ゲノム認識エピジェネティック変更化合物の開発

• Project Area: Analyses and regulation of germline epigenome
• Project/Area Number: 26112517
• 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: Chiba Cancer Center (Research Institute)
• Principal Investigator: 永瀬 浩喜 千葉県がんセンター(研究所), がん遺伝創薬研究室, 研究所長 (90322073)
• Project Period (FY): 2014-04-01 – 2016-03-31
• Project Status: Completed (Fiscal Year 2015)
• Budget Amount: ¥9,360,000 (Direct Cost: ¥7,200,000、Indirect Cost: ¥2,160,000); Fiscal Year 2015: ¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000); Fiscal Year 2014: ¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000)
• Keywords: ヒストン脱アセチル化酵素阻害剤 / エピゲノム変更 / DNA配列認識化合物 / ピロールイミダゾールポリアミド / ヒストンアセチル化酵素活性化剤 / リプログラミング / 多能性幹細胞誘導 / エピジェネティクス / エピゲノム / 化学バイオプローブ / ヒストン脱アセチル化阻害剤

Outline of Annual Research Achievements

本研究課題では、領域研究者の希望するゲノム領域に本技術による配列特異的な転写阻害剤PIP やエピゲノム変更薬PIP-SAHA、PIP-CBI等をデザイン・合成し供給していくことおよび他のヒストン修飾阻害剤とのコンジュゲートを合成し、新たな標的領域特異的なエピゲノム変更技術を確立することを目的とした。このことで、領域研究者による生殖細胞のエピゲノムの乱れの機能解析に役立て、エピゲノムを正常化する薬剤候補を開発・提供できるよう研究を進めた。
具体的にはDNA結合化合物（PIP）によるヒストン脱アセチル化酵素阻害剤（SAHA）を生殖細胞、幹細胞に送達させることの確認および作成したPIP-SAHAライブラリーによりヒト繊維芽細胞への化合物の効果を確認し、それぞれの化合物が誘導する遺伝子発現の変化を確認した。特に多能性幹細胞様の発現パターンをヒト皮膚繊維芽細胞で導く化合物E、F、H、Iについてその発現誘導とPIPのゲノム結合領域の予測による作用機序の解明を試み、幹細胞誘導因子と関連する遺伝子ネットワークの活性化を確認した。またPIP化合物が、マウス精巣細胞及び卵巣細胞内に送達されることも確認した。今後、生殖エピゲノムを化合物により変更する技術を開発し、学術領域の発展に貢献できれば幸いである。

