Chemical characterization of colibactin and understanding a mechanism of an onset of colorectal cancer by colibactin

• Project/Area Number: 19H02898
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Review Section: Basic Section 38040:Bioorganic chemistry-related
• Research Institution: University of Shizuoka
• Principal Investigator: Watanabe Kenji 静岡県立大学, 薬学部, 教授 (50360938)
• Project Period (FY): 2019-04-01 – 2022-03-31
• Project Status: Completed (Fiscal Year 2021)
• Budget Amount: ¥17,420,000 (Direct Cost: ¥13,400,000、Indirect Cost: ¥4,020,000); Fiscal Year 2021: ¥5,460,000 (Direct Cost: ¥4,200,000、Indirect Cost: ¥1,260,000); Fiscal Year 2020: ¥5,460,000 (Direct Cost: ¥4,200,000、Indirect Cost: ¥1,260,000); Fiscal Year 2019: ¥6,500,000 (Direct Cost: ¥5,000,000、Indirect Cost: ¥1,500,000)
• Keywords: コリバクチン / 天然物 / 生合成 / 大腸がん / 化学構造解析 / 遺伝毒性物質 / 大腸菌 / 天然物化学 / 大腸ガン / 細菌叢

Outline of Research at the Start

本研究では、E. coli-50を用いて、in vitro, in vivo における遺伝毒性、大腸発がん促進効果を検討するとともに、コリバクチン自体の化学構造を明らかにし、その遺伝毒性及びDNA付加体構造を解明する。

Outline of Final Research Achievements

In this research, using the high-colibactin-producing wild-type E. coli strain isolated from a clinical sample, we were able to identify colibactin 770, along with a more stable derivative, colibactin 788, which was further stabilized by converting the diketone moiety into quinoxaline in situ in the crude culture extract to form colibactin 860. The structure of colibactin 860 determined by NMR revealed a hydrolysis and rearrangement reactions performed to transfer the cyclopropanes in colibactin 770 to cyclobutanes in colibactin 788. Furthermore, colibactin 420 and 430 derived from alcoholysis of colibactin were also isolated and determined. The stereochemistry analysis of colibactin 860, 420, and 430 further confirmed the proposed mechanism of the rearrangement reaction.

Academic Significance and Societal Importance of the Research Achievements

本研究は物質科学としての理解が化学のみならず医学に対してもインパクトを与えるものとなったと考えられる.最近はMSデータのみの論文もよく見かけるが、NMRを駆使しての精密構造解析の「単離構造決定」が最先端科学であることを改めて示した研究成果と言える. それとは別に、構造解析の研究テーマ自体が,がん治療にまで影響を与えたことになり,これは医学へと直結する. 化学構造の解析データそのものは小さいことでもゲノム情報に対する知見に化学的なプラスアルファがあれば,合理的な理解から導き出される革新的な治療への道が拓かれると感じている.

