Generation of CYP3A4-expressing human ES/iPS cell-derived hepatocytes for drug screening

• Project/Area Number: 15H06365
• Research Category: Grant-in-Aid for Research Activity Start-up
• Allocation Type: Single-year Grants
• Research Field: Medical pharmacy
• Research Institution: Osaka University
• Principal Investigator: Takayama Kazuo 大阪大学, 薬学研究科, 特任助教(常勤) (10759509)
• Project Period (FY): 2015-08-28 – 2017-03-31
• Project Status: Completed (Fiscal Year 2016)
• Budget Amount: ¥2,860,000 (Direct Cost: ¥2,200,000、Indirect Cost: ¥660,000); Fiscal Year 2016: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2015: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
• Keywords: 肝細胞 / ヒトES/iPS細胞 / 薬物代謝 / CYP / 肝毒性 / 創薬試験 / CYP3A4 / ゲノム編集 / 薬物代謝酵素 / 相同組換え

Outline of Final Research Achievements

In this study, we aimed to establish an efficient differentiation method for highly functioning hepatocyte-like cells from human ES/iPS cells. First, we attempted to improve the hepatocyte differentiation method. Next, we developed a method to concentrate the highly functioning hepatocyte-like cells. The hepatic functions of human ES/iPS cell-derived hepatocyte-like cells were enhanced by culturing with conditioned medium of hepatocytes and cholangiocytes. In future, our group is going to establish the genetically engineered human ES/iPS cells, which carry puromycin resistant cassette under the CYP3A4.

