials: BR
authors.lastname: Ware
authors.affiliation: Hepregen Corporation, Medford, Massachusetts, USA.
authors.forename: Michael
authors.initials: M
authors.lastname: McVay
authors.affiliation: School of Biomedical Engineering, Colorado State University, Fort Collins, Colorado, USA.|Department of Chemical and Biological Engineering, Colorado State University, Fort Collins, Colorado, USA.
authors.forename: Wendy Y
authors.initials: WY
authors.lastname: Sunada
authors.affiliation: School of Biomedical Engineering, Colorado State University, Fort Collins, Colorado, USA.|Department of Bioengineering, University of Illinois at Chicago, Chicago, Illinois, USA.
authors.forename: Salman R
authors.initials: SR
authors.lastname: Khetani
mesh_terms: D016475:3T3 Cells; D000818:Animals; D002478:Cells, Cultured; D056486:Chemical and Drug Induced Liver Injury / Q000209:etiology* / Q000235:genetics; D002839:Chromans / Q000633:toxicity; D018920:Coculture Techniques; D004305:Dose-Response Relationship, Drug; D005347:Fibroblasts / Q000166:cytology / Q000187:drug effects*; D020869:Gene Expression Profiling; D022781:Hepatocytes / Q000166:cytology / Q000187:drug effects*; D006801:Humans; D008099:Liver / Q000187:drug effects*; D051379:Mice; D018384:Oxidative Stress / Q000187:drug effects / Q000235:genetics; D061251:Primary Cell Culture; D000077154:Rosiglitazone; D045162:Thiazolidinediones / Q000633:toxicity; D043922:Toxicogenetics; D059467:Transcriptome / Q000187:drug effects*; D000077288:Troglitazone
chemical_list: D002839:Chromans; D045162:Thiazolidinediones; D000077154:Rosiglitazone; D000077288:Troglitazone
keywords: drug-induced liver injury; micropatterned cocultures; micropatterning; primary human hepatocytes; toxicogenomics