染色体構造変化がもたらす持続的VCAM-1遺伝子発現機構解析

• Project Area: Systematic study of chromosome adaptation
• Project/Area Number: 25125707
• 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: 神吉 康晴 東京大学, アイソトープ総合センター, 助教 (00534869)
• 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: miRNA / ヒストン修飾 / 染色体構造 / 動脈硬化 / 接着因子 / 染色体立体構造 / ChIA-PET / ChIP-seq / VCAM-1 / TNFa / IL4

Outline of Annual Research Achievements

申請者はこれまでに、血管内皮細胞特異性に転写因子GATA2がその細胞特異的な染色体構造を形成することで寄与していること（Kanki Y et al 2011 EMBO J）、炎症性刺激で駆動される遺伝子発現に、活性型のmRNA転写と抑制型のmiRNA転写がカップリングして起こるマシナリーが介在すること（Papantonis A et al 2012 EMBO J）、動脈硬化の鍵となるVCAM-1 (Vascular Cell Adhesion Molecule-1)の持続的な発現に寄与している機能的なエンハンサーが存在すること（Kanki Y et al 2011 MCB）、等を見出し報告してきた。そこで、本研究では申請者のこれまでの染色体構造解析を更に発展させ、次世代シークエンサーにより網羅的に染色体構造を読み取る新しい手法ChIA-PET (Chromatin Interaction Analysis Using Paired End diTag Sequencing)、及びその可視化手法としての3D-FISHを用いて、VCAM-1の一過性の発現誘導、持続的な発現誘導、二つのパターンに関する染色体構造の違いを読み解くことを目的としている。
平成26年度は、特に2012年に報告したmiRNAを中心とする抑制性マシナリーに着目した。血管内皮細胞にTNF刺激を行った際に変動するmiRNAを捉えるためにmiRNAアレイを行い、VCAM1転写制御候補となるmiRNAを同定した。更に、このmiRNAの直接の標的を同定するためにAGO2抗体を用いたRIP解析を行い、染色体構造変化に寄与する候補因子を2個同定した。これらはそれぞれノックダウンすることで、VCAM1、ICAM1、E-selectinといった接着因子を抑制することから、動脈硬化初期病変の鍵であると考えられる。

