Analysis of RNA methylation in metastasis of human scirrhous gastric cancer

• Project/Area Number: 19K07698
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 50010:Tumor biology-related
• Research Institution: Aichi Gakuin University
• Principal Investigator: Hara Toshifumi 愛知学院大学, 薬学部, 講師 (80456219)
• Co-Investigator(Kenkyū-buntansha): 武井 佳史 愛知学院大学, 薬学部, 教授 (70362233)
• Project Period (FY): 2019-04-01 – 2022-03-31
• Project Status: Completed (Fiscal Year 2021)
• Budget Amount: ¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000); Fiscal Year 2021: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2020: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2019: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
• Keywords: 癌転移 / RNA修飾 / RNA / メチル化 / がん転移 / 癌転移の分子機構

Outline of Research at the Start

癌の転移は、癌による死の直接的な原因である。しかし、その分子機構は不明な点が多く転移を標的とした癌の治療法は開発されていない。申請者らは、スキルス胃がんの転移に注目し、その分子機構の解析を進める中で新たにRNAのメチル化酵素METTL3の発現が転移能獲得に伴って上昇することを見出した。そこで、本研究ではm6A修飾レベルを申請者らが構築している転移実験モデル系で変動させて、m6A修飾が癌転移で果たす役割を明らかにする。本研究は、遺伝子の機能を制御する上流マスター因子m6A修飾の役割を明らかにすることで、癌転移特異的なm6A修飾を標的とした新たな癌治療戦略の基盤構築を目指す。

Outline of Final Research Achievements

The patient prognosis of human scirrhous gastric cancers (SGCs) is very poor, mainly owing to the high frequency of peritoneal dissemination. In this study, we analyzed the functions of RNA methyltransferase METTL3 in the metastasis of SGCs. First, we generated the METTL3-knockdown cells in the metastatic model of SGCs. By the analysis of these cell lines, we found that METTL3 has a pivotal role in the cancer metastasis of SGCs. Next, we performed a comprehensive analysis of m6A modification on RNA by the Next Generation DNA Sequencer (NGS). As a result, we identified metastasis-specific m6A modifications in SGCs. Further, we generated a novel tool for gene-specific RNA demethylation based on CRISPR/Cas13b. Our findings indicate that METTL3-mediated m6A has an important role in SGCs metastasis and would be a key for understanding the molecular mechanisms of SGCs metastasis.

Academic Significance and Societal Importance of the Research Achievements

現在の癌治療において、癌転移を標的とする治療薬は開発されていない。その原因の一つは、癌転移の分子機構が未だ解明されていないことにある。本研究では、ヒトスキルス胃がんの腹膜転移細胞モデルを用いた解析により、これまで癌の転移で機能の知られていない数多くのRNAメチル化修飾が同定された。今後、RNAメチル化修飾が果たす役割の解明が進むことにより、癌転移の分子機構の一端を明らかにできる可能性を秘めている。加えて、本研究の成果は、RNAのメチル化修飾が癌転移を予測するための転移予測マーカーをして利用できることを示唆している。

Research Products

• [Journal Article] Elevated METTL9 is associated with peritoneal dissemination in human scirrhous gastric cancers (2022)
  • Author(s): ToshifumiHara, Yuuki Tominaga, Koji Ueda, Keichiro Mihara, KazuyoshiYanagihara, Yoshifumi Takei
  • Journal Title: Biochemistry and Biophysics Reports Volume: 30 Pages: 101255-101255
  • DOI: 10.1016/j.bbrep.2022.101255
  • Peer Reviewed / Open Access
• [Journal Article] Elevated Expression of CELSR1 is Associated with Peritoneal Metastasis in Human Scirrhous Gastric Cancers (2021)
  • Author(s): Toshifumi Hara, Yuri Nomura, Masaaki Hattori, Keichiro Mihara, Kazuyoshi Yanagihara, Yoshifumi Takei
  • Journal Title: BPB Reports Volume: 4 Issue: 4 Pages: 103-111
  • DOI: 10.1248/bpbreports.4.4_103
  • NAID: 130008074323
  • ISSN: 2434-432X
  • Peer Reviewed / Open Access
• [Journal Article] Promoter CpG methylation inhibits Kr?ppel-like factor 2 (KLF2)-Mediated repression of hTERT gene expression in human T-cells (2021)
  • Author(s): Mizuguchi Mariko、Hara Toshifumi、Yoshita-Takahashi Manami、Kohda Takashi、Tanaka Yuetsu、Nakamura Masataka
  • Journal Title: Biochemistry and Biophysics Reports Volume: 26 Pages: 100984-100984
  • DOI: 10.1016/j.bbrep.2021.100984
  • Peer Reviewed / Open Access
• [Journal Article] Long Noncoding RNA HOTAIR Promotes Epithelial-Mesenchymal Transition and Is a Suitable Target to Inhibit Peritoneal Dissemination in Human Scirrhous Gastric Cancers (2020)
  • Author(s): Takei Yoshifumi、Hara Toshifumi、Suzuki Akiko、Mihara Keichiro、Yanagihara Kazuyoshi
  • Journal Title: Pathobiology Volume: 87 Issue: 5 Pages: 277-290
  • DOI: 10.1159/000508350
  • Peer Reviewed / Open Access
• [Journal Article] USP10 is a critical factor for Tau-positive stress granule formation in neuronal cells (2019)
  • Author(s): Piatnitskaia S, Takahashi M, Kitaura H, Katsuragi Y, Kakihana T, Zhang L, Kakita A, Iwakura Y, Nawa H, Miura T, Ikeuchi T, Hara T, Fujii M
  • Journal Title: Scientific Reports Volume: 9 Issue: 1 Pages: 10591-10591
  • DOI: 10.1038/s41598-019-47033-7
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Presentation] スキルス胃癌の腹膜転移で発現増加するCELSR1の役割 (2021)
  • Author(s): 野村祐里、原敏文、森田あや美、武井佳史
  • Organizer: 第67回日本薬学会東海支部総会・大会
• [Presentation] Galectin 4 の発現変化とスキルス胃癌の腹膜転移能の関係 (2021)
  • Author(s): 森田あや美、岩砂浩樹、原敏文、武井佳史
  • Organizer: 第67回日本薬学会東海支部総会・大会
• [Presentation] The importance of methyltransferase like (METTL) genes on metastatic property in human scirrhous gastric cancer (2021)
  • Author(s): Toshifumi Hara, KazuyoshiYanagihara, Yoshifumi Takei
  • Organizer: 第80回日本癌学会学術総会
• [Presentation] A novel signature gene associated with peritoneal dissemination of human scirrhous gastric cancer (2020)
  • Author(s): Toshifumi Hara, Kazuyoshi Yanagihara, Yoshifumi Takei
  • Organizer: 日本癌学会
• [Presentation] Analysis of CELSR1 functions in peritoneal metastasis (2019)
  • Author(s): 原 敏文、柳原五吉、武井佳史
  • Organizer: 第78回日本癌学会学術総会