Identification of factors contributing to epigenetic alteration and maintenance in EBV infection.

• Project/Area Number: 16K08729
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Experimental pathology
• Research Institution: Chiba University
• Principal Investigator: 松坂 恵介 千葉大学, 大学院医学研究院, 講師 (40610150)
• Project Period (FY): 2016-04-01 – 2019-03-31
• Project Status: Completed (Fiscal Year 2018)
• Budget Amount: ¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000); Fiscal Year 2018: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000); Fiscal Year 2017: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000); Fiscal Year 2016: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
• Keywords: エピジェネティクス / DNAメチル化 / ウイルス / 胃癌 / EBV

Outline of Annual Research Achievements

本研究は胃癌を対象にEpstein-Barrウイルス(EBV)感染が誘導するゲノムワイドなエピゲノム改変に関わる因子の同定と、EBV感染脱落系を用いたエピゲノム維持機構の解明を目的としている。
1. EBV感染によるエピゲノム改変機構の解明：胃上皮細胞において感染早期の新規DNAメチル化形質誘導パターンを経時的に解析した結果、宿主細胞のDNAメチル化誘導に先駆けてウイルスゲノムDNAにメチル化が誘導されることを見いだした。DNAメチル化誘導時の宿主細胞とウイルスの遺伝子発現を次世代シーケンサーを用いたRNA-seq法にて包括的に解析した結果、種々のウイルス遺伝子の発現が認められ、複雑な機構の存在がうかがわれた。
2. EBV感染脱落系を用いたエピゲノム維持機構の解明：変異型EBNA1を用いたEBV脱落実験系において、単純な強制発現では十分な脱落効率が得られなかったが、蛍光ラベルで選別することで効率的にEBV脱落細胞を得る実験系を確立した。先行研究にてヒストンの活性化マークやDNA脱メチル化関連酵素TET2がEBV感染時の新規DNAメチル化の抵抗因子であることを見出しており、ウイルス脱落後の細胞においてそうしたエピゲノム修飾因子を変化させることで、EBV感染がDNAメチル化の維持に果たす役割について解析中である。
ウイルスと宿主との間の秩序だった相互作用をより詳細に解析することで、新規DNAメチル化誘導のメカニズムの解明が期待される。

