Establishment of a synthetic promoter-based system for sensing oncogenic alteration in human live thyroid cells at a single-cell level

• Project/Area Number: 15K09438
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Endocrinology
• Research Institution: Nagasaki University
• Principal Investigator: SAENKO Vladimir 長崎大学, 原爆後障害医療研究所, 准教授 (30343346)
• Co-Investigator(Kenkyū-buntansha): ログノビッチ タチアナ 長崎大学, 原爆後障害医療研究所, 助教 (30423643) ; 光武 範吏 長崎大学, 原爆後障害医療研究所, 准教授 (50404215)
• Project Period (FY): 2015-04-01 – 2018-03-31
• Project Status: Completed (Fiscal Year 2017)
• Budget Amount: ¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000); 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: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
• Keywords: 甲状腺癌 / 癌遺伝子 / 遺伝子プロモーター / 合成生物学 / 単一細胞レポーター

Outline of Final Research Achievements

Establishment of a system for sensing oncogene activation at a single-cell level in live cells was the purpose. The system employs a fluorescent reporter driven by a synthetic promoter consisting of transcription factor binding sites (TFBSs) that correlated with upregulation of gene expression after oncogenic challenge. The library of single- or multiple-copy TFBSs was prepared and screened for activity in a dual-luminescent assay. The best promoters were inserted into the fluorescent reporter plasmid and screened by cotransfection with small amounts of oncogene-encoding vectors. Several selected synthetic promoter-reporter plasmids were stably transfected into target cells, which after transfection with oncogene-encoding vectors displayed a fluorescent signal observed by live cell imaging.