Research Progress Status

27年度が最終年度であるため、記入しない。

Strategy for Future Research Activity

27年度が最終年度であるため、記入しない。

Research Products

• [Journal Article] Genetic alterations in Japanese extrahepatic biliary tract cancer. (2016)
  • Author(s): Noguchi R, Yamaguchi K, Ikenoue T, Terakado Y, Ohta Y, Yamashita N, Kainuma O, Yokoi S, Maru Y, Nagase H, Furukawa Y.
  • Journal Title: Oncology Letters Volume: -
  • Peer Reviewed
• [Journal Article] Targeting Oct1 genomic function inhibits androgen receptor signaling and castration-resistant prostate cancer growth (2016)
  • Author(s): Inoue S, Obinata D, Takayama K, Urano T, Suzuki T, Takahashi S, Fujimura T, Homma Y, Fujiwara K, Fukuda N, Nagase H, Aburatani H, and Tsutsumi S.
  • Journal Title: Oncogene Volume: -
  • Peer Reviewed
• [Journal Article] Selective Targeting of the KRAS Codon 12 Mutation Sequence by Pyrrole–Imidazole Polyamide seco-CBI Conjugates. (2015)
  • Author(s): Taylor RD, Chandran A, Kashiwazaki G, Hashiya K, Bando T, Nagase H and Sugiyama H.
  • Journal Title: Chemistry. Volume: 21 Issue: 42 Pages: 14996-5003
  • DOI: 10.1002/chem.201501870
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] The oncogenic role of GASC1 in chemically induced mouse skin cancer. (2015)
  • Author(s): Ozaki Y, Fujiwara K, Ikeda M, Ozaki T, Terui T, Soma M, Inazawa J, Nagase H.
  • Journal Title: Mammalian Genome Volume: 26 Pages: 591-597
  • Peer Reviewed
• [Journal Article] Silencing of RUNX2 enhances gemcitabine sensitivity of p53-deficient human pancreatic cancer AsPC-1 cells through the stimulation of TAp63-mediated cell death. (2015)
  • Author(s): Sugimoto H, Nakamura M, Yoda H, Hiraoka K, Shinohara K, Sang M, Fujiwara K, Shimozato O, Nagase H, Ozaki T.
  • Journal Title: Cell Death Discovery Volume: 6 Issue: 10 Pages: e1914-e1914
  • DOI: 10.1038/cddis.2015.242
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Preclinical Study of Novel Gene Silencer Pyrrole-Imidazole Polyamide Targeting Human TGF-β1 Promoter for Hypertrophic Scars in a Common Marmoset Primate Model. (2015)
  • Author(s): Igarashi J, Fukuda N, Inoue T, Nakai S, Saito K, Fujiwara K, Matsuda H, Ueno T, Matsumoto Y, Watanabe T, Nagase H, Bando T Sugiyama H Itoh T Soma M.
  • Journal Title: PlosOne. Volume: 10(5) Issue: 5 Pages: e0125295-e0125295
  • DOI: 10.1371/journal.pone.0125295
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Pyrrole-Imidazole (PI) polyamide-mediated silencing of KCNQ1OT1 expression induces cell death in Wilms tumor cells. (2015)
  • Author(s): Yoshizawa S, Fujiwara K, Sugito K, Uekusa S, Kawashima H, Hoshi R, Watanabe Y, Hirano T, Furuya T, Masuko T, Ueno T, Fukuda N, Soma M, Ozaki T, Koshinaga T, Nagase H.
  • Journal Title: International Journal of Oncology Volume: 47(1) Issue: 1 Pages: 115-121
  • DOI: 10.3892/ijo.2015.3018
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Runt-related transcription factor 2 (RUNX2) attenuates the transcriptional activity as well as DNA damage-mediated induction of pro-apoptotic TAp73 to regulate chemo-sensitivity (2015)
  • Author(s): Ozaki T, Sugimoto H, Nakamura M, Hiraoka K, Yoda H, Sang M, Fujiwara K, Nagase H.
  • Journal Title: FEBS Journal Volume: 82 Issue: 1 Pages: 114-28
  • DOI: 10.1111/febs.13108
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Identification of a novel E-box binding PI polyamide inhibiting MYC-driven cell-proliferation. (2015)
