低酸素適応システムによる巨核球への分化・成熟過程の解析

• Project Area: Molecular mechanisms of cell fate determination in the cells that undergo stepwise differentiation to multiple pathways
• Project/Area Number: 23118527
• 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: Keio University
• Principal Investigator: 田久保 圭誉 慶應義塾大学, 医学部, 講師 (50502788)
• Project Period (FY): 2011-04-01 – 2013-03-31
• Project Status: Completed (Fiscal Year 2012)
• Budget Amount: ¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000); Fiscal Year 2012: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000); Fiscal Year 2011: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000)
• Keywords: 巨核球 / 造血幹細胞 / ニッチ / 低酸素 / 血小板

Outline of Annual Research Achievements

2年度にわたる本研究では、巨核球の分化・成熟過程における新規シグナルの同定を目指した。特に巨核球前駆細胞から巨核球への分化、そして血小板産生に至る一連の分化過程における低酸素ニッチで機能する遺伝子セットの効果について検討した。また同時に未分化前駆細胞から巨核球への分化過程の経時的なトランスクリプトーム解析を行うことで新たな巨核球分化制御因子を同定することを目指した。本解析には新規に作成した純化した巨核球前駆細胞からin vitroで巨核球分化を高効率に誘導できるシステムを用いることで、巨核球分化過程の新規制御機構の解明を図った。
本年度は前年度に引き続いてFACSを用いて巨核球前駆細胞を単離し、ファイブロネクチンで表面をコートしたディッシュ上でサイトカインSCFとTPOを含む無血清培地で培養することで巨核球を誘導し、さらにproplateletも効率よく誘導するシステムの向上を図った。また、前年度までに同定した巨核球終末分化誘導遺伝子セットの解析を行い、定量的PCRや免疫染色、ウイルスベクターを用いた強制発現や低分子化合物によるシグナル抑制実験を行い、新たな巨核球終末分化で機能する関連シグナルの同定を行った。さらに、骨髄において、造血幹細胞と巨核球がニッチで共存していることを見出した。

