Development of human iPS cell-derived liver organoids for the discovery of target molecules for a therapy of liver fibrosis

• Project/Area Number: 18H02790
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Review Section: Basic Section 53010:Gastroenterology-related
• Research Institution: Tokyo Medical and Dental University
• Principal Investigator: KAKINUMA Sei 東京医科歯科大学, 大学院医歯学総合研究科, 教授 (30372444)
• Co-Investigator(Kenkyū-buntansha): 朝比奈 靖浩 東京医科歯科大学, 大学院医歯学総合研究科, 寄附講座教授 (00422692) ; 渡辺 守 東京医科歯科大学, 高等研究院, 特別栄誉教授 (10175127)
• Project Period (FY): 2018-04-01 – 2021-03-31
• Project Status: Completed (Fiscal Year 2020)
• Budget Amount: ¥17,160,000 (Direct Cost: ¥13,200,000、Indirect Cost: ¥3,960,000); Fiscal Year 2020: ¥5,460,000 (Direct Cost: ¥4,200,000、Indirect Cost: ¥1,260,000); Fiscal Year 2019: ¥5,460,000 (Direct Cost: ¥4,200,000、Indirect Cost: ¥1,260,000); Fiscal Year 2018: ¥6,240,000 (Direct Cost: ¥4,800,000、Indirect Cost: ¥1,440,000)
• Keywords: ヒトiPS細胞 / 肝前駆細胞 / 再生医学 / ゲノム編集 / 肝星細胞 / 疾患モデル / 肝線維化 / 胆管細胞 / 肝幹/前駆細胞 / iPS細胞

Outline of Final Research Achievements

Aim of this research project was to develop novel disease models that can mimic the pathophysiology of several liver diseases using human induced pluripotent stem cells (iPS cells). We established the original human iPS cell lines as a model of congenital hepatic fibrosis using genome-editing technology. Analysis of the iPS cell-derived cholangiocytic organoids revealed that interleukin-8 and CTGF are responsible for the pathogenesis of congenital hepatic fibrosis. We developed an original method for differentiation of human iPS cells into hepatic stellate-like cells (iPS-HSCs). Hepatocytic maturation in iPS-derived hepatic progenitor cells, such as their capacity for albumin production, was significantly increased in direct co-culture with iPS-HSCs overexpressing LHX2. Our research indicates that studies using genetically edited iPS-derived hepatic cells will be of value for research into pathophysiology of liver diseases.

Academic Significance and Societal Importance of the Research Achievements

本研究では、研究グループで独自に先天性肝線維症モデルとなるヒトiPS細胞培養系を開発した。研究結果では、本症でみられる胆管形成の異常と進行性の肝線維化の分子病態の一部が解明され、治療標的の提案がなされた。本疾患は肝移植を要することもある難治性疾患であり、その新規治療開発への応用が期待される結果だった。本成果は、国際的評価の高い専門医学誌への掲載、学会学術賞の受賞、各種メディア記事への掲載など、極めて高く評価された。また、ヒトiPS由来肝星細胞の誘導法の開発も、肝硬変を治療するための画期的創薬研究への基盤技術となることが期待され、今後の波及効果が高い研究として学会学術賞の受賞し、高く評価された。

Research Products

• [Int'l Joint Research] Stanford University School of Medicine(米国)
• [Int'l Joint Research] Stanford University School of Medicine(米国)
• [Journal Article] Intraepithelial Lymphocytes Suppress Intestinal Tumor Growth by Cell-to-Cell Contact via CD103/E-Cadherin Signal. (2021)
  • Author(s): Ryo Morikawa, Yasuhiro Nemoto, Yuki Yonemoto, Shohei Tanaka, Yuria Takei, Shigeru Oshima, Takashi Nagaishi, Kiichiro Tsuchiya, Kengo Nozaki , Tomohiro Mizutani, Tetsuya Nakamura, Mamoru Watanabe, Ryuichi Okamoto.
