Efficient generation of functional hepatocytes from human induced pluripotent stem cells and isolation of hepatic progenitor cells

• Project/Area Number: 23790193
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Medical pharmacy
• Research Institution: Tohoku Pharmaceutical University
• Principal Investigator: SASAKI Takamitsu 東北薬科大学, 薬学部, 助教 (20382674)
• Project Period (FY): 2012 – 2013
• Project Status: Completed (Fiscal Year 2013)
• Budget Amount: ¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000); Fiscal Year 2012: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000); Fiscal Year 2011: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000)
• Keywords: 薬物動態 / 代謝学 / iPS 細胞 / 肝分化誘導法 / CYP / 肝特異的転写因子 / HNF / iPS細胞 / トランスポーター / 転写因子

Research Abstract

The purpose of this study was to establish the generation of functional hepatocytes from human induced pluripotent stem cells (iPSCs). It revealed that the hepatic differentiation method using human HNF6-expressing adenovirus contributed to the efficient development of hepatocytes from iPSCs. HNF6 was introduced into the iPSHCs, CYP3A4 expression dramatically increased and the cells had the capability to metabolize testosterone. Our study proved for the first time that HNF6 transactivates the CYP3A4gene in hepatocyte-like cells differentiated from iPSCs.

Research Products

• [Journal Article] Hepatocyte Nuclear Factor 6 Activates the Transcription of <i>CYP3A4</i> in Hepatocyte-like Cells Differentiated from Human Induced Pluripotent Stem Cells (2013)
  • Author(s): Sasaki T, Takahashi S, Numata Y, Narita M, Tanaka Y, Kondo Y, Matsunaga T, Ohmori S, Nagata K
  • Journal Title: Drug Metabolism and Pharmacokinetics Volume: 28 Issue: 3 Pages: 250-259
  • DOI: 10.2133/dmpk.DMPK-12-RG-132
  • NAID: 130004463310
  • ISSN: 1347-4367, 1880-0920
  • Peer Reviewed
• [Journal Article] Effects of Hypoxia-inducible Factor-1&alpha; Chemical Stabilizer, CoCl₂ and Hypoxia on Gene Expression of CYP3As in Human Fetal Liver Cells (2012)
  • Author(s): Suzuki E
  • Journal Title: Drug Metabolism and Pharmacokinetics Volume: 27 Issue: 4 Pages: 398-404
  • DOI: 10.2133/dmpk.DMPK-11-RG-074
  • NAID: 130004463262
  • ISSN: 1347-4367, 1880-0920
  • Peer Reviewed
• [Journal Article] Polycyclic Aromatic Hydrocarbons Activate CYP3A4 Gene Transcription through Human Pregnane X Receptor (2012)
  • Author(s): Kumagai T, Suzuki H, Sasaki T, Sakaguchi S, Miyairi S, Yamazoe Y, Nagata K
  • Journal Title: Drug Metabolism and Pharmacokinetics Volume: 27 Issue: 2 Pages: 200-206
  • DOI: 10.2133/dmpk.DMPK-11-RG-094
  • NAID: 130004463240
  • ISSN: 1347-4367, 1880-0920
  • Peer Reviewed
• [Journal Article] Association between Cancer Risk and Drug-metabolizing Enzyme Gene (CYP2A6, CYP2A13, CYP4B1, SULT1A1, GSTM1, and GSTT1) Polymorphisms in Cases of Lung Cancer in Japan (2011)
  • Author(s): Tamaki, Y., Arai, T., Sugimura, H., Sasaki, T., Honda, M., Muroi, Y., Matsubara, Y., Kanno, S., Ishikawa, M., Hirasawa, N., Hiratsuka, M.
  • Journal Title: Drug Metabolism and Pharmacokinetics Volume: 26 Issue: 5 Pages: 516-522
  • DOI: 10.2133/dmpk.DMPK-11-RG-046
  • NAID: 130004463215
  • ISSN: 1347-4367, 1880-0920
  • Peer Reviewed
• [Journal Article] Progression of Alcoholic and Non-alcoholic Steatohepatitis: Common Metabolic Aspects of Innate Immune System and Oxidative Stress (2011)