  • Author(s): Mishra R, Watanabe T, Kimura M, Koshikawa N, Ikeda M, Uekusa S, Kawashima H, Wang X, Igarashi J, Choudhury D, Grandori C, Kemp C, Ohira M, Verma N, Kobayashi Y, Takeuchi J, Koshinaga T, Nemoto N, Fukuda N, Soma M, Kusafuka T, Fujiwara K, Nagase H.
  • Journal Title: Cancer Science Volume: 106(4) Issue: 4 Pages: 421-429
  • DOI: 10.1111/cas.12610
  • Peer Reviewed / Open Access
• [Journal Article] Inhibition of KRAS codon 12 mutants using a novel DNA-alkylating pyrrole-imidazole polyamide conjugate. (2014)
  • Author(s): Hiraoka K, Inoue T, Taylor RD, Watanabe T, Koshikawa N, Yoda H, Shinohara K, Takatori A, Sugimoto H, Maru Y, Denda T, Fujiwara K, Balmain A, Ozaki T, Bando T, Sugiyama H, Nagase H.
  • Journal Title: Nat. Commun. Volume: 6 Issue: 1 Pages: 6706-6706
  • DOI: 10.1038/ncomms7706
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Inhibition of malignant phenotypes of human osteosarcoma cells by a gene silencer, a pyrrole-imidazole polyamide, which targets E-box. (2014)
  • Author(s): 134. Taniguchi M, Nakai Y, Ozaki T, Koshikawa N, Kojima T, Kataba M, Oguni A, Matsuda H, Yoshida Y, Tokuhashi Y, Fukuda N, Ueno T, Soma T, Nagase H.
  • Journal Title: FEBS Open Bio Volume: 4 Issue: 1 Pages: 328-334
  • DOI: 10.1016/j.fob.2014.03.004
  • Peer Reviewed / Open Access
• [Journal Article] A novel gene regulator, pyrrole-imidazole polyamide targeting ABCA1 gene increases cholesterol efflux from macrophages and plasma HDL concentration (2014)
  • Author(s): Tsunemi A, Ueno T, Fukuda N, Watanabe T, Tahira K, Haketa A, Hatanaka Y, Tanaka S, Matsumoto T, Matsumoto Y, Nagase H, Soma M
  • Journal Title: Journal of Molecular Medicine Volume: 92 Issue: 5 Pages: 509-521
  • DOI: 10.1007/s00109-013-1118-x
  • Peer Reviewed
• [Journal Article] Inhibition of Human Osteosarcoma Cell Migration and Invasion by a Gene Silencer, Pyrrole–Imidazole Polyamide, Targeted at the Human MMP9 NF-κB Binding Site (2014)
  • Author(s): Kojima T, Wang X, Fujiwara K, Osakaa S, Yoshida Y, Osaka E, Taniguchi M, Ueno T, Fukuda N, Soma M, Tokuhashi Y and Nagase H.
  • Journal Title: Biological and Pharmaceutical Bulletin Volume: 37 Issue: 9 Pages: 1460-1465
  • DOI: 10.1248/bpb.b14-00147
  • NAID: 130004684785
  • ISSN: 0918-6158, 1347-5215
  • Peer Reviewed / Open Access
• [Journal Article] Identification of Frequent Differentially Methylated Region in Sporadic Bladder Cancers. (2014)
  • Author(s): Hasegawa R, Fujiwara K, Obinata D, Kawashima H, Shinojima Y, Igarashi J, Wang X, Ghosh S, Nagase H, Takahashi S.
  • Journal Title: Urol Int. Volume: 94(4) Issue: 4 Pages: 479-484
  • DOI: 10.1159/000365197
  • Peer Reviewed
• [Journal Article] Pyrrole-imidazole polyamide targeted to break fusion sites in TMPRSS2 and ERG gene fusion represses prostate tumor growth (2014)
  • Author(s): Obinata D, Ito Akiko, Fujiwara K, Takayama K, Ashikari D, Murata Y, Yamaguchi K, Urano T, Fujimura T, Fukuda N, Soma M, Watanabe T, Nagase H, Inoue S, Takahashi S
  • Journal Title: Cancer Sci Volume: 105 Issue: 10 Pages: 1272-1278
  • DOI: 10.1111/cas.12493
  • Peer Reviewed / Open Access
• [Journal Article] Genome-wide screening of aberrant DNA methylation which associated with gene expression in mouse skin cancers (2013)
  • Author(s): Fujiwara K, Ghosh S, Liang P, Morien E, Soma M, Nagase H
  • Journal Title: Molecular Carcinogenesis Volume: e-pub Issue: 3 Pages: 1-11
  • DOI: 10.1002/mc.22085
  • Peer Reviewed