  • Journal Title: Cell Mol Gastroenterol Hepatol. Volume: 11 Issue: 5 Pages: 1483-1503
  • DOI: 10.1016/j.jcmgh.2021.01.014
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Hepatocellular carcinoma recurrence after direct-acting antiviral therapy: an individual patient data meta-analysis (2021)
  • Author(s): Sapena V, Enea M, Torres F, Celsa C, Rios J, Rizzo GEM, Nahon P, Marino Z, Tateishi R, Minami T, Sangiovanni A, Forns X, Toyoda H, Brillanti S, Conti F, Degasperi E, Yu ML, Tsai PC, Jean K, El Kassas M, Shousha HI, Omar A, Zavaglia C, Nagata H, Nakagawa M, Asahina Y, et al
  • Journal Title: Gut Volume: in press Issue: 3 Pages: 323663-323663
  • DOI: 10.1136/gutjnl-2020-323663
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Mac-2 binding protein glycosylation isomer as a novel predictive biomarker for patient survival after hepatitis C virus eradication by DAAs (2020)
  • Author(s): Nakagawa M, Nawa N, Takeichi E, Shimizu T, Tsuchiya J, Sato A, Miyoshi M, Kawai-Kitahata F, Murakawa M, Nitta S, Itsui Y, Azuma S, Kakinuma S, Fujiwara T, Watanabe M, Tanaka Y, Asahina Y; Ochanomizu Liver Conference Study Group.
  • Journal Title: Journal of Gastroenterology Volume: 55 Issue: 10 Pages: 990-999
  • DOI: 10.1007/s00535-020-01715-6
  • Peer Reviewed / Open Access
• [Journal Article] Time course alterations of virus sequences and immunoglobulin titers in a chronic hepatitis E patient (2020)
  • Author(s): Nitta Sayuri、Takahashi Kazuaki、Kawai‐Kitahata Fukiko、Tsuchiya Jun、Sato Ayako、Miyoshi Masato、Murakawa Miyako、Itsui Yasuhiro、Nakagawa Mina、Azuma Seishin、Kakinuma Sei、Watanabe Mamoru、Asahina Yasuhiro
  • Journal Title: Hepatology Research Volume: 50 Issue: 4 Pages: 524-531
  • DOI: 10.1111/hepr.13480
  • Peer Reviewed
• [Journal Article] Relative dose intensity over the first four weeks of lenvatinib therapy is a factor of favorable response and overall survival in patients with unresectable hepatocellular carcinoma (2020)
  • Author(s): Kirino S, Tsuchiya K, Kurosaki M, Kaneko S, Inada K, Yamashita K, Osawa L, Hayakawa Y, Sekiguchi S, Okada M, Wang W, Higuchi M, Takaura K, Maeyashiki C, Tamaki N, Yasui Y, Nakanishi H, Itakura J, Takahashi Y, Asahina Y, Izumi N.
  • Journal Title: PLoS One Volume: 15 Issue: 4 Pages: e0231828-e0231828
  • DOI: 10.1371/journal.pone.0231828
  • Peer Reviewed / Open Access
• [Journal Article] Vasoactive Intestinal Peptide Derived From Liver Mesenchymal Cells Mediates Tight Junction Assembly in Mouse Intrahepatic Bile Ducts (2020)
  • Author(s): Sato A, Kakinuma S, Miyoshi M, Kamiya A, Tsunoda T, Kaneko S, Tsuchiya J, Shimizu T, Takeichi E, Nitta S, Kawai-Kitahata F, Murakawa M, Itsui Y, Nakagawa M, Azuma S, Koshikawa N, Seiki S, Nakauchi H, Asahina Y, Watanabe M.
  • Journal Title: Hepatology Communications Volume: 4 Issue: 2 Pages: 235-254
  • DOI: 10.1002/hep4.1459
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Loss of Fibrocystin Promotes Interleukin-8-Dependent Proliferation and CTGF Production of Biliary Epithelium. (2019)
  • Author(s): Tsunoda T, Kakinuma S, Miyoshi M, Kamiya A, Kaneko S, Sato A, Tsuchiya J, Nitta S, Kawai-Kitahata F, Murakawa M, Itsui Y, Nakagawa M, Azuma S, Sogo T, Komatsu H, Mukouchi R, Inui A, Fujisawa T, Nakauchi H, Asahina Y, Watanabe M.
