タンパク質メチル化を標的とした新規がんの分子標的治療薬の開発

• Project/Area Number: 22681030
• Research Category: Grant-in-Aid for Young Scientists (A)
• Allocation Type: Single-year Grants
• Research Field: Medical genome science
• Research Institution: The University of Tokyo
• Principal Investigator: 浜本 隆二 東京大学, 医科学研究所, 助教 (80321800)
• Project Period (FY): 2010-04-01 – 2014-03-31
• Project Status: Declined (Fiscal Year 2013)
• Budget Amount: ¥20,020,000 (Direct Cost: ¥15,400,000、Indirect Cost: ¥4,620,000); Fiscal Year 2013: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000); Fiscal Year 2012: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000); Fiscal Year 2011: ¥11,180,000 (Direct Cost: ¥8,600,000、Indirect Cost: ¥2,580,000); Fiscal Year 2010: ¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000)
• Keywords: タンパク質メチル化 / エピジェネティクス / がんの分子標的治療薬

Research Abstract

平成24年度においては、がん特異的にメチル化が亢進しているタンパク質を同定することを目的に、pan-methyl lysine抗体を用いて非腫瘍細胞と比して腫瘍細胞でメチル化が亢進しているタンパク質をマススペクトロメトリーで解析を行った結果、HSP70が同定された。同じくマススペクトロメトリーで詳細にメチル化部位を解析したところ、561番目のリジン残基がジメチル化していることが同定された。そこで、特異抗体を作製し臨床検体を用いて免疫組織学的解析を行った結果、このメチル化は肺がん組織、膀胱がん組織、腎がん組織において非腫瘍部と比して腫瘍部で著しくメチル化が亢進していることが分かった。興味深いことに、通常HSP70は細胞質に主に存在しているが、メチル化したHSP70は主に細胞核に局在していた。一方、このメチル化を司るメチル化酵素の探索を行ったところ、ヒストンメチル化酵素SETD1AがHSP70をメチル化していることを同定し、SETD1Aの発現もがん細胞特異的に亢進していることを突き止めた。
続いてがん細胞中でのメチル化HSP70の機能を解析する目的で、メチル化HSP70に結合するタンパク質を免疫沈降法で探索したところ、細胞分裂に必須なAurora Bキナーゼが同定された。メチル化HSP70がAurora Bキナーゼに結合するとその活性は増強されることから、メチル化HSP70が核内に局在することにより、細胞分裂が促進すると考えられた。実際に野生型HSP70に比して、メチル化非活性型HSP70を強制発現させることにより細胞の増殖が有意に低下することから、メチル化HSP70はAurora Bキナーゼを活性化させることにより、がん細胞の異常増殖及び悪性化に寄与していることが示唆された。
本研究によりヒストンメチル化酵素SETD1AがHSP70をメチル化し、メチル化HSP70はAurora Bキナーゼを介してがん細胞の異常増殖及び悪性化に寄与するという新しいメカニズムが明らかになった。SETD1Aの発現をノックダウンすることによりがん細胞の増殖が著しく抑制されることから、SETD1Aの機能を阻害しHSP70メチル化を抑制することは抗がん治療に直結することが期待される。

