Development of biliary and pancreatic tumor cell bank using organoid culture and its application to personalized therapy

• Project/Area Number: 17H03592
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Tumor diagnostics
• Research Institution: Keio University
• Principal Investigator: Saito Yoshimasa 慶應義塾大学, 薬学部(芝共立), 准教授 (90360114)
• Project Period (FY): 2017-04-01 – 2020-03-31
• Project Status: Completed (Fiscal Year 2019)
• Budget Amount: ¥17,940,000 (Direct Cost: ¥13,800,000、Indirect Cost: ¥4,140,000); Fiscal Year 2019: ¥5,850,000 (Direct Cost: ¥4,500,000、Indirect Cost: ¥1,350,000); Fiscal Year 2018: ¥6,110,000 (Direct Cost: ¥4,700,000、Indirect Cost: ¥1,410,000); Fiscal Year 2017: ¥5,980,000 (Direct Cost: ¥4,600,000、Indirect Cost: ¥1,380,000)
• Keywords: 胆道がん / 膵臓がん / オルガノイド / ドラッグ・リポジショニング / 抗真菌薬 / 胆道・膵臓がん / ドラッグリポジショニング / 既存薬 / オルガノイド培養 / 個別化治療 / がんの個性診断

Outline of Final Research Achievements

We successfully established organoids derived from biliary tract carcinoma (BTC) patients. These BTC organoids were cultured stably for over one year and closely recapitulated the histopathology, gene expression, and genetic alterations evident in the primary tumors. Gene expression profiling of the organoids and clinical data of patients revealed that SOX2, KLK6, and CPB2 could be a potential prognostic biomarker for patients with BTC. Moreover, we performed a drug screening with a compound library consisting of drugs employed clinically for their ability to suppress BTC organoids as a “drug repositioning” strategy. We discovered that the antifungal drugs amorolfine and fenticonalzole significantly suppressed the growth of BTC organoids. Antifungal drugs including amorolfine and fenticonalzole could be potentially applied for the prevention and treatment of BTC patients.

Academic Significance and Societal Importance of the Research Achievements

この研究成果についてプレスリリースを行なったところ、非常に大きな反響があり、多くの末期胆道がんの患者さんより、手紙やメールなどで直接問い合わせがあった。いずれも現行の治療で効果がなく、抗真菌薬の投与を試してみたい、または、もし治験が始まっているのであれば、是非エントリーしたいとのことであった。今後は、今回のスクリーニングにより特定された抗真菌薬や脂質異常症治療薬を基盤とした低分子化合物に関する基礎研究をさらに進め、動物実験、非臨床Proof of Concept (POC) の取得、医師主導治験を経て、可能な限り早く、胆道・膵臓がんに対する新たな治療薬の創出につなげていきたいと考えている。

