放射線被ばくが主要臓器に及ぼす影響に迫る「遺伝子改変動物を用いた時空間解析」

• Project Area: "LIVING IN SPACE" - Integral Understanding of life-regulation mechanism from "SPACE"
• Project/Area Number: 16H01640
• Research Category: Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area)
• Allocation Type: Single-year Grants
• Review Section: Complex systems
• Research Institution: Kyoto University
• Principal Investigator: 原田 浩 京都大学, 放射線生物研究センター, 教授 (80362531)
• Project Period (FY): 2016-04-01 – 2018-03-31
• Project Status: Completed (Fiscal Year 2017)
• Budget Amount: ¥6,240,000 (Direct Cost: ¥4,800,000、Indirect Cost: ¥1,440,000); Fiscal Year 2017: ¥3,120,000 (Direct Cost: ¥2,400,000、Indirect Cost: ¥720,000); Fiscal Year 2016: ¥3,120,000 (Direct Cost: ¥2,400,000、Indirect Cost: ¥720,000)
• Keywords: 放射線 / DNA損傷 / 遺伝子改変動物 / 宇宙放射線 / 光イメージング

Outline of Annual Research Achievements

１．概日リズム制御機構と放射線感受性/抵抗性制御機構のクロストーク：
細胞の放射線抵抗性を誘導する遺伝子としてhypoxia-inducible factor 1が、また生体の概日リズムを制御する遺伝子としてPeriod 2（PER2）が知られている。我々は、PER2がHIF-1を活性化するメカニズムを解明し、両機構の機能的クロストークの実態を捉えた（Kobayashi et al. FEBS J. 2017.）。従来より、HIF-1活性は主にその主要制御サブユニットHIF-1α蛋白質の安定性に依存していることが知られていた。しかし我々は、PER2がHIF-1α蛋白質の安定性には影響を及ぼさず、HIF-1αとHIF-1βで構成されるヘテロダイマーHIF-1を、DNA上のエンハンサー領域にリクルートする活性を持つことを見出した（Figure 1）。この成果は、HIF-1活性の制御における新たなメカニズムを明らかにしたのみならず、概日リズム（生体のリズム）が生体の放射線感受性を左右する可能性を示した点で大きな意義を持っている。
２．遺伝子改変マウスの作製：
①DNA二重鎖切断を可視化するための遺伝子改変マウスと、②細胞の放射線感受性を左右する組織内低酸素環境を可視化するための遺伝子改変マウスを樹立するため、2つのターゲティングベクターを構築した。そして各ベクターをRosa26用guide RNA、およびCas9発現ベクターと共にマイクロインジェクトし、genotypingを行うことによって、目的どおりの遺伝子改変マウスを作成することに成功した。

