腸管上皮－間質ニッチの包括的理解と自己補完的組織培養技術の確立

• Project/Area Number: 15J00981
• Research Category: Grant-in-Aid for JSPS Fellows
• Allocation Type: Single-year Grants
• Section: 国内
• Research Field: Gastroenterology
• Research Institution: Keio University
• Principal Investigator: 杉本 真也 慶應義塾大学, 医学研究科, 特別研究員(DC1)
• Project Period (FY): 2015-04-24 – 2018-03-31
• Project Status: Completed (Fiscal Year 2017)
• Budget Amount: ¥3,400,000 (Direct Cost: ¥3,400,000); Fiscal Year 2017: ¥1,100,000 (Direct Cost: ¥1,100,000); Fiscal Year 2016: ¥1,100,000 (Direct Cost: ¥1,100,000); Fiscal Year 2015: ¥1,200,000 (Direct Cost: ¥1,200,000)
• Keywords: オルガノイド / 腸管上皮幹細胞 / 炎症性腸疾患 / 粘膜再生治療 / 間質細胞

Outline of Annual Research Achievements

本年度は，前年度までに確立した無血清Wnt3a-Afaminを含む移植細胞培養系と，正常ヒト大腸オルガノイドを免疫不全マウスの大腸へ同所異種移植するマウスモデルを用いて，これまで実験モデルの欠如から困難であったin vivoでのヒト大腸幹細胞の時空間ダイナミクスの検討を行った．また，大動物における内視鏡的なオルガノイド自家移植系の移植手技最適化を行い，安全性を確認した．なお，間質細胞と上皮細胞の共培養系は，がん細胞への間質細胞からのニッチ因子供給を検討する実験にも応用できた（Seino et al. Cell Stem Cell 2018）．
大腸がんの遺伝学的細胞系譜解析において確立したCRISPR-Cas9システムによるLGR5遺伝子へのCreERT2のノックインとCre依存的に発現するRainbowレポーターを組み込んだオルガノイドを作成する手法を（Shimokawa et al. Nature 2017），正常大腸オルガノイドへ応用することでヒト腸管上皮のLGR5陽性細胞から産生される子孫細胞のクローンを異なる色で標識することが可能となった．このオルガノイドをマウス腸管へ同所異種移植することにより，ヒト大腸上皮細胞の遺伝学的細胞系譜解析が可能となり，ヒトLGR5陽性幹細胞の自己複製能および多分化能をin vivoにおいてはじめて実証することに成功した（Sugimoto et al. Cell Stem Cell 2018）．これにより，ヒト腸管上皮オルガノイドの移植細胞としての可能性が示され，in vivoでのヒト大腸幹細胞の機能的挙動の観察が可能となったのみならず，この移植モデルと遺伝子改変オルガノイドを組み合わせることで，腸内におけるヒトの腫瘍形成に関する病態理解を進展させる可能性をもつ有用なリソースになるものと考えられる．

