Generation of a bile salt export pump deficiency model using patient-specific induced pluripotent stem cell-derived hepatocyte-like cells.
Bile salt export pump (BSEP) plays an important role in hepatic secretion of bile acids and its deficiency results in severe cholestasis and liver failure. Mutation of the ABCB11 gene encoding BSEP induces BSEP deficiency and progressive familial intrahepatic cholestasis type 2 (PFIC2). Because liver transplantation remains standard treatment for PFIC2, the development of a novel therapeutic option is desired. However, a well reproducible model, which is essential for the new drug development for PFIC2, has not been established. Therefore, we attempted to establish a PFIC2 model by using iPSC technology. Human iPSCs were generated from patients with BSEP-deficiency (BD-iPSC), and were differentiated into hepatocyte-like cells (HLCs). In the BD-iPSC derived HLCs (BD-HLCs), BSEP was not expressed on the cell surface and the biliary excretion capacity was significantly impaired. We also identified a novel mutation in the 5'-untranslated region of the ABCB11 gene that led to aberrant RNA splicing in BD-HLCs. Furthermore, to evaluate the drug efficacy, BD-HLCs were treated with 4-phenylbutyrate (4PBA). The membrane BSEP expression level and the biliary excretion capacity in BD-HLCs were rescued by 4PBA treatment. In summary, we succeeded in establishing a PFIC2 model, which may be useful for its pathophysiological analysis and drug development.
Department of Child Health, Faculty of Medicine, University of Tsukuba, Ibaraki, Japan.|Laboratory of Biochemistry and Molecular Biology, Graduate School of Pharmaceutical Sciences, Osaka University, Osaka, Japan.|Laboratory of Hepatocyte Regulation, National Institutes of Biomedical Innovation, Health and Nutrition, Osaka, Japan.
Kazuo
K
Imagawa
Laboratory of Biochemistry and Molecular Biology, Graduate School of Pharmaceutical Sciences, Osaka University, Osaka, Japan.|Laboratory of Hepatocyte Regulation, National Institutes of Biomedical Innovation, Health and Nutrition, Osaka, Japan.|The Keihanshin Consortium for Fostering the Next Generation of Global Leaders in Research (K-CONNEX), Kyoto University, Kyoto, Japan.
Kazuo
K
Takayama
Department of Child Health, Faculty of Medicine, University of Tsukuba, Ibaraki, Japan.
Shigemi
S
Isoyama
Department of Diagnostic Pathology, Kurume University Hospital, Fukuoka, Japan.
Ken
K
Tanikawa
Department of Surgery, Kanagawa Children's Medical Center, Kanagawa, Japan.
Masato
M
Shinkai
Laboratory of Applied Environmental Biology, Graduate School of Pharmaceutical Sciences, Osaka University, Osaka, Japan.
Kazuo
K
Harada
Laboratory of Biochemistry and Molecular Biology, Graduate School of Pharmaceutical Sciences, Osaka University, Osaka, Japan.
Masashi
M
Tachibana
Laboratory of Biochemistry and Molecular Biology, Graduate School of Pharmaceutical Sciences, Osaka University, Osaka, Japan.|Laboratory of Regulatory Sciences for Oligonucleotide Therapeutics, Clinical Drug Development Project, Graduate School of Pharmaceutical Sciences, Osaka University, Osaka, Japan.
Fuminori
F
Sakurai
Department of Medical Genetics, Faculty of Medicine, University of Tsukuba, Ibaraki, Japan.
Emiko
E
Noguchi
Laboratory of Applied Environmental Biology, Graduate School of Pharmaceutical Sciences, Osaka University, Osaka, Japan.
Kazumasa
K
Hirata
Department of Diagnostic Pathology, Kurume University Hospital, Fukuoka, Japan.
Masayoshi
M
Kage
Laboratory of Stem Cell Regulation, National Institutes of Biomedical Innovation, Health and Nutrition, Osaka, Japan.|Laboratory of Biomedical Innovation, Graduate School of Pharmaceutical Sciences, Osaka University, Osaka, Japan.
Kenji
K
Kawabata
Department of Child Health, Faculty of Medicine, University of Tsukuba, Ibaraki, Japan.
Ryo
R
Sumazaki
Laboratory of Biochemistry and Molecular Biology, Graduate School of Pharmaceutical Sciences, Osaka University, Osaka, Japan.|Laboratory of Hepatocyte Regulation, National Institutes of Biomedical Innovation, Health and Nutrition, Osaka, Japan.|Global Center for Medical Engineering and Informatics, Osaka University, Osaka, Japan.
Hiroyuki
H
Mizuguchi
D000074020:ATP Binding Cassette Transporter, Subfamily B, Member 11 / Q000235:genetics / Q000378:metabolism*; D001647:Bile Acids and Salts / Q000378:metabolism*; D001692:Biological Transport; D015415:Biomarkers; D002454:Cell Differentiation; D002460:Cell Line; D002478:Cells, Cultured; D002648:Child; D002780:Cholestasis, Intrahepatic / Q000175:diagnosis / Q000235:genetics / Q000378:metabolism; D005260:Female; D015870:Gene Expression; D022781:Hepatocytes / Q000166:cytology* / Q000378:metabolism*; D006801:Humans; D007150:Immunohistochemistry; D057026:Induced Pluripotent Stem Cells / Q000166:cytology*; D008099:Liver / Q000378:metabolism / Q000473:pathology; D009154:Mutation; D010641:Phenotype; D012333:RNA, Messenger / Q000235:genetics; D017422:Sequence Analysis, DNA
D000074020:ATP Binding Cassette Transporter, Subfamily B, Member 11; D001647:Bile Acids and Salts; D015415:Biomarkers; D012333:RNA, Messenger