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] 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] Direct Evidence for Pitavastatin Induced Chromatin Structure Change in the KLF4 Gene in Endothelial Cells. (2014)
  • Author(s): Maejima T, Inoue T, Kanki Y, Kohro T, Li G, Ohta Y, Kimura H, Kobayashi M, Taguchi A, Tsutsumi S, Iwanari H, Yamamoto S, Aruga H, Dong S, Stevens JF, Poh HM, Yamamoto K, Kawamura T, Mimura I,
  • Journal Title: PLoS One. Volume: 9 Issue: 5 Pages: e96005-e96005
  • DOI: 10.1371/journal.pone.0096005
  • 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
  • DOI: 10.1186/gb-2014-15-4-r63
  • Peer Reviewed / Open Access
• [Journal Article] Direct evidence for pitavastatin induced chromatin structure change in the KLF4 gene in endothelial cells. (2014)
  • Author(s): Maejima T, Inoue T, Kanki Y,,,,, Kodama T, Wada Y
  • Journal Title: Plos One Volume: in press
  • NAID: 120006667792
  • Peer Reviewed
• [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: in press
  • Peer Reviewed
• [Journal Article] Revolution of nephrology research by deep sequencing : ChIP-seq and RNA-seq. (2014)
  • Author(s): Mimura I, Kanki Y, Kodama T, Nangaku M.
  • Journal Title: Kidney International Volume: 85 Issue: 1 Pages: 31-38
  • DOI: 10.1038/ki.2013.321
  • Peer Reviewed
• [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
• [Journal Article] Inhibition of histone demethylase JMJD1A improves anti-angiogenic therapy and reduces tumor associated macrophages (2013)
  • Author(s): Shimamura T, Wang F, Suehiro J, Kanki Y, Wada Y, Yuasa Y, Aburatani H, Miyano S, Minami T, Kodama T, Shibuya M
  • Journal Title: Cancer Res Volume: 73 Issue: 10 Pages: 3019-3028
  • DOI: 10.1158/0008-5472.can-12-3231
  • Peer Reviewed
• [Presentation] Three Dimensional Chromatin Architecture Correlates with Two Opposite Transcription Factors, which contributes the Cell Specificity. (2014)
  • Author(s): Yasuharu Kanki, Teppei Shimamura, Shuichi Tsutsumi, Hiroyuki Aburatani, Youichiro Wada
  • Organizer: The 4D Nucleome 2014
  • Place of Presentation: 安芸グランドホテル、広島県、日本
  • Year and Date: 2014-12-17 – 2014-12-20
• [Presentation] Three Dimensional Chromatin Architecture Correlates with Two Opposite Transcription Factors, which contributes the Cell Specificity. (2014)
  • Author(s): Yasuharu Kanki, Teppei Shimamura, Shuichi Tsutsumi, Hiroyuki Aburatani, Youichiro Wada
  • Organizer: 第37回日本分子生物学会年会
  • Place of Presentation: パシフィコ横浜、横浜市、日本
  • Year and Date: 2014-11-25 – 2014-11-27
• [Presentation] Three Dimensional Chromatin Architecture Correlates with Two Opposite Transcription Factors, which contributes the Cell Specificity. (2014)
  • Author(s): Yasuharu Kanki, Teppei Shimamura, Shuichi Tsutsumi, Hiroyuki Aburatani, Youichiro Wada
  • Organizer: 第87回日本生化学会大会
  • Place of Presentation: 京都国際会議場、京都市、日本
  • Year and Date: 2014-10-15 – 2014-10-18
• [Presentation] Three Dimensional Chromatin Architecture Correlates with Two Opposite Transcription Factors, which contributes to the Cell Specificity (2014)
  • Author(s): Yasuharu Kanki
  • Organizer: 11th EMBL Conference Transcription and Chromatin
  • Place of Presentation: EMBL, Heidelberg, Germany
  • Year and Date: 2014-08-23 – 2014-08-26
• [Presentation] GENETIC AND EPIGENETIC LANDSCAPE OF ENDOTHELIAL CELLS DIFFERENTITATION REVEAL THE TRANSCRIPTIONAL FACTORS NETWORK (2014)
  • Author(s): Kanki, Yasuharu, Matsunaga, Taichi, Ryo, Nakaki, Yamamizu, Kohei, Shimamura, Teppei, Miyano, Satoru, Aburatani, Hiroyuki, Kodama, Tatsuhiko, Wada, Youichiro, Yamashita, Jun, Minami, Takashi
  • Organizer: International Society For Stem Cell Research the 12th annual meeting
  • Place of Presentation: The Convention Center, Vancouver, Canada
  • Year and Date: 2014-06-18 – 2014-06-21
• [Presentation] Transcription Factor GATA2 is indispensable for the Differentiation and Maintenance of Vascular Endothelial Cells (2014)
  • Author(s): Yasuharu Kanki, Taichi Matsunaga, Hiroyuki Aburatani, Youichiro Wada, Tatsuhiko Kodama, Jun Yamashita, Takashi Minami
  • Organizer: the 18th International Vascular Biology Meeting
  • Place of Presentation: みやこメッセ、京都市、日本
  • Year and Date: 2014-04-14 – 2014-04-17
• [Presentation] 染色体立体構造網羅解析からの細胞特異性解読へのアプローチ (2014)
  • Author(s): 神吉康晴
  • Organizer: 京都大学再生医科学研究所平成25年度共同研究報告会
  • Place of Presentation: 京都大学再生医科学研究所、京都市、日本
  • Invited
• [Presentation] エピゲノム制御からの 新規血管新生因子の同定 (2014)
  • Author(s): 神吉康晴
  • Organizer: 第2回生活習慣病の分子細胞病態学研究会
  • Place of Presentation: ホテル椿山荘東京、東京都、日本
• [Presentation] Transcription factor GATA2 is indispensable for the differentiation and maintenance of vascular endothelial cells (2013)
  • Author(s): Yasuharu Kanki, Taichi Matsunaga, Hiroyuki Aburatani, Youichiro Wada, Tatsuhiko Kodama, Jun Yamashita, Takashi Minami
  • Organizer: The 11th Japan-Korea Joint Symposium on Vascular Biology
  • Place of Presentation: Hyatt Regency jeju Hotel, Jeju Island, Korea
• [Presentation] Transcription factor GATA2 is indispensable for the differentiation and maintenance of vascular endothelial cells (2013)
  • Author(s): 神吉康晴、興梠貴英、堤修一、三村維真理、松永太一、山水康平、片山志織、油谷浩幸、児玉龍彦、和田洋一郎、山下潤、南敬
  • Organizer: 第86回日本生化学会大会
  • Place of Presentation: パシフィコ横浜、横浜市、日本
• [Presentation] A novel epigenetic mechanism revealed tumor associated angiogenesis (2013)
  • Author(s): Yasuharu Kanki, Jun-ichi Suehiro, Tsuyoshi Osawa, Imari Mimura, Youichiro Wada, Hiroyuki Aburatani, Tatsuhiko Kodama, Takashi Minami
  • Organizer: 第72回日本癌学会学術総会
  • Place of Presentation: パシフィコ横浜、横浜市、日本
• [Book] 血管医学 (2013)
  • Author(s): 神吉康晴
  • Total Pages: 9
  • Publisher: メディカルレビュー社