Research Progress Status

29年度が最終年度であるため、記入しない。

Strategy for Future Research Activity

29年度が最終年度であるため、記入しない。

Research Products

• [Int'l Joint Research] オックスフォード大学(英国)
• [Int'l Joint Research] University of Oxford/Imperial College London(英国)
• [Journal Article] HIF-1 maintains a functional relationship between pancreatic cancer cells and stromal fibroblasts by upregulating expression and secretion of Sonic hedgehog (2018)
  • Author(s): Katagiri Tomohiro、Kobayashi Minoru、Yoshimura Michio、Morinibu Akiyo、Itasaka Satoshi、Hiraoka Masahiro、Harada Hiroshi
  • Journal Title: Oncotarget Volume: 9 Issue: 12 Pages: 10525-10535
  • DOI: 10.18632/oncotarget.24156
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Regulatory mechanisms of hypoxia-inducible factor 1 activity: Two decades of knowledge (2018)
  • Author(s): Koyasu Sho、Kobayashi Minoru、Goto Yoko、Hiraoka Masahiro、Harada Hiroshi
  • Journal Title: Cancer Science Volume: 109 Issue: 3 Pages: 560-571
  • DOI: 10.1111/cas.13483
  • NAID: 120006454672
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] A circadian clock gene, PER2, activates HIF-1 as an effector molecule for recruitment of HIF-1α to promoter regions of its downstream genes (2017)
  • Author(s): Kobayashi Minoru、Morinibu Akiyo、Koyasu Sho、Goto Yoko、Hiraoka Masahiro、Harada Hiroshi
  • Journal Title: The FEBS Journal Volume: 284 Issue: 22 Pages: 3804-3816
  • DOI: 10.1111/febs.14280
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] The emerging roles of the ubiquitination/deubiquitination system in tumor radioresistance regarding DNA damage responses, cell cycle regulation, hypoxic responses, and antioxidant properties: Insight into the development of novel radiosensitizing strategies (2017)
  • Author(s): Goto Yoko、Koyasu Sho、Kobayashi Minoru、Harada Hiroshi
  • Journal Title: Mutation Research/Fundamental and Molecular Mechanisms of Mutagenesis Volume: 803-805 Pages: 76-81
  • DOI: 10.1016/j.mrfmmm.2017.07.007
  • Peer Reviewed
• [Journal Article] UCHL1-HIF-1 axis-mediated antioxidant property of cancer cells as a therapeutic target for radiosensitization (2017)
  • Author(s): Nakashima Ryota、Goto Yoko、Koyasu Sho、Kobayashi Minoru、Morinibu Akiyo、Yoshimura Michio、Hiraoka Masahiro、Hammond Ester M.、Harada Hiroshi
  • Journal Title: Scientific Reports Volume: 7 Issue: 1 Pages: 6879-6879
  • DOI: 10.1038/s41598-017-06605-1
  • NAID: 120006355750
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] pH-Responsive near-infrared fluorescent cyanine dyes for molecular imaging based on pH sensing (2017)
  • Author(s): Miki Koji、Kojima Kentaro、Oride Kazuaki、Harada Hiroshi、Morinibu Akiyo、Ohe Kouichi
  • Journal Title: Chemical Communications Volume: 53 Issue: 55 Pages: 7792-7795
  • DOI: 10.1039/c7cc03035e
  • Peer Reviewed
• [Journal Article] HIF-1-mediated suppression of mitochondria electron transport chain function confers resistance to lidocaine-induced cell death (2017)
  • Author(s): Okamoto Akihisa、Sumi Chisato、Tanaka Hiromasa、Kusunoki Munenori、Iwai Teppei、Nishi Kenichiro、Matsuo Yoshiyuki、Harada Hiroshi、Takenaga Keizo、Bono Hidemasa、Hirota Kiichi
  • Journal Title: Scientific Reports Volume: 7 Issue: 1 Pages: 3816-3816
  • DOI: 10.1038/s41598-017-03980-7
  • NAID: 120006364104
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] HIF-1とがんの代謝 (2017)
