Analysis of a novel molecular mechanism in bile duct carcinoma using a human iPS cell-derived cholangiocyte culture system

• Project/Area Number: 19K08461
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 53010:Gastroenterology-related
• Research Institution: Tokyo Medical and Dental University
• Principal Investigator: Kobayashi Masanori 東京医科歯科大学, 東京医科歯科大学病院, 特任助教 (10825459)
• Co-Investigator(Kenkyū-buntansha): 柿沼 晴 東京医科歯科大学, 大学院医歯学総合研究科, 教授 (30372444) ; 土屋 輝一郎 東京医科歯科大学, 医学部附属病院, 准教授 (40376786) ; 大島 茂 東京医科歯科大学, 医学部附属病院, 講師 (50376787)
• Project Period (FY): 2019-04-01 – 2022-03-31
• Project Status: Completed (Fiscal Year 2021)
• Budget Amount: ¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000); Fiscal Year 2021: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000); Fiscal Year 2020: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000); Fiscal Year 2019: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
• Keywords: ヒトiPS細胞 / 胆管癌 / 疾患モデル / 胆管上皮細胞 / iPS細胞 / 胆汁 / 発癌機序

Outline of Research at the Start

本研究は1)ヒト胆管上皮に惹起される細胞内シグナルの差異，2)長期暴露による癌形質獲得の有無，3)胆汁中の発癌物質の同定を中心課題とし，胆汁暴露から胆管上皮内部で起きる細胞生理学的な変化そして発癌までの病態を明らかにし，発癌物質を同定することを目的とする．申請者らは健常者由来ヒトiPS細胞から，ヒトiPS細胞由来肝前駆様細胞を樹立し維持培養を可能としており，この系を使用することでヒト由来の正常胆管細胞を，胆管癌患者由来の胆汁に暴露させるという極めて生態に近い環境での胆管細胞の挙動が詳細に観察できる．正常細胞が癌化するまでの形質変化を検討することで発癌機序および発癌物質の同定を試みる．

Outline of Final Research Achievements

We constructed a bile database derived from patients of bile duct cancer and those without cancers, and established a novel co-culture system of bile duct epithelial organoids derived from human iPS cells and bile. We also analyzed molecular signals in human iPS-derived bile duct epithelium. We improved the endoscopic procedure for patients with biliary and pancreatic diseases during bile collection (Pancreas 2021, Video GIE 2022). We founded that vasoactive intestinal peptide (VIP) plays an important role in formation of biliary tract. VIP promoted luminal formation of bile duct derived from human iPS-hepatic progenitor cells. VIP promoted pump function and formation of tight junction in cholangiocytes. VIP also promoted bile duct formation during liver injury in vivo (Hepatol Commun, 2020).

Academic Significance and Societal Importance of the Research Achievements

胆管癌は、本邦では年間2万人以上が新たに診断されており、死亡率も欧米と比較して高い傾向にある。発癌機序の詳細について不明な点が多いが、胆管癌患者が高齢者に多いことからも発癌能を有する化学物質の微量な摂取が蓄積されて発癌に寄与している可能性を考え、本研究では胆汁とヒトiPS細胞由来胆管上皮オルガノイドの形質との関係性に着目して研究を進めた。本研究課題の成果として、内視鏡的検査手技の改善により、臨床における改善点が得られたこと、さらに胆管の管腔構造形成にVasoactive intestinal peptideが直接作用することが示された点に、社会的、学術的な意義が高いと考えられる。

