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

Yohan Kim, Kyojin Kang, Seung Bum Lee, Daekwan Seo, Sangtae Yoon, Sung Joo Kim, Kiseok Jang, Yun Kyung Jung, Kyeong Geun Lee, Valentina M. Factor, Jaemin Jeong, Dongho Choi

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)97-107
Number of pages11
JournalJournal of Hepatology
Volume70
Issue number1
DOIs
StatePublished - 2019 Jan 1

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Hepatocytes
Stem Cells
Liver
Gene Expression Profiling
Liver Diseases
Epithelial Cells
Regenerative Medicine
MAP Kinase Signaling System
Cell- and Tissue-Based Therapy
Karyotype
Epidermal Growth Factor
Regeneration
Rodentia
Signal Transduction
Therapeutics
Transplantation
Tissue Donors
Population
In Vitro Techniques
Chir 99021

Keywords

  • Human chemically derived hepatic progenitors
  • Human hepatocytes
  • Reprogramming
  • Small molecules

Cite this

Kim, Yohan ; Kang, Kyojin ; Lee, Seung Bum ; Seo, Daekwan ; Yoon, Sangtae ; Kim, Sung Joo ; Jang, Kiseok ; Jung, Yun Kyung ; Lee, Kyeong Geun ; Factor, Valentina M. ; Jeong, Jaemin ; Choi, Dongho. / Small molecule-mediated reprogramming of human hepatocytes into bipotent progenitor cells. In: Journal of Hepatology. 2019 ; Vol. 70, No. 1. pp. 97-107.
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abstract = "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.",
keywords = "Human chemically derived hepatic progenitors, Human hepatocytes, Reprogramming, Small molecules",
author = "Yohan Kim and Kyojin Kang and Lee, {Seung Bum} and Daekwan Seo and Sangtae Yoon and Kim, {Sung Joo} and Kiseok Jang and Jung, {Yun Kyung} and Lee, {Kyeong Geun} and Factor, {Valentina M.} and Jaemin Jeong and Dongho Choi",
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Kim, Y, Kang, K, Lee, SB, Seo, D, Yoon, S, Kim, SJ, Jang, K, Jung, YK, Lee, KG, Factor, VM, Jeong, J & Choi, D 2019, 'Small molecule-mediated reprogramming of human hepatocytes into bipotent progenitor cells', Journal of Hepatology, vol. 70, no. 1, pp. 97-107. https://doi.org/10.1016/j.jhep.2018.09.007

Small molecule-mediated reprogramming of human hepatocytes into bipotent progenitor cells. / Kim, Yohan; Kang, Kyojin; Lee, Seung Bum; Seo, Daekwan; Yoon, Sangtae; Kim, Sung Joo; Jang, Kiseok; Jung, Yun Kyung; Lee, Kyeong Geun; Factor, Valentina M.; Jeong, Jaemin; Choi, Dongho.

In: Journal of Hepatology, Vol. 70, No. 1, 01.01.2019, p. 97-107.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Small molecule-mediated reprogramming of human hepatocytes into bipotent progenitor cells

AU - Kim, Yohan

AU - Kang, Kyojin

AU - Lee, Seung Bum

AU - Seo, Daekwan

AU - Yoon, Sangtae

AU - Kim, Sung Joo

AU - Jang, Kiseok

AU - Jung, Yun Kyung

AU - Lee, Kyeong Geun

AU - Factor, Valentina M.

AU - Jeong, Jaemin

AU - Choi, Dongho

PY - 2019/1/1

Y1 - 2019/1/1

N2 - 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.

AB - 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.

KW - Human chemically derived hepatic progenitors

KW - Human hepatocytes

KW - Reprogramming

KW - Small molecules

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