  • Author(s): 原田浩
  • Journal Title: 実験医学 Volume: 35 Pages: 1586-1592
• [Journal Article] がん特異的グルコース代謝経路による放射線抵抗性と放射線治療増感への展開 (2017)
  • Author(s): 後藤容子、中島良太、原田浩
  • Journal Title: 放射線生物研究 Volume: 52 Pages: 277-290
  • NAID: 40021362518
  • Peer Reviewed
• [Journal Article] Hypoxia-inducible factor 1 promotes chemoresistance of lung cancer by inducing carbonic anhydrase IX espression (2017)
  • Author(s): Sowa T, Menju T, Chen-Yoshikawa TF, Takahashi K, Nishikawa S, Nakanishi T, Shikuma K, Motoyama H, Hijiya K, Aoyama A, Sato T, Sonobe M, Harada H, Date H
  • Journal Title: Cancer Medicine Volume: 6 Issue: 1 Pages: 288-297
  • DOI: 10.1002/cam4.991
  • Peer Reviewed / Open Access
• [Journal Article] Cigarette smoke reversibly activates hypoxia-inducible factor 1 in a reactive oxygen species-dependent manner (2016)
  • Author(s): Daijo H, Hoshino Y, Kai S, Suzuki K, Nishi K, Matsuo Y, Harada H, Hirota K
  • Journal Title: Scientific Reports Volume: 6 Issue: 1 Pages: 34424-34424
  • DOI: 10.1038/srep34424
  • NAID: 120005850269
  • Peer Reviewed / Open Access
• [Journal Article] LY6E: a conductor of malignant tumor growth through modulation of the PTEN/PI3K/Akt/HIF-1 axis (2016)
  • Author(s): Harada H et al.
  • Journal Title: Oncotarget Volume: 7(40) Issue: 40 Pages: 65837-65848
  • DOI: 10.18632/oncotarget.11670
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] VHL-deficient renal cancer cells gain resistance to mitochondria-activating apoptosis inducers by activating AKT through the IGF1R-PI3K pathway (2016)
  • Author(s): Yamaguchi R, Harada H, Hirota K
  • Journal Title: Tumor Biology Volume: 37 Issue: 10 Pages: 13295-13306
  • DOI: 10.1007/s13277-016-5260-2
  • Peer Reviewed / Open Access
• [Journal Article] Hypoxia-inducible factor 1-mediated characteristic features of cancer cells for tumor radiaresistance. (2016)
  • Author(s): *Harada H.
  • Journal Title: J Radiat Res Volume: 印刷中 Issue: S1 Pages: 99-99
  • DOI: 10.1093/jrr/rrw012
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Potent anti-tutor effects of EGFR-targeted hybrid peptide on mice bearing liver metastases (2016)
  • Author(s): Gaowa A, Horibe T, Kohno M, Harada H, Hiraoka M, Kawakami K
  • Journal Title: Clin Exp Metastasis Volume: 33 Issue: 1 Pages: 87-95
  • DOI: 10.1007/s10585-015-9760-z
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Evaluation of Tumor-associated Stroma and Its Relationship with Tumor Hypoxia Using Dynamic Contrast-enhanced CT and 18F Misonidazole PET in Murine Tumor Models. (2016)
  • Author(s): Koyasu, S., Tsuji, Y., Harada, H., Nakamoto, Y., Nobashi, T., Kimura, H., Sano, K., Koizumi, K., Hamaji, M., Togashi, K.
  • Journal Title: Radiology. Volume: 278 Issue: 3 Pages: 734-41
  • DOI: 10.1148/radiol.2015150416
  • Peer Reviewed / Open Access
• [Journal Article] H3K9me3 facilitates hypoxia-induced p53-dependent apoptosis through repression of APAK. (2016)
  • Author(s): Olcina MM, Leszczynska K, Senra JM, Isa N, Harada H, Hammond EM.
  • Journal Title: Oncogene Volume: 35 Issue: 6 Pages: 793-799
  • DOI: 10.1038/onc.2015.134
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Presentation] 照準を合わせろ！ がんに対する放射線治療の現状と課題. (2018)
  • Author(s): 原田浩
  • Organizer: 第13回京都大学附置研究所・センターシンポジウム
  • Invited