Research Products

• [Journal Article] A convenient and reliable method for endoscopic mapping biopsy of cholangiocarcinoma using a double-lumen cytology device. (2022)
  • Author(s): Kobayashi Masanori、Ryozawa Shomei、Ohtsuka Kazuo、Tanabe Minoru、Okamoto Ryuichi
  • Journal Title: VideoGIE Volume: in press
  • Peer Reviewed / Open Access
• [Journal Article] Caenorhabditis elegans as a Diagnostic Aid for Pancreatic Cancer (2021)
  • Author(s): Kobayashi Masanori、Fujita Akashi、Ogawa Tomoya、Tanisaka Yuki、Mizuide Masafumi、Kondo Nobuaki、Imaizumi Yuki、Hirotsu Takaaki、Ryozawa Shomei
  • Journal Title: Pancreas Volume: 50 Issue: 5 Pages: 673-678
  • DOI: 10.1097/mpa.0000000000001814
  • Peer Reviewed
• [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] Oral administration of D-serine prevents the onset and progression of colitis in mice. (2021)
  • Author(s): Takehito Asakawa, Michio Onizawa, Chikako Saito, Rie Hikichi, Daiki Yamada, Ai Minamidate, Tomoaki Mochimaru, Shun-ichiro Asahara, Yoshiaki Kido, Shigeru Oshima, Takashi Nagaishi, Kiichiro Tsuchiya, Hiromasa Ohira, Ryuichi Okamoto, Mamoru Watanabe.
  • Journal Title: Journal of Gastroenterolog. Volume: 56 Issue: 8 Pages: 732-745
  • DOI: 10.1007/s00535-021-01792-1
  • Peer Reviewed / Open Access
• [Journal Article] A rare case of hepatocellular carcinoma with bile duct invasion diagnosed by peroral cholangioscopy (2021)
  • Author(s): Ito Risako、Kobayashi Masanori、Ohtsuka Kazuo、Tanabe Minoru、Okamoto Ryuichi
  • Journal Title: VideoGIE Volume: 6 Issue: 8 Pages: 354-357
  • DOI: 10.1016/j.vgie.2021.04.007
  • Peer Reviewed / Open Access
• [Journal Article] Caenorhabditis Elegans/ as a Diagnostic Aid for Pancreatic Cancer (2021)
  • Author(s): Kobayashi M, Fujita A, Ogawa T, Yanisaka Y, Mizuide M, Kondo N, Imaizumi Y, Hirotsu T, Ryozawa S
  • Journal Title: Pancreas Volume: in press
  • Peer Reviewed
• [Journal Article] Colorectal and gastric metastases from lobular breast cancer that resembled superficial neoplastic lesions (2020)
  • Author(s): Kobayashi Masanori、Tashima Tomoaki、Nagata Koji、Sakuramoto Shinichi、Osaki Akihiko、Ryozawa Shomei
  • Journal Title: Clinical Journal of Gastroenterology Volume: 14 Issue: 1 Pages: 103-108
  • DOI: 10.1007/s12328-020-01285-3
  • Peer Reviewed
• [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] Receptor-Interacting Protein Kinase 3 (RIPK3) inhibits autophagic flux during necroptosis in intestinal epithelial cells. (2020)
  • Author(s): Otsubo K, Maeyashiki C, Nibe Y, Tamura A, Aonuma E, Matsuda H, Kobayashi M, Onizawa M, Nemoto Y, Nagaishi T, Okamoto R, Tsuchiya K, Nakamura T, Torii S, Itakura E, Watanabe M, Oshima S.
  • Journal Title: FEBS letters Volume: 594 Issue: 10 Pages: 1586-1595
  • DOI: 10.1002/1873-3468.13748
  • Peer Reviewed / Open Access
• [Journal Article] Nickel ions attenuate autophagy flux and induce transglutaminase 2 (TG2) mediated post-translational modification of SQSTM1/p62. (2020)
  • Author(s): Emi Aonuma, Akiko Tamura, Hiroki Matsuda, Takehito Asakawa, Yuriko Sakamaki, Kana Otsubo, Yoichi Nibe, Michio Onizawa, Yasuhiro Nemoto, Takashi Nagaishi, Kiichiro Tsuchiya, Tetsuya Nakamura, Motohiro Uo, Mamoru Watanabe, Ryuichi Okamoto, Shigeru Oshima.
  • Journal Title: Biochem Biophys Res Commun. Volume: 542 Pages: 17-23
