Human induced-pluripotent stem cell-derived hepatocyte-like cells as an in vitro model of human hepatitis B virus infection

Sakurai, Fuminori; Mitani, Shigeru; Yamamoto, Takaya; Takayama, Kazuo; Tachibana, Masashi; Watashi, Koichi; Wakita, Takaji; Iijima, Sayuki; Tanaka, Yasuhito; Mizuguchi, Hiroyuki

doi:10.1038/srep45698

Cited by 45 publications

(38 citation statements)

References 58 publications

(79 reference statements)

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“…Based on the human hepatoma cell lines, several cell lines were generated by stable expressing hNTCP, but they lack the properties of PHHs and are malignant, which limits their applications in HBV research ( Nkongolo et al, 2014 ; Watashi et al, 2014 ; Xia et al, 2017 ). To solve the shortcomings of the PHHs and hepatoma cell lines, HLCs derived from human stem cells, especially from hiPSCs have been developed and demonstrated capable to support in vitro HBV infection ( Paganelli et al, 2013 ; Shlomai et al, 2014 ; Sakurai et al, 2017 ; Xia et al, 2017 ). In the present study, we differentiate the hiPSCs to hiPSC-HLCs.…”

Section: Discussionmentioning

confidence: 99%

A Chimeric Humanized Mouse Model by Engrafting the Human Induced Pluripotent Stem Cell-Derived Hepatocyte-Like Cell for the Chronic Hepatitis B Virus Infection

Yuan

Zhang

et al. 2018

Front. Microbiol.

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Humanized mouse model generated by grafting primary human hepatocytes (PHHs) to immunodeficient mouse has contributed invaluably to understanding the pathogenesis of hepatitis B virus (HBV). However, the source of PHHs is limited, which necessitates the search for alternatives. Recently, hepatocyte-like cells (HLCs) generated from human induced pluripotent stem cells (hiPSCs) have been used for in vitro HBV infection. Herein, we developed a robust human liver chimeric animal model to study in vivo HBV infection by engrafting the hiPSC-HLCs to Fah-/-Rag2-/-IL-2Rγc-/- SCID (FRGS) mice. After being optimized by a small molecule, XMU-MP-1, the hiPSC-HLCs engrafted FRGS (hHLC-FRGS) mice displayed approximately 40% liver chimerism at week 6 after engraftment and maintained at this level for at least 14 weeks. Viremia and HBV infection markers include antigens, RNA, DNA, and covalently closed circular DNA were detectable in HBV infected hHLC-FRGS mice. Furthermore, hiPSC-HLCs and hHLC-FRGS mice were successfully used to evaluate different antivirals. Therefore, we established a humanized mouse model for not only investigating HBV pathogenesis but also testing the effects of the anti-HBV drugs.Highlights: (1) The implanted hiPSC-HLCs established a long-term chimerism in FRGS mice liver. (2) hHLC-FRGS mice are adequate to support chronic HBV infection with a full viral life cycle. (3) hiPSC-HLCs and hHLC-FRGS mice are useful tools for evaluation of antivirals against HBV infection in vitro and in vivo.Research in Context To overcome the disadvantages of using primary human hepatocytes, we induced human pluripotent stem cells to hepatocyte-like cells (hiPSC-HLCs) that developed the capability to express important liver functional markers and critical host factors for HBV infection. The hiPSC-HLCs were permissive for the HBV infection and supported a full HBV replication. The hiPSC-HLCs were then engrafted to immunodeficient mouse to establish a chimeric liver mouse model, which was capable of supporting HBV infection in vivo and evaluating the effects of antiviral drugs. Our results shed light into improving the cellular and animal models for studying HBV and other hepatotropic viruses.

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Section: Discussionmentioning

confidence: 99%

A Chimeric Humanized Mouse Model by Engrafting the Human Induced Pluripotent Stem Cell-Derived Hepatocyte-Like Cell for the Chronic Hepatitis B Virus Infection

Yuan

Zhang

et al. 2018

Front. Microbiol.