  • Journal Title: Journal of Heptatology Volume: 印刷中 Issue: 1 Pages: 143-152
  • DOI: 10.1016/j.jhep.2019.02.024
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Analysis of the mechanism underlying liver diseases using human induced pluripotent stem cells (2019)
  • Author(s): Kakinuma S and Watanabe M
  • Journal Title: Immunological Medicine Volume: 42 Issue: 2 Pages: 71-78
  • DOI: 10.1080/25785826.2019.1657254
  • Peer Reviewed / Open Access
• [Journal Article] Comprehensive genetic analysis of cholangiolocellular carcinoma with a coexistent hepatocellular carcinoma‐like area and metachronous hepatocellular carcinoma (2019)
  • Author(s): Kawai-Kitahata F, Asahina Y, Kaneko S, Tsuchiya J, Sato A, Miyoshi M, Tsunoda T, Inoue-Shinomiya E, Murakawa M, Nitta S, Itsui Y, Nakagawa M, Azuma S, Kakinuma S, Tanabe M, Sugawara E, Takemoto A, Ojima H, Sakamoto M, Muraoka M, Takano S, Maekawa S, Enomoto N and Watanabe M.
  • Journal Title: Hepatology Research Volume: 49 Issue: 12 Pages: 1466-1474
  • DOI: 10.1111/hepr.13403
  • Peer Reviewed / Open Access
• [Journal Article] LIM homeobox 2 promotes interaction between human iPS-derived hepatic progenitors and iPS-derived hepatic stellate-like cells. (2019)
  • Author(s): Miyoshi M, Kakinuma S, Kamiya A, Tsunoda T, Tsuchiya J, Sato A, Kaneko S, Nitta S, Kawai-Kitahata F, Murakawa M, Itsui Y, Nakagawa M, Azuma S, Nakauchi H, Asahina Y, Watanabe M.
  • Journal Title: Scientific Reports Volume: 9 Issue: 1 Pages: 2072-2072
  • DOI: 10.1038/s41598-018-37430-9
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Impact of novel NS5A resistance-associated substitutions of hepatitis C virus detected in treatment-experienced patients (2019)
  • Author(s): Nitta S, Asahina Y, Kato T, Tsuchiya J, Inoue-Shinomiya E, Sato A, Tsunoda T, Miyoshi M, Kawai-Kitahata F, Murakawa M, Itsui Y, Nakagawa M, Azuma S, Kakinuma S, Hikita H, Takehara T, Watanabe M.
  • Journal Title: Scientific Reports Volume: 9 Issue: 1 Pages: 5722-5722
  • DOI: 10.1038/s41598-019-42114-z
  • Peer Reviewed / Open Access
• [Journal Article] .Association of serum IFN-λ3 levels with hepatocarcinogenesis in chronic hepatitis C patients treated with direct-acting antiviral agents. (2019)
  • Author(s): Emi Inoue-Shinomiya, Miyako Murakawa, Yasuhiro Asahina, Mina Nakagawa, Jun Tsuchiya, Ayako Sato, Tomoyuki Tsunoda, Masato Miyoshi, Sayuri Nitta, Fukiko Kawai-Kitahata, Yasuhiro Itsui, Seishin Azuma, Sei Kakinuma, Kazumoto Murata, Masashi Mizokami, Mamoru Watanabe.
  • Journal Title: Hepatology Research Volume: 印刷中 Issue: 5 Pages: 500-511
  • DOI: 10.1111/hepr.13307
  • Peer Reviewed / Open Access
• [Journal Article] Diabetic Retinopathy as a Risk Factor Associated with the Development of Hepatocellular Carcinoma in Nonalcoholic Fatty Liver Disease. (2019)
  • Author(s): Azuma S, Asahina Y, Kakinuma S, Azuma K, Miyoshi M, Inoue E, Tsunoda T, Sato A, Kaneko S, Nagata H, Kawai-Kitahata F, Murakawa M, Nitta S, Itsui Y, Tomita M, Nakagawa M, Watanabe M.