Research Products

• [Int'l Joint Research] Olivia Newton-John Institute(オーストラリア)
• [Int'l Joint Research] Medical University of Vienna(オーストリア)
• [Journal Article] Establishment of Patient-Derived Organoids and Drug Screening for Biliary Tract Carcinoma. (2019)
  • Author(s): Saito Y, Muramatsu T, Kanai Y, Ojima H, Sukeda A, Hiraoka N, Arai E, Sugiyama Y, Matsuzaki J, Uchida R, Yoshikawa N, Furukawa R, Saito H.
  • Journal Title: Cell Reports Volume: 27 Pages: 1265-1276
  • DOI: 10.1016/j.celrep.2019.03.088
  • Peer Reviewed / Open Access
• [Journal Article] Genomic Profiling of Biliary Tract Cancer Cell Lines Reveals Molecular Subtypes and Actionable Drug Targets. (2019)
  • Author(s): Lau DK, Mouradov D, Wasenang W, Luk IY, Scott CM, Williams DS, Yeung YH, Limpaiboon T, Iatropoulos GF, Jenkins LJ, Reehorst CM, Chionh F, Nikfarjam M, Croagh D, Dhillon AS, Weickhardt AJ, Muramatsu T, Saito Y, Tebbutt NC, Sieber OM, Mariadason JM.
  • Journal Title: iScience Volume: 21 Pages: 624-637
  • DOI: 10.1016/j.isci.2019.10.044
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Generation of human hepatic progenitor cells with regenerative and metabolic capacities from primary hepatocytes. (2019)
  • Author(s): Katsuda T, Matsuzaki J, Yamaguchi T, Yamada Y, Prieto-Vila M, Hosaka K, Takeuchi A, Saito Y, Ochiya T.
  • Journal Title: Elife Volume: 8
  • DOI: 10.7554/elife.47313
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Establishment of an organoid bank of biliary tract and pancreatic cancers and its application for personalized therapy and future treatment. (2019)
  • Author(s): Saito Y.
  • Journal Title: Journal of Gastroenterology and Hepatology Volume: 34 Pages: 1906-1910
  • DOI: 10.1111/jgh.14773
  • Peer Reviewed / Open Access
• [Journal Article] Glucose Depletion Enhances the Stem Cell Phenotype and Gemcitabine Resistance of Cholangiocarcinoma Organoids through AKT Phosphorylation and Reactive Oxygen Species. (2019)
  • Author(s): Yoshikawa N, Saito Y, Manabe H, Nakaoka T, Uchida R, Furukawa R, Muramatsu T, Sugiyama Y, Kimura M, Saito H.
  • Journal Title: Cancers Volume: 11
  • DOI: 10.3390/cancers11121993
  • Peer Reviewed / Open Access
• [Journal Article] Circulating microRNA-1246 as a possible biomarker for early tumor recurrence of hepatocellular carcinoma. (2019)
  • Author(s): Chuma M, Toyoda H, Matsuzaki J, Saito Y, Kumada T, Tada T, Kaneoka Y, Maeda A, Yokoo H, Ogawa K, Kamiyama T, Taketomi A, Matsuno Y, Yazawa K, Takeda K, Kunisaki C, Ogushi K, Moriya S, Hara K, Nozaki A, Kondo M, Fukuda H, Numata K, Tanaka K, Maeda S, Sakamoto N.
  • Journal Title: Hepatology Research Volume: 49 Pages: 810-822
  • DOI: 10.1111/hepr.13338
  • Peer Reviewed / Open Access
• [Journal Article] Establishment of patient-derived organoids and drug screening for biliary tract carcinoma (2019)
  • Author(s): Saito Yoshimasa, Muramatsu Toshihide, Kanai Yae, Ojima Hidenori, Sukeda Aoi, Hiraoka Nobuyoshi, Arai Eri, Sugiyama Yuko, Matsuzaki Juntaro, Uchida Ryoei, Yoshikawa Nao, Furukawa Ryo, Saito Hidetsugu
  • Journal Title: Cell Reports Volume: 印刷中
  • Peer Reviewed / Open Access
• [Journal Article] Epigenetic silencing of Lgr5 induces senescence of intestinal epithelial organoids during the process of aging (2019)
  • Author(s): Uchida Ryoei、Saito Yoshimasa、Nogami Kazuki、Kajiyama Yohei、Suzuki Yukana、Kawase Yasuhiro、Nakaoka Toshiaki、Muramatsu Toshihide、Kimura Masaki、Saito Hidetsugu
  • Journal Title: npj Aging and Mechanisms of Disease Volume: 5 Pages: 1-1