  • Author(s): Sakaguchi S, Takahashi S, Sasaki T, Kumagai T, Nagata K
  • Journal Title: Drug Metabolism and Pharmacokinetics Volume: 26 Issue: 1 Pages: 30-46
  • DOI: 10.2133/dmpk.DMPK-10-RV-087
  • NAID: 130004463160
  • ISSN: 1347-4367, 1880-0920
  • Peer Reviewed
• [Presentation] Hepatic differentiation of human induced pluripotent stem cells by using factors involved in liverfunction and development (2013)
  • Author(s): Sasaki T
  • Organizer: International Symposium for Neurosciences
  • Place of Presentation: 仙台
  • Year and Date: 2013-02-26
• [Presentation] Hepatic differentiation of human induced pluripotent stem cells by using factors involved in liver function and development (2013)
  • Author(s): Takamitsu Sasaki
  • Organizer: International Symposium for Neurosciences
  • Place of Presentation: 仙台
• [Presentation] microRNA 導入による肝薬物代謝酵素発現への影響 (2012)
  • Author(s): 菅野 高弘
  • Organizer: 第51回日本薬学会東北支部大会
  • Place of Presentation: 青森
  • Year and Date: 2012-10-07
• [Presentation] HNF6 導入時期による肝分化iPS 細胞の薬物代謝酵素発現への影響 (2012)
  • Author(s): 沼田 喜弘
  • Organizer: 第51回日本薬学会東北支部大会
  • Place of Presentation: 青森
  • Year and Date: 2012-10-07
• [Presentation] 健康食品によるシトクロムP450 活性阻害の検討 (2012)
  • Author(s): 齋藤 詩奈子
  • Organizer: 第51回日本薬学会東北支部大会
  • Place of Presentation: 青森
  • Year and Date: 2012-10-07
• [Presentation] 肝特異的転写因子 HNF-6 による薬物代謝酵素発現誘導:新規薬物代謝研究ツールを志向した肝分化 iPS 細胞樹立への応用 (2012)
  • Author(s): 佐々木 崇光
  • Organizer: 第1回東北薬科大学創薬研究センターシンポジウム
  • Place of Presentation: 仙台
  • Year and Date: 2012-05-19
• [Presentation] Identification of a novel transactivation region of the MRP3 gene (2012)
  • Author(s): Sasaki T
  • Organizer: International Symposium on PPF Molecular Pharmacokinetics
  • Place of Presentation: 東京
  • Year and Date: 2012-01-17
• [Presentation] MicroRNA enhances the expression of CYP genes in HepG2 cells and hepatocyte-like cells differentiatedfromhumaninducedpluripotentstemcells (2012)
  • Author(s): Sasaki T
  • Organizer: 27 th, JSSX annual meeting
  • Place of Presentation: 千葉
• [Presentation] Small Molecule Compounds Enhance Differentiation to Hepatocytes from induced Pluripotent Stem Cells (2012)
  • Author(s): Kondo Y
  • Organizer: 27 th, JSSX annual meeting
  • Place of Presentation: 千葉
• [Presentation] Identification of a novel transactivation mechanism of the MRP3 gene (2012)
  • Author(s): Numata Y
  • Organizer: 27 th, JSSX annual meeting
  • Place of Presentation: 千葉
• [Presentation] 健康食品と医薬品における薬物相互作用解明を目指した健康食品使用実態調査 (2012)
  • Author(s): 小西 麻美子
  • Organizer: 第20回クリニカルファーマシーシンポジウム
  • Place of Presentation: 福岡
• [Presentation] Enhanced expression of cytochrome P450 genes by hepatocyte nuclear factor-6 in hepatocyte-likecells differentiated from human induced pluripotent stem cells (2012)
  • Author(s): Takahashi S
  • Organizer: 19th International Symposium on MDO/12th European regional ISSX meeting
  • Place of Presentation: Noordwijik aan Zee, The Netherlands
• [Presentation] MicroRNA enhances the expression of CYP genes in HepG2 cells and hepatocyte-like cells differentiated from human induced pluripotent stem cells (2012)
  • Author(s): Takamitsu Sasaki
  • Organizer: 27 th, JSSX annual meeting
  • Place of Presentation: 千葉