  • Journal Title: Digestive Diseases Volume: 37 Issue: 3 Pages: 247-254
  • DOI: 10.1159/000493580
  • Peer Reviewed
• [Presentation] 【シンポジウム臓器領域別再生医療の最前線】肝・胆道：ヒトiPS細胞由来肝臓構成細胞による肝線維症の病態解析 (2021)
  • Author(s): 柿沼 晴
  • Organizer: 第20回 日本再生医療学会総会
  • Invited
• [Presentation] ヒトiPS細胞由来肝臓構成細胞の疾患モデルへの応用 (2020)
  • Author(s): 柿沼 晴
  • Organizer: 第27回 肝細胞研究会
  • Invited
• [Presentation] 肝間葉系細胞によるvasoactive intestinal peptideを介した胆管形成調節機構 (2020)
  • Author(s): 柿沼 晴、佐藤綾子、三好正人、紙谷聡英、土屋 淳、志水太郎、北畑富貴子、村川美也子、新田沙由梨、井津井康浩、中川美奈、東 正新、朝比奈靖浩、渡辺 守.
  • Organizer: 第27回 肝細胞研究会
• [Presentation] Vasoactive intestinal peptideの肝内胆管形成および胆汁うっ滞性肝障害への寄与 (2020)
  • Author(s): 佐藤綾子、柿沼 晴、志水太郎、土屋 淳、三好正人、北畑富貴子、村川美也子、新田沙由梨、井津井康浩、中川美奈、東 正新、朝比奈靖浩.
  • Organizer: JDDW2020 第24回 日本肝臓学会大会
• [Presentation] 【シンポジウム肝疾患と微小環境】肝間葉系細胞由来vasoactive intestinal peptideによる胆管形成の促進機構 (2020)
  • Author(s): 佐藤綾子、柿沼 晴、朝比奈靖浩
  • Organizer: 第56回日本肝臓学会総会
• [Presentation] Comprehensive analysis of cancer-related genes and AAV/Hepatitis B virus integration into genome on development of hepatocellular carcinoma in patients with prior Hepatitis B virus infection. (2020)
  • Author(s): Fukiko Kawai-Kitahata, Yasuhiro Asahina , Sei Kakinuma, Miyako Murakawa, Sayuri Nitta, Masato Miyoshi, Ayako Sato, Jun Tsuchiya, Taro Shimizu, Eiko Takeichi, Mina Nakagawa, Yasuhiro Itsui, Seishin Azuma, Shinji Tanaka , Minoru Tanabe, Shinya Maekawa, Nobuyuki Enomoto and Mamoru Watanabe.
  • Organizer: EASL The Digital International Liver Congress 2020
  • Int'l Joint Research
• [Presentation] Poor improvement of on-treatment FIB-4 index after initiation of nucleos(t)ide analogs is associated with development of hepatocellular carcinoma in both cirrhotic and non-cirrhotic chronic hepatitis B patients. (2020)
  • Author(s): Miyako Murakawa, Taro Shimizu, Eiko Takeichi, Jun Tsuchiya, Ayako Sato, Masato Miyoshi, Fukiko Kawai-Kitahata, Sayuri Nitta, Yasuhiro Itsui, Mina Nakagawa, Seishin Azuma, Sei Kakinuma, Yasuhiro Asahina.
  • Organizer: EASL The Digital International Liver Congress 2020
  • Int'l Joint Research
• [Presentation] 【シンポジウム消化器疾患における最先端のトランスレーショナルリサーチ】ヒトiPS細胞を用いた肝線維症における分子標的の探索 (2020)
  • Author(s): 柿沼 晴、角田知之、朝比奈靖浩
  • Organizer: 第106回日本消化器病学会総会
• [Presentation] 【シンポジウム 1:技術革新がもたらした消化器研究のパラダイムシフト】ヒト iPS 細胞を用いた疾患モデルによる肝線維化の病態研究. (2019)
  • Author(s): 角田知之、柿沼 晴、渡辺 守
  • Organizer: 第105回 日本消化器病学会総会
• [Presentation] 【ワークショップ:肝構成細胞の病態生理】転写因子 LHX2のヒトiPS細胞由来肝星細胞様細胞における機能の解析. (2019)
  • Author(s): 三好正人、柿沼 晴、渡辺 守
  • Organizer: 第55 回日本肝臓学会総会
• [Presentation] Comprehensive analysis of cancer gene mutations and viral integration in hepatocellular carcinoma arising from non-fibrotic liver. (2019)
  • Author(s): Asahina Y, Kawai-Kitahata F, Murakawa M, Nitta S, Itsui Y, Nakagawa M, Azuma S, Kakinuma S.