Research Progress Status

29年度が最終年度であるため、記入しない。

Strategy for Future Research Activity

29年度が最終年度であるため、記入しない。

Research Products

• [Journal Article] Reconstruction of the Human Colon Epithelium In Vivo. (2018)
  • Author(s): Sugimoto S, Ohta Y, Fujii M, Matano M, Shimokawa M, Nanki K, Date S, Nishikori S, Nakazato Y, Nakamura T, Kanai T, Sato T.
  • Journal Title: Cell Stem Cell. Volume: 22 Issue: 2 Pages: 171-176
  • DOI: 10.1016/j.stem.2017.11.012
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Human Pancreatic Tumor Organoids Reveal Loss of Stem Cell Niche Factor Dependence during Disease Progression. (2018)
  • Author(s): Seino T, Kawasaki S, Shimokawa M, Tamagawa H, Toshimitsu K, Fujii M, Ohta Y, Matano M, Nanki K, Kawasaki K, Takahashi S, Sugimoto S, Iwasaki E, Takagi J, Itoi T, Kitago M, Kitagawa Y, Kanai T, Sato T.
  • Journal Title: Cell Stem Cell. Volume: 22 Issue: 3 Pages: 454-467
  • DOI: 10.1016/j.stem.2017.12.009
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Fecal microbiota transplantation for recurrent Clostridium difficile infection in a patient with ulcerative colitis (2018)
  • Author(s): Kosaku Nanki, Shinta Mizuno, Katsuyoshi Matsuoka, Keiko Ono, Shinya Sugimoto, Hiroki Kiyohara, Mari Arai, Moeko Nakashima, Kozue Takeshita, Keiichiro Saigusa, Mitsutoshi Senoh, Tadashi Fukuda, Makoto Naganuma, Haru Kato, Wataru Suda, Masahira Hattori, Takanori Kanai
  • Journal Title: Intestinal Research Volume: 16 Issue: 1 Pages: 142-146
  • DOI: 10.5217/ir.2018.16.1.142
  • Peer Reviewed / Open Access
• [Journal Article] Visualization and Targeting of LGR5+ Human Colon Cancer Stem Cells (2017)
  • Author(s): Shimokawa M, Ohta Y, Nishikori S, Matano M, Takano A, Fujii M, Date S, Sugimoto S, Kanai T, Sato T
  • Journal Title: Nature Volume: 印刷中 Issue: 7653 Pages: 13-14
  • DOI: 10.1038/nature22081
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Efficacy of Indigo naturalis in a Multicenter Randomized Controlled Trial of Patients with Ulcerative Colitis (2017)
  • Author(s): .Naganuma M, Sugimoto S, Mitsuyama K, Kobayashi T, Yoshimura N, Ohi H, Tanaka S, Andoh A, Ohmiya N, Saigusa K, Yamamoto T, Morohoshi Y, Ichikawa H, Matsuoka K, Hisamatsu T, Watanabe K, Mizuno S, Suda W, Hattori M, Fukuda S, Hirayama A, Abe T, Watanabe M, Hibi T, Suzuki Y, Kanai T
  • Journal Title: Gastroenterology Volume: 4 Issue: 4 Pages: 935-947
  • DOI: 10.1053/j.gastro.2017.11.024
  • Peer Reviewed / Open Access
• [Journal Article] Endocytoscopy can be used to assess histological healing in ulcerative colitis patients with clinical and endoscopic remission (2017)
  • Author(s): Yoshihiro Nakazato, Makoto Naganuma, Shinya Sugimoto, Rieko Bessho, Mari Arai, Hiroki Kiyohara, Keiko Ono, Kosaku Nanki, Makoto Mutaguchi, Shinta Mizuno, Taku Kobayashi, Naoki Hosoe, Masayuki Shimoda, Takayuki Abe, Nagamu Inoue, Haruhiko Ogata, Yasushi Iwao, Takanori Kanai
  • Journal Title: Endoscopy Volume: 49 Issue: 06 Pages: 560-563
  • DOI: 10.1055/s-0043-106556
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] The risk factor of clinical relapse in ulcerative colitis patients with low dose 5-aminosalicylic acid as maintenance therapy: A report from the IBD registry (2017)
  • Author(s): Tomohiro Fukuda, Makoto Naganuma, Shinya Sugimoto, Kosaku Nanki, Shinta Mizuno, Makoto Mutaguchi, Yoshihiro Nakazato, Nagamu Inoue, Haruhiko Ogata, Yasushi Iwao, Takanori Kanai
  • Journal Title: PLoS One Volume: 12 Issue: 11 Pages: 935-47
  • DOI: 10.1371/journal.pone.0187737
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Single fecal microbiota transplantation failed to change intestinal microbiota and had limited effectiveness against ulcerative colitis in Japanese patients (2017)
