チェルノブイリ小児甲状腺癌のパラフィン組織バンク設立と新規癌遺伝子解析

2013 Fiscal Year Annual Research Report

• Project/Area Number: 24406031
• Section: 海外学術
• Research Institution: Nagasaki University
• Principal Investigator: 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)
• Co-Investigator(Kenkyū-buntansha): SAENKO Vladimir 長崎大学, 原爆後障害医療研究所, 准教授 (30343346) ; 中島 正洋 長崎大学, 原爆後障害医療研究所, 教授 (50284683) ; 柴田 義貞 長崎大学, 医歯(薬)学総合研究科, 客員教授 (40010954)
• Project Period (FY): 2012-04-01 – 2015-03-31
• Keywords: 小児甲状腺乳頭癌 / パラフィン組織バンク / 次世代シーケンシング / 新規癌遺伝子

Research Abstract

During 25年 we accomplished collection of paraffin-embedded tissues and nearly finished to collect blood samples from the most aggressive (young age of exposure and short-latent period) pediatric PTCs (operated from 1990 and until October 1998), which are not included in the Chernobyl Tissue Bank by dispatching two specialists to affected after Chernobyl fallout areas of Belarus and Ukraine. For all collected PTC samples we made data base of patients, which includes demographic, clinical and morphological information.
From all collected blood samples we extracted DNA and from all Normal and Tumor counterparts of paraffin-embedded PTC samples we extracted DNA and RNA from some of them. We performed genetic analysis of all extracted Normal and Tumor PTC samples for known mutations (BRAF, H-, K-, N-RAS) and rearrangements (RET/PTC1, RET/PTC3 and AKAP9/BRAF).
For five pairs of PTC samples (Normal and Tumor) we performed DNA mutational analysis (all exons) using next-generation sequencing HiSeq system (Illumina). Within these five PTC samples, two had RET/PTC rearrangements (RET/PTC3 and RET/PTC1), but three others do not harbor any other known mutations and rearrangements.
For two additional Tumor PTC samples we performed mRNA whole genome fusion gene and mutational analysis using next-generation sequencing HiSeq system (Illumina). We chose two PTC samples without any known mutations and rearrangement, with solid and solid-trabecullar histological structures and with the same stage (T1N1bMO).

Current Status of Research Progress

2: Research has progressed on the whole more than it was originally planned.

Reason

ほぼ、当初の計画道り進行している。

Strategy for Future Research Activity

During 26年 we plan to accomplish collection of blood samples by dispatching specialists to affected after Chernobyl fallout areas, further development of database and performing DNA extraction from all collected blood samples.
We plan to perform biostatistical analysis of obtained data of DNA-exome sequence (point mutations) and mRNA whole genome sequence data (expressed rearrangements and point mutations) to narrow the spectrum of putative new cancer “driver” rearrangement and/or point mutation followed by verification of detected mutation/rearrangements by capillary sequencing, and frequency determination in all DNA samples extracted from paraffin-embedded tissues, and functional study.

Expenditure Plans for the Next FY Research Funding

Originally we were going to analyze more PTC cases using a next-generation sequencing HiSeq system (Illumina), however we decided to enroll a smaller number of PTCs.
We plan to use the remaining money for functional analysis to find new “driver” oncogenes.