Research Products

• [Journal Article] The histone methyltransferase Wolf-Hirschhorn syndrome candidate 1-like 1 (WHSC1L1) is involved in human carcinogenesis. (2013)
  • Author(s): Kang D, Cho HS, Toyokawa G, Kogure M, Yamane Y, Iwai Y, Hayami S, Tsunoda T, Field HI, Matsuda K, Neal DE, Ponder BA, Maehara Y, Nakamura Y, Hamamoto R.
  • Journal Title: Genes Chromosomes Cancer Volume: 52 Issue: 2 Pages: 126-139
  • DOI: 10.1002/gcc.22012
  • Peer Reviewed
• [Journal Article] Histone Lysine Methyltransferase SETD8 Promotes Carcinogenesis by Deregulating PCNA Expression (2012)
  • Author(s): Masashi Takawa, Hyun-Soo Cho, Shinya Hayami, Gouji Toyokawa, Masaharu Kogure, Yuka Yamane, Yukiko Iwai, Kazuhiro Maejima, Koji Ueda, Akiko Masuda, Naoshi Dohmae, Helen I. Field, Tatsuhiko Tsunoda, Takaaki Kobayashi, Takayuki Akasu, Masanori Sugiyama, Shin-ichi Ohnuma, Yutaka Atomi, Bruce A. J. Ponder, Yusuke Nakamura, Ryuji Hamamoto
  • Journal Title: Cancer Research Volume: 72 Pages: 3217-3227
  • Peer Reviewed
• [Journal Article] B1 Methylation by SMYD2 Enhances Cell Cycle Progression through an Increase of RB1 Phosphorylation (2012)
  • Author(s): Hyun-Soo Cho. Shinya Hayami, Gouji Toyokawa, Kazuhiro Maejima, Yuka Yamane, Takehiro Suzuki, Naoshi Dohmae, Masaharu Kogure, Daechun Kang, David E. Neal, Bruce A. J. Ponder, Hiroki Yamaue, Yusuke Nakamura, Ryuji Hamamoto
  • Journal Title: Neoplasia Volume: 14 Pages: 476-486
  • Peer Reviewed
• [Journal Article] Recognition of modification status on a histone H3 tail by linked histone reader modules of UHRF1 (2012)
  • Author(s): Kyohei Arita, Shin Isogai, Takashi Oda, Motoko Unoki, Kazuya Sugita, Naotaka Sekiyama, Keiko Kuwata, Ryuji Hamamoto Hidehito Tochio, Mamoru Sato, Mariko Ariyoshi, Masahiro
  • Journal Title: Proc Natl Acad Sci USA Volume: 109 Pages: 12950-12955
  • Peer Reviewed
• [Journal Article] Enhanced HSP70 lysine methylation promotes proliferation of cancer cells through activation of aurora kinase B (2012)
  • Author(s): Hyun-Soo Cho, Shinya Hayami, Gouji Toyokawa, Kazuhiro Maejima, Yuka Yamane, Takehiro Suzuki, Naoshi Dohmae, Masaharu Kogure, Daechun Kang, David E, Neal, Bruce, A. J. Ponder, Hiroki Yamaue, Yusuke Nakamura, Ryuji Hamamoto
  • Journal Title: Nature Communications Volume: 3 Issue: 1 Pages: 1072-1072
  • DOI: 10.1038/ncomms2074
  • Peer Reviewed
• [Journal Article] Histone lysine methyltransferase Wolf-Hirschhorn syndrome candidate 1 is involved in human carcinogenesis through regulation of the Wnt pathway (2011)
  • Author(s): Toyokawa G, Cho HS, Masuda K, Yamane Y, Yoshimatsu M, Hayami S, Takawa M, Iwai Y, Daigo Y, Tsuchiya E, Tsunoda T, Field HI, Kelly JD, Neal DE, Maehara Y, Ponder BA, Nakamura Y, Hamamoto R
  • Journal Title: Neoplasia Volume: 13 Pages: 887-898
  • Peer Reviewed
• [Journal Article] The JmjC domain-containing histone demethylase KDM3A is a positive regulator of the G(1)/S transition in cancer cells via transcriptional regulation of the HOXA1 gene (2011)
  • Author(s): Cho HS, Toyokawa G, Daigo Y, Hayami S, Masuda K, Ikawa N, Yamane Y, Maejima K, Tsunoda T, Field HI, Kelly JD, Neal DE, Ponder BA, Maehara Y, Nakamura Y, Hamamoto R
  • Journal Title: Int J Cancer Volume: (in press) Issue: 3
  • DOI: 10.1002/ijc.26501
  • Peer Reviewed
• [Journal Article] The histone demethylase JMJD2B plays an essential role in human carcinogenesis through positive regulation of cyclin-dependent kinase 6 (2011)
  • Author(s): Toyokawa G, Cho HS, Iwai Y, Yoshimatsu M, Takawa M, Hayami S, Maejima K, Shimizu N, Tanaka H, Tsunoda T, Field HI, Kelly JD, Neal DE, Ponder BA, Maehara Y, Nakamura Y, Hamamoto R
  • Journal Title: Cancer Prev Res (Phila) Volume: 4 Pages: 2051-2061
  • Peer Reviewed
• [Journal Article] Enhanced expression of EHMT2 is involved in the proliferation of cancer cells through negative regulation of SIAH1 (2011)
  • Author(s): Cho HS, Kelly JD, Hayami S, Toyokawa G, Takawa M, Yoshimatsu M, Tsunoda T, Field HI, Neal DE, Ponder BA, Nakamura Y, Hamamoto R
  • Journal Title: Neoplasia Volume: 13 Pages: 676-684