  • Organizer: AASLD The Liver Meeting 2019
  • Int'l Joint Research
• [Presentation] Vasoactive intestinal peptide による胆管形成調節の分子機序. (2019)
  • Author(s): 佐藤綾子、柿沼 晴、土屋 淳、三好正人、角田知之、井津井康浩、中川美奈、東 正新、朝比奈靖浩、渡辺 守.
  • Organizer: 第26回肝細胞研究会
• [Presentation] [会長賞受賞講演]ヒトiPS細胞による疾患モデルを利用した先天性肝線維症分子標的の探索. (2019)
  • Author(s): 柿沼 晴.
  • Organizer: JDDW2019
  • Invited
• [Presentation] 【統合シンポジウム:肝移植と肝再生の現状と展望】ヒト iPS 細胞由来肝星細胞様細胞による肝前駆細胞機能の調節. (2019)
  • Author(s): 三好正人、柿沼 晴、渡辺 守
  • Organizer: JDDW2019
• [Presentation] ヒトiPS 細胞を用いた先天性肝線維症病態モデルの 樹立と分子標的の探索 (2018)
  • Author(s): 角田知之、柿沼 晴、渡辺 守
  • Organizer: 第54回日本肝臓学会総会
• [Presentation] ヒトiPS細胞由来星細胞様細胞における転 写因子LHX2の機能解析 (2018)
  • Author(s): 三好正人、柿沼晴、金子俊、紙谷聡英、佐藤綾子、角田知之、四宮恵美、北畑富貴子、新田沙由梨、村川美也子、井津井康浩、中川美奈、東正新、朝比奈靖浩、渡辺 守
  • Organizer: 第54回日本肝臓学会総会
• [Presentation] Gene Mutational Profile and Viral Integration in Hepatocellular Carcinoma with or without HBV/HCV Suppression. (2018)
  • Author(s): Asahina Y, Kawai-Kitahata F, Murakawa M, Nitta S, Nakagawa M, Kakinuma S, Watanabe M
  • Organizer: AASLD The Liver Meeting 2018
  • Int'l Joint Research
• [Presentation] Post-Treatment M2BPGi Level Is Useful for Predicting HCC Occurrence and Recurrence after Viral Eradication in Chronic Hepatitis C Patients. (2018)
  • Author(s): Nakagawa M, Asahina Y, Kawai-Kitahata F, Murakawa M, Nitta S, Itsui Y, Azuma S, Kakinuma S, Tomita M, Watanabe M
  • Organizer: AASLD The Liver Meeting 2018
  • Int'l Joint Research
• [Presentation] iPS細胞先天 性肝線維症病態モデルによる胆管細胞異常に起因する肝線維化における免疫応答の解析 (2018)
  • Author(s): 角田知之、柿沼 晴、朝比奈靖浩
  • Organizer: 第42回日本肝臓学会東部会
• [Book] A Rodent Model for Cell Transplantation of Hepatic Progenitor Cells (Hepatic Stem Cells) (2019)
  • Author(s): Kakinuma S., Kamiya A (In: Tanimizu N)
  • Total Pages: 248
  • Publisher: Springer Nature
  • ISBN: 9781493989614
• [Remarks] ヒトiPS細胞を利用した肝疾患モデル（肝細胞研究会・研究交流）
  • URL: http://hepato.umin.jp/kouryu/kouryu54.html
• [Remarks] Press release
  • URL: http://www.tmd.ac.jp/english/press-release/20190416_1/index.html
• [Remarks] ヒトiPS細胞由来肝星細胞の作製と細胞間相互作用を介した肝細胞の成熟化に成功
  • URL: http://www.tmd.ac.jp/archive-tmdu/kouhou/20190215_1.pdf
• [Remarks] ヒトiPS細胞を用いた疾患モデルを開発し、難治性肝疾患の病態解明に成功
  • URL: http://www.tmd.ac.jp/archive-tmdu/kouhou/20190319_1.pdf