並列タイトル等オルガノイド イショク ノ スケール アップ シュホウ ノ カクリツ
Oruganoido ishoku no sukēru appu shuhō no kakuritsu
Establishment of a scale-up method for organoid transplantation
一般注記type:text
小腸は栄養吸収を担う主要な臓器であり，その広範な切除では十分な消化吸収が行われず，短腸症候群と呼ばれる栄養不良状態に至る．重症の短腸症候群患者では小腸移植が唯一の根治的な治療法となりうるが，他臓器に比べて著しく高い拒絶率のため普及しておらず，短腸症候群に対する新規治療法の開発が求められている．小腸の構造の複雑さゆえに，腸管上皮オルガノイドを用いた再生医療の小腸疾患への応用はこれまで現実的なものではなかった．最近，我々は免疫不全マウスの大腸上皮を剥離し，ヒト上皮に置換することで、マウスにヒト正常大腸上皮を構築する技術の開発に成功した（Sugimoto et al. Cell Stem Cell 2018）。さらに，大腸構造を利用して上皮のみをドナー細胞と置き換えるこの移植技術を小腸疾患に応用し，機能的な小腸化大腸の作製に小動物において成功している（Sugimoto et al. Nature 2021）．
本研究では小動物において確立した小腸オルガノイドの培養，移植をスケールアップする手法を確立し，大動物へと展開する基盤を構築することを目的とする．研究計画初年度である今年度は，ラットおよびブタ大腸の切除腸管を用いた粘膜剥離手法の選定結果に基づいて，ブタのin vivoにおいて大腸粘膜剥離手法を試みた．様々な手法の検討の末，上皮の剥離に成功している．また，ラットにおいて達成した術式をブタで行う際の問題点を抽出した．ブタの小腸オルガノイドを樹立し，蛍光標識することにも成功しており，大動物レベルでの移植実験を行う基盤が構築された．
The small intestine is the primary site for nutrient absorption. Extensive small intestinal resection leads to inadequate digestion and malabsorption, a condition known as short bowel syndrome (SBS). Intestinal transplantation is currently the only potentially-curative treatment option for severe SBS patients, but severe allogenic reaction reduces the graft survival rate and has prevented the widespread use this treatment. Therefore, development of a novel therapeutic strategy for SBS is warranted. The structural complexity of the small intestine has precluded the application of organoid-based regenerative medicine to small intestinal diseases. Recently, we have successfully developed a technique to construct normal human colonic epithelium in mice by removing the colonic epithelium from immunodeficient mice and replacing it with human epithelium (Sugimoto et al. Cell Stem Cell 2018). Furthermore, this transplantation technique, in which only the epithelium is replaced by donor cells using the colonic structure, has been applied to small intestinal diseases, and we have successfully generated a functional small intestinalized colon in small animals (Sugimoto et al. Nature 2021).
The aim of this study is to establish a method to scale up the culture and transplantation of small intestinal organoids, which has been established in small animals, and to establish the platform to expand the method to large animals. This year, the first year of the research plan, we tried in vivo colonic mucosa removal in pigs based on the results of the selected mucosa removal methods using resected intestine of rats and pigs. After examining various procedures, the epithelium was successfully removed. In addition, problems in performing the technique achieved in rats were extracted when it was performed in pigs. We have also succeeded in establishing and fluorescently labeling small intestinal organoids from pigs, establishing the basis for transplantation experiments at the large animal level.
申請種類 : 福澤基金研究補助
連携機関・データベース国立情報学研究所 : 学術機関リポジトリデータベース（IRDB）（機関リポジトリ）