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J. Hepatol., Available online 19 September 2018 | https://doi.org/10.1016/j.jhep.2018.09.007 Small molecule-mediated reprogramming of human hepatocytes into bipotent progenitor cells

Yohan Kim^1,2,†, Kyojin Kang^1,2,†, Seung Bum Lee^3,†, Daekwan Seo^4,†, Sangtae Yoon^1,2, Heung Mo Yang⁵, Sung Joo Kim⁵, Kiseok Jang⁶, Yun Kyung Jung¹, Kyeong Geun Lee¹, Valentina M Factor⁷, Jaemin Jeong^1,2,*, Dongho Choi^1,2,*¹Department of Surgery, Hanyang University College of Medicine, Seoul 04763, Korea

²HY Indang Center of Regenerative Medicine and Stem Cell Research, Hanyang University, Seoul 04763, Korea

³Laboratory of Radiation Exposure & Therapeutics, National Radiation Emergency Medical Center, Korea Institute of Radiological & Medical Science, Seoul 01812, Korea

⁴Macrogen Corporation, Rockville, MD 20850, USA

⁵Department of Surgery, Samsung Medical Center, Sungkyunkwan University College of Medicine, Seoul 03063, Korea

⁶Department of Pathology, Hanyang University College of Medicine, Seoul 04763, Korea

⁷Laboratory of Molecular Pharmacology, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892, USA
^†Authors contributed equally to this study as co-first authors.
^*Corresponding authors

Abstract

Background & Aims

Currently, much effort is directed towards the development of new cell sources for clinical therapy using cell fate conversion approaches 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 two 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 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 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 toward 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 a 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 two 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 allows to generate patient-specific hepatic progenitors and provides a platform for personal and stem cell-based regenerative medicine.

Keywords : Human hepatocytes, human chemically derived hepatic progenitors, reprogramming, small molecules

논문정보

형식Research article
게재일2018년 09월 (BRIC 등록일 2018-10-05)
연구진국내(교신)+국외 연구진
분야 의약학 > 분자세포의학

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