Establishment of a bio-assay for DNA homologous recombination ability using breast cancer cells to catalog responsible genetic alterations

• Project/Area Number: 18K19585
• Research Category: Grant-in-Aid for Challenging Research (Exploratory)
• Allocation Type: Multi-year Fund
• Review Section: Medium-sized Section 55:Surgery of the organs maintaining homeostasis and related fields
• Research Institution: Nagasaki University
• Principal Investigator: MITSUTAKE Norisato 長崎大学, 原爆後障害医療研究所, 教授 (50404215)
• Co-Investigator(Kenkyū-buntansha): 中沢 由華 名古屋大学, 環境医学研究所, 助教 (00533902)
• Project Period (FY): 2018-06-29 – 2021-03-31
• Project Status: Completed (Fiscal Year 2020)
• Budget Amount: ¥6,240,000 (Direct Cost: ¥4,800,000、Indirect Cost: ¥1,440,000); Fiscal Year 2019: ¥2,860,000 (Direct Cost: ¥2,200,000、Indirect Cost: ¥660,000); Fiscal Year 2018: ¥3,380,000 (Direct Cost: ¥2,600,000、Indirect Cost: ¥780,000)
• Keywords: 乳癌 / 相同組換え修復 / 相同組み換え修復 / DNA修復

Outline of Final Research Achievements

We have established a system to measure the ability of DNA homologous recombination repair using primary breast cancer cells directly grown from breast cancer tissues. The system uses fluorescence immunostaining for EdU, CENP-F, and 53BP1 after olaparib treatment and scores by the number of DNA double-strand breaks in the G2 phase before and after 24 h treatment. This system was validated using siRNA-based BRCA1 knockdown cells.
Using the system, we selected five cases in which homologous recombination seemed to be impaired and performed whole genome sequencing. We did not identify any known mutations which have been suggested to be associated with hereditary beast cancers; however, we identified some alterations that fit the hereditary model. The functional analysis of these genes is necessary.

Academic Significance and Societal Importance of the Research Achievements

遺伝子変異解析では、病的意義が判明している既知の遺伝子変異を検出した場合のみDNA相同組み換え修復能欠損があるかどうか、ひいては特定の薬剤の適応となるかが分かるが、いまだ全ての変異が明らかにされているわけではない。しかし、実際に乳癌組織より細胞を培養し、本研究で開発した手法を用いれば、DNA相同組み換え修復能を予想でき、薬剤を選択できる可能性が示唆された。

