Clarification of low-dose radiation-induced cellular responses by multimodal analyses

• Project/Area Number: 15K00543
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Risk sciences of radiation and chemicals
• Research Institution: Fukushima Medical University
• Principal Investigator: Tsuyama Naohiro 福島県立医科大学, 医学部, 准教授 (10335747)
• Co-Investigator(Kenkyū-buntansha): 阿部 悠 福島県立医科大学, 医学部, 助教 (00722472)
• Project Period (FY): 2015-10-21 – 2019-03-31
• Project Status: Completed (Fiscal Year 2018)
• Budget Amount: ¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000); Fiscal Year 2017: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2016: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2015: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
• Keywords: 低線量放射線 / 細胞応答 / メタボローム / 蛍光分子解析 / 蛍光分子分析

Outline of Final Research Achievements

For the purpose of multidimensional understandings of intra-cellular environmental changes after low-dose ionizing radiation, we made multimodal analyses using molecular species-specific fluorescent probes and liquid chromatography-mass spectrometry.
After the knockdown of DNA damage response genes in human fibroblasts, we found the loss of alteration of nucleic acids metabolites and lipid derivatives which we reported previously, and loss of response in mitochondrial activity and redox state by low-dose irradiation.
In addition, we made a system of DNA sequence specific digestion using CRISPR/Cas9, and confirmed DNA damages and induction of mutation which corresponds to low dose irradiation.

Academic Significance and Societal Importance of the Research Achievements

低線量放射線照射による細胞応答について、正常細胞の系統的なノックダウンによる同一の実験系を使い、DNA損傷応答シグナル伝達機構からの細胞内環境や代謝調節への関与を、複数の解析モードにより結果を得ることができた。特定のシグナル分子と細胞内環境を並行して解析できたことから、今後シグナルのクロストーク解析に有用であると考えられた。
また電離放射線を使わず、DNA二本鎖切断を誘導できるCRISPR/Cas9系を作成したことで、低線量で生じる反応を高感度に定量的に解析できる可能性が示された。