  • DOI: 10.1038/s41514-019-0035-9
  • Peer Reviewed / Open Access
• [Journal Article] Vonoprazan-Based Third-Line Therapy Has a Higher Eradication Rate against Sitafloxacin-Resistant Helicobacter pylori (2019)
  • Author(s): Saito Yoshimasa、Konno Kaho、Sato Moeka、Nakano Masaru、Kato Yukako、Saito Hidetsugu、Serizawa Hiroshi
  • Journal Title: Cancers Volume: 11 Pages: 116-116
  • DOI: 10.3390/cancers11010116
  • Peer Reviewed / Open Access
• [Journal Article] CAPZA1 determines the risk of gastric carcinogenesis by inhibiting Helicobacter pylori CagA-degraded autophagy (2019)
  • Author(s): Tsugawa H., Mori H., Matsuzaki J., Sato A., Saito Y., Imoto M., Suematsu M., Suzuki H.
  • Journal Title: Autophagy. Volume: 15 Pages: 1-17
  • DOI: 10.1080/15548627.2018.1515530
  • Peer Reviewed / Open Access
• [Journal Article] Nrf2-mediated anti-oxidant effects contribute to suppression of non-alcoholic steatohepatitis-associated hepatocellular carcinoma in murine model (2018)
  • Author(s): Yamada Shoji、Kimura Masaki、Saito Yoshimasa、Saito Hidetsugu
  • Journal Title: J. Clin. Biochem. Nutr. Volume: 63 Issue: 2 Pages: 123-128
  • DOI: 10.3164/jcbn.17-125
  • NAID: 130007480019
  • ISSN: 0912-0009, 1880-5086
  • Peer Reviewed / Open Access
• [Journal Article] Dual effects of the Nrf2 inhibitor for inhibition of hepatitis C virus and hepatic cancer cells. (2018)
  • Author(s): Murakami Y, Sugiyama K, Ebinuma H, Nakamoto N, Ojiro K, Chu PS, Taniki N, Saito Y, Teratani T, Koda Y, Suzuki T, Saito K, Fukasawa M, Ikeda M, Kato N, Kanai T, Saito H.
  • Journal Title: BMC Cancer Volume: 3 Pages: 680-680
  • DOI: 10.1186/s12885-018-4588-y
  • Peer Reviewed / Open Access
• [Journal Article] Bile acid metabolism regulated by the gut microbiota promotes non-alcoholic steatohepatitis-associated hepatocellular carcinoma in mice. (2018)
  • Author(s): Yamada S, Takashina Y, Watanabe M, Nagamine R, Saito Y, Kamada N, Saito H.
  • Journal Title: Oncotarget Volume: 9 Pages: 9925-9939
  • DOI: 10.18632/oncotarget.24066
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Induction of differentiation of intrahepatic cholangiocarcinoma cells to functional hepatocytes using an organoid culture system (2018)
  • Author(s): Saito Yoshimasa、Nakaoka Toshiaki、Muramatsu Toshihide、Ojima Hidenori、Sukeda Aoi、Sugiyama Yuko、Uchida Ryoei、Furukawa Ryo、Kitahara Aya、Sato Toshiro、Kanai Yae、Saito Hidetsugu
  • Journal Title: Sci Rep Volume: 8 Pages: 2821-2821
  • DOI: 10.1038/s41598-018-21121-6
  • Peer Reviewed / Open Access
• [Journal Article] Gut microbiota-mediated generation of saturated fatty acids elicits inflammation in the liver in murine high-fat diet-induced steatohepatitis. (2017)
  • Author(s): Yamada S, Kamada N, Amiya T, Nakamoto N, Nakaoka T, Kimura M, Saito Y, Ejima C, Kanai T, Saito H.
  • Journal Title: BMC Gastroenterology Volume: 17 Pages: 136-136
  • DOI: 10.1186/s12876-017-0689-3
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Aberrant DNA Methylation as a Biomarker and a Therapeutic Target of Cholangiocarcinoma. (2017)
  • Author(s): Nakaoka T, Saito Y, Saito H.
  • Journal Title: International Journal of Molecular Science Volume: 18 Pages: 1111-1111
  • DOI: 10.3390/ijms18061111
  • Peer Reviewed / Open Access
• [Journal Article] Cluster microRNAs miR-194 and miR-215 suppress the tumorigenicity of intestinal tumor organoids. (2017)
  • Author(s): Nakaoka T, Saito Y, Shimamoto Y, Muramatsu T, Kimura M, Kanai Y, Saito H.