Research Progress Status

24年度が最終年度であるため、記入しない。

Strategy for Future Research Activity

24年度が最終年度であるため、記入しない。

Research Products

• [Journal Article] Induction of pluripotent stem cells by each of the known reprogramming factors from primordial germ cells. (2013)
  • Author(s): Nagamatsu G
  • Journal Title: Stem Cells Volume: 31 Issue: 3 Pages: 479-487
  • DOI: 10.1002/stem.1303
  • Peer Reviewed
• [Journal Article] Regulation of glycolysis by Pdk functions as a metabolic checkpoint for cell cycle quiescence in hematopoietic stem cells. (2013)
  • Author(s): Takubo K
  • Journal Title: Cell Stem Cell Volume: 12 Issue: 1 Pages: 49-61
  • DOI: 10.1016/j.stem.2012.10.011
  • Peer Reviewed
• [Journal Article] Prostaglandin E2 regulates murine hematopoietic stem/progenitorcells directly via EP4 receptor and indirectly through mesenchymal progenitor cells. (2013)
  • Author(s): Ikushima YM. Arai F, Hosokawa K, Toyama H, Takubo K, Furuyashiki T, Narumiya S, Suda T
  • Journal Title: Blood Volume: 121(11) Issue: 11 Pages: 1995-2007
  • DOI: 10.1182/blood-2012-06-437889
  • Peer Reviewed
• [Journal Article] Tracing the conversion process for primordial germ cells to pluripotent (2012)
  • Author(s): Nagamatsu G
  • Journal Title: Biol Reprod Volume: 86 Issue: 6 Pages: 182-182
  • DOI: 10.1095/biolreprod.111.096792
  • Peer Reviewed
• [Journal Article] Roles of the hypoxia response system in hematopoietic and leukemic stem cells. (2012)
  • Author(s): Keiyo Takubo
  • Journal Title: Int J Hematol. Volume: 95 Issue: 5 Pages: 478-483
  • DOI: 10.1007/s12185-012-1071-4
  • NAID: 10030632958
  • Peer Reviewed
• [Journal Article] Optimal ratio of transcription factors for somatic cell reprogramming. (2012)
  • Author(s): Nagamatsu G
  • Journal Title: J Biol Chem Volume: 287 Issue: 43 Pages: 36273-36282
  • DOI: 10.1074/jbc.m112.380683
  • Peer Reviewed
• [Journal Article] TAK1 (MAP3K7) signaling regulates hematopoietic stem cells through TNF-dependent and -independent mechanisms (2012)
  • Author(s): Takaesu G, Inagaki M, Takubo K, Mishina Y, Hess PR, Dean GA, Yoshimura A, Matsumoto K, Suda T, Ninomiya-Tsuji J
  • Journal Title: PLoS One Volume: 7(11) Issue: 11 Pages: e51073-e51073
  • DOI: 10.1371/journal.pone.0051073
  • Peer Reviewed
• [Journal Article] Dynamic regulation of Th17 differentiation by oxygen concentrations (2012)
  • Author(s): Ai Ikejiri
  • Journal Title: International Immunology Volume: 24 Issue: 3 Pages: 137-146
  • DOI: 10.1093/intimm/dxr111
  • Peer Reviewed
• [Journal Article] In vivo behavior of multiple myeloma cells and their niche in immunodeficient mice (2012)
  • Author(s): Iriuchishima H, Kizaki M, et al
  • Journal Title: PLoS ONE Volume: 7(2) Issue: 2 Pages: e30557-e30557
  • DOI: 10.1371/journal.pone.0030557
  • Peer Reviewed
• [Journal Article] Ataxia-telangiectasia mutated (ATM) deficiency decreases reprogramming efficiency and leads to genomic instability in iPS cells (2011)
  • Author(s): Taisuke Kinoshita
  • Journal Title: Biochemical and Biophysical Research Communications Volume: 407 Issue: 2 Pages: 321-326
  • DOI: 10.1016/j.bbrc.2011.03.013
  • Peer Reviewed
• [Journal Article] Telomerase reverse transcriptase protects ATM-deficient hematopoietic stem cells from ROS-induced apoptosis through a telomere independent mechanism (2011)
  • Author(s): Nitta Eriko
  • Journal Title: Blood Volume: 117 Issue: 16 Pages: 4169-80
  • DOI: 10.1182/blood-2010-08-297390
  • Peer Reviewed
• [Journal Article] Isolation and function of mouse tissue resident vascular precursors marked by myelin protein zero (2011)
  • Author(s): Kubota Y, Takubo K, Hirashima M, Nagoshi N, Kishi K, Sano K, Murakami M, Ema M, Omatsu Y, Takahashi S, Nagasawa T, Shibuya M, Okano H, Suda T
  • Journal Title: J Exp Med Volume: 208 Issue: 5 Pages: 949-960
  • DOI: 10.1084/jem.20102187
  • Peer Reviewed
• [Journal Article] Metabolic regulation of hematopoietic stem cells in the hypoxic niche (2011)
  • Author(s): Suda T, et al
  • Journal Title: Cell Stem Cell Volume: 9 Issue: 4 Pages: 298-310
  • DOI: 10.1016/j.stem.2011.09.010
  • Peer Reviewed
• [Presentation] 巨核球分化シグナルと造血幹細胞機能の連関 (2012)
  • Author(s): 田久保 圭誉
  • Organizer: 新学術領域研究「細胞運命制御」班会議
  • Place of Presentation: 広島
  • Year and Date: 2012-06-06
• [Presentation] Glycolysis by Pdk maintains the quiescence of hematopoietic stem cells (2011)
  • Author(s): Keiyo Takubo
  • Organizer: DiSCUSS Meeting
  • Place of Presentation: MPI-CBG(ドイツ ドレスデン)
  • Year and Date: 2011-12-06
• [Presentation] 骨髄腫ニッチの解析 (2011)
  • Author(s): 田久保圭誉
  • Organizer: 第14回癌と骨病変研究会
  • Place of Presentation: 千代田放送会館(東京)
  • Year and Date: 2011-11-18
• [Presentation] Glycolytic Energy Metabolism by Pdk Maintains Quiescence of Hematopoietic Stem Cells (2011)
  • Author(s): 田久保圭誉
  • Organizer: 第73回日本血液学会学術集会
  • Place of Presentation: 名古屋国際会議場(名古屋)
  • Year and Date: 2011-10-15
• [Presentation] NOGマウスを用いた多発性骨髄腫と骨髄腫ニッチへのBendamustineの効果の解析 (2011)
  • Author(s): 田久保圭誉
  • Organizer: 第70回日本癌学会学術総会
  • Place of Presentation: 名古屋国際会議場(名古屋)
  • Year and Date: 2011-10-04
• [Presentation] 造血幹細胞の新たな幹細胞性としてのエネルギー代謝制御機構 (2011)
  • Author(s): 田久保圭誉
  • Organizer: 国立感染症研究所血液・安全性研究部セミナー
  • Place of Presentation: 国立感染症研究所(東京)
  • Year and Date: 2011-09-28
• [Presentation] Pdkを介した代謝プログラムによる造血幹細胞維持とその操作 (2011)
  • Author(s): 田久保圭誉
  • Organizer: 新学術領域「細胞運命制御」若手の会
  • Place of Presentation: 軽井沢プリンスホテル(軽井沢)
  • Year and Date: 2011-09-24
• [Presentation] 低酸素応答系による造血幹細胞の代謝プログラム制御機構 (2011)
  • Author(s): 田久保圭誉
  • Organizer: 第84回日本生化学会大会
  • Place of Presentation: 国立京都国際会館(京都)
  • Year and Date: 2011-09-22
• [Presentation] Anaerobic energy metabolism regulates hematopoietic stem cell quiescence (2011)
  • Author(s): Keiyo Takubo
  • Organizer: 3rd Else Kroner--Fresenius Symposium on Molecular Mechanisms of Adult Stem Cell Aging
  • Place of Presentation: Reisensburg Castle(ドイツ ギュンツブルグ)
  • Year and Date: 2011-05-20
• [Presentation] Glycolysis maintains quiescence of hematopoietic stem cells (2011)
  • Author(s): Keiyo Takubo
  • Organizer: Keio University School of Medicine Global COE Program International Symposium 2011 "Cutting Edge of Stem Cell Medicine"
  • Place of Presentation: 慶應義塾大学医学部(東京)
  • Year and Date: 2011-05-16
• [Presentation] A protective metabolic program in hematopoietic stem cells (2011)
  • Author(s): Keiyo Takubo
  • Organizer: 第9回幹細胞シンポジウム
  • Place of Presentation: 泉ガーデンギャラリー(東京)
  • Year and Date: 2011-05-14
• [Book] Advances in Hematopoietic Stem Cell Research(Edited by Rosana Pelayo) (2012)
  • Author(s): Keiyo Takubo
  • Total Pages: 464
  • Publisher: InTech
• [Remarks]
  • URL: http://web.sc.itc.keio.ac.jp/celldiff/
• [Remarks]
  • URL: http://web.sc.itc.keio.ac.jp/celldiff/taisya.html