Research Products

• [Journal Article] Dose-response curves for analyzing of dicentric chromosomes and chromosome translocation following doses of 1,000 mGy or less based on irradiated peripheral blood samples from 5 healthy individuals. (2018)
  • Author(s): Abe Y, Yoshida MA, Fujioka K, Kurosu Y, Ujiie R, Yanagi A, Tsuyama N, Miura T, Inaba T, Kamiya K, Sakai A
  • Journal Title: Journal of Radiation Research Volume: 59 Issue: 1 Pages: 35-42
  • DOI: 10.1093/jrr/rrx052
  • Peer Reviewed / Open Access
• [Journal Article] 放射線メタボローム解析 (2017)
  • Author(s): 津山尚宏
  • Journal Title: 放射線生物研究 Volume: 52 Pages: 291-306
  • NAID: 40021362524
  • Peer Reviewed
• [Journal Article] The great importance of normalization of LC-MS data for highly-accurate non-targeted metabolomics. (2017)
  • Author(s): Mizuno H, Ueda K, Kobayashi Y, Tsuyama N, Todoroki K, Min JZ, Toyo'oka T.
  • Journal Title: Biomed Chromatogr. Volume: 31 Issue: 1 Pages: 1-7
  • DOI: 10.1002/bmc.3864
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Analysis of chromosome translocation frequency after a single CT scan in adults (2016)
  • Author(s): Yu Abe, Tomisato Miura, Mitsuaki A. Yoshida, et al.
  • Journal Title: Journal of Radiation Research, Volume: 57 Issue: 3 Pages: 1-7
  • DOI: 10.1093/jrr/rrv090
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Direct metabolomics for plant cells by live single-cell mass spectrometry. (2015)
  • Author(s): Fujii T, Matsuda S, Tejedor ML, Esaki T, Sakane I, Mizuno H, Tsuyama N, Masujima T.
  • Journal Title: Nature Protocols Volume: 10 Issue: 9 Pages: 1445-56
  • DOI: 10.1038/nprot.2015.084
  • Peer Reviewed
• [Journal Article] Increase in dicentric chromosome formation after a single CT scan in adults. (2015)
  • Author(s): Abe Y, Miura T, Yoshida MA, Ujiie R, Kurosu Y, Kato N, Katafuchi A, Tsuyama N, Ohba T, Inamasu T, Shishido F, Noji H, Ogawa K, Yokouchi H, Kanazawa K, Ishida T, Muto S, Ohsugi J, Suzuki H, Ishikawa T, Kamiya K, Sakai A.
  • Journal Title: Scinetific Reports Volume: 9 Issue: 1 Pages: 13882-13882
  • DOI: 10.1038/srep13882
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Presentation] Analysis of the number of chromosome aberrations induced by three consecutive CT examinations. (2019)
  • Author(s): Yu Abe, Hideyoshi Noji, Misaki Sugai, Yumiko Kurosu, Naohiro Tsuyama, Aki Yanagi, Yukari Yanai, Takashi Ohba, Tetsuo Ishikawa, Tomisato Miura, Kenji Kamiya, Mitsuaki A. Yoshida, Akia Sakai
  • Organizer: 放射線災害・医科学研究拠点 第３回国際シンポジウム
  • Int'l Joint Research
• [Presentation] Induction of t(11;14) IgH enhancer/promoter-cyclin D1 gene translocation using CRISPR/Cas9. (2019)
  • Author(s): Naohiro Tsuyama, Yu Abe, Aki Yanagi, Misaki Sugai, Kenji Kamiya, Akira Sakai
  • Organizer: 放射線災害・医科学研究拠点 第３回国際シンポジウム.
  • Int'l Joint Research
• [Presentation] CRISPR/Cas9を用いたt(11; 14)染色体転座誘発 (2018)
  • Author(s): 津山尚宏、阿部悠、柳亜紀、菅井美咲、神谷研二、坂井晃
  • Organizer: 第61回日本放射線影響学会
• [Presentation] Investigation of the cumulative number of chromosome aberrations induced by three consecutive CT scans. (2018)
  • Author(s): Yu Abe, Hideyoshi Noji, Misaki Sugai, Yumiko Kurosu, Takashi Ohba, Naohiro Tsuyama, Aki Yanagi, Yukari Yanai, Tetsuo Ishikawa, Tomisato Miura, Kenji Kamiya, Mitsuaki A Yoshida, Akira Sakai
  • Organizer: American Association for Cancer Research Annual Meeting 2018
  • Int'l Joint Research
• [Presentation] Construction of dose response curves for cytogenetic biodosimetry in the low dose range based on five persons. (2018)
  • Author(s): Yu Abe, Mitsuaki A Yoshida, Kurumi Fujioka, Yumiko Kurosu, Risa Ujiie, Aki Yanagi, Naohiro Tsuyama, Tomisato Miura, Toshiya Inaba, Kenji Kamiya, Akira Sakai
  • Organizer: EPRBioDose 2018
  • Int'l Joint Research
• [Presentation] Analysis of the number of chromosome aberrations induced by three consecutive CT examinations. (2018)
  • Author(s): Abe Y, Noji H, Sugai M, Kurosu Y, Tsuyama N, Yanagi A, Yanai Y, Ohba T, Ishikawa T, Miura T, Kamiya K, Yoshida MA, Sakai A
  • Organizer: 第61回日本放射線影響学会
• [Presentation] Attempt to make abnormal B lymphocytes as myeloma-initiating cells from iPSs. (2018)
  • Author(s): 柳 亜希、津山尚宏、柳井祐佳里、阿部 悠、菅井美咲、太田明伸、シバスンダラン カルナン、笹谷めぐみ、神谷研二、花村一朗、池添隆之、小野寺雅史、坂井 晃
  • Organizer: 第80回日本血液学会学術集会
• [Presentation] Attempt to prove the existence of abnormal B lymphocyte as myeloma-initiating cells from B cell-derived induced pluripotent stem cells. (2018)
  • Author(s): Yanagi A, Tsuyama N, Yanai Y, Abe Y, Sugai M, Ota A, Sivasundaram K, Shigemura T, Sasatani M, Kamiya K, Hanamura I, Ikezoe T, Onodera M, Sakai A
  • Organizer: 60th ASH Annual Meeting
  • Int'l Joint Research
• [Presentation] 低線量放射線被ばくによる血清中代謝変動解析と循環器疾患関連分子探索 (2018)
  • Author(s): 山本健太、上田一樹、水野 初、津山尚宏、三角宗近、村上秀子、大石和佳、長町安希子、稲葉俊哉、豊岡利正、高橋規郎、轟木堅一郎
  • Organizer: 日本薬学会第138年会
• [Presentation] 高血圧自然発症ラット（SHR）血漿/血清を用いた放射線誘発高血圧の経時変動指標分子探索 (2017)
  • Author(s): 津山尚宏、水野初、山本健太、上田一樹、三角宗近、村上秀子、大石和佳、長町安希子、稲葉俊哉、児玉和紀、高橋規郎
  • Organizer: 日本放射線影響学会第第60回大会
• [Presentation] 高血圧自然発症ラット（SHR）血漿/血清を用いた放射線と相関する高血圧症の経時変動指標分子探索 (2017)
  • Author(s): 山本健太、上田一樹、水野 初、津山尚宏、三角宗近、村上秀子、大石和佳、長町安希子、稲葉俊哉、豊岡利正、高橋規郎、轟木堅一郎
  • Organizer: 新アミノ酸分析研究会第7回学術講演会
• [Presentation] 高血圧自然発症ラット（SHR）を用いた放射線に相関した高血圧における代謝物変動追跡 (2016)
  • Author(s): 津山尚宏、三角宗近、村上秀子、大石和佳、長町安希子、稲葉俊哉、丹羽保晴、児玉和紀、高橋規郎
  • Organizer: 日本放射線影響学会第59回大会
  • Place of Presentation: 広島アステールプラザ
  • Year and Date: 2016-10-26
• [Presentation] N標識化クロレラを用いた多成分高精度分析法の開発 (2015)
  • Author(s): 小林勇太，上田一樹，水野初，轟木堅一郎，閔俊哲，津山尚宏，豊岡利正
  • Organizer: 新アミノ酸分析研究会 第5回学術講演会
  • Place of Presentation: 東京大学武田先端知ビル
  • Year and Date: 2015-12-07