  • Peer Reviewed
• [Journal Article] Minichromosome Maintenance Protein 7 is a potential therapeutic target in human cancer and a novel prognostic marker of non-small cell lung cancer (2011)
  • Author(s): Toyokawa G, Masuda K, Daigo Y, Cho HS, Yoshimatsu M, Takawa M, Hayami S, Maejima K, Chino M, Field HI, Neal DE, Tsuchiya E, Ponder BA, Maehara Y, Nakamura Y, Hamamoto R
  • Journal Title: Mol Cancer Volume: 13 Pages: 65-65
  • Peer Reviewed
• [Journal Article] Validation of the histone methyltransferase EZH2 as a therapeutic target for various types of human cancer and as a prognostic marker (2011)
  • Author(s): Takawa M, Masuda K, Kunizaki M, Daigo Y, Takagi K, Iwai Y, Cho HS, Toyokawa G, Yamane Y, Maejima K, Field HI, Kobayashi T, Akasu T, Sugiyama M, Tsuchiya E, Atomi Y, Ponder BA, Nakamura Y, Hamamoto R.
  • Journal Title: Cancer Sci Volume: 102 Issue: 7 Pages: 1298-1305
  • DOI: 10.1111/j.1349-7006.2011.01958.x
  • Peer Reviewed
• [Journal Article] Demethylation of RB regulator MYPT1 by histone demethylase LSD1 promotes cell cycle progression in cancer cells (2011)
  • Author(s): Cho HS, Suzuki T, Dohmae N, Hayami S, Unoki M, Yoshimatsu M, Toyokawa G, Takawa M, Chen T, Kurash JK, Field HI, Ponder BA, Nakamura Y, Hamamoto R
  • Journal Title: Cancer Research Volume: 71 Pages: 655-660
  • Peer Reviewed
• [Journal Article] Dysregulation of PRMT1 and PRMT6, Type I arginine methyltransferases, is involved in various types of human cancers (2011)
  • Author(s): Yoshimatsu M, Toyokawa G, Hayami S, Unoki M, Tsunoda T, Field HI, Kelly JD, Neal DE, Maehara Y, Ponder BA, Nakamura Y, Hamamoto R
  • Journal Title: International Journal of Cancer Volume: 128 Pages: 562-573
  • Peer Reviewed
• [Journal Article] Overexpression of LSD1 contributes to human carcinogenesis through chromatin regulation in various cancers (2011)
  • Author(s): Hayami S, Kelly JD, Cho HS, Yoshimatsu M, Unoki M, Tsunoda T, Field HI, Neal DE, Yamaue H, Ponder BA, Nakamura Y, Hamamoto R
  • Journal Title: International Journal of Cancer Volume: 128 Pages: 574-586
  • Peer Reviewed
• [Journal Article] UHRF1 is a novel diagnostic marker of lung cancer. (2010)
  • Author(s): Unoki M, Daigo Y, Koinuma J, Tsuchiya E, Hamamoto R, Nakamura Y
  • Journal Title: British Journal of Cancer Volume: 103 Pages: 217-222
  • Peer Reviewed
• [Journal Article] Involvement of the tubulin tyrosine ligase-like family member 4 polyglutamylase in PELP1 polyglutamylation and chromatin remodeling in pancreatic cancer cells (2010)
  • Author(s): Kashiwaya K, Nakagawa H, Hosokawa M, Mochizuki Y, Ueda K, Piao L, Chung S, Hamamoto R, Eguchi H, Ohigashi H, Ishikawa O, Janke C, Shinomura Y, Nakamura Y
  • Journal Title: Cancer Research Volume: 70 Pages: 4024-4033
  • Peer Reviewed
• [Journal Article] Overexpression of the JmjC histone demethylase KDM5B in human carcinogenesis : involvement in the proliferation of cancer cells through the E2F/RB pathway (2010)
  • Author(s): Hayami S, Yoshimatsu M, Veerakumarasivam A, Unoki M, Iwai Y, Tsunoda T, Field HI, Kelly JD, Neal DE, Yamaue H, Ponder BA, Nakamura Y, Hamamoto R
  • Journal Title: Molecular Cancer Volume: 13 Pages: 59-59
  • Peer Reviewed
• [Presentation] RBI methylation by SMYD2 enhances cell cycle progression through an increase of RBI phosphorylation (2012)
  • Author(s): Shinya Hayami, Hyun-Soo Cho, Hiroki Yamaue, Yusuke Nakamura, Ryuii Hamamoto
  • Organizer: 第71回日本癌学会学術集会
  • Place of Presentation: 札幌
  • Year and Date: 2012-09-20
• [Presentation] A Project Developing a New Cancer Therapy Targeting Histone Methyltransferase and Demethylase. (2010)
  • Author(s): Ryuji Hamamoto
  • Organizer: 日本癌学会
  • Place of Presentation: 大阪国際会議場
  • Year and Date: 2010-09-21
• [Remarks]
  • URL: http://www.u-tokyo.ac.jp/ja/todai-research/research-information/heat-shock-protein-70-hsp70-or-hsp1a1-is-a-ubiquitous-molecular-chaperone-that-functions-in-a-myriad-of-biological-processes-including-modulatin-of-polypeptide-folding-protein-degradation-translo/