造血幹細胞の自己複製と分化の誘導を掛け分ける分子基盤

• Project Area: Molecular mechanisms of cell fate determination in the cells that undergo stepwise differentiation to multiple pathways
• Project/Area Number: 25118717
• Research Category: Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area)
• Allocation Type: Single-year Grants
• Review Section: Biological Sciences
• Research Institution: Hiroshima University
• Principal Investigator: 瀧原 義宏 広島大学, 原爆放射線医科学研究所, 教授 (60226967)
• Project Period (FY): 2013-04-01 – 2015-03-31
• Project Status: Completed (Fiscal Year 2014)
• Budget Amount: ¥11,960,000 (Direct Cost: ¥9,200,000、Indirect Cost: ¥2,760,000); Fiscal Year 2014: ¥5,980,000 (Direct Cost: ¥4,600,000、Indirect Cost: ¥1,380,000); Fiscal Year 2013: ¥5,980,000 (Direct Cost: ¥4,600,000、Indirect Cost: ¥1,380,000)
• Keywords: 造血幹細胞 / 自己複製 / 分化 / Geminin / ポリコーム複合体1 / Hoxb4 / Hoxa9

Outline of Annual Research Achievements

研究室ではまず造血幹細胞の活性を支持する２大内的因子であるポリコーム複合体1やHoxb4が供にGemininに対するE3ユビキチンリガーゼとして機能していることを独自に明らかにした。Gemininは細胞の増殖と分化を同時に調節することから、Gemininが造血幹細胞の自己複製と分化の誘導を掛け分ける中核因子として機能していることが推測される。そこで、本研究では造血幹細胞におけるGemininの発現動態を詳細に明らかにすることを目的として、Geminin遺伝子に蛍光タンパク質遺伝子EYFPをノックインし、Geminin-EYFP融合タンパク質が発現するようにしたGeminin-EYFPノックインマウスを作製し、Geminin分子を可視化した。ホモ接合型Geminin-EYFPノックインマウスにおいてEYFPの蛍光強度が充分高く、内在性のGemininの発現をよく反映していることや、発生や妊孕性に特に問題のないことを確かめた。そして、G0/G1期の未分化造血細胞の中で長期骨髄再構築能を有する造血幹細胞においてGemininの発現が高いことを見い出した。一方、FGF4のmembrane translocating motifを組み込んだcell-penetrating Gemininを作製し、Gemininタンパク質を直接細胞内に導入する実験系の構築を行った。これらの研究成果は、造血幹細胞が自己複製する際にGemininの発現がどのように調節され、未分化性が保たれつつ細胞増殖するのかについて理解を深めることと、Gemininの発現量を調節することによって造血幹細胞の活性を操作できる技術の開発に道を開くことが期待される。