Research Products

• [Journal Article] The frequency of promoter DNA hypermethylation is decreased in colorectal neoplasms of familial adenomatous polyposis. (2018)
  • Author(s): Takane K, Fukuyo M, Matsusaka K, Ota S, Rahmutulla B, Matsushita K, Miyauchi H, Nakatani Y, Matsubara H, Kaneda A.
  • Journal Title: Oncotarget Volume: 9 Pages: 32653-32666
  • DOI: 10.18632/oncotarget.25987
  • Peer Reviewed / Open Access
• [Journal Article] Synergistic cytotoxic effect on gastric cancer cells of an immunotoxin cocktail in which antibodies recognize different epitopes on CDH17. (2018)
  • Author(s): Kusano-Arai O, Iwanari H, Kudo S, Akiba H, Matsusaka K, Kaneda A, Fukayama M, Tsumoto K, Hamakubo T.
  • Journal Title: Monoclon Antib Immunodiagn Immunother Volume: 37 Pages: 1-11
  • DOI: 10.1089/mab.2017.0043
  • Peer Reviewed
• [Journal Article] Epstein-Barr virus infection induces genome-wide de novo DNA methylation in non-neoplastic gastric epithelial cells. (2017)
  • Author(s): Matsusaka K, Funata S, Fukuyo M, Seto Y, Aburatani H, Fukayama M, Kaneda A.
  • Journal Title: J Pathol Volume: 242 Pages: 391-399
  • DOI: 10.1002/path.4909
  • Peer Reviewed / Open Access
• [Journal Article] Frequent promoter hypermethylation associated with human papillomavirus infection in pharyngeal cancer (2017)
  • Author(s): Nakagawa T, Matsusaka K, Misawa K, Ota S, Takane K, Fukuyo M, Rahmutulla B, Shinohara K, Kunii N, Sakurai D, Hanazawa T, Matsubara H, Nakatani Y, Okamoto Y, Kaneda A.
  • Journal Title: Cancer Letter Volume: 407 Pages: 21-31
  • DOI: 10.1016/j.canlet.2017.08.008
  • Peer Reviewed / Open Access
• [Journal Article] Regulation of tumour related genes by dynamic epigenetic alteration at enhancer regions in gastric epithelial cells infected by Epstein-Barr virus (2017)
  • Author(s): Okabe A, Funata S, Matsusaka K, Namba H, Fukuyo M, Rahmutulla B, Oshima M, Iwama A, Fukayama M, Kaneda A.
  • Journal Title: Scientific Reports Volume: 7 Pages: 7924-7924
  • DOI: 10.1038/s41598-017-08370-7
  • Peer Reviewed / Open Access
• [Journal Article] Histone modification alteration coordinated with acquisition of promoter DNA methylation during Epstein-Barr virus infection. (2017)
  • Author(s): Funata S, Matsusaka K, Yamanaka R, Yamamoto S, Okabe A, Fukuyo M, Aburatani H, Fukayama M, Kaneda A.
  • Journal Title: Oncotarget Volume: 8 Pages: 55265-55279
  • DOI: 10.18632/oncotarget.19423
  • Peer Reviewed / Open Access
• [Journal Article] Tumor Content Chart-Assisted HER2/CEP17 Digital PCR Analysis of Gastric Cancer Biopsy Specimens (2016)
  • Author(s): Matsusaka K, Ishikawa S, Nakayama A, Ushiku T, Nishimoto A, Urabe M, Kaneko N, Kunita A, Kaneda A, Aburatani H, Fujishiro M, Seto Y, Fukayama M.
  • Journal Title: PLoS One. Volume: 11
  • DOI: 10.1371/journal.pone.0154430
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Two subtypes of colorectal tumor with distinct molecular features in familial adenomatous polyposis (2016)
  • Author(s): Matsusaka K, Ushiku T, Urabe M, Fukuyo M, Abe H, Ishikawa S, Seto Y, Aburatani H, Hamakubo T, Kaneda A, Fukayama M.
  • Journal Title: Oncotarget. Volume: 7 Pages: 64168-64181
  • DOI: 10.18632/oncotarget.11510
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Coupling CDH17 and CLDN18 Markers for Comprehensive Membrane-targeted Detection of Human Gastric Cancer. (2016)
  • Author(s): K. Matsusaka, T. Ushiku, M. Urabe, M. Fukuyo, H. Abe, S. Ishikawa, Y. Seto, H. Aburatani, T. Hamakubo, A. Kaneda, M. Fukayama
  • Journal Title: Oncotarget, Volume: 7 Pages: 64168-64181
  • DOI: 10.18632/oncotarget.11638
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] TET2 functions as a resistance factor against DNA methylation acquisition during Epstein-Barr virus infection (2016)
  • Author(s): Namba-Fukuyo H, Funata S, Matsusaka K, Fukuyo M, Rahmutulla B, Mano Y, Fukayama M, Aburatani H, Kaneda A
  • Journal Title: Oncotarget Volume: 7 Pages: 81512-26
  • DOI: 10.18632/oncotarget.13130
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Presentation] 胃癌におけるTP53変異とは独立したゲノム・エピゲノム相互作用の同定 (2018)
  • Author(s): 松坂恵介, 眞野恭伸, 福世真樹, 浦辺雅之, 喜多和子, 根本哲宏, バハテヤリ・ラムヒトラ, 瀬戸泰之, 深山正久, 金田篤志.
  • Organizer: 第107回日本病理学会総会
• [Presentation] Identification of genetic/epigenetic alterations based on wild-type TP53 in high methylation gastric cancer. (2018)
  • Author(s): Keisuke Matsusaka, Yasunobu Mano, Masaki Fukuyo, Masayuki Urabe, Rahmutulla Bahityar, Eriko Ikeda, Kazuko Kita, Hiroyuki Abe, Tetsuhiro Nemoto, Yasuyuki Seto, Masashi Fukayama, Atsushi Kaneda.
  • Organizer: The 77th Annual Meeting of the Japanese Cancer Association
• [Presentation] 高メチル化胃癌におけるTP53変異非依存性ゲノム・エピゲノム異常の相乗効果． (2018)
  • Author(s): 松坂恵介、眞野恭伸、福世真樹、松田和暁、井上貴登、浦辺雅之、バハテヤリ・ラヒムトラ、池田英里子、阿部浩幸、喜多和子、瀬戸泰之、根本哲宏、深山正久、金田篤志．
  • Organizer: 第29回日本消化器癌発生学会総会
• [Presentation] メチル化胃癌における特異的なゲノム・エピゲノム変異の同定 (2017)
  • Author(s): 松坂恵介、眞野恭伸、福世真樹、浦辺雅之、油谷浩幸、深山正久、瀬戸泰之、金田篤志.
  • Organizer: 第76回日本癌学会総会
• [Presentation] Identification of unique genetic/epigenetic alteration in gastric cancer with high DNA methylation. (2017)
  • Author(s): 松坂恵介, 眞野恭伸、福世真樹、浦辺雅之、バハテヤリ・ラムヒトラ、油谷浩幸、瀬戸泰之、深山正久、金田篤志
  • Organizer: 第106回日本病理学会学術総会
• [Presentation] 発がんに関わるエピゲノム異常誘導因子としての病原体の意義 (2016)
  • Author(s): 松坂恵介
  • Organizer: 日本環境変異原学会 第45回大会
  • Place of Presentation: つくば国際会議場（茨城県、つくば市）
  • Year and Date: 2016-11-17
• [Presentation] Unique genetic and epigenetic features of gastric cancer subtypes with high-methylation or Epstein-Barr virus infection (2016)
  • Author(s): Matsusaka K, Mano Y, Fukuyo M, Urabe M, Rahmutulla B, Sakai E, Aburatani H, Seto Y, Fukayama M, Kaneda A.
  • Organizer: The75th Annual Meeting of the Japanese Cancer Association
  • Place of Presentation: パシフィコ横浜（神奈川県、横浜市）
  • Year and Date: 2016-10-06
• [Book] DNA and Histone Methylation as Cancer Targets (2017)
  • Author(s): Matsusaka K, Kaneda A.
  • Total Pages: 624
  • Publisher: Springer
• [Book] 病理と臨床 (2016)
  • Author(s): 松坂恵介、宇於崎宏
  • Total Pages: 4
  • Publisher: 文光堂
• [Remarks] 千葉大学大学院医学研究院分子腫瘍学
  • URL: http://www.m.chiba-u.ac.jp/class/moloncol/