Research Products

• [Journal Article] Radiation-associated small cell neuroendocrine carcinoma of the thyroid:a case report with molecular analyses (2014)
  • Author(s): Mussazhanova Z, Miura S, Stanojevic B, Rougounovitch T, Saenko V, Shiraishi T, Kurashige T, Shichijo K, Kaneko K, Takahashi H, Ito M, Nakashima M.
  • Journal Title: Thyroid Volume: 24(3) Pages: 593-598
  • DOI: doi: 10.1089/thy.2013.0214.
  • Peer Reviewed
• [Journal Article] Patterns of FOXE1 expression in papillary thyroid carcinoma by immunohistochemistry (2013)
  • Author(s): Bychkov A,, Saenko V, Nakashima M, Mitsutake N, Rogounovitch T, Nikitski A, Orim F, Yamashita S.
  • Journal Title: Thyroid Volume: 23(7) Pages: 817-828
  • DOI: doi: 10.1089/thy.2012.0466.
  • Peer Reviewed
• [Journal Article] Significance of p53-binding protein 1 (53BP1) expression in thyroid papillary microcarcinoma: association with BRAFV600E mutation status (2013)
  • Author(s): Mussazhanova Z, Matsuda K, Naruke Y, Mitsutake N, Stanojevic B, Rougounovitch T, Saenko V, Suzuki K, Nishihara E, Hirokawa M, Ito M, Nakashima M.
  • Journal Title: Histopathology Volume: 63(5) Pages: 726-734
  • DOI: doi: 10.1111/his.12233.
  • Peer Reviewed
• [Presentation] Genetic polymorphism on chromosome 14q13.3 associates with risk for thyroid follicular adenoma in Japanese patients (2013)
  • Author(s): ログノビッチ タチアナ, サエンコ ウラジミール, ビチコブ アンドレイ, ニキツキー アリャクサンドル, 高橋めい子, 中島正洋, 林徳眞吉, 廣川満良, 宮内昭, 石垣克, 重松和人, 光武範吏, 伊藤正博, 松田文彦, 山下俊一
  • Organizer: 第56回日本甲状腺学会学術集会
  • Place of Presentation: 和歌山県民文化会館 (和歌山市)
  • Year and Date: 20131114-20131116
• [Presentation] Association of three polymorphisms of the FOXE1 gene with papillary thyroid carcinoma in Belarussian and Japanese populations (2013)
  • Author(s): サエンコ ウラジミル,高橋めい子, ログノビッチ タチアナ, ドロズド ワレンチナ, ビチコフ アンドレイ, 川口喬久, 光武範吏, 廣川満良, 西原永潤, 宮内昭, 吉浦孝一郎, 松田文彦, 山下俊一
  • Organizer: 第56回日本甲状腺学会学術集会
  • Place of Presentation: 和歌山県民文化会館 (和歌山市)
  • Year and Date: 20131114-20131116
• [Presentation] Transcriptional activities of FOXE1 polyalanine tract variants (2013)
  • Author(s): ニキツキー アリャクサンドル,サエンコ ウラジミール, ビチコブ アンドレイ, ログノビッチ タチアナ, 光武範吏, 山下俊一
  • Organizer: 第56回日本甲状腺学会学術集会
  • Place of Presentation: 和歌山県民文化会館 (和歌山市)
  • Year and Date: 20131114-20131116
• [Presentation] Rare allele of rs944289 (NKX2-1(TTF1)) associated with increasing risk of both malignant (PTC) and benign (FA) tumors in Japanese population (2013)
  • Author(s): Rogounovitch TI, Saenko VA, Bychkov A, Nikitski AV, Takahashi M, Nakashima M, Hayashi T, Hirokawa M, Miyauchi A, Shigematsu K, Mitsutake N, Matsuda F, Yamashita S
  • Organizer: European Thyroid Association 37th Annual Meeting
  • Place of Presentation: Leiden, The Netherlands
  • Year and Date: 20130907-20130911
• [Presentation] Molecular epidemiology study of Chernobyl thyroid cancer from Belarus and Ukraine (2013)
  • Author(s): Vladimir Saenko, M.Takahashi, T.Rogounovitch, V.Drozd, Yu.Demidchik, T.Bogdanova, M.Tronko, N.Mitsutake, N.Takamura, F.Matsuda, S.Yamashita
  • Organizer: European Thyroid Association 37th Annual Meeting
  • Place of Presentation: Leiden, The Netherlands
  • Year and Date: 20130907-20130911
• [Presentation] MAY HIGH NITRATE LEVELS IN DRINKING WATER INCREASE THE PREVALENCE OF RADIATION-RELATED THYROID CANCER IN BELARUS? (2013)
  • Author(s): Drozd Valentina, Demidchik Yuri, Danilova Larisa, Kudelsky Alexander, Branovan Igor, Saenko Vladimir, Rogounovitch Tatiana, Yamashita Shunichi, Biko Johannes, Reiners Christoph
  • Organizer: 2nd World Congress on Thyroid Cancer
  • Place of Presentation: Toronto, Canada
  • Year and Date: 20130710-20130714
• [Presentation] Identification of genes cooperating with mutant BRAF in thyroid (2013)
  • Author(s): ログノビッチ タチアナ, サエンコ ウラジミル, 光武範吏, ビチコフ アンドレイ,永山雄二,山下俊一
  • Organizer: 第86回日本内分泌学会学術総会
  • Place of Presentation: 仙台国際センター (仙台)
  • Year and Date: 20130425-20130427
• [Presentation] Association of the NRG1 gene with risk for developing thyroid cancer after Chernobyl (2013)
  • Author(s): サエンコ ウラジミル,高橋めい子, ログノビッチ タチアナ, ドロズド ワレンチナ, デミドチク ユーリ, ボグダノワ タチアナ, トロンコ ミコラ, 光武範吏, 高村昇, 松田文彦 , 山下俊一
  • Organizer: 第86回日本内分泌学会学術総会
  • Place of Presentation: 仙台国際センター (仙台)
  • Year and Date: 20130425-20130427
• [Remarks] 長崎大学 原爆後障害医療研究所 原研医療
  • URL: http://www-sdc.med.nagasaki-u.ac.jp/drms/