Research Products

• [Journal Article] Digenic mutations in ALDH2 and ADH5 impair formaldehyde clearance and cause a multisystem disorder, AMeD syndrome (2020)
  • Author(s): Oka Y, Hamada M, Nakazawa Y, ..., Tanaka M, Suganami T, Nakada S, Mitsutake N, Hara Y, Kato K, ..., Kaneko H, Matsumoto N, Matsuda F, Matsuo K, Takahashi Y, Mashimo T, Kojima S, Ogi T
  • Journal Title: Science Advances Volume: 6 Issue: 51 Pages: 7197-7197
  • DOI: 10.1126/sciadv.abd7197
  • Peer Reviewed / Open Access
• [Journal Article] Topoisomerase I-driven repair of UV-induced damage in NER-deficient cells (2020)
  • Author(s): Saha Liton Kumar、Wakasugi Mitsuo、Akter Salma、Prasad Rajendra、Wilson Samuel H.、Shimizu Naoto、Sasanuma Hiroyuki、Huang Shar-yin Naomi、Agama Keli、Pommier Yves、Matsunaga Tsukasa、Hirota Kouji、Iwai Shigenori、Nakazawa Yuka、Ogi Tomoo、Takeda Shunichi
  • Journal Title: Proceedings of the National Academy of Sciences Volume: 117 Issue: 25 Pages: 14412-14420
  • DOI: 10.1073/pnas.1920165117
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Biallelic VPS35L pathogenic variants cause 3C/Ritscher-Schinzel-like syndrome through dysfunction of retriever complex. (2020)
  • Author(s): Kato K, Okuno Y, Vasilev FF, Otomo T, Oishi H, Muramatsu H, Kawano Y, Oka Y, Nakazawa Y, Ogi T, Takahashi Y, Saitoh S.
  • Journal Title: Journal of Medical genetics Volume: 57 Issue: 4 Pages: 245-253
  • DOI: 10.1136/jmedgenet-2019-106213
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Papillary thyroid carcinoma in Ukraine after Chernobyl and in Japan after Fukushima: different histopathological scenarios. (2020)
  • Author(s): Bogdanova TI, Saenko VA, Hashimoto Y, Hirokawa M, Zurnadzhy LY, Hayashi T, Ito M, Iwadate M, Mitsutake N, Rogounovitch T, Sakamoto A, Naganuma H, Miyauchi A, Tronko MD, Thomas G, Yamashita S, Suzuki S:
  • Journal Title: Thyroid Volume: 308 Issue: 9 Pages: 1-13
  • DOI: 10.1089/thy.2020.0308
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] The Clinicopathological Results of Thyroid Cancer With BRAFV600E Mutation in the Young Population of Fukushima. (2020)
  • Author(s): Iwadate M, Mitsutake N, Matsuse M, Fukushima T, Suzuki S, Matsumoto Y, Ookouchi C, Mizunuma H, Nakamura I, Nakano K, Sakamoto A, Hirokawa M, Ito M, Naganuma H, Hashimoto Y, Shimura H, Yamashita S, Suzuki S
  • Journal Title: J Clin Endocrinol Metab Volume: 105 Issue: 12 Pages: 573-573
  • DOI: 10.1210/clinem/dgaa573
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Ubiquitination of DNA Damage-Stalled RNAPII Promotes Transcription-Coupled Repair (2020)
  • Author(s): Nakazawa Y, Hara Y, Oka Y, Komine O, van den Heuvel D, Guo C, Daigaku Y, Isono M, He Y, Shimada M, Kato K, Jia N, Hashimoto S, Kotani Y, Miyoshi Y, Tanaka M, Sobue A, Mitsutake N, Suganami T, Masuda A, Ohno K, Nakada S, Mashimo T, Yamanaka K, Luijsterburg MS, Ogi T
  • Journal Title: Cell Volume: 19 Issue: 6 Pages: 1228-1244
  • DOI: 10.1016/j.cell.2020.02.010
  • NAID: 120006871293
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] TERT mRNA Expression as a Novel Prognostic Marker in Papillary Thyroid Carcinomas (2019)
  • Author(s): Tanaka Aya、Matsuse Michiko、Saenko Vladimir、Nakao Tomoe、Yamanouchi Kosho、Sakimura Chika、Yano Hiroshi、Nishihara Eijun、Hirokawa Mitsuyoshi、Suzuki Keiji、Miyauchi Akira、Eguchi Susumu、Yoshiura Ko-ichiro、Yamashita Shunichi、Nagayasu Takeshi、Mitsutake Norisato
