Elucidation of pathological mechanism caused by defects in the repair of TOPII-induced DNA double-strand breaks

• Project/Area Number: 16H02953
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Risk sciences of radiation and chemicals
• Research Institution: Kyoto University
• Principal Investigator: SASANUMA Hiroyuki 京都大学, 医学研究科, 准教授 (00531691)
• Project Period (FY): 2016-04-01 – 2019-03-31
• Project Status: Completed (Fiscal Year 2018)
• Budget Amount: ¥18,070,000 (Direct Cost: ¥13,900,000、Indirect Cost: ¥4,170,000); Fiscal Year 2018: ¥5,590,000 (Direct Cost: ¥4,300,000、Indirect Cost: ¥1,290,000); Fiscal Year 2017: ¥6,240,000 (Direct Cost: ¥4,800,000、Indirect Cost: ¥1,440,000); Fiscal Year 2016: ¥6,240,000 (Direct Cost: ¥4,800,000、Indirect Cost: ¥1,440,000)
• Keywords: ゲノム / ゲノム編集 / 相同組換え / DNA損傷修復 / DNA二重鎖切断 / ステロイドホルモン / トポイソメラーゼ2 / トポイソメラーゼ２ / トポイソメラーゼII型 / エトポシド / オーキシンデグロン

Outline of Final Research Achievements

The aim of this study is to characterize the DNA damages induced by estrogen and clarify the molecular mechanism of how estrogen-induced DNA damages are repaired. To this end, we generated BRCA1 and TOP2b deficient cells using breast cancer MCF-7 cells. We found that estrogen induced Top2b; covalently bound to DSB ends, named TOP2cc, in serum starved G1-phase cells. The loss of BRCA1 caused the accumulation of TOP2cc induced by estrogen, indicating an important role of BRCA1 in the repair of estrogen-induced DSBs. We have previously demonstrated that MRE11 nuclease also functions in the removal of Top2cc from DSB ends. BRCA1 promotes the recruitment of MRE11 onto Top2cc sites for subsequent removal of Top2cc. Our results suggest the loss of BRCA1 enhances estrogen’s genotoxicity. This function of BRCA1 may help explain the female organ-specific carcinogenesis of BRCA1-mutation carriers.

Academic Significance and Societal Importance of the Research Achievements

家族性乳がん卵巣がん症候群は、30代ぐらいから発がん率が上昇します。BRCA変異がなぜ女性臓器特異的がんを発症するかはわかっていませんでした。エストロゲンは、細胞増殖促進作用があることは知られていましたが、DNAを傷つける毒性については知られていませんでした。本研究では、BRCA1が欠損することによってエストロゲンのDNA毒性が増強し、発がんに繋がることを明らかにしました。

