Elucidating the mechanism underlying the mesenchymal polarization of the trachea through Wnt signaling

• Project/Area Number: 19K16156
• Research Category: Grant-in-Aid for Early-Career Scientists
• Allocation Type: Multi-year Fund
• Review Section: Basic Section 44020:Developmental biology-related
• Research Institution: Institute of Physical and Chemical Research
• Principal Investigator: Kishimoto Keishi 国立研究開発法人理化学研究所, 生命機能科学研究センター, 研究員 (70700029)
• Project Period (FY): 2019-04-01 – 2022-03-31
• Project Status: Completed (Fiscal Year 2021)
• Budget Amount: ¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000); Fiscal Year 2021: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000); Fiscal Year 2020: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000); Fiscal Year 2019: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
• Keywords: 気管 / 極性化 / 間充織 / Wnt / 上皮 / 幹細胞 / オルガノイド / 細胞極性 / Wnt signaling / マウス気管

Outline of Research at the Start

臓器のかたちは、細胞の振る舞いや細胞間の協調的な運動によって決定づけられる。細胞の極性化は、細胞や組織に方向性や協調性を与えるイベントであり、臓器が正しい三次元構造を作るために必須の過程である。
申請者は、間充織細胞が上皮組織方向に極性を同調させながら、マウス気管管腔の長さを決定することを発見した。これらの結果は、臓器の形態形成における間充織細胞の極性化の重要性を示している。本研究計画では、気管発生における間充織細胞の極性化を制御する上皮―間充織相互作用を理解することを目的とする。さらに、幹細胞を用いた再構成論的アプローチにより、ヒト気管発生における間充織細胞の極性化過程の可視化にも取り組む。

Outline of Final Research Achievements

In this study, we attempted to explore the Wnt-related genes which are expected to be involved in mesenchymal polarization of the trachea. scRNA-seq transcriptomics and its validation with in situ hybidization have identified several Wnt signaling components during mesenchymal polarization. We also tried to establish the system in which the process of mesenchymal polarization can be analyzed in vitro. We have succeeded in generating tracheal mesenchyme from human/mouse embryonic stem cells. We are currently combining these mesenchyme with epithelial cells to recapitulate the process of tracheal mesenchymal polarization.

Academic Significance and Societal Importance of the Research Achievements

我々は先行研究において、気管が正しい長さと太さに成長するためには、間充織細胞の極性化が必要であることを見出した。一方、これまでの当該分野の研究の多くは上皮細胞に焦点をあっており、間充織組織の発生についてはあまり研究されていなかった。本研究では、呼吸器間充織細胞をヒト幹細胞から作成することを世界に先駆けて成功した。これらの細胞を上皮細胞と混合培養することで、ヒト臓器の形成過程を培養皿上で再構築する技術の確立に繋がる。これらの技術はヒト臓器の正常な形成機構の解明・病態の理解につながると期待される。

Research Products

• [Int'l Joint Research] CincinnatiChildrensHospitalMedicalCenter(米国)
• [Int'l Joint Research] Cincinnati Children Hospital(米国)
• [Journal Article] Functional human gastrointestinal organoids can be engineered from three primary germ layers derived separately from pluripotent stem cells (2022)
  • Author(s): Eicher Alexandra K.、Kechele Daniel O.、Sundaram Nambirajan、Berns H. Matthew、Poling Holly M.、Haines Lauren E.、Sanchez J. Guillermo、Kishimoto Keishi、Krishnamurthy Mansa、Han Lu、Zorn Aaron M.、Helmrath Michael A.、Wells James M.
  • Journal Title: Cell Stem Cell Volume: 29 Issue: 1 Pages: 36-51.e6
  • DOI: 10.1016/j.stem.2021.10.010
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Directed differentiation of human pluripotent stem cells into diverse organ-specific mesenchyme of the digestive and respiratory systems. (2022)
  • Author(s): Keishi Kishimoto, Kentaro Iwasawa, Alice Sorel, Carlos Ferran-Heredia, Lu Han, Mitsuru Morimoto, James M Wells, Takanori Takebe and Aaron M Zorn
  • Journal Title: nature protocols Volume: -
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Mammalian tracheal development and reconstruction: insights from <i>in vivo</i> and <i>in vitro</i> studies (2021)
  • Author(s): Kishimoto Keishi、Morimoto Mitsuru
  • Journal Title: Development Volume: 148 Issue: 13
  • DOI: 10.1242/dev.198192
  • Peer Reviewed / Open Access
• [Journal Article] Single cell transcriptomics identifies a signaling network coordinating endoderm and mesoderm diversification during foregut organogenesis. (2020)
  • Author(s): Han L, Chaturvedi P, Kishimoto K, Koike H, Nasr T, Iwasawa K, Giesbrecht K, Witcher PC, Eicher A, Haines L, Lee Y, Shannon JM, Morimoto M, Wells JM, Takebe T, Zorn AM.
  • Journal Title: Nature communications Volume: 11(1) Issue: 1 Pages: 4158-4158
  • DOI: 10.1038/s41467-020-17968-x
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Bidirectional Wnt signaling between endoderm and mesoderm confers tracheal identity in mouse and human cells. (2020)
  • Author(s): Kishimoto K, Furukawa KT, Luz-Madrigal A, Yamaoka A, Matsuoka C, Habu M, Alev C, Zorn AM, Morimoto M.
  • Journal Title: Nature communications Volume: 11(1) Issue: 1 Pages: 4159-4159
  • DOI: 10.1038/s41467-020-17969-w
  • NAID: 120006882430
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Bidirectional Wnt signaling between endoderm and mesoderm confer tracheal identity in mouse and human (2020)
  • Author(s): Keishi Kishimoto, Kana T. Furukawa, Agustin Luz Madrigal, Akira Yamaoka, Chisa Matsuoka, Masanobu Habu, Cantas Alev, Aaron M. Zorn, Mitsuru Morimoto
  • Journal Title: Biorxiv Volume: 756825 Pages: 1-31
  • DOI: 10.1101/758235
  • Open Access / Int'l Joint Research
• [Journal Article] Single cell transcriptomics reveals a signaling roadmap coordinating endoderm and mesoderm diversification during foregut organogenesis (2020)
  • Author(s): Lu Han, Praneet Chaturvedi, Keishi Kishimoto, Hiroyuki Koike, Talia Nasr, Kentaro Iwasawa, Kirsten Giesbrecht, Phillip C Witcher, Alexandra Eicher, Lauren Haines, Yarim Lee, John M Shannon, Mitsuru Morimoto, James M Wells, Takanori Takebe, Aaron M Zorn
  • Journal Title: Biorxiv Volume: 861781 Pages: 1-49
  • DOI: 10.1101/756825
  • Open Access / Int'l Joint Research
• [Presentation] Induction of organ specific splanchnic mesoderm from human embryonic stem cell (2020)
  • Author(s): Keishi Kishimoto
  • Organizer: CLEAR consortium 2020
  • Int'l Joint Research