  • DOI: 10.1016/j.bbrc.2021.01.023
  • 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] Investigation of Factors Affecting the Sensitivity of Bile Duct Brush Cytology (2019)
  • Author(s): Kobayashi Masanori、Ryozawa Shomei、Araki Ryuichiro、Nagata Koji、Tanisaka Yuki、Fujita Akashi、Kobatake Tsutomu
  • Journal Title: Internal Medicine Volume: 58 Issue: 3 Pages: 329-335
  • DOI: 10.2169/internalmedicine.1551-18
  • NAID: 130007590231
  • ISSN: 0918-2918, 1349-7235
  • Year and Date: 2019-02-01
  • Peer Reviewed / Open Access
• [Journal Article] The usefulness of wire-guided endoscopic snare papillectomy for tumors of the major duodenal papilla (2019)
  • Author(s): Kobayashi Masanori、Ryozawa Shomei、Iwano Hirotoshi、Araki Ryuichiro、Tanisaka Yuki、Fujita Akashi、Kobatake Tsutomu
  • Journal Title: PLOS ONE Volume: 14 Issue: 1 Pages: e0211019-e0211019
  • DOI: 10.1371/journal.pone.0211019
  • 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] 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
• [Presentation] The usefulness of endoscopic ultrasound-fine needle aspiration in the grading of pancreatic neuroendocrine neoplasms (2021)
  • Author(s): Kobayashi M, Ohtsuka K, Kirimura S, Tanabe M, Okamoto R.
  • Organizer: APDW2021
  • Int'l Joint Research
• [Presentation] Factors associated with HCC development and patients’ survival in patients with an SVR (2021)
  • Author(s): Nakagawa M, Kakinuma S, Asahina Y, et al.
  • Organizer: JSH International Liver Conference 2021
  • Int'l Joint Research
• [Presentation] 膵神経内分泌腫瘍のgradingにおける超音波内視鏡下穿刺生検の有用性の検討 (2021)
  • Author(s): 小林正典, 大塚和朗, 桐村進, 工藤篤, 田邉稔, 岡本隆一
  • Organizer: 第101回 消化器内視鏡学会総会
• [Presentation] ヒトiPS細胞を利用したHBV関連肝細胞癌におけるMLL4の機能的意義の解析 (2021)
  • Author(s): 土屋 淳、柿沼 晴、朝比奈靖浩
  • Organizer: 第107回日本消化器病学会総会
• [Presentation] 疾患モデルの応用を目指したヒトiPS細胞を利用した肝細胞オルガノイドの樹立 (2021)
  • Author(s): 志水太郎、柿沼 晴、朝比奈靖浩
  • Organizer: 第57回日本肝臓学会総会
• [Presentation] 新規ヒトiPS細胞由来肝細胞オルガノイド系の構築と肝疾患研究 (2021)
  • Author(s): 三好正人、柿沼 晴、朝比奈靖浩
  • Organizer: 第25回日本肝臓学会大会
• [Presentation] 当院におけるIPMN診断治療の現状 (2020)
  • Author(s): 小林正典，大塚和朗，伴大輔，田邉稔
  • Organizer: 第106回日本消化器病学会総会
• [Presentation] 当院におけるEST後出血への対応 (2020)
  • Author(s): 小林正典，大塚和朗，土屋輝一郎
  • Organizer: 第99回日本消化器内視鏡学会総会
• [Presentation] IPMN術前精査における超音波内視鏡検査の役割 (2020)
  • Author(s): 小林正典，大塚和朗，田邉稔
  • Organizer: JDDW2020
• [Presentation] 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] 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] 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] 胆管カニュレーション困難症例に対する細胞診ブラシのカニュラへの応用 (2019)
  • Author(s): 小林正典，谷坂優樹，水出雅文，良沢昭銘，大塚和朗，渡辺守
  • Organizer: 第97回 日本消化器内視鏡学会総会
• [Presentation] 総胆管に穿破した 主膵管型IPMNの1例 (2019)
  • Author(s): 小林正典，大塚和朗，小川康介，伴大輔，田邉稔
  • Organizer: 第55回胆道学会総会
• [Presentation] Effectiveness of Endoscopic Ultrasonography-Guided Fine-Needle Aspiration for the Diagnosis of Pancreatic Neuroendocrine Tumors (2019)
  • Author(s): Masanori Kobayashi, Kazuo Ohtsuka, Daisuke Ban, Atsushi Kudo, Minoru Tanabe
  • Organizer: American Pancreatic Association 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] 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、角田知之、三好正人、紙谷聡英、佐藤綾子、土屋 淳、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: JDDW2019