心筋細胞分化における時期特異的クロマチンリモデリング制御に関する研究

• Project Area: Molecular mechanisms of cell fate determination in the cells that undergo stepwise differentiation to multiple pathways
• Project/Area Number: 25118709
• 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: The University of Tokyo
• Principal Investigator: 油谷 浩幸 東京大学, 先端科学技術研究センター, 教授 (10202657)
• Project Period (FY): 2013-04-01 – 2015-03-31
• Project Status: Completed (Fiscal Year 2014)
• Budget Amount: ¥11,180,000 (Direct Cost: ¥8,600,000、Indirect Cost: ¥2,580,000); Fiscal Year 2014: ¥5,590,000 (Direct Cost: ¥4,300,000、Indirect Cost: ¥1,290,000); Fiscal Year 2013: ¥5,590,000 (Direct Cost: ¥4,300,000、Indirect Cost: ¥1,290,000)
• Keywords: 細胞系譜 / エピゲノム / 分化

Outline of Annual Research Achievements

多能性細胞から心筋細胞への分化に伴う過程で、DNAメチル化、ヒストン修飾等のエピゲノム標識データに加えて、ヒドロキシメチル化シトシンの局在をhmeDIP-seq法によって調べたところ、時期特異的に増加する領域を同定した。先に得られたFAIRE陽性領域とも重なることが判明し、developmental enhancer領域においてDNA脱メチル化が生じていると考えられた。すなわち、転写因子が安定的に結合するためには、ヌクレオソームを除去し、脱DNAメチル化を行い、HATなどをリクルートしてdevelopmental enhancerを活性化することが明らかとなった。
さらに一細胞のトランスクリプトーム解析を各タイムポイントで実施した。トランスクリプトーム解析は1細胞を単離後、得られた全RNAからoligo(dT)プライマーを用いてSMARTer 法でcDNA合成を行い、トランスポゼースを用いてライブラリーを作成して次世代シーケンサーでRNA-seqを実施した。各データポイント内においての細胞毎の遺伝子発現の不均一性が示された一方で、それぞれの細胞を分化の時間軸に沿って配置することができ、β-cateninやBrachyury などの転写因子のChIP-seqデータから想定される遺伝子転写カスケードをよく反映していた。
すなわち、細胞分化のプロセスにおいてdevelopmental enhancerが転写因子により認識され、安定的に活性化されるためにはクロマチンリモデリング複合体やDNA脱メチル化関連酵素、ヒストンアセチル化酵素などをリクルートしていることをゲノムワイドに示した。

