Human induced pluripotent stem cell-derived hepatic cell lines as a new model for host interaction with hepatitis B virus

Abstract

Hepatitis B virus (HBV) is not eradicated by current antiviral therapies due to persistence of HBV covalently closed circular DNA (cccDNA) in host cells, and thus development of novel culture models for productive HBV infection is urgently needed, which will allow the study of HBV cccDNA eradication. To meet this need, we developed culture models of HBV infection using human induced pluripotent stem cell-derived hepatocyte lineages, including immature proliferating hepatic progenitor-like cell lines (iPS-HPCs) and differentiated hepatocyte-like cells (iPS-Heps). These cells were susceptible to HBV infection, produced HBV particles, and maintained innate immune responses. The infection efficiency of HBV in iPS-HPCs predominantly depended on the expression levels of sodium taurocholate cotransporting polypeptide (NTCP), and was low relative to iPS-Heps: however, long-term culture of iPS-Heps was difficult. To provide a model for HBV persistence, iPS-HPCs overexpressing NTCP were established. The long-term persistence of HBV cccDNA was detected in iPS-HPCs overexpressing NTCP, and depended on the inhibition of the Janus-kinase signaling pathway. In conclusion, this study provides evidence that iPS-derived hepatic cell lines can be utilized for novel HBV culture models with genetic variation to investigate the interactions between HBV and host cells and the development of anti-HBV strategies.

Analysis of the mechanism underlying liver diseases using human induced pluripotent stem cells

“…Previous studies including our data showed that iPS-Heps can respond to anti-viral IFN-a stimulation, which is susceptible to HBV [19,52,53]. Immature proliferating hepatic progenitor-like cell lines derived from iPS cells (iPS-HPCs) at a relatively immature stage of hepatic differentiation can be infected with HBV in vitro [19].…”

“…Hepatocyte-like cells derived from iPS cells (iPS-Heps) exhibit properties of hepatocytes, such as albumin secretion and metabolic functional properties. Several pathophysiological disorders can be reproduced using human iPS-Heps; on the contrary, the phenotypes of iPS-Heps are immature compared with adult mature hepatocytes with respect to albumin production, cytochrome P450 (CYP) activity and metabolic functions [18,19]. Recent reports have shown that patientderived iPS-Heps have been used to investigate the pathophysiology of genetic disorders, such as a1antitrypsin deficiency, glycogen storage disease type 1, Crigler-Najjar syndrome, hereditary tyrosinemia type 1, Gaucher disease, Niemann-Pick disease type C and Wilson's disease [14][15][16]20,21].…”

“…Sodium taurocholate cotransporting polypeptide (NTCP) was reported to be an entry receptor for HBV, and the overexpression of NTCP in hepatoma cell lines rendered them susceptible to HBV infection [51]. However, hepatoma cell lines lack several cellular pathways, including innate immune responses, and the utilization of iPS cells as a model of hepatitis B virus infection for drug development has been reported [19,52,53].…”

“…Immature proliferating hepatic progenitor-like cell lines derived from iPS cells (iPS-HPCs) at a relatively immature stage of hepatic differentiation can be infected with HBV in vitro [19]. We have shown that iPS-HPCs overexpressing NTCP can be used as a model for HBV persistence and that these cells maintain innate immune responses against HBV [19]. These studies demonstrated that iPS-Heps and iPS-HPCs can be utilized as an experimental model to investigate HBV-host interactions, develop novel therapies and elucidate the association between HBV persistence and cell differentiation.…”

Vasoactive Intestinal Peptide Derived From Liver Mesenchymal Cells Mediates Tight Junction Assembly in Mouse Intrahepatic Bile Ducts

“…Lentiviruses expressing shRNA were produced in 293T cells, as described. (26) HPPL (a mouse hepatoblast cell line) (27) or primary hepatoblasts were infected with the lentiviruses at five multiplicity-of-infection at day 3 of 2D preculture before cholangiocytic differentiation. Cells were collected on day 7 of preculture and subjected to 3D culture for cholangiocytic differentiation.…”