転写開始点マッピングによる分化特異的・時期特異的発現遺伝子の同定

2016 Fiscal Year Annual Research Report

• Project/Area Number: 16H05159
• Research Institution: Nagasaki University
• Principal Investigator: 吉浦 孝一郎 長崎大学, 原爆後障害医療研究所, 教授 (00304931)
• Project Period (FY): 2016-04-01 – 2019-03-31
• Keywords: 5'末端 / switching / concatamerization

Outline of Annual Research Achievements

転写量の少ないRNAの５’末端部を効率的にクローニングすることが目的である。利用テクノロジーは，M-MLV （マウス白血病ウイルス）由来の逆転写酵素を利用したswitching 法による5’末端へのリンカー付加（SMART法）である。本年度は，逆転写反応時の5’末端へのリンカー付加の効率化とconcatamer形成（concatamerization）の阻害について，検討をおこなった。
当初1年目の予定は，5’末端へのリンカー付加の効率化のみを考えていたが，商品化されたキットではなく自作プライマー等でSwitching 法を行った際に，concatamer形成が大きな問題であることがわかった。本年度は，concatamer形成をおこさないオリゴヌクレオチド改良や修飾法を試して，技術的な問題の克服を目指した。concatamer形成阻害のために isocytosine，isoguanocine を含んだオリグヌクレオチドの利用が，論文化されているが，それらの使用は非常に高価であり，今後，本法をさまざまなアプリケーションに使っていくには自由度が少ないと考え，その他の修飾を探った。isocytosine，isoguanocine を含んだオリグヌクレオチドでない修飾によって，concatamer形成を減じることが出来つつある。

Current Status of Research Progress

3: Progress in research has been slightly delayed.

Reason

5’末端へのリンカー付加時のconcatamer形成（concatamerization）が起こり，この技術的解決に時間がかかっている。

Strategy for Future Research Activity

5’末端へのリンカー付加時のconcatamer形成（concatamerization）について，解決策がほぼ見つかっている。技術的な問題が解決できるので，「転写量の少ないRNAの５’末端部を効率的にシーケンスし，新規遺伝子末端部を同定する」当初の目的に向かって研究を進められる。