  • Author(s): Mizuno S, Nanki K, Matsuoka K, Saigusa K, Ono K, Arai M, Sugimoto S, Kiyohara H, Nakashima M, Takeshita K, Naganuma M, Suda W, Hattori M, Kanai T.
  • Journal Title: Intest Res Volume: 15(1) Issue: 1 Pages: 68-74
  • DOI: 10.5217/ir.2017.15.1.68
  • Peer Reviewed / Open Access
• [Journal Article] Ulcerative Colitis Endoscopic Index of Severity Is Associated with Long-term Prognosis in Ulcerative Colitis Patients Treated with Infliximab (2016)
  • Author(s): Keiichiro Saigusa, Katsuyoshi Matsuoka, Shinya Sugimoto, Mari Arai, Hiroki Kiyohara, Kozue Takeshita, Shinta Mizuno, Kiyoto Mori, Kosaku Nanki, Tatsuya Takeshita, Yoshihiro Nakazato, Tomoharu Yajima, Makoto Naganuma, Tadakazu Hisamatsu, Haruhiko Ogata, Yasushi Iwao, Takanori Kanai
  • Journal Title: Digestive Endsoscopy Volume: 印刷中 Issue: 6 Pages: 665-670
  • DOI: 10.1111/den.12655
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] The Ulcerative Colitis Endoscopic Index of Severity (UCEIS) is useful to predict medium to long-term prognosis in ulcerative colitis patients with clinical remission. (2016)
  • Author(s): Arai M, Naganuma M, Sugimoto S, Kiyohara H, Ono K, Mori K, Saigusa K, Nanki K, Mutaguchi M, Mizuno S, Bessho R, Nakazato Y, Hosoe N, Matsuoka K, Inoue N, Ogata H, Iwao Y, Kanai T.
  • Journal Title: J Crohns Colitis Volume: 10(11) Issue: 11 Pages: 1303-1309
  • DOI: 10.1093/ecco-jcc/jjw104
  • Peer Reviewed
• [Journal Article] Endoscopic morphologic features of ulcerative colitis-associated dysplasia classified according to the SCENIC consensus statement (2016)
  • Author(s): Sugimoto S, Naganuma M, Iwao Y, Matsuoka K, Shimoda M, Mikami S, Mizuno S, Nakazato Y, Nanki K, Inoue N, Ogata H, Kanai T.
  • Journal Title: Gastrointest Endosc. Volume: 85(3) Issue: 3 Pages: 639-646
  • DOI: 10.1016/j.gie.2016.11.013
  • Peer Reviewed
• [Journal Article] Indole compounds may be promising medicines for ulcerative colitis. (2016)
  • Author(s): Sugimoto S, Naganuma M, Kanai T.
  • Journal Title: J Gastroenterol. Volume: 51(9) Issue: 9 Pages: 853-61
  • DOI: 10.1007/s00535-016-1220-2
  • Peer Reviewed
• [Journal Article] Recent trends and future directions for the medical treatment of ulcerative colitis. (2016)
  • Author(s): Makoto Naganuma, Shinta Mizuno, Kosaku Nanki, Shinya Sugimoto, Takanori Kanai
  • Journal Title: Clinical Journal of Gastroenterology Volume: 9 Issue: 6 Pages: 329-336
  • DOI: 10.1007/s12328-016-0686-z
  • Peer Reviewed / Open Access
• [Journal Article] Establishment of 3D intestinal organoid cultures from intestinal stem cells. (2016)
  • Author(s): Shinya Sugimoto, Toshiro Sato
  • Journal Title: Methods in Molecular Biology Volume: 未定
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] 腸管幹細胞 (2015)
  • Author(s): 中里 圭宏, 杉本 真也, 南木 康作, 佐藤 俊朗
  • Journal Title: 日本臨牀 Volume: 73 Pages: 232-237
• [Presentation] オルガノイドを用いた細胞系譜解析によるヒト正常大腸幹細胞挙動の可視化 (2018)
  • Author(s): 佐藤 俊朗, 杉本 真也, 太田 悠木, 下川 真理子
  • Organizer: 第17回日本再生医療学会総会
  • Invited
• [Presentation] Efficacy of therapeutic intervention for ulcerative colitis patients with the Mayo endoscopic score of 1 (2018)
  • Author(s): Tomohiro Fukuda, Makoto Naganuma, Shinya Sugimoto, Keiko Ono, Kosaku Nanki, Shinta Mizuno, Kayoko Kimura, Makoto Mutaguchi, Kaoru Takabayashi, Nagamu Inoue, Haruhiko Ogata, Yasushi Iwao, Takanori Kanai
  • Organizer: 13th Congress of European Crohn’s and Colitis Organization (ECCO)
  • Int'l Joint Research
• [Presentation] Indigo naturalis induces mucosal healing in patients with ulcerative colitis: A multicentre randomised controlled trial (2018)
  • Author(s): Makoto Naganuma, Shinya Sugimoto, Keiichi Mitsuyama, Taku Kobayashi, Naoki Yoshimura, Hidehisa Ohi, Katsuyoshi Matsuoka, Tadakazu Hisamatsu, Kenji Watanabe, Takayuki Abe, Mamoru Watanabe, Toshifumi Hibi, Yasuo Suzuki, Takanori Kanai, INDIGO study group