Research Progress Status

26年度が最終年度であるため、記入しない。

Strategy for Future Research Activity

26年度が最終年度であるため、記入しない。

Research Products

• [Journal Article] MK3 modulation affects BMI1-dependent and independent cell cycle check-points (2015)
  • Author(s): Prickaerts, P., Niessen, H. E. C., Dahlmans, V. E. H., Salvaing, J., Vanhove, J., Spaapen, F., Geijselaers, C., Partouns, I., Bartels, S. J. J., Neumann, D., Speel, E. J., Takihara, Y., Wouters, B. G., Voncken, J. W.
  • Journal Title: PLoS ONE Volume: 10 Issue: 4 Pages: e0118840-e0118840
  • DOI: 10.1371/journal.pone.0118840
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Transcription of the Geminin gene is regulated by a negative-feedback loop. (2014)
  • Author(s): Ohno, Y., Saeki, K., Yasunaga, S., Kurogi. T., Suzuki-Takedachi, K., Shirai, M., Mihara K., Yoshida, K., Voncken J. W, Ohtsubo, M. & Takihara Y.
  • Journal Title: Mol. Biol. Cel Volume: 25(8) Issue: 8 Pages: 1374-1383
  • DOI: 10.1091/mbc.e13-09-0534
  • Peer Reviewed / Open Access
• [Journal Article] Simple diagnosis of STAT1 gain-of-function alleles in patients with chronic mucocutaneous candidiasis. (2014)
  • Author(s): Mizoguchi Y, Tsumura M, Okada S, Hirata O, Minegishi S, Imai K, Hyakuna N, Muramatsu H, Kojima S, Ozaki Y, Imai T, Takeda S, Okazaki T, Ito T, Yasunaga S, Takihara Y, Bryant VL, Kong XF, Cypowyj S, Boisson-Dupuis S, Puel A, Casanova JL, Morio T, Kobayashi M.
  • Journal Title: J. Leukoc. Biol. Volume: 95 Issue: 4 Pages: 667-76
  • DOI: 10.1189/jlb.0513250
  • Peer Reviewed / Open Access
• [Journal Article] Heterozygosity for the Y701C STAT1 mutation in a multiplex kindred with multifocal osteomyelitis (2013)
  • Author(s): Hirata, O.
  • Journal Title: Haematologica Volume: 98 Issue: 10 Pages: 1641-1649
  • DOI: 10.3324/haematol.2013.083741
  • Peer Reviewed
• [Presentation] Geminin-EYFP knock-in mice provide a powerful tool for dissecting a molecular mechanism regulating self-renewal or differentiation of hematopoietic stem cells (2014)
  • Author(s): Suzuki-Takedachi, K., Yasunaga, S., Ohno, Y., Kurogi, T., Santo, M., Ohtsubo, M., Takihara, Y.
  • Organizer: 第37回日本分子生物学会年会
  • Place of Presentation: 横浜、パシフィコ横浜
  • Year and Date: 2014-11-25 – 2014-11-27
• [Presentation] Cell-penetrating recombinant Geminin protein may provide a new strategy for regulating self-renewal or cellular differentiation in hematopoietic stem cells (2014)
  • Author(s): Kurogi, T., Ohno, Y., Yasunaga, S., Suzuki-Takedachi, K.,, Santo, M., Masuhiro, Y., Hanazawa, S., Ohtsubo, M., Takihara, Y.
  • Organizer: 第37回日本分子生物学会年会
  • Place of Presentation: 横浜、パシフィコ横浜
  • Year and Date: 2014-11-25 – 2014-11-27
• [Presentation] Geminin aids detailed molecular analysis of self-renewing hematopoietic stem cells (2014)
  • Author(s): Yasunaga, S., Ohno, S., Suzuki-Takedachi, K., Kurogi, T., Santo, M., Ohtsubo, M. Takihara, Y.
  • Organizer: 第76回日本血液学会学術集会
  • Place of Presentation: 大阪、大阪国際会議場
  • Year and Date: 2014-10-31 – 2014-11-02
• [Presentation] Direct transmission of a cell-penetrating form of Geminin, a regulator for hematopoietic stem cells (2014)
  • Author(s): Ohno, S., Yasunaga, S., Kurogi, T., Suzuki-Takedachi, K., Santo, M., Ohtsubo, M. Takihara, Y.
  • Organizer: 第76回日本血液学会学術集会
  • Place of Presentation: 大阪、大阪国際会議場
  • Year and Date: 2014-10-30 – 2014-11-02
• [Presentation] Molecular role for Geminin in governing hematopoietic stem cell activity (2014)
  • Author(s): Takihara, Y.
  • Organizer: Views into nuclear function: A See Drug Workshop
  • Place of Presentation: Patras, Greece
  • Year and Date: 2014-09-11 – 2014-09-13
  • Invited
• [Presentation] Direct transduction of a cell-penetrating form of Geminin, a molecule regulating DNA replication and chromatin remodeling (2014)
  • Author(s): Takihara, Y., Yasunaga, S., Ohno, Y., Kurogi, T., Suzuki-Takedachi, K., Santo, M., Ohtsubo, M.
  • Organizer: The 43rd Annual Scientific Meeting of the ISEH-International Society of Experimental Hematology
  • Place of Presentation: Montreal, Canada
  • Year and Date: 2014-08-21 – 2014-08-24
• [Presentation] Geminin regulates transcription via chromatin remodeling to govern cellular proliferation and differentiation (2014)
  • Author(s): Ohno, Y., Yasunaga, S., Kurogi, T., Suzuki-Takedachi, K., Santo, M., Ohtsubo, M., Takihara, Y.
  • Organizer: The 12nd Stem Cell Research Symposium
  • Place of Presentation: 福岡、九州大学医学部百年講堂
  • Year and Date: 2014-05-30 – 2014-05-31
• [Presentation] Identification and functional analysis of self-renewing hematopoietic stem cells by using Geminin-EYFP knock-in mice (2014)
  • Author(s): Yasunaga, S., Ohno, S., Suzuki-Takedachi, K., Kurogi, T., Santo, M., Ohtsubo, M., Takihara, Y.
  • Organizer: The 12nd Stem Cell Research Symposium
  • Place of Presentation: 福岡、九州大学医学部百年講堂
  • Year and Date: 2014-05-30
• [Presentation] Visualization and functional dissection of Geminin, a molecule governing cellular proliferation and differentiation in hematopoietic stem cells (2013)
  • Author(s): Takihara, Y.
  • Organizer: 42nd annual scientific meeting of the international society for hematology and stem cells
  • Place of Presentation: Viena
• [Presentation] Visualization and functional analysis of Geminin in the hematopoietic system (2013)
  • Author(s): Takihara, Y.
  • Organizer: th annual meeting of Korean Society for Stem Cell Research
  • Place of Presentation: Seoul
  • Invited
• [Presentation] Expression and molecular function of Geminin, which governs hematopoietic stem cell activity (2013)
  • Author(s): Takihara, Y.
  • Organizer: 第86回日本生化学会大会 インターナショナルセッション
  • Place of Presentation: 横浜
  • Invited
• [Presentation] olecular role for Geminin in switching self-renewal and differentiation of hematopoietic stem cells (2013)
  • Author(s): Yasunaga, S.
  • Organizer: 第74回日本血液学会学術集会
  • Place of Presentation: 札幌
• [Presentation] Negative feed-back loop regulates transcription of the Geminin gene, which governs HSC activity (2013)
  • Author(s): Ohno, S.
  • Organizer: 第74回日本血液学会学術集会
  • Place of Presentation: 札幌
• [Presentation] Molecular analysis of a role for Geminin in regulating proliferation and differentiation of hematopoietic stem/progenitor cells by visualizing Geminin expression (2013)
  • Author(s): Suzuki-Takedachi, K
  • Organizer: 第36回日本分子生物学会年会
  • Place of Presentation: 神戸
• [Presentation] Negative feed-back loop regulates E2F-mediated transcriptional activation of the Geminin gene (2013)
  • Author(s): Kurogi, T.
  • Organizer: 第36回日本分子生物学会年会
  • Place of Presentation: 神戸