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“…They showed that HBsAg was efficiently produced in the supernatant after infection; however, the HBsAg levels gradually dropped to background levels. In their study, Sakurai and coworkers observed an increase in HBsAg secretion in culture supernatant up to 17 days following HBV infection in iPS-HLCs [87]. This indicated that iPS-HLCs could support long-term HBV infection.…”

Section: In Vitro Model Systems Based On Inducible Pluripotent Stem Cmentioning

confidence: 93%

In Vitro Systems for Studying Different Genotypes/Sub-Genotypes of Hepatitis B Virus: Strengths and Limitations

Kinge

Bhoola

Kramvis

2020

Viruses

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Hepatitis B virus (HBV) infects the liver resulting in end stage liver disease, cirrhosis, and hepatocellular carcinoma. Despite an effective vaccine, HBV poses a serious health problem globally, accounting for 257 million chronic carriers. Unique features of HBV, including its narrow virus–host range and its hepatocyte tropism, have led to major challenges in the development of suitable in vivo and in vitro model systems to recapitulate the HBV replication cycle and to test various antiviral strategies. Moreover, HBV is classified into at least nine genotypes and 35 sub-genotypes with distinct geographical distributions and prevalence, which have different natural histories of infection, clinical manifestation, and response to current antiviral agents. Here, we review various in vitro systems used to study the molecular biology of the different (sub)genotypes of HBV and their response to antiviral agents, and we discuss their strengths and limitations. Despite the advances made, no system is ideal for pan-genotypic HBV research or drug development and therefore further improvement is required. It is necessary to establish a centralized repository of HBV-related generated materials, which are readily accessible to HBV researchers, with international collaboration toward advancement and development of in vitro model systems for testing new HBV antivirals to ensure their pan-genotypic and/or customized activity.

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“…Accordingly, protocols to differentiate ES/iPS cells into hepatocytes, hereafter iHep, has also evolved substantially over time; however, the degree of maturity is still far from that of freshly isolated PHH. [284][285][286][287] For example, transcriptome analysis of iHep demonstrated a highly distinctive gene expression profile compared with that of PHH. 284 These important observations collectively indicate that there remains a significant gap in terms of the degree of differen-tiation/maturation between genuine hepatocytes and iHeps.…”

Section: Regenerative Biologymentioning

confidence: 99%

Art of Making Artificial Liver: Depicting Human Liver Biology and Diseases in Mice

et al. 2020

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Advancement in both bioengineering and cell biology of the liver led to the establishment of the first-generation humanized liver chimeric mouse (HLCM) model in 2001. The HLCM system was initially developed to satisfy the necessity for a convenient and physiologically representative small animal model for studies of hepatitis B virus and hepatitis C virus infection. Over the last two decades, the HLCM system has substantially evolved in quality, production capacity, and utility, thereby growing its versatility beyond the study of viral hepatitis. Hence, it has been increasingly employed for a variety of applications including, but not limited to, the investigation of drug metabolism and pharmacokinetics and stem cell biology. To date, more than a dozen distinctive HLCM systems have been established, and each model system has similarities as well as unique characteristics, which are often perplexing for end-users. Thus, this review aims to summarize the history, evolution, advantages, and pitfalls of each model system with the goal of providing comprehensive information that is necessary for researchers to implement the ideal HLCM system for their purposes. Furthermore, this review article summarizes the contribution of HLCM and its derivatives to our mechanistic understanding of various human liver diseases, its potential for novel applications, and its current limitations.

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Human induced-pluripotent stem cell-derived hepatocyte-like cells as an in vitro model of human hepatitis B virus infection

Cited by 45 publications

References 58 publications

A Chimeric Humanized Mouse Model by Engrafting the Human Induced Pluripotent Stem Cell-Derived Hepatocyte-Like Cell for the Chronic Hepatitis B Virus Infection

A Chimeric Humanized Mouse Model by Engrafting the Human Induced Pluripotent Stem Cell-Derived Hepatocyte-Like Cell for the Chronic Hepatitis B Virus Infection

In Vitro Systems for Studying Different Genotypes/Sub-Genotypes of Hepatitis B Virus: Strengths and Limitations

Art of Making Artificial Liver: Depicting Human Liver Biology and Diseases in Mice

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