Research Products

• [Journal Article] Targeted Foxe1 overexpression in mouse thyroid causes the development of multinodular goiter but does not promote carcinogenesis. (2016)
  • Author(s): Nikitski A, Saenko V, Shimamura M, Nakashima M, Matsuse M, Suzuki K, Rogounovitch T, Bogdanova T, Shibusawa N, Yamada M, Nagayama Y, Yamashita S, Mitsutake N
  • Journal Title: Endocrinology Volume: 印刷中 Issue: 5 Pages: 2182-2195
  • DOI: 10.1210/en.2015-2066
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Major Factors Affecting Incidence of Childhood Thyroid Cancer in Belarus after the Chernobyl Accident: Do Nitrates in Drinking Water Play a Role? (2015)
  • Author(s): Drozd VM, Saenko VA, Brenner AV, Drozdovitch V, Pashkevich VI, Kudelsky AV, Demidchik YE, Branovan I, Shiglik N, Rogounovitch TI, Yamashita S, Biko J, Reiners C
  • Journal Title: PLoS One Volume: 10 Issue: 9 Pages: e0137226-e0137226
  • DOI: 10.1371/journal.pone.0137226
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] TCUKIN and CORGI Consortiums, Carmona LG; JTCMS Consortium: The 8q24 rs6983267G variant is associated with increased thyroid cancer risk. (2015)
  • Author(s): Sahasrabudhe R, Estrada A, Lott P, Martin L, Polanco Echeverry G, Velez A, Neta G, Takahasi M, Saenko V, Mitsutake N, Jaeguer E, Duque CS, Rios A, Bohorquez M, Prieto R, Criollo A, Echeverry M, Tomlinson I
  • Journal Title: Endocr Relat Cancer Volume: 22 Issue: 5 Pages: 841-849
  • DOI: 10.1530/erc-15-0081
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] BRAFV600E mutation is highly prevalent in thyroid carcinomas in the young population in Fukushima: a different oncogenic profile from Chernobyl. (2015)
  • Author(s): Mitsutake N, Fukushima T, Matsuse M, Rogounovitch T, Saenko V, Uchino S, Ito M, Suzuki K, Suzuki S, Yamashita S
  • Journal Title: Sci Rep Volume: 5 Issue: 1 Pages: 16976-16976
  • DOI: 10.1038/srep16976
  • NAID: 120006986823
  • Peer Reviewed / Open Access
• [Presentation] Genetic signatures of thyroid cancer according to radiation dose level (2016)
  • Author(s): サエンコ ウラジミール
  • Organizer: Joint symposium of KTA-KHNP "Thyroid cancer"(Korean Thyroid Association-Korean Hydro & Nuclear Power Co., Ltd)
  • Place of Presentation: ソウル（韓国）
  • Year and Date: 2016-12-02
  • Int'l Joint Research
• [Presentation] 甲状腺がん発症分子機構解明の為の特異的転写応答合成プロモーター活性を指標とした高感度細胞内シグナル活性経路検出アッセイ系の確立 (2016)
  • Author(s): サエンコ ウラジミール
  • Organizer: 第59回 日本甲状腺学会学術集会
  • Place of Presentation: 虎ノ門ヒルズフォーラム（東京都港区）
  • Year and Date: 2016-11-03
• [Presentation] Novel RET/PTC rearrangement identified by RNA-NGS in a sporadic pediatric PTC (2016)
  • Author(s): ログノビッチ タチアナ、サエンコ ウラジミール、マンコフスカヤ スベトラナ、フリドマン ミハイル、デミドチック ユリ、山下俊一
  • Organizer: 第89回 日本内分泌学会学術総会
  • Place of Presentation: 国立京都国際会館（京都府京都市）
  • Year and Date: 2016-04-21
• [Presentation] The combined influence of nitrates in drinking water and radiation on incidence of childhood thyroid cancer in Belarus after the Chernobyl accident (2015)
  • Author(s): Valentina M. Drozd, Vladimir A. Saenko, Alina V. Brenner, Vladimir Drozdovitch, Vasilii I. Pashkevich, Anatoliy V. Kudelsky, Yuri E. Demidchik, Igor Branovan, Nikolay Shiglik, Tatiana I. Rogounovitch, Shunichi Yamashita, Johannes Biko, Christoph Reiners
  • Organizer: Global Summit and Medicare Expo on Head & Neck Surgery
  • Place of Presentation: サンアントニオ（アメリカ）
  • Year and Date: 2015-11-30
  • Int'l Joint Research
• [Presentation] Screening for Thyroid Nodular Disease in Brest Region of Belarus: 29 Years after The Chernobyl Catastrophy (2015)
  • Author(s): M. Lushchyk, V. Drozd, L. Danilova, A. Grigorovich, V. Sivuda, A. Romanovskyj, Y. Demidchik, T. Rogunovich, V. Saenko, I.D. Branovan, N. Takamura, S. Yamashita
  • Organizer: 15th International Thyroid Congress and 85th Annual Meeting of the ATA
  • Place of Presentation: オーランド（アメリカ）
  • Year and Date: 2015-10-18
  • Int'l Joint Research
• [Presentation] Influence of Nitrate in Drinking Water in Open Wells on Childhood Thyroid Cancer Rates in Belarus and Ukraine after The Chernobyl Accident (2015)
  • Author(s): V. Drozd, V. Saenko, A. Brenner, V. Drozdovitch, M. Tronko, V. Shpak, T. Bogdanova, A. Serduk, I. Chernychenko, S. Tsymbaliuk, V. Pashkevich, A. Kudelsky, D. Branovan, N. Shiglik, T. Rogounovitch, S. Yamashita, J. Biko, C. Reiners
  • Organizer: 15th International Thyroid Congress and 85th Annual Meeting of the ATA
  • Place of Presentation: オーランド（アメリカ）
  • Year and Date: 2015-10-18
  • Int'l Joint Research
• [Presentation] Gene rearrangements in radiation-induced thyroid cancer (2015)
  • Author(s): Vladimir Saenko
  • Organizer: The 15th International Congress of Radiation Research
  • Place of Presentation: 国立京都国際会館（京都府・京都市）
  • Year and Date: 2015-05-25
  • Int'l Joint Research
• [Presentation] Foxe1 Overexpression and Radiation Effects on Mouse Thyroid (2015)
  • Author(s): Alyaksandr Nikitski, Norisato Mitsutake, Keiji Suzuki, Vladimir Saenko, Tatiana Rogounovitch, Shunichi Yamashita
  • Organizer: The 15th International Congress of Radiation Research
  • Place of Presentation: 国立京都国際会館（京都府・京都市）
  • Year and Date: 2015-05-25
  • Int'l Joint Research
• [Presentation] Novel RET/PTC rearrangement identified by RNA-NGS in pediatric papillary thyroid carcinoma from Belarus (2015)
  • Author(s): T. Rogounovitch, V. Saenko, S. Mankouskaya, M. Fridman, Yu. Demidchik, S. Yamashita
  • Organizer: The 15th International Congress of Radiation Research
  • Place of Presentation: 国立京都国際会館（京都府・京都市）
  • Year and Date: 2015-05-25
  • Int'l Joint Research
• [Presentation] Potential Joint Effect of Nitrates in Drinking Water and Radiation Dose to the Thyroid on Incidence of Pediatric Thyroid Cancer in Belarus (2015)
  • Author(s): Valentina M Drozd, Vladimir A Saenko, Alina V Brenner, Vladimir Drozdovitch, Vasilii I Pashkevich, Alexander V Kudelsky, Yuri E Demidchik, Igor Branovan, Nikolay Shiglik, Tatiana I Rogounovitch, Shunichi Yamashita, Tamara Platonova, Johannes Biko, Christoph Reiners
  • Organizer: The 15th International Congress of Radiation Research
  • Place of Presentation: 国立京都国際会館（京都府・京都市）
  • Year and Date: 2015-05-25
  • Int'l Joint Research
• [Remarks] 長崎大学 原研疫学ホームページ
  • URL: http://www-sdc.med.nagasaki-u.ac.jp/dhrc/
• [Remarks] 長崎大学 原研疫学ホームページ
  • URL: http://www-sdc.med.nagasaki-u.ac.jp/dhrc
• [Remarks] 長崎大学原研疫学ホームページ
  • URL: http://www-sdc.med.nagasaki-u.ac.jp/dhrc/