• [Presentation] microRNA導入による肝薬物代謝酵素発現への影響 (2012)
  • Author(s): 菅野 高弘
  • Organizer: 第51回日本薬学会東北支部大会
  • Place of Presentation: 青森
• [Presentation] HNF6導入時期による肝分化iPS細胞の薬物代謝酵素発現への影響 (2012)
  • Author(s): 沼田 喜弘
  • Organizer: 第51回日本薬学会東北支部大会
  • Place of Presentation: 青森
• [Presentation] 健康食品によるシトクロムP450活性阻害の検討 (2012)
  • Author(s): 齋藤詩奈子
  • Organizer: 第51回日本薬学会東北支部大会
  • Place of Presentation: 青森
• [Presentation] Enhanced expression of cytochrome P450 genes by hepatocyte nuclear factor-6 in hepatocyte-like cells differentiated from human induced pluripotent stem cells (2012)
  • Author(s): Shogo Takahashi
  • Organizer: 19th International Symposium on MDO/12th European regional ISSX meeting
  • Place of Presentation: Noordwijik aan Zee, The Netherlands
• [Presentation] 肝特異的転写因子HNF-6による薬物代謝酵素発現誘導：新規薬物代謝研究ツールを志向した肝分化iPS細胞樹立への応用 (2012)
  • Author(s): 佐々木崇光
  • Organizer: 第1回東北薬科大学創薬研究センターシンポジウム
  • Place of Presentation: 仙台
• [Presentation] Identification of a novel transactivation region of the MRP3 gene (2012)
  • Author(s): Sasaki T, Numata Y, Narita M, Matsui S, Toriyabe T, Yamazoe Y, Nagata K
  • Organizer: International Symposium on PPF Molecular Pharmacokinetics
  • Place of Presentation: 一橋記念講堂（東京都）
• [Presentation] 凍結 P450 発現アデノウイルス感染細胞を用いた薬物代謝評価系の構築 (2011)
  • Author(s): 安藤 寛美
  • Organizer: 第50回日本薬学会東北支部大会
  • Place of Presentation: 仙台
  • Year and Date: 2011-10-30
• [Presentation] 板藍根による CYP3A4 活性誘導の検討 (2011)
  • Author(s): 菅原 亮輔
  • Organizer: 第50回日本薬学会東北支部大会
  • Place of Presentation: 仙台
  • Year and Date: 2011-10-30
• [Presentation] iPS 細胞を用いた肝分化誘導法の検討 (2011)
  • Author(s): 山田 健太
  • Organizer: 第50回日本薬学会東北支部大会
  • Year and Date: 2011-10-30
• [Presentation] Indirubin, a compornent of Ban-Lan-Gen, activates CYP3A4 gene transcription through human pregnane X receptor (2011)
  • Author(s): Kumagai T
  • Organizer: 26 th, JSSX annual meeting
  • Place of Presentation: 広島
• [Presentation] Hepatocyte nuclear factor-6 enhances expression of CYP3A4 in HepG2 cells and hepatocyte-like cells differentiated from human induced pluripotent stem cells (2011)
  • Author(s): Sasaki T
  • Organizer: 26 th, JSSX annual meeting
  • Place of Presentation: 広島
• [Presentation] iPS細胞を用いた肝分化誘導法の検討 (2011)
  • Author(s): 山田健太、佐々木崇光、高橋昌悟、松永民秀、永田清
  • Organizer: 第50回日本薬学会東北支部大会
  • Place of Presentation: 東北薬科大学（仙台市）
• [Presentation] 板藍根によるCYP3A4活性誘導の検討 (2011)
  • Author(s): 菅原亮輔、熊谷健、三浦真知、高橋昌悟、佐々木崇光、坂口修平、宮入伸一、永田清
  • Organizer: 第50回日本薬学会東北支部大会
  • Place of Presentation: 東北薬科大学（仙台市）
• [Presentation] 凍結P450発現アデノウイルス感染細胞を用いた薬物代謝評価系の構築 (2011)
  • Author(s): 安藤寛美、佐藤裕、高橋昌悟、佐々木崇光、永田清
  • Organizer: 第50回日本薬学会東北支部大会
  • Place of Presentation: 東北薬科大学（仙台市）
• [Presentation] Hepatocyte nuclear factor-6 enhances expression of CYP3A4 in HepG2 cells and hepatocyte-like cells differentiated from human induced pluripotent stem cells (2011)
  • Author(s): Sasaki T, Tanaka Y, Takahashi S, Kumagai T, Sakaguchi S, Matsunaga T, Nagata K
  • Organizer: 日本薬物動態学会第26回年会
  • Place of Presentation: 広島国際会議場（広島市）
• [Presentation] Indirubin, a compornent of Ban-Lan-Gen, activates CYP3A4 gene transcription through human pregnane X receptor (2011)
  • Author(s): Kumagai T, Sugawara R, Miura M, Sasaki T, Miyairi S, Nagata K
  • Organizer: 日本薬物動態学会第26回年会
  • Place of Presentation: 広島国際会議場（広島市）