Elucidation of super-undifferentiated state of mouse ES cells and application to reprogramming of ES/iPS cells of other species

• Project/Area Number: 16H04683
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Laboratory animal science
• Research Institution: Nara Medical University
• Principal Investigator: Horie Kyoji 奈良県立医科大学, 医学部, 教授 (30333446)
• Project Period (FY): 2016-04-01 – 2019-03-31
• Project Status: Completed (Fiscal Year 2019)
• Budget Amount: ¥18,070,000 (Direct Cost: ¥13,900,000、Indirect Cost: ¥4,170,000); Fiscal Year 2018: ¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000); Fiscal Year 2017: ¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000); Fiscal Year 2016: ¥9,620,000 (Direct Cost: ¥7,400,000、Indirect Cost: ¥2,220,000)
• Keywords: ES細胞 / iPS細胞 / 遺伝子 / 未分化状態 / 分化 / CRISPR / 再生医学 / 発生・分化 / 多能性 / ゲノム編集

Outline of Final Research Achievements

We identified a novel heterogeneity in mouse ES cells. By comprehensive analysis of gene expression, we identified a highly undifferentiated cell population. We also showed that chimeric mice could be produced from this cell population with high efficiency, demonstrating the high pluripotency of this cell population. Furthermore, the transcription factor network that regulates this heterogeneity was identified by measuring the transcription factor activity at the one-cell level and by disrupting transcription factors using CRISPR/Cas9. The manipulation of this network may lead to producing high-quality ES/iPS cells in species other than mice.

Academic Significance and Societal Importance of the Research Achievements

本研究では、マウスES細胞の解析を通じて、一見して均一に見えるマウスES細胞の多能性が、個々の細胞ごとに異なることを明らかにした。さらに、多能性状態の不均一性を制御する遺伝子群を同定し、これらの遺伝子群の発現レベルを変えることで、多能性状態を人為的に操作できる可能性を示した。この遺伝子群は、ヒトをはじめとする他の生物種でも保存されている。よって、今回の知見は、様々な生物種のES/iPS細胞へも適用できると考えられ、再生医学や発生生物学の発展への寄与を期待できる。

Research Products

• [Presentation] Identifying the heterogeneity of ground state pluripotency in mouse embryonic stem cells and elucidating its regulatory mechanism (2019)
  • Author(s): Kyoji Horie, Hitomi Watanabe, Yosuke Nishimura, Hikaru Watanabe, Masahide Seki, Akio Seita, Kagayaki Kato, Yuichi Wakayama, Jun Sese, Yutaka Suzuki, Takuji Yamada, Gen Kondoh, Junko Yoshida
  • Organizer: 第42回日本分子生物学会年会
  • Invited
• [Presentation] Identifying heterogeneity of ground state pluripotency in mouse embryonic stem cells (2018)
  • Author(s): Kyoji Horie, Junko Yoshida
  • Organizer: International Society for Stem Cell Research Annual Meeting
  • Int'l Joint Research
• [Presentation] Ground stateにおけるマウスES細胞の不均一性の同定 (2018)
  • Author(s): 堀江恭二, 渡邊仁美, 西村陽介, 渡邊日佳流, 関真秀, 清田晃央, 加藤輝, 若本祐一, 鈴木穣, 山田拓司, 近藤玄, 吉田純子
  • Organizer: 第41回日本分子生物学会年会