Research Products

• [Journal Article] 大腸菌が産生する発がんリスク因子「コリバクチン」の化学構造の全容解明 (2022)
  • Author(s): 平山 裕一郎、渡辺 賢二
  • Journal Title: 化学と生物 Volume: 3 Pages: 123-130
  • Peer Reviewed
• [Journal Article] Isolation of New Colibactin Metabolites from Wild-Type Escherichia coli and In Situ Trapping of a Mature Colibactin Derivative (2021)
  • Author(s): Zhou Tao、Hirayama Yuichiro、Tsunematsu Yuta、Suzuki Nanami、Tanaka Seiji、Uchiyama Nahoko、Goda Yukihiro、Yoshikawa Yuko、Iwashita Yuji、Sato Michio、Miyoshi Noriyuki、Mutoh Michihiro、Ishikawa Hideki、Sugimura Haruhiko、Wakabayashi Keiji、Watanabe Kenji
  • Journal Title: Journal of the American Chemical Society Volume: 143 Issue: 14 Pages: 5526-5533
  • DOI: 10.1021/jacs.1c01495
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Isolation of New Colibactin Metabolites from Wild-Type Escherichia coli and In Situ Trapping of a Mature Colibactin Derivative (2021)
  • Author(s): Tao Zhou, Yuichiro Hirayama, Yuta Tsunematsu, Nanami Suzuki, Seiji Tanaka, Nahoko Uchiyama, Yukihiro Goda, Yuko Yoshikawa, Yuji Iwashita, Michio Sato, Noriyuki Miyoshi, Michihiro Mutoh, Hideki Ishikawa, Haruhiko Sugimura, Keiji Wakabayashi, and Kenji Watanabe*
  • Journal Title: JACS Volume: 1 Pages: 1-8
  • Peer Reviewed
• [Journal Article] o-Anisidine Dimer, 2-Methoxy-N4-(2-methoxyphenyl) Benzene-1,4-diamine, in Rat Urine Associated with Urinary bladder Carcinogenesis (2021)
  • Author(s): Kobayashi Takuma、Toyoda Takeshi、Tajima Yuya、Kishimoto Shinji、Tsunematsu Yuta、Sato Michio、Matsushita Kohei、Yamada Takanori、Shimamura Yuko、Masuda Shuichi、Ochiai Masako、Ogawa Kumiko、Watanabe Kenji、Takamura-Enya Takeji、Totsuka Yukari、Wakabayashi Keiji、Miyoshi Noriyuki
  • Journal Title: Chemical Research in Toxicology Volume: 34 Issue: 3 Pages: 912-919
  • DOI: 10.1021/acs.chemrestox.0c00536
  • Peer Reviewed
• [Journal Article] Artesunate Inhibits Intestinal Tumorigenesis through Inhibiting Wnt Signaling. (2020)
  • Author(s): Hamoya T, Fujii G, Iizumi Y, Narita T, Komiya M, Matsuzawa Y, Miki K, Kondo T, Kishimoto S, Watanabe K, Wakabayashi K, Sakai T, Toshima J, Mutoh M
  • Journal Title: Carcinogenesis Volume: 42 Issue: 1 Pages: 148-58
  • DOI: 10.1093/carcin/bgaa084
  • Peer Reviewed / Open Access
• [Journal Article] Genotyping of a gene cluster for production of colibactin and in vitro genotoxicity analysis of Escherichia coli strains obtained from the Japan Collection of Microorganisms (2020)
  • Author(s): Kawanishi Masanobu、Shimohara Chiaki、Oda Yoshimitsu、Hisatomi Yuuta、Tsunematsu Yuta、Sato Michio、Hirayama Yuichiro、Miyoshi Noriyuki、Iwashita Yuji、Yoshikawa Yuko、Sugimura Haruhiko、Mutoh Michihiro、Ishikawa Hideki、Wakabayashi Keiji、Yagi Takashi、Watanabe Kenji
  • Journal Title: Genes and Environment Volume: 42 Issue: 1 Pages: 12-17
  • DOI: 10.1186/s41021-020-00149-z
  • Peer Reviewed / Open Access
• [Journal Article] Activity-Based Probe for Screening of High-Colibactin Producers from Clinical Samples (2019)
  • Author(s): Hirayama Yuichiro、Tsunematsu Yuta、Yoshikawa Yuko、Tamafune Ryota、Matsuzaki Nobuo、Iwashita Yuji、Ohnishi Ippei、Tanioka Fumihiko、Sato Michio、Miyoshi Noriyuki、Mutoh Michihiro、Ishikawa Hideki、Sugimura Haruhiko、Wakabayashi Keiji、Watanabe Kenji
  • Journal Title: Organic Letters Volume: 21 Issue: 12 Pages: 4490-4494
  • DOI: 10.1021/acs.orglett.9b01345
  • Peer Reviewed
• [Journal Article] <i>In vitro</i> genotoxicity analyses of colibactin-producing <i>E. coli</i> isolated from a Japanese colorectal cancer patient (2019)
  • Author(s): Kawanishi Masanobu、Hisatomi Yuuta、Oda Yoshimitsu、Shimohara Chiaki、Tsunematsu Yuta、Sato Michio、Hirayama Yuichiro、Miyoshi Noriyuki、Iwashita Yuji、Yoshikawa Yuko、Sugimura Haruhiko、Mutoh Michihiro、Ishikawa Hideki、Wakabayashi Keiji、Yagi Takashi、Watanabe Kenji
  • Journal Title: The Journal of Toxicological Sciences Volume: 44 Issue: 12 Pages: 871-876
  • DOI: 10.2131/jts.44.871
  • NAID: 130007759775
  • ISSN: 0388-1350, 1880-3989
  • Peer Reviewed / Open Access
• [Presentation] 反応メカニズムの理解に基づく天然物生合成リデザイン (2022)
  • Author(s): 渡辺賢二
  • Organizer: 新学術領域研究 「生合成リデザイン」 取りまとめ公開シンポジウム
  • Invited
• [Presentation] Structural elucidation of natural product colibactin isolated from Escherichia coli in tumor tissues of clinical samples (2021)
  • Author(s): Kenji Watanabe
  • Organizer: Pacifichem2021
  • Int'l Joint Research / Invited
• [Presentation] 大腸がんリスク因子コリバクチンの化学構造解析 (2021)
  • Author(s): 周 韜、平山 裕一郎、恒松 雄太、鈴木 七海、佐藤 道大、 田中 誠司、内山 奈穂子、合田 幸広、渡辺 賢二
  • Organizer: 第63回天然有機化合物討論会
• [Presentation] 大腸がんリスク因子コリバクチン産生菌の増殖抑制を目的としたパイロット介入試験 (2019)
  • Author(s): 渡辺賢二
  • Organizer: 日本がん予防学会学
  • Int'l Joint Research / Invited
• [Remarks] WATANABE RESEARCH GROUP
  • URL: https://sweb.u-shizuoka-ken.ac.jp/~kenji55-lab/
• [Remarks] https://sweb.u-shizuoka-ken.ac.jp/~kenji55-lab/
• [Patent(Industrial Property Rights)] PCT出願 (2022)
  • Inventor(s): 周韜、渡辺賢二
  • Industrial Property Rights Holder: 周韜、渡辺賢二
  • Industrial Property Rights Type: 特許
  • Filing Date: 2022
  • Overseas