  • Journal Title: Cancer Science Volume: 108 Pages: 678-684
  • DOI: 10.1111/cas.13165
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Presentation] A new 3D co-culture model of intrahepatic cholangiocarcinoma organoids and hepatic stellate cells in a microfluidic device. (2019)
  • Author(s): 須田雄大、齋藤義正、齋藤英胤
  • Organizer: 第78回日本癌学会学術総会
• [Presentation] Organoid培養法により樹立した腸管上皮幹細胞に対するNMN (Nicotinamide mononucleotide) による抗老化効果の検討 (2019)
  • Author(s): 野上和幹、内田諒英、梶山洋平、鈴木裕佳水、齋藤義正、木村真規、齋藤英胤
  • Organizer: 第19回日本抗加齢医学会総会
• [Presentation] オルガノイド培養による難治性がんの前臨床モデルの確立と創薬研究への応用 (2019)
  • Author(s): 齋藤 義正， 村松 俊英，齋藤 英胤
  • Organizer: 第105回日本消化器病学会総会
• [Presentation] オルガノイド培養による難治性がんの前臨床モデルの構築とその創薬への応用 (2019)
  • Author(s): 齋藤義正
  • Organizer: 細胞アッセイ研究会シンポジウム
  • Invited
• [Presentation] Establishment and long-term in vitro culture of organoids derived from human biliary tract carcinomas (2018)
  • Author(s): Saito Yoshimasa, Muramatsu Toshihide, Sugiyama Yuko, Uchida Ryoei, Furukawa Ryo, Yoshikawa Nao, Kimura Masaki, Saito Hidetsugu
  • Organizer: Annual Meeting of American Association for Cancer Research (AACR) 2018
  • Int'l Joint Research
• [Presentation] Establishment of preclinical models for refractory cancers by organoid culture and its application for drug discovery (2018)
  • Author(s): Saito Yoshimasa
  • Organizer: 2018 International Meeting on 22nd MDO and 33rd JSSX
  • Int'l Joint Research / Invited
• [Presentation] Establishment of an organoid bank of biliary tract and pancreatic cancers and its application for personalized therapy (2018)
  • Author(s): Saito Yoshimasa
  • Organizer: Asian Pacific Digestive Week 2018
  • Int'l Joint Research / Invited
• [Presentation] 肝内胆管がんオルガノイドの樹立と分化誘導による肝細胞性の獲得 (2018)
  • Author(s): 齋藤義正、中岡俊哉、村松俊英、杉山優子、木村真規、齋藤 英胤
  • Organizer: 第104回日本消化器病学会総会
• [Presentation] 胆道系悪性腫瘍由来オルガノイドの長期培養と創薬研究への応用 (2018)
  • Author(s): 齋藤義正、齋藤英胤
  • Organizer: 第54回日本肝臓学会総会
• [Presentation] Intrahepatic cholangiocarcinoma cells can be converted into functional hepatocytes by inhibition of Wnt signaling pathway. (2017)
  • Author(s): Saito Y, Nakaoka T, Muramatsu T, Kimura M, Saito H.
  • Organizer: American Association for Cancer Research (AACR) Annual Meeting 2017
  • Int'l Joint Research
• [Presentation] オルガノイド培養技術を用いたがん研究 (2017)
  • Author(s): 齋藤 義正、中岡 哉彰、村松 俊英、木村 真規、齋藤 英胤
  • Organizer: 第17回日本抗加齢医学会総会
  • Invited
• [Presentation] オルガノイド培養技術による難治性がんのin vitroモデルの構築と創薬研究への応用 (2017)
  • Author(s): 齋藤 義正
  • Organizer: 第77回分析化学討論会
  • Invited
• [Presentation] 胆管がんオルガノイドにおける糖欠乏下での幹細胞性の増強 (2017)
  • Author(s): 吉川 直、齋藤 義正、齋藤 英胤
  • Organizer: 第76回日本癌学会学術総会
• [Presentation] DNAメチル化阻害による抗ウイルス免疫応答の活性化を介した胆道がんオルガノイドの増殖抑制 (2017)
  • Author(s): 古川 遼、齋藤 義正、齋藤 英胤
  • Organizer: 第76回日本癌学会学術総会
• [Remarks] 慶應義塾大学プレスリリース
  • URL: https://www.keio.ac.jp/ja/press-releases/2019/4/24/28-52550/
• [Remarks] Keio Research Highlights
  • URL: https://research-highlights.keio.ac.jp/article/99/ancient-antiviral-response-could-help-treat-cancer
• [Remarks] がん幹細胞におけるエピゲノム異常を標的とした革新的な創薬研究
  • URL: https://www.youtube.com/watch?v=QgmysbH_6_A&t
• [Patent(Industrial Property Rights)] 癌治療用医薬組成物 (2018)
  • Inventor(s): 齋藤 義正、齋藤 英胤、杉山 優子、村松 俊英
  • Industrial Property Rights Holder: 学校法人慶應義塾
  • Industrial Property Rights Type: 特許
  • Filing Date: 2018
• [Patent(Industrial Property Rights)] 胆道癌オルガノイド又は膵臓癌オルガノイドの培養用培地 (2018)
  • Inventor(s): 齋藤義正、金井弥栄、村松俊英
  • Industrial Property Rights Holder: 齋藤義正、金井弥栄、村松俊英
  • Industrial Property Rights Type: 特許
  • Filing Date: 2018