Human iPSC-derived hepatocyte system models cholestasis with tight junction protein 2 deficiency

Chao Zheng Li, Hiromi Ogawa, Soon Seng Ng, Xindi Chen, Eriko Kishimoto, Kokoro Sakabe, Aiko Fukami, Yueh Chiang Hu, Christopher N. Mayhew, Jennifer Hellmann, Alexander Miethke, Nahrin L. Tasnova, Samuel J.I. Blackford, Zu Ming Tang, Adam M. Syanda, Liang Ma, Fang Xiao, Melissa Sambrotta, Oliver Tavabie, Filipa SoaresOliver Baker, Davide Danovi, Hisamitsu Hayashi, Richard J. Thompson, S. Tamir Rashid, Akihiro Asai*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

4 Citations (Scopus)


Background & Aims: The truncating mutations in tight junction protein 2 (TJP2) cause progressive cholestasis, liver failure, and hepatocyte carcinogenesis. Due to the lack of effective model systems, there are no targeted medications for the liver pathology with TJP2 deficiency. We leveraged the technologies of patient-specific induced pluripotent stem cells (iPSC) and CRISPR genome-editing, and we aim to establish a disease model which recapitulates phenotypes of patients with TJP2 deficiency. Methods: We differentiated iPSC to hepatocyte-like cells (iHep) on the Transwell membrane in a polarized monolayer. Immunofluorescent staining of polarity markers was detected by a confocal microscope. The epithelial barrier function and bile acid transport of bile canaliculi were quantified between the two chambers of Transwell. The morphology of bile canaliculi was measured in iHep cultured in the Matrigel sandwich system using a fluorescent probe and live-confocal imaging. Results: The iHep differentiated from iPSC with TJP2 mutations exhibited intracellular inclusions of disrupted apical membrane structures, distorted canalicular networks, altered distribution of apical and basolateral markers/transporters. The directional bile acid transport of bile canaliculi was compromised in the mutant hepatocytes, resembling the disease phenotypes observed in the liver of patients. Conclusions: Our iPSC-derived in vitro hepatocyte system revealed canalicular membrane disruption in TJP2 deficient hepatocytes and demonstrated the ability to model cholestatic disease with TJP2 deficiency to serve as a platform for further pathophysiologic study and drug discovery. Lay summary: We investigated a genetic liver disease, progressive familial intrahepatic cholestasis (PFIC), which causes severe liver disease in newborns and infants due to a lack of gene called TJP2. By using cutting-edge stem cell technology and genome editing methods, we established a novel disease modeling system in cell culture experiments. Our experiments demonstrated that the lack of TJP2 induced abnormal cell polarity and disrupted bile acid transport. These findings will lead to the subsequent investigation to further understand disease mechanisms and develop an effective treatment.

Original languageEnglish
Article number100446
JournalJHEP Reports
Issue number4
Early online date1 Feb 2022
Publication statusPublished - Apr 2022


  • Bile acid transport
  • Cellular polarity
  • PFIC (progressive familial intrahepatic cholestasis)


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