• [Presentation] Hypoxic Microenvironments in Malignant Solid Tumors (2018)
  • Author(s): Harada H
  • Organizer: International Mini-symposium on Organs under Stress
  • Int'l Joint Research / Invited
• [Presentation] がん・低酸素・HIF-1 (2018)
  • Author(s): 原田浩
  • Organizer: 国立京都医療センター臨床研究センター特別講演会
  • Invited
• [Presentation] 腫瘍内微小環境と放射線感受性・抵抗性 (2018)
  • Author(s): 原田浩
  • Organizer: JASTRO第8回放射線生物学セミナー
  • Invited
• [Presentation] 細胞の低酸素応答を担う新規遺伝子の作用機序と機能 (2017)
  • Author(s): 原田浩
  • Organizer: ConBio2017
  • Invited
• [Presentation] がんに対する放射線治療の現状・課題・私たちの取り組み；低酸素バイオロジーの視点から (2017)
  • Author(s): 原田浩
  • Organizer: 富山大学医歯薬総合研究科特別講演会
  • Invited
• [Presentation] How can we overcome tumor radioresistance; lessons from hypoxia biology (2017)
  • Author(s): Harada H
  • Organizer: The 1st isRTB
  • Int'l Joint Research / Invited
• [Presentation] がんに対する放射線治療の現状と課題 (2017)
  • Author(s): 原田浩
  • Organizer: 知の拠点セミナー
  • Invited
• [Presentation] Radioresistance of Cancer Cells; Lessons from HIF-1 Biology (2017)
  • Author(s): 原田浩
  • Organizer: 放射線生物学セミナー in Gifu
  • Invited
• [Presentation] HIF-1バイオロジーで迫るがんの放射線抵抗性 (2017)
  • Author(s): 原田浩
  • Organizer: 若手放射線生物研究専門研究会
  • Invited
• [Presentation] HIF-1の生物学：基礎研究から創薬研究への展開 (2017)
  • Author(s): 原田浩
  • Organizer: 新規分子標的治療薬創薬に向けた大学発ベンチャー基盤の確立シンポジウム
  • Invited
• [Presentation] がんの悪性進展と治療抵抗性を担うHIF-1を中心とする遺伝子ネットワーク (2017)
  • Author(s): 原田浩
  • Organizer: 第5回低酸素研究会
  • Invited
• [Presentation] がん細胞のグルコース代謝 (2017)
  • Author(s): 原田浩
  • Organizer: 第19回京都大学大学院生命科学研究科シンポジウム
  • Invited
• [Presentation] Tumor hypoxia; an imaging and therapeutic target of interest for innovative radiation therapy (2017)
  • Author(s): Harada H
  • Organizer: The 3rd Bordeaux-Kyoto Symposium
  • Int'l Joint Research / Invited
• [Presentation] 悪性固形腫瘍内の低酸素環境 -放射線生物学の視点から- (2017)
  • Author(s): 原田浩
  • Organizer: 日本放射線腫瘍学会 第46回 放射線による制癌シンポジウム
  • Invited
• [Presentation] HIF-1を中心とする活性酸素ストレス応答の分子基盤と機能 (2016)
  • Author(s): 原田浩
  • Organizer: 日本放射線影響学会第59回大会
  • Place of Presentation: 広島
  • Year and Date: 2016-10-26
  • Invited
• [Presentation] Function of the UCHL1-HIF-1 Axis in Distant Metastasis and Radioresistance of Cancer Cells (2016)
  • Author(s): Hiroshi Harada
  • Organizer: International Symposium of Radiation Biology Center, Kyoto University
  • Place of Presentation: Kyoto
  • Year and Date: 2016-09-01
  • Int'l Joint Research / Invited
• [Presentation] Radioresistance of Cells; Lessons from HIF-1 Biology (2016)
  • Author(s): Hiroshi Harada
  • Organizer: Annual Meeting of Association for Radiation Research 2016
  • Place of Presentation: Leicester, UK
  • Year and Date: 2016-06-27
  • Int'l Joint Research / Invited
• [Presentation] Hypoxia; a Potent Inducer of Radioresistance of Cells (2016)
  • Author(s): Hiroshi Harada
  • Organizer: The 2nd KU RBC-CEA Joint Workshop
  • Place of Presentation: Kyoto
  • Year and Date: 2016-04-11
  • Int'l Joint Research / Invited
• [Book] 放射線医科学 (2016)
  • Author(s): 原田浩
  • Total Pages: 189
  • Publisher: 医療科学社
• [Remarks] 京都大学放射線生物研究センターがん細胞生物学分野ホームページ
  • URL: http://www.rbc.kyoto-u.ac.jp/cancer_biology/
• [Remarks] 京都大学放射線生物研究センターがん細胞生物学分野ホームページ
  • URL: http://radiotherapy.kuhp.kyoto-u.ac.jp/biology/index.html
• [Funded Workshop] The 33rd International Symposium of Radiation Biology Center, Kyoto University (2017)