Development of primitive-gut-derived organ bioengineering methods based on spheroid designing.

• Project/Area Number: 20K21597
• Research Category: Grant-in-Aid for Challenging Research (Exploratory)
• Allocation Type: Multi-year Fund
• Review Section: Medium-sized Section 53:Organ-based internal medicine and related fields
• Research Institution: Tokyo Medical and Dental University
• Principal Investigator: Okamoto Ryuichi 東京医科歯科大学, 大学院医歯学総合研究科, 教授 (50451935)
• Project Period (FY): 2020-07-30 – 2022-03-31
• Project Status: Completed (Fiscal Year 2021)
• Budget Amount: ¥6,500,000 (Direct Cost: ¥5,000,000、Indirect Cost: ¥1,500,000); Fiscal Year 2021: ¥2,600,000 (Direct Cost: ¥2,000,000、Indirect Cost: ¥600,000); Fiscal Year 2020: ¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000)
• Keywords: iPS細胞 / 原腸由来臓器 / スフェロイド / 原腸 / 胚体内胚葉 / 腸 / スフェロイド・デザイニング / 前腸スフェロイド

Outline of Research at the Start

ヒトiPS細胞より任意デザインの原腸由来臓器を作成するための基盤技術を確立するため、以下３つの課題を計画期間2年で実施する
1) iPS細胞由来前腸スフェロイドの高効率作成法の開発
2) iPS細胞由来前腸スフェロイドの融合・分化能の解析
3)生体内移植による前腸スフェロイド由来臓器の特異的機能獲得の検証
これらで得られた結果に応じ、各臓器の欠損モデルに対する同所移植も試みる事により、特異的機能獲得の有無について検証を行う。

Outline of Final Research Achievements

Through our development of a highly efficient iPS cell-derived primitive-gut spheroid production and the analysis of their fusion/differentiation potential, we have optimized the spheroid-forming method. Also, we have succeeded in developing a method to produce a highly uniform and abundant number of primitive-gut spheroids under the control of their final size and regulate the differentiation of those spheroids. In addition, we have confirmed that those or spheroids can engraft within a certain period after transplantation to severely immunodeficient mice, and also acquire tissue-specific functions and increase their tissue size in vivo. From these results, we have succeeded in developing methods and essential knowledge to induce differentiation from human iPS cells to primitive-gut-derived tissue via the definitive endoderm with the help of both in vitro and in vivo experimental environments.

Academic Significance and Societal Importance of the Research Achievements

高い効率で任意のサイズを有する原腸スフェロイドを作成可能であるばかりでなく、同法で作成したスフェロイドが極めて高い融合指向性と体外環境における原腸由来臓器への分化能を兼ね備えていることを利用し、任意の形状・サイズを有する原腸由来前駆体から原腸由来組織を分化誘導する「スフェロイド・デザイニング」を可能とするための基盤となる知見が確立された。

Research Products

• [Journal Article] Transplantation of intestinal organoids into a mouse model of colitis (2022)
  • Author(s): Watanabe Satoshi、Kobayashi Sakurako、Ogasawara Nobuhiko、Okamoto Ryuichi、Nakamura Tetsuya、Watanabe Mamoru、Jensen Kim B.、Yui Shiro
  • Journal Title: Nature Protocols Volume: 17 Issue: 3 Pages: 649-671
  • DOI: 10.1038/s41596-021-00658-3
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Nickel particles are present in Crohn's disease tissue and exacerbate intestinal inflammation in IBD susceptible mice. (2022)
  • Author(s): Hiroki Matsuda, Yoichi Nibe-Shirakihara, Akiko Tamura, Emi Aonuma, Satoko Arakawa, Kana Otsubo, Yasuhiro Nemoto, Takashi Nagaishi, Kiichiro Tsuchiya, Shigeomi Shimizu, Averil Ma, Mamoru Watanabe, Motohiro Uo, Ryuichi Okamoto, Shigeru Oshima.
  • Journal Title: Biochemical and Biophysical Research Communications. Volume: 592 Pages: 74-80
  • DOI: 10.1016/j.bbrc.2021.12.111
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] A potent endocytosis inhibitor Ikarugamycin up-regulates TNF production. (2021)
  • Author(s): Ai Minamidate, Michio Onizawa, Chikako Saito, Rie Hikichi, Tomoaki Mochimaru, Mai Murakami, Chiharu Sakuma, Takehito Asakawa, Yuichi Hiraoka, Shigeru Oshima, Takashi Nagaishi, Kiichiro Tsuchiya, Hiromasa Ohira, Ryuichi Okamoto, Mamoru Watanabe.
  • Journal Title: Biochemistry and Biophysics Reports. Volume: 27 Pages: 101065-101065
  • DOI: 10.1016/j.bbrep.2021.101065
  • Peer Reviewed / Open Access
• [Journal Article] Importance of Telomere Shortening in the Pathogenesis of Ulcerative Colitis: A New Treatment From the Aspect of Telomeres in Intestinal Epithelial Cells (2021)
  • Author(s): Watanabe Sho、Hibiya Shuji、Katsukura Nobuhiro、Kitagawa Sayuki、Sato Ayako、Okamoto Ryuichi、Watanabe Mamoru、Tsuchiya Kiichiro
  • Journal Title: Journal of Crohn's and Colitis Volume: 16 Issue: 1 Pages: 109-121
  • DOI: 10.1093/ecco-jcc/jjab115
  • Peer Reviewed
• [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] Functional analysis of isoflavones using patient-derived human colonic organoids (2021)
  • Author(s): Tsuchiya Mao、Ito Go、Hama Minami、Nagata Sayaka、Kawamoto Ami、Suzuki Kohei、Shimizu Hiromichi、Anzai Sho、Takahashi Junichi、Kuno Reiko、Takeoka Sayaka、Hiraguri Yui、Sugihara Hady Yuki、Mizutani Tomohiro、Yui Shiro、Oshima Shigeru、Tsuchiya Kiichiro、Watanabe Mamoru、Okamoto Ryuichi
  • Journal Title: Biochemical and Biophysical Research Communications Volume: 542 Pages: 40-47
  • DOI: 10.1016/j.bbrc.2021.01.021
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Notch and TNF-α signaling promote cytoplasmic accumulation of OLFM4 in intestinal epithelium cells and exhibit a cell protective role in the inflamed mucosa of IBD patients (2021)
  • Author(s): Kuno Reiko、Ito Go、Kawamoto Ami、Hiraguri Yui、Sugihara Hady Yuki、Takeoka Sayaka、Nagata Sayaka、Takahashi Junichi、Tsuchiya Mao、Anzai Sho、Mizutani Tomohiro、Shimizu Hiromichi、Yui Shiro、Oshima Shigeru、Tsuchiya Kiichiro、Watanabe Mamoru、Okamoto Ryuichi
  • Journal Title: Biochemistry and Biophysics Reports Volume: 25 Pages: 100906-100906
  • DOI: 10.1016/j.bbrep.2020.100906
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Presentation] 1.東京医科歯科大学における内視鏡を用いた基礎研究の取り組み (2021)
  • Author(s): 岡本隆一
  • Organizer: 第101回日本消化器内視鏡学会総会
• [Presentation] Organoid-based regenerative and precision medicine for IBD (2021)
  • Author(s): Okamoto R
  • Organizer: KDDW2021
  • Int'l Joint Research