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Small molecule-mediated reprogramming of human hepatocytes into bipotent progenitor cells

  • Author Footnotes
    † Authors contributed equally to this study as co-first authors.
    Yohan Kim
    Footnotes
    † Authors contributed equally to this study as co-first authors.
    Affiliations
    Department of Surgery, Hanyang University College of Medicine, Seoul 04763, Republic of Korea

    HY Indang Center of Regenerative Medicine and Stem Cell Research, Hanyang University, Seoul 04763, Republic of Korea
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  • Author Footnotes
    † Authors contributed equally to this study as co-first authors.
    Kyojin Kang
    Footnotes
    † Authors contributed equally to this study as co-first authors.
    Affiliations
    Department of Surgery, Hanyang University College of Medicine, Seoul 04763, Republic of Korea

    HY Indang Center of Regenerative Medicine and Stem Cell Research, Hanyang University, Seoul 04763, Republic of Korea
    Search for articles by this author
  • Author Footnotes
    † Authors contributed equally to this study as co-first authors.
    Seung Bum Lee
    Footnotes
    † Authors contributed equally to this study as co-first authors.
    Affiliations
    Laboratory of Radiation Exposure & Therapeutics, National Radiation Emergency Medical Center, Korea Institute of Radiological & Medical Science, Seoul 01812, Republic of Korea
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  • Author Footnotes
    † Authors contributed equally to this study as co-first authors.
    Daekwan Seo
    Footnotes
    † Authors contributed equally to this study as co-first authors.
    Affiliations
    Macrogen Corporation, Rockville, MD 20850, USA
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  • Sangtae Yoon
    Affiliations
    Department of Surgery, Hanyang University College of Medicine, Seoul 04763, Republic of Korea

    HY Indang Center of Regenerative Medicine and Stem Cell Research, Hanyang University, Seoul 04763, Republic of Korea
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  • Sung Joo Kim
    Affiliations
    Department of Surgery, Samsung Medical Center, Sungkyunkwan University College of Medicine, Seoul 03063, Republic of Korea
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  • Kiseok Jang
    Affiliations
    Department of Pathology, Hanyang University College of Medicine, Seoul 04763, Republic of Korea
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  • Yun Kyung Jung
    Affiliations
    Department of Surgery, Hanyang University College of Medicine, Seoul 04763, Republic of Korea
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  • Kyeong Geun Lee
    Affiliations
    Department of Surgery, Hanyang University College of Medicine, Seoul 04763, Republic of Korea
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  • Valentina M. Factor
    Affiliations
    Laboratory of Molecular Pharmacology, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892, USA
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  • Jaemin Jeong
    Correspondence
    Corresponding authors. Address: Department of Surgery, Hanyang University College of Medicine, Seoul 04763, Republic of Korea; Tel.: +82 2 2290 8449; fax: +82 2 2281 0224 (D. Choi), or Department of Surgery, Hanyang University College of Medicine, Seoul 04763, Republic of Korea; Tel.: +82 2 2220 0647; fax: +82 2 2281 0224. (J. Jeong).
    Affiliations
    Department of Surgery, Hanyang University College of Medicine, Seoul 04763, Republic of Korea

    HY Indang Center of Regenerative Medicine and Stem Cell Research, Hanyang University, Seoul 04763, Republic of Korea
    Search for articles by this author
  • Dongho Choi
    Correspondence
    Corresponding authors. Address: Department of Surgery, Hanyang University College of Medicine, Seoul 04763, Republic of Korea; Tel.: +82 2 2290 8449; fax: +82 2 2281 0224 (D. Choi), or Department of Surgery, Hanyang University College of Medicine, Seoul 04763, Republic of Korea; Tel.: +82 2 2220 0647; fax: +82 2 2281 0224. (J. Jeong).
    Affiliations
    Department of Surgery, Hanyang University College of Medicine, Seoul 04763, Republic of Korea

    HY Indang Center of Regenerative Medicine and Stem Cell Research, Hanyang University, Seoul 04763, Republic of Korea
    Search for articles by this author
  • Author Footnotes
    † Authors contributed equally to this study as co-first authors.
Published:September 18, 2018DOI:https://doi.org/10.1016/j.jhep.2018.09.007

      Highlights

      • Human hepatic progenitors (hCdHs) are generated from adult hepatocytes.
      • HGF is required for chemical reprogramming induced by A83-01 and CHIR99021.
      • hCdHs proliferate for at least 10 passages without losing differentiation potential in vitro.
      • Bipotent hCdHs can repopulate injured liver and acquire functional properties.

      Background & Aims

      Currently, much effort is directed towards the development of new cell sources for clinical therapy using cell fate conversion by small molecules. Direct lineage reprogramming to a progenitor state has been reported in terminally differentiated rodent hepatocytes, yet remains a challenge in human hepatocytes.

      Methods

      Human hepatocytes were isolated from healthy and diseased donor livers and reprogrammed into progenitor cells by 2 small molecules, A83-01 and CHIR99021 (AC), in the presence of EGF and HGF. The stemness properties of human chemically derived hepatic progenitors (hCdHs) were tested by standard in vitro and in vivo assays and transcriptome profiling.

      Results

      We developed a robust culture system for generating hCdHs with therapeutic potential. The use of HGF proved to be an essential determinant of the fate conversion process. Based on functional evidence, activation of the HGF/MET signal transduction system collaborated with A83-01 and CHIR99021 to allow a rapid expansion of progenitor cells through the activation of the ERK pathway. hCdHs expressed hepatic progenitor markers and could self-renew for at least 10 passages while retaining a normal karyotype and potential to differentiate into functional hepatocytes and biliary epithelial cells in vitro. Gene expression profiling using RNAseq confirmed the transcriptional reprogramming of hCdHs towards a progenitor state and the suppression of mature hepatocyte transcripts. Upon intrasplenic transplantation in several models of therapeutic liver repopulation, hCdHs effectively repopulated the damaged parenchyma.

      Conclusion

      Our study is the first report of successful reprogramming of human hepatocytes to a population of proliferating bipotent cells with regenerative potential. hCdHs may provide a novel tool that permits expansion and genetic manipulation of patient-specific progenitors to study regeneration and the repair of diseased livers.

      Lay summary

      Human primary hepatocytes were reprogrammed towards hepatic progenitor cells by a combined treatment with 2 small molecules, A83-01 and CHIR99021, and HGF. Chemically derived hepatic progenitors exhibited a high proliferation potential and the ability to differentiate into hepatocytes and biliary epithelial cells both in vitro and in vivo. This approach enables the generation of patient-specific hepatic progenitors and provides a platform for personal and stem cell-based regenerative medicine.

      Graphical abstract

      Keywords

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