  • Organizer: 13th Congress of European Crohn’s and Colitis Organization (ECCO)
  • Int'l Joint Research
• [Presentation] The impact of establishment of organoid-based orthotopic xenograft model on intestinal stem cell research (2017)
  • Author(s): Shinya Sugimoto, Yuki Ohta, Takanori Kanai, Toshiro Sato
  • Organizer: The 54th Annual Meeting of the Japan Society for Mucosal Immunology
  • Invited
• [Presentation] 寛解期潰瘍性大腸炎における腸内・口腔内細菌叢の構成と予後との関連の検討 (2017)
  • Author(s): 大野 恵子, 水野 慎大, 清原 裕貴, 杉本 真也, 南木 康作, 牟田口 真, 中里 圭宏, 筋野 智久, 長沼 誠, 金井 隆典
  • Organizer: 第54回日本消化器免疫学会総会
• [Presentation] 潰瘍性大腸炎症例の散発性大腸腫瘍の臨床像 (2017)
  • Author(s): 牟田口 真, 長沼 誠, 杉本 真也, 南木 康作, 水野 慎大, 緒方 晴彦, 岩男 泰, 金井 隆典
  • Organizer: 第94回消化器内視鏡学会総会（JDDW2017）
• [Presentation] Indigo naturalis is effective for inducing clinical remission and mucosal healing in patients with ulcerative colitis (INDIGO study) (2017)
  • Author(s): Makoto Naganuma, Shinya Sugimoto, Keiichi Mitsuyama, Taku Kobayashi, Naoki Yoshimura, Katsuyoshi Matsuoka, Tadakazu Hisamatsu, Kenji Watanabe, Takayuki Abe, Yasuo Suzuki, Toshifumi Hibi, Takanori Kanai
  • Organizer: UEG Week 2017
  • Int'l Joint Research
• [Presentation] Clinical characteristics in ulcerative colitis patients with colitis associated dysplasia/cancer and sporadic tumor (2017)
  • Author(s): Makoto Mutaguchi, Makoto Naganuma, Yasushi Iwao, Tomohiro Fukuda, Shinya Sugimoto Kosaku Nanki, Shinta Mizuno, Haruhiko Ogata, Takanori Kanai
  • Organizer: UEG Week 2017
  • Int'l Joint Research
• [Presentation] Stem Cell Research and Treatment: From Bench to Clinical Practice. (2017)
  • Author(s): Shinya Sugimoto
  • Organizer: The 1st Basic Science Research Workshop: Evolving Basic Science in Intestinal Diseases
  • Place of Presentation: Seoul, Korea
  • Int'l Joint Research / Invited
• [Presentation] 腸管上皮オルガノイドを用いた幹細胞移植技術の確立 (2016)
  • Author(s): 杉本 真也, 金井 隆典, 佐藤 俊朗
  • Organizer: 第58回日本消化器病学会大会（JDDW2016）
  • Place of Presentation: 神戸コンベンションセンター（兵庫県神戸市）
• [Presentation] Efficacy and safety of fecal microbiota transplantation on ulcerative colitis. (2016)
  • Author(s): Shinta Mizuno, Kosaku Nanki, Katsuyoshi Matsuoka, Keiko Ono, Makoto Mutaguchi, Mari Arai, Shinya Sugimoto, Hiroki Kiyohara, Kozue Takeshita, Makoto Naganuma, Takanori Kanai
  • Organizer: The 4th Annual Meeting of Asian Organization for Crohn's and Colitis (AOCC)
  • Place of Presentation: Kyoto International Conference Center（京都府左京区）
  • Int'l Joint Research
• [Presentation] Gut environments and human colorectal carcinogenesis. (2016)
  • Author(s): Toshiro Sato, Shinya Sugimoto, Mami Matano
  • Organizer: The 75th Annual Meeting of the Japanese Cancer Association.
  • Place of Presentation: パシフィコ横浜（神奈川県横浜市）
• [Presentation] Cultivation of human norovirus using human duodenal organoids. (2016)
  • Author(s): Akira Fujimoto, Kei Haga, Shinya Sugimoto, Toshiro Sato, Yen Hai Doan, Motohiro Miki, Reiko Takai-Todaka, Kazuhiko Katayama
  • Organizer: The 64th Annual Meeting of the Japanese Society for Virology.
  • Place of Presentation: Sapporo Convention Center（北海道札幌市）
• [Presentation] The Investigation of Medium- to Long-Term Prognosis of Ulcerative Colitis Patients Using Ulcerative Colitis Endoscopic Index of Severity (UCEIS). (2016)
  • Author(s): Mari Arai, Makoto Naganuma, Shinya Sugimoto, Hiroki Kiyohara, Keiko Ohno, Kiyoto Mori, Kosaku Nanki, Makoto Mutaguchi, Shinta Mizuno, Rieko Bessho, Yoshihiro Nakazato, Nagamu Inoue, Haruhiko Ogata, Yasushi Iwao, Takanori Kanai
  • Organizer: Digestive Disease Week (DDW) 2016
  • Place of Presentation: San Diego, USA
  • Int'l Joint Research