• [Presentation] Chemical genomics approaches to modify biological processes and pathological state (2015)
  • Author(s): Hiroki Nagase
  • Organizer: 横浜港湾記念会館
  • Place of Presentation: 横浜港湾記念会館
  • Year and Date: 2015-11-12
  • Int'l Joint Research / Invited
• [Presentation] Inhibition of KRAS mutant colon cancer using a novel DNA-alkylating Pyrrole-Imidazole polyamide conjugate targeting KRAS Codon12 Mutant DNA (2015)
  • Author(s): ・Hiroki Nagase, Kiriko Hiraoka, Takahiro Inoue, Takayoshi Watanabe, Ken-Ichi Shinohara, Nobuko Koshikawa, Ozaki Toshinori.
  • Organizer: 3rd International Conference of Federation of Asian Clinical Oncology(FACO)
  • Place of Presentation: 京都国際会議場
  • Year and Date: 2015-10-29
  • Int'l Joint Research / Invited
• [Presentation] Tumor accumulation of a novel alkylating pyrrole-imidazole polyamide (KR12) targeting KRAS codon 12 mutations in murine xenograft models of human colon cancer (2015)
  • Author(s): ・Takahiro Inoue, Kiriko Hiraoka, Yusei Suzuki, Hiroyuki Yoda, Takayoshi Watanabe, Atsushi Takatori, Nobuko Koshikawa, Toshinori Ozaki, Hiroki Nagase.
  • Organizer: AACR American Association for Cancer Research ANUAL MEETING 2015 ミニシンポジウム“Exploiting the MAPK pathway in cancer”
  • Place of Presentation: フィラデルフィア 米国
  • Year and Date: 2015-04-18
  • Int'l Joint Research / Invited
• [Presentation] A novel alkylating pyrrole-imidazole polyamide, KR12, specifically recognizes mutant KRAS genes and potently induces cell death (2015)
  • Author(s): ・Kiriko Hiraoka, Takahiro Inoue, Hiroyuki Yoda, Atsushi Takatori, Takayoshi Watanabe, Nobuko Koshikawa, Toshinori Ozaki, Hiroki Nagase.
  • Organizer: AACR American Association for Cancer Research ANUAL MEETING 2015 ミニシンポジウム“Exploiting the MAPK pathway in cancer”
  • Place of Presentation: フィラデルフィア 米国
  • Year and Date: 2015-04-18
  • Int'l Joint Research / Invited
• [Presentation] Targeting specific DNA sequences by N-methylpyrrole and N-methylimidazole polyamides provides insights for the development of novel diagnostic and therapeutic drugs. (2014)
  • Author(s): Atsushi Takatori, Hiroyuki Yoda, Kiriko Hiraoka, Ken ichi Shinohara, Takayoshi Watanabe, Nobuko Koshikawa, and Hiroki Nagase.
  • Organizer: The 28th International Mammalian Genome Conference.
  • Place of Presentation: Bar Harbor, USA
  • Year and Date: 2014-10-26 – 2014-10-29
• [Presentation] ZEB1/E-cadherin 転写制御を標的とした新規ピロールイミダゾールポリアミドの開発 (2014)
  • Author(s): 養田裕行、高取敦志、渡部隆義、篠原憲一、井上貴博、平岡桐子、越川信子、尾崎俊文、永瀬浩喜
  • Organizer: 第73回日本癌学会学術総会
  • Place of Presentation: 横浜
  • Year and Date: 2014-09-25 – 2014-09-27
• [Presentation] MMP-9を標的とした脂肪族/芳香族アミノ酸ペアを有するPIポリアミドによる遺伝子抑制 (2014)
  • Author(s): 渡部隆義、越川信子、尾崎俊文、板東俊和、杉山弘、永瀬浩喜
  • Organizer: 第73回日本癌学会学術総会
  • Place of Presentation: 横浜
  • Year and Date: 2014-09-25 – 2014-09-27
• [Presentation] Precision Medicineを目指したオンコジェニックドライバーを標的とした治療 (2014)
  • Author(s): 永瀬浩喜、平岡桐子、井上隆博、渡部隆義、越川信子、尾崎俊文
  • Organizer: 第23回癌病態治療研究会
  • Place of Presentation: 岐阜
  • Year and Date: 2014-06-12 – 2014-06-13
• [Book] 「がんのエピジェネティクス研究の進歩」臨床免疫・アレルギー科，65（4）：345 - 350 (2016)
  • Author(s): 永瀬浩喜
  • Total Pages: 5
  • Publisher: 化学評論社
• [Remarks] 千葉県がんセンター研究所がん遺伝創薬研究室
  • URL: http://www.pref.chiba.lg.jp/gan/kenkyujo/soshiki/iden/index.html
• [Patent(Industrial Property Rights)] ピロールイミダゾール含有ポリアミドを用いた標的二本鎖核酸分子の濃縮方法およびキット (2015)
  • Inventor(s): 永瀬 浩喜、塚本 圭、北野 史朗
  • Industrial Property Rights Holder: 永瀬 浩喜、塚本 圭、北野 史朗
  • Industrial Property Rights Type: 特許
  • Filing Date: 2015-10-08