Development and clinical evaluation of hepatitis onset model of chimeric mice with iPS-derived human hepatocytes

• Project/Area Number: 25293176
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Partial Multi-year Fund
• Section: 一般
• Research Field: Gastroenterology
• Research Institution: Nagoya City University
• Principal Investigator: Tanaka Yasuhito 名古屋市立大学, 医学(系)研究科(研究院), 教授 (90336694)
• Co-Investigator(Kenkyū-buntansha): MATSUNAGA Tamihide 名古屋市立大学, 大学院薬学研究科, 教授 (40209581)
• Project Period (FY): 2013-04-01 – 2016-03-31
• Project Status: Completed (Fiscal Year 2015)
• Budget Amount: ¥18,590,000 (Direct Cost: ¥14,300,000、Indirect Cost: ¥4,290,000); Fiscal Year 2015: ¥5,850,000 (Direct Cost: ¥4,500,000、Indirect Cost: ¥1,350,000); Fiscal Year 2014: ¥5,850,000 (Direct Cost: ¥4,500,000、Indirect Cost: ¥1,350,000); Fiscal Year 2013: ¥6,890,000 (Direct Cost: ¥5,300,000、Indirect Cost: ¥1,590,000)
• Keywords: 肝炎ウイルス / HBV / iPS細胞 / キメラマウス / HCV

Outline of Final Research Achievements

In the differentiation to hepatocytes from human iPS cells, it was revealed that the HDAC inhibitor valproic acid in combination with cytokines induce differentiation cells into hepatic progenitor cells followed by maturation into functional hepatocytes. The hepatocytes formed a spheroid by the three-dimensional culture, and the gene expression of CYP3A which is hepatocytes marker and drug metabolizing enzyme is markedly higher than that in 2D cultures. The 3D-cultured iPS-hepatocyte spheroids were transplanted into the portal vein of highly immunodeficient NOG mice (TK-NOG) that is induced liver injury by ganciclovir treatment. At one week post-injection of hepatic spheroids, human albumin levels elevated temporarily.

Research Products

• [Int'l Joint Research] Suez Canal University(エジプト)
• [Journal Article] Virological characteristics of hepatitis B genotype G/A2 recombination virus in Japan. (2016)
  • Author(s): Tsuzuki Y, Watanabe T, Iio E, Fujisaki S, Ibe S, Kani S, Hamada-Tsutsumi S, Yokomaku Y, Iwatani Y, Sugiura W, Okuse C, Okumura A, Sato Y, Tanaka Y.
  • Journal Title: Hepatol Res. Volume: 印刷中 Issue: 8 Pages: 75-783
  • DOI: 10.1111/hepr.12612
  • Peer Reviewed
• [Journal Article] Characterization of novel entecavir resistance mutations. (2015)
  • Author(s): Hayashi S, Murakami S, Omagari K, Matsui T, Iio E, Isogawa M, Watanabe T, Karino Y, Tanaka Y.
  • Journal Title: J Hepatol. Volume: 63(3) Issue: 3 Pages: 546-53
  • DOI: 10.1016/j.jhep.2015.03.020
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Hepatitis B surface gene variants isolated from blood donors with overt and occult HBV infection in north eastern Egypt. (2015)
  • Author(s): Kishk R, Nemr N, Elkady A, Mandour M, Aboelmagd M, Ramsis N, Hassan M, Soliman N, Iijima S, Murakami S, Tanaka Y, Ragheb M.
  • Journal Title: Virol J. Volume: 12 Issue: 1
  • DOI: 10.1186/s12985-015-0389-y
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] 個別化（オーダーメイド）医療を志向した薬物動態研究および毒性試験へのヒトiPS細胞の利用 (2015)
  • Author(s): 岩尾岳洋，松永民秀
  • Journal Title: Organ Biology Volume: 22 Pages: 39-48
  • NAID: 130005098698
• [Journal Article] Postexposure Prophylactic Effect of Hepatitis B Virus (HBV)-Active Antiretroviral Therapy against HBV Infection. (2015)
  • Author(s): Watanabe T, Hamada-Tsutsumi S, Yokomaku Y, Imamura J, Sugiura W, Tanaka Y.