Research Products

• [Journal Article] SMCHD1 mutations associated with a rare muscular dystrophy can also cause isolated arhinia and Bosma arhinia microphthalmia syndrome. (2017)
  • Author(s): Shaw ND, Brand H, Kupchinsky ZA, Sato D, Okamoto N, Jacobsen C, Macarthur D, Moore SA, Yoshiura KI, Gusella JF, Marsh JA, Graham JM Jr, Lin AE, Katsanis N, Jones PL, Crowley WF Jr, Davis EE, FitzPatrick DR, Talkowski ME. その他
  • Journal Title: Nature Genetics Volume: 49(2) Pages: 238-248
  • DOI: 10.1038/ng.3743
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] De novo mutations in SMCHD1 abrogate nasal development. (2017)
  • Author(s): Gordon CT, Xue S, Yigit G, Filali H, Chen K, Rosin N, Yoshiura K-i, Oufadem M, Beck TJ, McGowan R, Teo ASM, Cunningham ML, Sefiani A, Kayserili H, Murphy JM, Chatdokmaiprai C, Hillmer AM, Wattanasirichaigoon D, Lyonnet S, Magdinier F, Javed A, Blewitt ME, Amiel J, Wollnik B, Reversade B. その他
  • Journal Title: Nature Genetics Volume: 49(2) Pages: 249-255
  • DOI: 10.1038/ng.3765
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Genetic variants in oxytocin receptor and arginine-vasopressin receptor 1A are associated with the neural correlates of maternal and paternal affection towards their child. (2017)
  • Author(s): Nishitani S, Ikematsu K, Takamura T, Honda S, Yoshiura KI, Shinohara K.
  • Journal Title: Horm Behav. Volume: 87 Pages: 47-56
  • DOI: 10.1016/j.yhbeh.2016.09.010.
  • Peer Reviewed
• [Journal Article] Identification of 11p14.1-p15.3 deletion probably associated with short stature, relative macrocephaly, and delayed closure of the fontanelles. (2017)
  • Author(s): Dateki S, Watanabe S, Kinoshita F, Yoshiura KI, Moriuchi H.
  • Journal Title: Am J Med Genet. A. Volume: 173(1) Pages: 217-220
  • DOI: 10.1002/ajmg.a.37978.
  • Peer Reviewed
• [Journal Article] Rapid growth of mitotically active cellular fibroma of the ovary: a case report and review of the literature. (2016)
  • Author(s): Matsuda K, Tateishi S, Akazawa Y, Kinoshita A, Yoshida S, Morisaki S, Fukushima A, Matsuwaki T, Yoshiura KI, Nakashima M.
  • Journal Title: Diagn Pathol. Volume: 11(1) Pages: 101
  • DOI: 10.1186/s13000-016-0554-7
  • Peer Reviewed
• [Journal Article] A significant association between rs8067378 at 17q12 and invasive cervical cancer originally identified by a genome-wide association study in Han Chinese is replicated in a Japanese population. (2016)
  • Author(s): Miura K, Mishima H, Yasunami M, Kaneuchi M, Kitajima M, Abe S, Higashijima A, Fuchi N, Miura S, Yoshiura KI, Masuzaki H.
  • Journal Title: J Hum Genet. Volume: 61(9) Pages: 793-796
  • DOI: 10.1038/jhg.2016.50
  • Peer Reviewed
• [Journal Article] Clinical and histological findings of autosomal dominant renal-limited disease with LMX1B mutation. (2016)
  • Author(s): Konomoto T, Imamura H, Orita M, Tanaka E, Moritake H, Sato Y, Fujimoto S, Harita Y, Hisano S, Yoshiura KI, Nunoi H.
  • Journal Title: Nephrology (Carlton) Volume: 21(9) Pages: 765-773
  • DOI: 10.1111/nep.12666
  • Peer Reviewed
• [Journal Article] Expression of Somatostatin Receptor Type 2A and PTEN in Neuroendocrine Neoplasms Is Associated with Tumor Grade but Not with Site of Origin. (2016)
  • Author(s): Wada H, Matsuda K, Akazawa Y, Yamaguchi Y, Miura S, Ueki N, Kinoshita A, Yoshiura K, Kondo H, Ito M, Nagayasu T, Nakashima M.
  • Journal Title: Endocr Pathol. Volume: 27(3) Pages: 179-187
  • DOI: 10.1007/s12022-016-9436-5
  • Peer Reviewed
• [Journal Article] Familial Mediterranean fever is no longer a rare disease in Japan. (2016)