  • Journal Title: Thyroid Volume: 29 Issue: 8 Pages: 1105-1114
  • DOI: 10.1089/thy.2018.0695
  • NAID: 120006988431
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] The JAK/STAT3 and NF-kappa B signaling pathways regulate cancer stem cell properties in anaplastic thyroid cancer cells (2019)
  • Author(s): K. Shiraiwa, M. Mitsutake, Y. Nakazawa, T. Ogi, K. Suzuki, V. Saenko, K. Umezawa, S. Yamashita, K. Tsukamoto
  • Journal Title: Thyroid Volume: in press Issue: 5 Pages: 674-682
  • DOI: 10.1089/thy.2018.0212
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Functional and clinical relevance of novel mutations in a large cohort of patients with Cockayne syndrome. (2018)
  • Author(s): Calmels N, Botta E, Jia N, Fawcett H, Nardo T, Nakazawa Y, Lanzafame M, Moriwaki S, Sugita K, Kubota M, Obringer C, Spitz MA, Stefanini M, Laugel V, Orioli D, Ogi T, Lehmann A.
  • Journal Title: Journal of Medical Genetics Volume: 55 Issue: 5 Pages: 329-343
  • DOI: 10.1136/jmedgenet-2017-104877
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Presentation] 乳がんにおけるDNA修復能の測定法の確立 (2021)
  • Author(s): 田中 彩, 鈴木 啓司, 松瀬 美智子, 森田 道, 大坪 竜太, 松本 恵, 久芳 さやか, 及川 将弘, 矢野 洋, 江口 晋, 光武 範吏, 永安 武
  • Organizer: 第29回 日本乳癌学会学術総会
• [Presentation] 増悪する甲状腺乳頭癌を予測できる分子マーカー：さらなる高精度化と細胞診への応用 (2020)
  • Author(s): 松瀬 美智子, Vladimir Saenko, 中尾 朋恵, 山下 俊一, 光武 範吏
  • Organizer: 第63回 日本甲状腺学会学術総会
• [Presentation] 遺伝子変異からみた甲状腺がんの診断：穿刺吸引細胞診におけるTERTプロモーター変異の検出 (2020)
  • Author(s): 光武 範吏
  • Organizer: 第30回 臨床内分泌代謝Update
  • Invited
• [Presentation] チェルノブイリと福島の甲状腺がんにおける様々な病理学的シナリオ (2020)
  • Author(s): サエンコ ウラジミール, ボグダノワ タチアナ, 橋本 優子, 廣川 満良, リュドミラ ズルナドジ, 林 俊哲, 伊東 正博, 岩館 学, 光武 範吏, ログノビッチ タチアナ, 坂本 穆彦, 長沼 廣, 宮内 昭, トロンコ ミコラ, トマス ジュラルディン, 山下 俊一, 鈴木 眞一
  • Organizer: 第63回 日本放射線影響学会 年次大会
  • Invited
• [Presentation] チェルノブイリ原発事故後と福島県における小児・若年者甲状腺癌における遺伝子変異 (2020)
  • Author(s): 光武 範吏, 松瀬 美智子, 岩館 学, 山下 俊一, サエンコ ウラジミール, 鈴木 眞一
  • Organizer: 第63回 日本放射線影響学会 年次大会
  • Invited
• [Presentation] 乳頭癌の遺伝子異常 (2020)
  • Author(s): 光武 範吏
  • Organizer: 第32回 日本内分泌外科学会総会
  • Invited
• [Presentation] 小児・若年者甲状腺がんと成人甲状腺がんの遺伝子変異の相違 (2020)
  • Author(s): 光武 範吏, 松瀬 美智子, 岩館 学, サエンコ ウラジミール, 鈴木 眞一, 山下 俊一
  • Organizer: 第93回 日本内分泌学会学術総会
  • Invited
• [Presentation] 転写共役ヌクレオチド除去修復機構に重要なRNAポリメラーゼユビキチン化部位の同定 (2019)
  • Author(s): 中沢由華, 原雄一郎, 岡泰由, 小峯起, Diana, van, den, Heuvel, 郭朝万, 大学保一, 磯野真由, 何予希, 嶋田繭子, 加藤香奈, 賈楠, 橋下悟, 小谷祐子, 三好由夏, 田中都, 祖父江顕, 光武範吏, 菅波孝祥, 増田章男, 大野欽司, 中田慎一郎, 真下知士, 山中宏二, Martijn, S., Luijsterburg, 荻朋男
  • Organizer: 第42回日本分子生物学会年会
• [Presentation] ChIP-seqを利用したDNA損傷およびヌクレオチド除去修復のモニタリング (2019)
  • Author(s): 原雄一郎, 中沢由華, 岡泰由, 小峯起, Diana, van, den, Heuvel, 郭朝万, 大学保一, 磯野真由, 何予希, 嶋田繭子, 加藤香奈, 賈楠, 橋下悟, 小谷祐子, 三好由夏, 田中都, 祖父江顕, 光武範吏, 菅波孝祥, 増田章男, 大野欽司, 中田慎一郎, 真下知士, 山中宏二, Martijn, S., Luijsterburg, 荻朋男
  • Organizer: 第42回日本分子生物学会年会
• [Presentation] ゲノム不安定性を示す遺伝性疾患の分子病態 (2018)
  • Author(s): 中沢由華
  • Organizer: 第3回名大医薬系3部局交流シンポジウム～岐阜薬科大学・岐阜大学G-CHAIN・ラクオリア創薬合同シンポジウム～
• [Presentation] Alterations in RNA polymerase IIo ubiquitination cause Cockayne syndrome-like premature aging phenotype in mice due to TC-NER defect (2018)
  • Author(s): Nakazawa Y
  • Organizer: International Symposium on XP and other Nucleotide Excision Repair Disorders
  • Int'l Joint Research