  • Journal Title: Antimicrob Agents Chemother. Volume: 59 Issue: 2 Pages: 1292-1298
  • DOI: 10.1128/aac.04459-14
  • Peer Reviewed / Open Access
• [Journal Article] Hepatitis virus infection affects DNA methylation in mice with humanized livers. (2014)
  • Author(s): Okamoto Y, Shinjo K, et al.
  • Journal Title: Gastroenterology Volume: 146 Issue: 2 Pages: 562-572
  • DOI: 10.1053/j.gastro.2013.10.056
  • Peer Reviewed / Open Access
• [Journal Article] Reactivation of hepatitis B virus in patients with undetectable HBsAg undergoing chemotherapy for malignant lymphoma or multiple myeloma. (2013)
  • Author(s): Matsui T, Kang JH, Nojima M, Tomonari A, Aoki H, Yamazaki H, Yane K, Tsuji K, Andoh S, Andoh S, Sakai H, Maemori M, Maguchi H, Tanaka Y.
  • Journal Title: J Med Virol. Volume: 85(11) Issue: 11 Pages: 1900-1906
  • DOI: 10.1002/jmv.23694
  • Peer Reviewed
• [Journal Article] Application of a newly developed high-sensitivity HBsAg chemiluminescent enzyme immunoassay for hepatitis B patients with HBsAg seroclearance. (2013)
  • Author(s): Shinkai N, Matsuura K, Sugauchi F, Watanabe T, Murakami S, Iio E, Ogawa S, Nojiri S, Joh T, Tanaka Y.
  • Journal Title: J Clin Microbiol. Volume: 51(11) Issue: 11 Pages: 3484-3491
  • DOI: 10.1128/jcm.00726-13
  • Peer Reviewed
• [Presentation] Novel 4'-modified nucleoside analogs exert antiviral replication against hepatitis B virus with drug resistance mutations (2016)
  • Author(s): Hayashi S, Takamatsu Y, Maeda K, Murakami S, Omagari K, Matsui T, Isogawa M, Watanabe T, Karino Y, Kohgo S, Mitsuya H, Tanaka Y.
  • Organizer: 25th Conference of the Asian Pasific Association for the Study of the Liver
  • Place of Presentation: 新高輪 国際館パミール(東京都・港区）
  • Year and Date: 2016-02-20
  • Int'l Joint Research
• [Presentation] ヒトiPS細胞から肝細胞への低分子化合物を用いた分化誘導 (2016)
  • Author(s): 松永民秀, 栗木駿輔, 坂本栄, 岩尾岳洋
  • Organizer: シンポジウム 細胞アッセイ技術の現状と将来
  • Place of Presentation: 東京大学生産技術研究所コンベンションホール（東京都・目黒区））
  • Year and Date: 2016-01-19
• [Presentation] Differentiation of human pluripotent stem cells to hepatocytes by suspension culture using a novel medium for a three-dimensional culture (2015)
  • Author(s): 阿武志保, 岩尾岳洋, 金井達郎, 松永民秀
  • Organizer: 第38回日本分子生物学会
  • Place of Presentation: 神戸ポートピアホテル(兵庫県・神戸市）
  • Year and Date: 2015-12-01
• [Presentation] A novel entecavir resistance mutation rtA186T causes viral breakthrough in chronic hepatitis B patients (2015)
  • Author(s): Hayashi S, Murakami S, Omagari K, Matsui T, Iio E, Isogawa M, Watanabe T, Karino Y, Takamatsu Y, Kohgo S, Maeda K, Mitsuya H, Tanaka Y.