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] A C-terminal mutant of CCAAT-enhancer-binding protein α (C/EBPα-C(m)) downregulates Csf1r, a potent accelerator in the progression of acute myeloid leukemia with C/EBPα-C(m). (2015)
  • Author(s): Togami K, Kitaura J, Uchida T, Inoue D, Nishimura K, Kawabata KC, Nagase R, Horikawa S, Izawa K, Fukuyama T, Nakahara F, Oki T, Harada Y, Harada H, Aburatani H, Kitamura T.
  • Journal Title: Exp Hematol. Volume: 43(4) Issue: 4 Pages: 300-308
  • DOI: 10.1016/j.exphem.2014.11.011
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Genome-wide comprehensive analysis reveals critical cooperation between Smad and c-Fos in RANKL-induced osteoclastogenesis (2015)
  • Author(s): Omata Y, Yasui T, Hirose J, Izawa N, Imai Y, Matsumoto T, Masuda H, Tokuyama N, Nakamura S, Tsutsumi S, Yasuda H, Okamoto K, Takayanagi H, Hikita A, Imamura T, Matsuo K, Saito T, Kadono Y, Aburatani H, Tanaka S
  • Journal Title: J Bone Miner Res. Volume: 30 Issue: 5 Pages: 869-77
  • DOI: 10.1002/jbmr.2418
  • Peer Reviewed / Open Access
• [Journal Article] Genome-wide approaches reveal functional vascular endothelial growth factor (VEGF)-inducible nuclear factor of activated T cells (NFAT) c1 binding to angiogenesis-related genes in endothelium. (2014)
  • Author(s): Suehiro JI, Kanki Y, Makihara C, Schadler K, Miura M, Manabe Y, Aburatani H, Kodama T, Minami T
  • Journal Title: J Biol Chem Volume: 289 Issue: 42 Pages: 29044-59
  • DOI: 10.1074/jbc.m114.555235
  • Peer Reviewed / Open Access
• [Journal Article] Cross-enhancement of ANGPTL4 transcription by HIF1 alpha and PPAR beta/delta is the result of the conformational proximity of two response elements. (2014)
  • Author(s): Inoue T, Kohro T, Tanaka T, Kanki Y, Li G, Poh HM, Mimura I, Kobayashi M, Taguchi A, Maejima T, Suehiro JI, Sugiyama A, Kaneki K, Aruga H, Dong S, Stevens JF, Yamamoto S, Tsutsumi S, Fujita T, Ruan X, Aburatani H, Nangaku M, Ruan Y, Kodama T, Wada Y.
  • Journal Title: Genome Biology Volume: 15 Issue: 4 Pages: R63-R63
  • DOI: 10.1186/gb-2014-15-4-r63
  • Peer Reviewed / Open Access
• [Journal Article] Genome-wide analysis of the chromatin composition of histone H2A and H3 variants in mouse embryonic stem cells. (2014)
  • Author(s): Yukawa, M. et al.
  • Journal Title: PLOS One Volume: 9 Issue: 3 Pages: e92689-e92689
  • DOI: 10.1371/journal.pone.0092689
  • Peer Reviewed / Open Access
• [Journal Article] A novel cell-cycle indicator, mVenus-p27K-, identifies quiescent cells and visualizes G0-G1 transition. (2014)
  • Author(s): Oki, T., Nishimura, K., Kitaura, J., Togami, K., Maehara, A., Izawa, K., Sakaue-Sawano. A., Niida, A., Miyano, S., Aburatani, H., Kiyonari, H., Miyawaki, A. and Kitamura, T.
  • Journal Title: Scientific Reports Volume: 4 Issue: 1 Pages: 4012-4012
  • DOI: 10.1038/srep04012
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] The miR-17/106-p38 axis is a key regulator of the neurogenic-to-gliogenic transition in developing neural stem/progenitor cells (2014)
  • Author(s): Hayato Naka-Kanedaa, Shiho Nakamuraa, Mana Igarashi, Hisashi Aoid, Hiroaki K ankib, Jun Tsuyama, Shuichi Tsutsumie, Hiroyuki Aburatanie, Takuya Shimazakib, and Hideyuki Okano.
  • Journal Title: Proceedings of the National Academy of Sciences Volume: 111(4) Issue: 4 Pages: 1604-1609
  • DOI: 10.1073/pnas.1315567111
  • Peer Reviewed
• [Journal Article] Noninvasive and quantitative live imaging reveals a potential stress-responsive enhancer in the failing heart. (2014)
  • Author(s): Matsuoka K, Asano Y, Higo S, Tsukamoto O, Yan Y, Yamazaki S, Matsuzaki T, Kioka H, Kato H, Uno Y, Asakura M, Asanuma H, Minamino T, Aburatani H, Kitakaze M, Komuro I, Takashima S.
  • Journal Title: FASEB J. Volume: 28(4) Issue: 4 Pages: 1870-9
  • DOI: 10.1096/fj.13-245522
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] GATA factor switching from GATA2 to GATA1 contributes to erythroid differentiation (2013)
  • Author(s): Suzuki M, Kobayashi-Osaki M, Tsutsumi S, Pan X, Ohmori S, Takai J, Moriguchi T, Ohneda O, Ohneda K, Shimizu R, Kanki Y, Kodama T, Aburatani H, Yamamoto M
  • Journal Title: Genes Cells Volume: 18 Issue: 11 Pages: 921-933
  • DOI: 10.1111/gtc.12086
  • Peer Reviewed
• [Presentation] Single cell transcriptome analysis reveals cellular heterogeneity and transcriptional networks in cardiomyocyte differentiation (2015)
  • Author(s): 油谷浩幸
  • Organizer: AGBT2015
  • Place of Presentation: Marco Island (USA)
  • Year and Date: 2015-02-25 – 2015-02-28
• [Presentation] Coordinated chromatin regulation by developmental signals during cardiomyocyte differentiation (2015)
  • Author(s): 油谷浩幸
  • Organizer: Joint symposium on TGF-b family and cancer
  • Place of Presentation: Tukuba International Conference Center（茨城県、筑波市）
  • Year and Date: 2015-01-13
  • Invited
• [Presentation] Cooperative epigenomic switch during cardiomyocyte differentiation (2014)
  • Author(s): 油谷浩幸
  • Organizer: Cold Spring Harbor Asia Conference on Systems Biology of gene regulation & genome editing
  • Place of Presentation: 蘇州（中国）
  • Year and Date: 2014-09-09
  • Invited
• [Presentation] Cooperative epigenomic switch during cardiomyocyte differentiation (2014)
  • Author(s): 油谷浩幸
  • Organizer: 国立遺伝学研究所研究会
  • Place of Presentation: 国立遺伝学研究所（静岡県、三島市）
  • Year and Date: 2014-06-27
  • Invited
• [Presentation] Chromatin remodeling in cardiomyocyte differentiation
  • Author(s): 油谷浩幸
  • Organizer: The Fukuoka International Symposium on Genomics & Epigenomics 2013
  • Place of Presentation: 福岡
  • Invited
• [Presentation] 心筋細胞分化における協調的エピゲノム転換
  • Author(s): 油谷浩幸
  • Organizer: 第36回日本分子生物学会年会
  • Place of Presentation: 神戸
  • Invited
• [Presentation] Single cell RNA sequencing reveals transition of cell populations with epigenomic switch in cell fate determination along cardiomyocyte differentiation
  • Author(s): 油谷浩幸
  • Organizer: AGBT2014
  • Place of Presentation: Marco Island, Florida, USA
• [Presentation] Epigenomic Switch in Cardiomyocyte Differentiation
  • Author(s): 油谷浩幸
  • Organizer: 第７８回日本循環器学会学術総会
  • Place of Presentation: 東京
  • Invited
• [Remarks] 東京大学先端科学技術研究センターゲノムサイエンス分野
  • URL: http://www.genome.rcast.u-tokyo.ac.jp/
• [Remarks] 東京大学先端科学技術研究センター ゲノムサイエンス分野
  • URL: http://www.genome.rcast.u-tokyo.ac.jp/