Research Products

• [Journal Article] Human iPS Cell-Derived Hepatocyte-Like Cells Achieve Zone-Specific Hepatic Properties by Modulation of WNT Signaling (2017)
  • Author(s): Mitani S., Takayama K., Nagamoto Y., Imagawa K., Sakurai F., Tachibana M., Sumazaki R., Mizuguchi H.
  • Journal Title: Molecular Therapy Volume: 25 Pages: 1420-1433
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Isolation and Expansion of Human Pluripotent Stem Cell-derived Hepatic Progenitor Cells by Growth Factor Defined Serum-free Culture Conditions (2017)
  • Author(s): Fukuda T., Takayama K., Hirata M., Liu Y., Yanagihara K., Suga M., Mizuguchi H., Furue M.
  • Journal Title: Experimental Cell Research Volume: 352 Issue: 2 Pages: 333-345
  • DOI: 10.1016/j.yexcr.2017.02.022
  • Peer Reviewed / Open Access
• [Journal Article] Highly efficient biallelic genome editing of human ES/iPS cells using a CRISPR/Cas9 or TALEN systems (2017)
  • Author(s): Takayama K., Igai K., Hagihara Y., Hashimoto R., Hanawa M., Sakuma T., Tachibana M., Sakurai F., Yamamoto T., Mizuguchi H
  • Journal Title: Nucleic Acids Research Volume: 45 Issue: 9 Pages: 5198-5207
  • DOI: 10.1093/nar/gkx130
  • Peer Reviewed / Open Access
• [Journal Article] Hepatocyte Nuclear Factor 4 Alpha Promotes Definitive Endoderm Differentiation From Human Induced Pluripotent Stem Cells (2017)
  • Author(s): Hanawa M., Takayama K., Sakurai F., Tachibana M., Mizuguchi H.
  • Journal Title: Stem Cells Reviews & Reports Volume: 13 Issue: 4 Pages: 542-551
  • DOI: 10.1007/s12015-016-9709-x
  • Peer Reviewed
• [Journal Article] Generation of a bile salt export pump deficiency model using patient-specific induced pluripotent stem cell-derived hepatocyte-like cells (2017)
  • Author(s): Imagawa K. 1, Takayama K. 1, Isoyama S., Tanikawa K., Shinkai M., Harada K., Tachibana M., Sakurai F., Noguchi E., Hirata K., Kage M., Kawabata K., Sumazaki R., Mizuguchi H.
  • Journal Title: Scientific Reports Volume: 7 Issue: 1 Pages: 41806-41806
  • DOI: 10.1038/srep41806
  • NAID: 120007135051
  • Peer Reviewed / Open Access
• [Journal Article] Prediction of drug-induced liver injury by using human pluripotent stem cell-derived hepatocyte-like cells. (2017)
  • Author(s): Takayama K., Mizuguchi H.
  • Journal Title: Drug Metabolism and Pharmacokinetics Volume: 32 Issue: 1 Pages: 12-20
  • DOI: 10.1016/j.dmpk.2016.10.408
  • Peer Reviewed
• [Journal Article] Prediction of differentiation tendency toward hepatocytes from gene expression in undifferentiated human pluripotent stem cells. (2016)
  • Author(s): Yanagihara K., Liu Y., Kanie K., Takayama K., Okamura M., Hirata M., Fukuda T., Suga M., Nikawa H., Mizuguchi H., Kato R., Furue M.
  • Journal Title: Stem Cells and Development Volume: 25 Pages: 1884-1897
  • Peer Reviewed
• [Journal Article] Laminin 411 and 511 promote the cholangiocyte differentiation of human induced pluripotent stem cells (2016)
  • Author(s): Takayama K., Mitani S., Nagamoto Y., Sakurai F., Tachibana M., Taniguchi Y., Sekiguchi K., Mizuguchi H.
  • Journal Title: Biochem Biophys Res Commun. Volume: 474 Issue: 1 Pages: 91-96
  • DOI: 10.1016/j.bbrc.2016.04.075
  • Peer Reviewed
• [Journal Article] Modeling of drug-mediated CYP3A4 induction by using human iPS cell-derived enterocyte-like cells (2016)
  • Author(s): Negoro R., Takayama K., Nagamoto Y, Sakurai F., Tachibana M, Mizuguchi H.
  • Journal Title: Biochem Biophys Res Commun. Volume: 472 Issue: 4 Pages: 631-636
  • DOI: 10.1016/j.bbrc.2016.03.012
  • Peer Reviewed / Open Access
• [Journal Article] Hepatic Maturation of Human iPS Cell-Derived Hepatocyte-Like Cells by ATF5, c/EBPα, and PROX1 Transduction (2016)
  • Author(s): Nakamori D., Takayama K., Nagamoto Y., Mitani S., Sakurai F., Tachibana M., Mizuguchi H.
  • Journal Title: Biochem Biophys Res Commun Volume: 469 Issue: 3 Pages: 424-429
  • DOI: 10.1016/j.bbrc.2015.12.007
  • Peer Reviewed / Open Access
• [Journal Article] Generation of enterocyte-like cells from human induced pluripotent stem cells for drug absorption and metabolism studies in human small intestine (2015)
  • Author(s): Ozawa T., Takayama K., Okamoto R., Negoro R., Sakurai F., Tachibana M., Kawabata K., Mizuguchi H.
  • Journal Title: Scientific Reports Volume: 12 Issue: 1 Pages: 16479-16479
  • DOI: 10.1038/srep16479
  • Peer Reviewed / Open Access
• [Presentation] ヒトES/iPS細胞における高効率相同組換え技術の開発 (2017)
  • Author(s): 高山和雄、井貝圭佑、櫻井文教、水口裕之
  • Organizer: 日本薬学会第137年会
  • Place of Presentation: 仙台
  • Year and Date: 2017-03-25
• [Presentation] ヒトES/iPS細胞由来肝臓細胞の作製とその創薬研究への利用 (2017)
  • Author(s): 高山和雄
  • Organizer: 第6回超異分野学会
  • Place of Presentation: 東京
  • Year and Date: 2017-03-02
  • Invited
• [Presentation] ヒトES/iPS細胞由来肝細胞の三次元培養による機能向上 (2016)
  • Author(s): 高山和雄
  • Organizer: 細胞凝集研究会2016
  • Place of Presentation: 札幌
  • Year and Date: 2016-09-09
  • Invited
• [Presentation] ヒトiPS細胞由来肝細胞の創薬・再生医療への応用 (2016)
  • Author(s): 高山和雄
  • Organizer: K-CONNEXスタートアップシンポジウム
  • Place of Presentation: 大阪
  • Year and Date: 2016-01-29
• [Presentation] Generation of enterocyte-like cells from human iPS cells for drug absorption and metabolism studies in small intestine (2015)
  • Author(s): Takayama K., Ozawa T., Sakurai F., Tachibana M., Mizuguchi H.
  • Organizer: 日本薬物動態学会第30回年会
  • Place of Presentation: 東京
  • Year and Date: 2015-11-12
• [Presentation] ヒト多能性幹細胞から肝細胞様細胞への高効率分化誘導技術の開発―創薬応用を目指して― (2015)
  • Author(s): 高山和雄、三村菜摘、萩原康子、立花雅史、櫻井文教、神田勝弘、川端健二、水口裕之
  • Organizer: 第65回日本薬学会近畿支部総会・大会
  • Place of Presentation: 大阪
  • Year and Date: 2015-10-17
• [Book] ヒトiPS細胞由来肝細胞の創薬研究・非臨床試験への応用を目指して (2015)
  • Author(s): 高山和雄、水口裕之
  • Total Pages: 5
  • Publisher: 薬剤学
• [Book] ヒト多能性幹細胞由来肝細胞を用いた次世代型創薬の実現を目指した基盤技術創成 (2015)
  • Author(s): 高山和雄
  • Total Pages: 6
  • Publisher: 薬学雑誌