  • Organizer: The 66th Annual Meeting of the American Association for the Study of Liver Diseases
  • Place of Presentation: サンフランシスコ（アメリカ）
  • Year and Date: 2015-11-13
  • Int'l Joint Research
• [Presentation] ヒト人工多能性幹細胞から肝細胞様細胞への分化促進機構の解明と毒性試験への応用 (2015)
  • Author(s): 坂本 栄, 栗木駿輔, 近藤祐樹, 土井孝良, 岩尾岳洋, 鈴木孝禎, 宮田直樹, 松永民秀
  • Organizer: 第42回日本毒性学会学術年会
  • Place of Presentation: 石川県立音楽堂（石川県・金沢市）
  • Year and Date: 2015-06-29
• [Presentation] エンテカビル治療中にviral breakthroughをきたしたB型慢性肝炎の1例～新規耐性変異rtl163V/A 186Tの同定 (2015)
  • Author(s): 林佐奈衣, 村上周子, 尾曲克己, 松居剛志, 飯尾悦子, 五十川正記, 渡邊綱正, 狩野吉康, 田中靖人
  • Organizer: 第25回抗ウイルス療法学会総会
  • Place of Presentation: 国立感染症研究所(東京都・新宿区）
  • Year and Date: 2015-05-22
• [Presentation] 新規エンテカビル耐性株の同定とその特徴 (2015)
  • Author(s): 林佐奈衣, 村上周子, 尾曲克己, 松居剛志, 飯尾悦子, 五十川正記, 渡邊綱正, 狩野吉康, 田中靖人
  • Organizer: 第51回日本肝臓学会総会
  • Place of Presentation: ホテル日航熊本(熊本県・熊本市）
  • Year and Date: 2015-05-21
• [Presentation] ヒトiPS細胞から成熟した肝細胞への分化誘導法の最適化 (2015)
  • Author(s): 権田 革達、近藤 祐樹、栗木 駿輔、岩尾 岳洋、中村 克徳、松永民秀
  • Organizer: 日本薬学会第135年会
  • Place of Presentation: 神戸学院大学他（兵庫県・神戸市）
  • Year and Date: 2015-03-25 – 2015-03-28
• [Presentation] Post-exposure prophylactic effect of nucleot(s)ide analogues demonstrated using primary human hepatocytes isolated from chimeric mice. (2014)
  • Author(s): Hamada-Tsutsumi S, Watanabe T, Murakami S, Iio E, Matsunami K, Iijima S, Omagari K, Shinkai N, Sugiura W, Tanaka Y.
  • Organizer: The 11th JSH Single Topic Conference.
  • Place of Presentation: ホテルグランヴィア広島（広島県・広島市）
  • Year and Date: 2014-11-20 – 2014-11-21
• [Presentation] Cross-Presentation by Bone Marrow Derived Cells is Required, but not Sufficient, for the Induction of HBV-specific CD8+ T Cell Responses. (2014)
  • Author(s): Isogawa M, Murata Y, Sheikh K, Tanaka Y, Chisari FV.
  • Organizer: The 11th JSH Single Topic Conference.
  • Place of Presentation: ホテルグランヴィア広島（広島県・広島市）
  • Year and Date: 2014-11-20 – 2014-11-21
• [Presentation] HBV 特異的CD8＋T 細胞の増殖と細胞傷害能は肝細胞による内因性抗原提示に依存する (2014)
  • Author(s): 五十川 正記、村田 泰洋、田中 靖人
  • Organizer: 第62回日本ウイルス学会学術集会
  • Place of Presentation: パシフィコ横浜会議場センター（神奈川県・横浜市）
  • Year and Date: 2014-11-10 – 2014-11-12
• [Presentation] Pre- and Post-Exposure Prophylaxis against Hepatitis B Virus Infection by HBV-active Antiretroviral Therapy. (2014)
  • Author(s): Watanabe T, Hamada-Tsutsumi S, Shinkai N, Iio E, Matsunami K, Iijima S, Murakami S, Omagari K, Isogawa M, Sugiura W, Tanaka Y.
  • Organizer: The 65th Annual Meeting of the American Association for the Study of Liver Diseases.