Research Products

• [Journal Article] PDIP38/PolDIP2 controls the DNA damage tolerance pathways by increasing the relative usage of translesion DNA synthesis over template switching. (2019)
  • Author(s): Tsuda M, Ogawa S, Ooka M, Kobayashi K, Hirota K, Wakasugi M, Matsunaga T, Sakuma T, Yamamoto T, Chikuma S, Sasanuma H, Debatisse M, Doherty AJ, Fuchs RP, Takeda S
  • Journal Title: PLoS One Volume: 14 Issue: 3 Pages: e0213383-e0213383
  • DOI: 10.1371/journal.pone.0213383
  • NAID: 120006822506
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] BRCA1 Haploinsufficiency Is Masked by RNF168-Mediated Chromatin Ubiquitylation (2019)
  • Author(s): Zong D, Adam S, Wang Y, Sasanuma H, Callen E, Murga M, Day A, Kruhlak MJ, Wong N, Munro M, Ray Chaudhuri A, Karim B, Xia B, Takeda S, Johnson N, Durocher D, Nussenzweig A
  • Journal Title: Molecular Cell Volume: 73 Issue: 6 Pages: 1267-1281
  • DOI: 10.1016/j.molcel.2018.12.010
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] SUMOylation of PCNA by PIAS1 and PIAS4 promotes template switch in the chicken and human B cell lines (2018)
  • Author(s): Mohiuddin Mohiuddin、Evans Terry John、Rahman Md Maminur、Keka Islam Shamima、Tsuda Masataka、Sasanuma Hiroyuki、Takeda Shunichi
  • Journal Title: Proceedings of the National Academy of Sciences Volume: 115 Issue: 50 Pages: 12793-12798
  • DOI: 10.1073/pnas.1716349115
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] BRCA1 ensures genome integrity by eliminating estrogen-induced pathological topoisomerase II-DNA complexes (2018)
  • Author(s): Sasanuma Hiroyuki、Tsuda Masataka、Morimoto Suguru、Saha Liton Kumar、Rahman Md Maminur、Kiyooka Yusuke、Fujiike Haruna、Cherniack Andrew D.、Itou Junji、Callen Moreu Elsa、Toi Masakazu、Nakada Shinichiro、Tanaka Hisashi、Tsutsui Ken、Yamada Shintaro、Nussenzweig Andre、Takeda Shunichi
  • Journal Title: Proceedings of the National Academy of Sciences Volume: 115 Issue: 45
  • DOI: 10.1073/pnas.1803177115
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Disruption of Hif-1α enhances cytotoxic effects of metformin in murine squamous cell carcinoma. (2018)
  • Author(s): Sanada Y, Sasanuma H, Takeda S, Tano K, Masunaga S.
  • Journal Title: Int J Radiat Biol Volume: 94 Issue: 1 Pages: 88-96
  • DOI: 10.1080/09553002.2018.1409443
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] The dominant role of proofreading exonuclease activity of replicative polymerase ε in cellular tolerance to cytarabine (Ara-C). (2017)
  • Author(s): Tsuda M, Terada K, Ooka M, Kobayashi K, Sasanuma H, Fujisawa R, Tsurimoto T, Yamamoto J, Iwai S, Kadoda K, Akagawa R, Huang SN, Pommier Y, Sale JE, Takeda S, Hirota K.
  • Journal Title: Oncotarget Volume: in press Issue: 20 Pages: 33457-33474
  • DOI: 10.18632/oncotarget.16508
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] TDP1 is critical for the repair of DNA breaks induced by sapacitabine, a nucleoside also targeting ATM- and BRCA-deficient tumors. (2017)
  • Author(s): Abo MA, Sasanuma H, Liu X, Rajapakse VN, Huang SN, Kiselev E, Takeda S, Plunkett W, Pommier Y
  • Journal Title: Mol Cancer Ther. Volume: 16 Issue: 11 Pages: 2543-2551
  • DOI: 10.1158/1535-7163.mct-17-0110
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Selective cytotoxicity of the anti-diabetic drug, metformin, in glucose-deprived chicken DT40 cells (2017)
  • Author(s): Kadoda K, Moriwaki T, Tsuda M, Sasanuma H, Ishiai M, Takata M, Ide H, Masunaga S, Takeda S and Tano K
  • Journal Title: PLOS ONE Volume: 12 Issue: 9 Pages: e0185141-e0185141
  • DOI: 10.1371/journal.pone.0185141
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Complementation of aprataxin deficiency by base excision repair enzymes in mitochondrial extracts (2017)
  • Author(s): Caglayan M, Prasad R, Krasich R, Longley MJ, Kadoda K, Tsuda M, Sasanuma H, Takeda S, Tano K, Copeland WC and Wilson SH
  • Journal Title: Nucleic Acids Res. Volume: 45 Issue: 17 Pages: 10079-10088
  • DOI: 10.1093/nar/gkx654
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Mre11 Is Essential for the Removal of Lethal Topoisomerase 2 Covalent Cleavage Complexes. (2016)
  • Author(s): Hoa NN, Shimizu T, Zhou ZW, Wang ZQ, Deshpande RA, Paull TT, Akter S, Tsuda M, Furuta R, Tsusui K, Takeda S, Sasanuma H
  • Journal Title: Mol Cell Volume: 64 Issue: 3 Pages: 580-592
  • DOI: 10.1016/j.molcel.2016.10.011
  • Peer Reviewed / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Deazaflavin Inhibitors of Tyrosyl-DNA Phosphodiesterase 2 (TDP2) Specific for the Human Enzyme and Active against Cellular TDP2. (2016)
  • Author(s): Marchand C, Abdelmalak M, Kankanala J, Huang SY, Kiselev E, Fesen K, Kurahashi K, Sasanuma H, Takeda S, Aihara H, Wang Z, Pommier Y
  • Journal Title: ACS Chem Biol. Volume: 11 Issue: 7 Pages: 1925-1933
  • DOI: 10.1021/acschembio.5b01047
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Poor recognition of O6-isopropyl dG by MGMT triggers double strand break-mediated cell death and micronucleus induction in FANC-deficient cells. (2016)
  • Author(s): Hashimoto K, Sharma V, Sasanuma H, Tian X, Takata M, Takeda S, Swenberg JA, Nakamura J
  • Journal Title: Oncotarget Volume: 7 Issue: 37 Pages: 59795-59808
  • DOI: 10.18632/oncotarget.10928
  • NAID: 120005980805
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Presentation] BRCA1-Mre11 ensures genome integrity by eliminating estrogen-induced pathological Topoisomerase II-DNA complexes (2019)
  • Author(s): Hiroyuki Sasanuma
  • Organizer: 5th HiHA International Symposium
  • Int'l Joint Research / Invited
• [Presentation] BRCA1 ensures genome integrity by eliminating estrogen-induced pathological Topoisomerase II-DNA complexes (2018)
  • Author(s): Hiroyuki Sasanuma
  • Organizer: 第41回分子生物学会
  • Invited
• [Presentation] BRCA1は、静止期細胞においてTop2依存的なDNA二重鎖切断修復に関与する (2018)
  • Author(s): 笹沼博之
  • Organizer: 第90回遺伝学会
  • Invited
• [Presentation] BRCA1 ensures genome integrity by eliminating estrogen-induced pathological Topoisomerase II-DNA complexes (2018)
  • Author(s): Hiroyuki Sasanuma
  • Organizer: IPR international seminar
  • Int'l Joint Research / Invited
• [Presentation] Mre11蛋白質は、DNAに共有結合したTopIIを除去するのに必須である。 (2017)
  • Author(s): 笹沼博之
  • Organizer: 分子生物学会
  • Invited
• [Presentation] The essential roleof Mre11-Rad50-Nbs1 complex in the removal of abortive top2-DNA complex (2017)
  • Author(s): Hiroyuki Sasanuma
  • Organizer: 33rd International Symposium of Radiation Biology Center, Kyoto University
  • Int'l Joint Research / Invited
• [Presentation] Mre11 Is Essential for the Removal of Lethal Topoisomerase 2 Covalent Cleavage Complexes (2016)
  • Author(s): Hiroyuki Sasanuma
  • Organizer: 3R symposium
  • Place of Presentation: 島根県松江
  • Year and Date: 2016-11-13
  • Int'l Joint Research
• [Remarks] TK6 consortium
  • URL: http://www.nihs.go.jp/dgm/tk6.html