Development of intestinal graft production and transplantation method using human iPS cell-derived self-organization intestinal organoids.

• Project/Area Number: 20K22920
• Research Category: Grant-in-Aid for Research Activity Start-up
• Allocation Type: Multi-year Fund
• Review Section: 0903:Organ-based internal medicine and related fields
• Research Institution: Tokyo Medical and Dental University
• Principal Investigator: Mizutani Tomohiro 東京医科歯科大学, 医学部附属病院, 助教 (80632588)
• Project Period (FY): 2020-09-11 – 2022-03-31
• Project Status: Completed (Fiscal Year 2021)
• Budget Amount: ¥2,860,000 (Direct Cost: ¥2,200,000、Indirect Cost: ¥660,000); Fiscal Year 2021: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2020: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
• Keywords: 腸管スフェロイド / iPS細胞 / 腸管グラフト / 自己組織化 / 短腸症候群

Outline of Research at the Start

近年の幹細胞研究、再生医療の発展に伴い、免疫拒絶反応を回避し得る自家腸管組織移植への期待は高まっているが、未だ全層性の管腔構造として腸管機能を有する「iPS 細胞由来の腸管グラフト」を作出する技術は達成されてない。本研究では、iPS 細胞由来の「自己組織化腸管スフェロイド」を用い、実際の移植治療に応用可能な腸管グラフトへと成熟させる技術の開発、および臨床応用への検証としてマウスを用いた移植、二期的な成熟腸管グラフト吻合技術の開発を目指す。本研究の遂行により臨床応用可能な技術開発の基盤のみならず、腸上皮幹細胞による体外での自律的な3次元臓器形成、機能獲得に関する重要な知見を創出を期待する。

Outline of Final Research Achievements

Self-organization intestinal spheroids derived from human iPS cells were successfully auto-fused into intestine-like tubular spheroids on a special low-attachment micro groove plate. Efficient culture conditions to maintain a large structure of tubular intestinal spheroids were also found by introducing rotational suspension culture using a bioreactor. The established intestinal spheroids were transplanted into immunodeficient mice for further tissue maturation in vivo. We were able to establish a stable transplantation method of intestinal tubular spheroids to the mesenteric region. The intestinal spheroids grew and matured into intestinal graft tissue after transplantation.

Academic Significance and Societal Importance of the Research Achievements

iPS 細胞由来の「自己組織化腸管スフェロイド」を用い、実際の移植治療に応用可能な腸管グラフトへと成熟させる技術の開発を目指す本研究は、世界で初めての管腔構造を有する腸管グラフトを作出し、免疫不全マウス実験系を用いたグラフト成熟技術の確立により、体外で作出した腸管グラフトを移植することで腸機能を回復させるという新規治療のPOC (Proof Of Concept) 取得を目指したものである。本研究の成果を発展させることで、広範な腸機能の消失を伴う短腸症候群に対する根本的治療の基盤技術開発のみならず、腸上皮幹細胞による体外での自律的な3次元臓器形成、機能獲得に関する重要な知見の創出が期待される。

Research Products

• [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
• [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] 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