  • Place of Presentation: Boston(USA)
  • Year and Date: 2014-11-07 – 2014-11-11
• [Presentation] Utility of iPS Cells for Drug Metabolizing Enzyme Expression: Differentiation of human iPS cells into hepatocytes and enterocytes. (2014)
  • Author(s): Tamihide Matsunaga
  • Organizer: 19th North American ISSX Meeting and 29th JSSX Annual Meeting
  • Place of Presentation: San Francisco(USA)
  • Year and Date: 2014-10-19 – 2014-10-23
• [Presentation] ヒトiPS細胞の創薬研究への応用 (2014)
  • Author(s): 松永 民秀
  • Organizer: 製剤機械技術学会 第24回大会
  • Place of Presentation: 名古屋ウインクあいち（愛知県・名古屋市）
  • Year and Date: 2014-10-08 – 2014-10-09
• [Presentation] Endogenous antigen presentation by hepatocytes plays an essential role in the induction of HBV-specific CD8+ T cell responses. (2014)
  • Author(s): Isogawa M, Murata Y, Sheikh K, Tanaka Y, Chisari F.
  • Organizer: 2014 International Meeting on the Molecular Biology of Hepatitis B Viruses.
  • Place of Presentation: Los Angeles(USA)
  • Year and Date: 2014-09-03 – 2014-09-06
• [Presentation] Evaluating hepatitis B virus lifecycle and screening anti-viral drugs using primary human hepatocytes isolated from chimeric mice. (2014)
  • Author(s): Tsutsumi S, Watanabe T, Murakami S, Iijima S, Hayashi S, Omagari K, Isogawa M, Tanaka Y.
  • Organizer: 2014 International Meeting on the Molecular Biology of Hepatitis B Viruses.
  • Place of Presentation: Los Angeles(USA)
  • Year and Date: 2014-09-03 – 2014-09-06
• [Presentation] 大量調整可能なヒト肝細胞を用いたHBV in vitro 感染培養系を利用した創薬探索の可能性 (2014)
  • Author(s): 堤 進、渡邊 綱正、村上 周子、飯島 沙幸、飯尾 悦子、松波 加代子、新海 登、松浦 健太郎、五十川 正記、田中 靖人
  • Organizer: 第50回日本肝臓学会総会
  • Place of Presentation: ホテルニューオータニ（東京都・千代田区）
  • Year and Date: 2014-05-29 – 2014-05-30
• [Presentation] Incomplete prophylactic effect of nucleot(s)ide analogues using in vitro hepatitis B virus infection model. (2014)
  • Author(s): Watanabe T, Hamada-Tsutsumi S, Murakami S, Iio E, Matsunami K, Iijima S, Omagari K, Shinkai N, Sugiura W, Tanaka Y.
  • Organizer: The Asian Pacific Association for the Study of the Liver 2014.
  • Place of Presentation: Brisbane Convention & Exhibition Center(オーストラリア、ブリスベン）
  • Invited
• [Presentation] Clinical evaluation of a newly developed high-sensitive detection of hepatitis B virus surface antigen by a semi-automated immune complex transfer chemiluminescent enzyme immunoassay. (2014)
  • Author(s): Shinkai N, Oone K, Ogawa S, Iio E, Watanabe T, Matsuura K, Endo M, Fujiwara K, Nojiri S, Joh T, Tanaka Y.
  • Organizer: The Asian Pacific Association for the Study of the Liver 2014.
  • Place of Presentation: Brisbane Convention & Exhibition Center(オーストラリア、ブリスベン）
  • Invited
• [Presentation] 当院におけるB型肝炎ウイルス感染患者の HBV ジェノタイプと病態について． (2014)
  • Author(s): 新海登，松浦健太郎，田中靖人．
  • Organizer: 第99回日本消化器病学会総会．
  • Place of Presentation: 城山観光ホテル、かごしま県民交流センター（鹿児島県鹿児島市）
  • Invited
• [Presentation] A novel three-dimensional long-term culture system of primary human hepatocytes isolated from chimeric mice with humanized liver for hepatitis B virus infection. (2013)
  • Author(s): Murakami S, Watanabe T, Omagari K, Inoue T, Iijima S, Hamada-Tsutsumi S, Hayashi S, Tajiri K, Kishi H, Tanaka Y.
  • Organizer: 2013 International Meeting on the Molecular Biology of Hepatitis B Viruses.
  • Place of Presentation: Shanghai Medical College Fudan University（中国、上海）
  • Invited
• [Book] 薬剤学実験法 必携マニュアル Pharmaceutical Scientistのために (2014)
  • Author(s): 松永 民秀、岩尾 岳洋
  • Total Pages: 423
  • Publisher: 南江堂