title: Functional human induced hepatocytes (hiHeps) with bile acid synthesis and transport capacities: A novel in vitro cholestatic model.
abstract: Drug-induced cholestasis is a leading cause of drug withdrawal. However, the use of primary human hepatocytes (PHHs), the gold standard for predicting cholestasis in vitro, is limited by their high cost and batch-to-batch variability. Mature hepatocyte characteristics have been observed in human induced hepatocytes (hiHeps) derived from human fibroblast transdifferentiation. Here, we evaluated whether hiHeps could biosynthesize and excrete bile acids (BAs) and their potential as PHH alternatives for cholestasis investigations. Quantitative real-time PCR (qRT-PCR) and western blotting indicated that hiHeps highly expressed BA synthases and functional transporters. Liquid chromatography tandem mass spectrometry (LC-MS/MS) showed that hiHeps produced normal intercellular unconjugated BAs but fewer conjugated BAs than human hepatocytes. When incubated with representative cholestatic agents, hiHeps exhibited sensitive drug-induced bile salt export pump (BSEP) dysfunction, and their response to cholestatic agent-mediated cytotoxicity correlated well with that of PHHs (r2= 0.8032). Deoxycholic acid (DCA)-induced hepatotoxicity in hiHeps was verified by elevated aspartate aminotransferase (AST) and γ-glutamyl-transferase (γ-GT) levels. Mitochondrial damage and cell death suggested DCA-induced toxicity in hiHeps, which were attenuated by hepatoprotective drugs, as in PHHs. For the first time, hiHeps were reported to biosynthesize and excrete BAs, which could facilitate predicting cholestatic hepatotoxicity and screening potential therapeutic drugs against cholestasis.
authors.affiliation: Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai 201203, China.|University of Chinese Academy of Sciences, No. 19A Yuquan Road, Beijing, 100049, China.
authors.forename: Xuan
authors.initials: X
authors.lastname: Ni
authors.affiliation: State Key Laboratory of Cell Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, 200031, China.
authors.forename: Yimeng
authors.initials: Y
authors.lastname: Gao
authors.affiliation: Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai 201203, China.|University of Chinese Academy of Sciences, No. 19A Yuquan Road, Beijing, 100049, China.
authors.forename: Zhitao
authors.initials: Z
authors.lastname: Wu
authors.affiliation: Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai 201203, China.|University of Chinese Academy of Sciences, No. 19A Yuquan Road, Beijing, 100049, China.
authors.forename: Leilei
authors.initials: L
authors.lastname: Ma
authors.affiliation: Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai 201203, China.|University of Chinese Academy of Sciences, No. 19A Yuquan Road, Beijing, 100049, China.
authors.forename: Chen
authors.initials: C
authors.lastname: Chen
authors.affiliation: Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai 201203, China.|University of Chinese Academy of Sciences, No. 19A Yuquan Road, Beijing, 100049, China.
authors.forename: Le
authors.initials: L
authors.lastname: Wang
authors.affiliation: Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai 201203, China.|University of Chinese Academy of Sciences, No. 19A Yuquan Road, Beijing, 100049, China.
authors.forename: Yunfei
authors.initials: Y
authors.lastname: Lin
authors.affiliation: State Key Laboratory of Cell Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, 200031, China.
authors.forename: Lijian
authors.initials: L
authors.lastname: Hui
authors.affiliation: Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai 201203, China.|University of Chinese Academy of Sciences, No. 19A Yuquan Road, Beijing, 100049, China.
authors.forename: Guoyu
authors.initials: G
authors.lastname: Pan
mesh_terms: D000074020:ATP Binding Cassette Transporter, Subfamily B, Member 11 / Q000378:metabolism; D001647:Bile Acids and Salts / Q000009:adverse effects / Q000096:biosynthesis / Q000378:metabolism*; D001692:Biological Transport; D015415:Biomarkers; D002470:Cell Survival / Q000187:drug effects; D002779:Cholestasis / Q000209:etiology / Q000378:metabolism / Q000473:pathology; D004305:Dose-Response Relationship, Drug; D015971:Gene Expression Regulation, Enzymologic; D022781:Hepatocytes / Q000187:drug effects / Q000378:metabolism*; D006801:Humans; D066298:In Vitro Techniques; D020011:Protective Agents / Q000494:pharmacology
chemical_list: D000074020:ATP Binding Cassette Transporter, Subfamily B, Member 11; D001647:Bile Acids and Salts; D015415:Biomarkers; D020011:Protective Agents