Establishment of stem cell competition analysis system utilizing somatic cell mosaicism

• Project/Area Number: 18K19476
• Research Category: Grant-in-Aid for Challenging Research (Exploratory)
• Allocation Type: Multi-year Fund
• Review Section: Medium-sized Section 50:Oncology and related fields
• Research Institution: Osaka University
• Principal Investigator: FUJIWARA Tomoko 大阪大学, 放射線科学基盤機構附属ラジオアイソトープ総合センター, 特任研究員(常勤) (70402922)
• Project Period (FY): 2018-06-29 – 2021-03-31
• Project Status: Completed (Fiscal Year 2020)
• Budget Amount: ¥6,370,000 (Direct Cost: ¥4,900,000、Indirect Cost: ¥1,470,000); Fiscal Year 2020: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000); Fiscal Year 2019: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000); Fiscal Year 2018: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
• Keywords: stem cell competition / 細胞間相互作用 / ゲノム編集技術 / メダカ / 損傷応答 / 発がん / 高発がん個体 / 体細胞モザイク

Outline of Final Research Achievements

Mutagenesis is a terminal biological effect of various DNA damage stresses and a critical factor of carcinogenesis. It has been reported that carcinogenesis can be suppressed even if the Driver mutation required for carcinogenesis occurs. Stem cells competition has been proposed as one of the factors of tumor suppression. We have tried to establish the detection system for genes involved in Stem cells competition. We established three mutants required for this system, tumor suppressor gene (Bap1), regulation factor for cell differentiation signal (Notch1a), cell adhesion factor (Fat1a), by CRISPR. We have also constructed a BAC vector which enable to introduce deletion mutation of the target gene in mosaic expression pattern, and have confirmed that the vector can generate proper deletion mutation in vitro.

Academic Significance and Societal Importance of the Research Achievements

がんは我が国を含む先進国において常に死因の上位を占める疾患で、stem cell competitionはその発症の成否に関与する重要な現象である。本研究では、これに関与する遺伝子の検出系確立のために必要な遺伝子変異体をメダカにおいて作製し、さらにこれら遺伝子の正常、異常アリルを同一個体内でモザイク状に発現させることを可能にするBAC vectorを作製することができた。今後、本系の完成により当該分野のみならず広く生物・医学分野で極めて大きな役割を果たすと考えられる。

Research Products

• [Journal Article] Involvement of Rev1 in alkylating agent-induced loss of heterozygosity in Oryzias latipes. (2020)
  • Author(s): Fujikawa Y, Ishikawa-Fujiwara T, Kuo T, Shinkai N, Shoji T, Kawasaki T, Kamei Y, Sakuraba Y, Sato A, Kinoshita M, Gondo Y, Yuba S, Tsujimura T, Sese J, Todo T
  • Journal Title: Genes to Cells Volume: 25 Issue: 2 Pages: 124-138
  • DOI: 10.1111/gtc.12746
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Coulomb and CH-π interactions in (6-4) photolyase-DNA complex dominate DNA binding and repair abilities (2018)
  • Author(s): Terai Yuma、Sato Ryuma、Yumiba Takahiro、Harada Ryuhei、Shimizu Kohei、Toga Tatsuya、Ishikawa-Fujiwara Tomoko、Todo Takeshi、Iwai Shigenori、Shigeta Yasuteru、Yamamoto Junpei
  • Journal Title: Nucleic Acids Research Volume: 46 Issue: 13 Pages: 6761-6772
  • DOI: 10.1093/nar/gky364
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Presentation] Genome analysis of Rev3l deficient medaka mutant (2020)
  • Author(s): Tomoko (Ishikawa) Fujiwara, Yoshihiro Fujikawa, Andres Canela, Tatsuma Shoji, Fei Sun, Jun Sese, Takeshi Todo
  • Organizer: 日本放射線影響学会第63回大会
• [Presentation] Characteristic feature of genomic instability in rev1 and rev3 mutants (2019)
  • Author(s): Tomoko Ishikawa-Fujiwara, Yoshihiro Fujikawa, Takeshi Todo
  • Organizer: 16th International Congress of Radiation Research
  • Int'l Joint Research
• [Presentation] A unique phenotype of medaka rev3l mutant (2019)
  • Author(s): Yoshihiro Fujikawa, Tomoko Ishikawa-Fujiwara, Ayuko Sato, Tetsushi Sakuma,Tsutomu Shimura, Takashi Yamamoto, Seiji Kodama, Tohru Tsujimura, Takeshi Todo
  • Organizer: 16th International Congress of Radiation Research
  • Int'l Joint Research
• [Presentation] Involvement of Trans Lesion Synthesis DNA polymerase on maintenance of Genome stability (2019)
  • Author(s): Tomoko Fujiwara, Yoshihiro Fujikawa, Takeshi Todo
  • Organizer: 日本放射線影響学会第62回大会
  • Invited
• [Presentation] Characteristic feature of genomic instability in rev1 and rev3 mutants (2019)
  • Author(s): Yoshihiro Fujikawa, Tomoko Fujiwara, Ayuko Sato, Tsutomu Shimura, Seiji Kodama, Tohru Tsujimura, Takeshi Todo
  • Organizer: 日本放射線影響学会第62回大会
• [Presentation] 消化管腫瘍を高頻度に発がんするメダカrev3L変異体を用いた発がんメカニズムの解析 (2019)
  • Author(s): 藤川 芳宏 、藤原 智子 、佐藤 鮎子 、志村 勉 、 児玉 靖司 、辻村 亨 、藤堂 剛
  • Organizer: 第6回アジア環境変異原学会日本環境変異原学会第48回大会合同大会
• [Presentation] メダカBAP1 遺伝子変異体の作製 (2018)
  • Author(s): 藤原 智子 、藤川 芳宏 、藤堂 剛
  • Organizer: 日本放射線影響学会第61回大会
• [Presentation] 年齢・組織依存的高発がんモデル：メダカ rev3l 変異体 (2018)
  • Author(s): 藤川 芳宏 、藤原 智子 、佐藤 鮎子 、佐久間 哲史 、山本 卓 、 児玉 靖司 、辻村 亨 、藤堂 剛
  • Organizer: 日本放射線影響学会第61回大会
• [Presentation] メダカrev3l変異体における染色体不安定性の消化管腫瘍への関与 (2018)
  • Author(s): 藤川 芳宏 、藤原 智子 、佐藤 鮎子 、佐久間 哲史 、山本 卓 、 児玉 靖司 、辻村 亨 、藤堂 剛
  • Organizer: 日本環境変異原学会第47回大会