  • Author(s): Migita K, Izumi Y, Jiuchi Y, Iwanaga N, Kawahara C, Agematsu K, Yachie A, Masumoto J, Fujikawa K, Yamasaki S, Nakamura T, Ubara Y, Koga T, Nakashima Y, Shimizu T, Umeda M, Nonaka F, Yasunami M, Eguchi K, Yoshiura K, Kawakami A.
  • Journal Title: Arthritis Res Ther. Volume: 30;18 Pages: 175
  • DOI: 10.1186/s13075-016-1071-5
  • Peer Reviewed
• [Journal Article] Association between p53-binding protein 1 expression and genomic instability in oncocytic follicular adenoma of the thyroid. (2016)
  • Author(s): Mussazhanova Z, Akazawa Y, Matsuda K, Shichijo K, Miura S, Otsubo R, Oikawa M, Yoshiura KI, Mitsutake N, Rogounovitch T, Saenko V, Kozykenova Z, Zhetpisbaev B, Shabdarbaeva D, Sayakenov N, Amantayev B, Kondo H, Ito M, Nakashima M.
  • Journal Title: Endocr J. Volume: 63(5) Pages: 457-467
  • DOI: 10.1507/endocrj.EJ15-0629
  • Peer Reviewed
• [Journal Article] Genetic background of hyperphenylalaninemia in Nagasaki, Japan. (2016)
  • Author(s): Dateki S, Watanabe S, Nakatomi A, Kinoshita E, Matsumoto T, Yoshiura K, Moriuchi H.
  • Journal Title: Pediatr Int. Volume: 58(5) Pages: 431-433
  • DOI: 10.1111/ped.12924
  • Peer Reviewed
• [Journal Article] Detailed analysis of 26 cases of 1q partial duplication/triplication syndrome. (2016)
  • Author(s): Watanabe S, Shimizu K, Ohashi H, Kosaki R, Okamoto N, Shimojima K, Yamamoto T, Chinen Y, Mizuno S, Dowa Y, Shiomi N, Toda Y, Tashiro K, Shichijo K, Minatozaki K, Aso S, Minagawa K, Hiraki Y, Shimokawa O, Matsumoto T, Fukuda M, Moriuchi H, Yoshiura K, Kondoh T.
  • Journal Title: Am J Med Genet A. Volume: 170(4) Pages: 908-917
  • DOI: 10.1002/ajmg.a.37496
  • Peer Reviewed
• [Journal Article] Multiple Serum Cytokine Profiling to Identify Combinational Diagnostic Biomarkers in Attacks of Familial Mediterranean Fever. (2016)
  • Author(s): Koga T, Migita K, Sato S, Umeda M, Nonaka F, Kawashiri SY, Iwamoto N, Ichinose K, Tamai M, Nakamura H, Origuchi T, Ueki Y, Masumoto J, Agematsu K, Yachie A, Yoshiura K, Eguchi K, Kawakami A.
  • Journal Title: Medicine (Baltimore) Volume: 95(16) Pages: e3449
  • DOI: 10.1097/MD.0000000000003449
  • Peer Reviewed
• [Presentation] IRUD-P 解析プログラムで判明したFG症候群を疑われた家族発症例の原因変異． (2016)
  • Author(s): 前川隆太，佐藤智生，吉浦孝一郎，近藤達郎
  • Organizer: 2.第二十三回遺伝性疾患に関する出生前診断研究会
  • Place of Presentation: 長崎大学良順会館ボードインホール（長崎県，長崎市）
  • Year and Date: 2016-09-24 – 2016-09-24
• [Presentation] 長崎大学における Initiative on Rare and Undiagnosed Disease in Pediatrics (IRUD-P)解析についての報告． (2016)
  • Author(s): 吉浦孝一郎，木下 晃，三嶋博之，林田知佐，近藤達郎，渡邊順子，伊達木澄人，要 匡，松原洋一
  • Organizer: 1.第一回放射線災害・医科学研究拠点カンファランス
  • Place of Presentation: 長崎大学良順会館専斎ホール（長崎県，長崎市）
  • Year and Date: 2016-06-04 – 2016-06-04
• [Presentation] Evaluation of genome-eide association study-identified SNPs at 4q12, 17q12, and 6p21.32 with cervical cancer susceptibility in a Japanese population. (2016)
  • Author(s): Kiyonori Miura, Hiroyuki Mishima, Shuhei Abe, Yuko Murakami, Naoki Fuchi, Ai Higashijima, Yuri Hasegawa, Shoko Miura, Masako Masuzaki, Masanori Kaneuchi, Koh-ichiro Yoshiura, Hideaki Masuzaki.
  • Organizer: The international Congress of Human Genetics (ICHG2016)
  • Place of Presentation: Kyoto International Conference Center, Japan. （京都府，京都市）
  • Year and Date: 2016-04-03 – 2016-04-07
  • Int'l Joint Research
• [Presentation] Bio-Virtuso: A package of Docker containers for multiple source data retrieval, RDF conversion, and triplestore deployment in a simplified manner. (2016)
  • Author(s): Hiroyuki Mishima, Koh-ichiro Yoshiura.
  • Organizer: The international Congress of Human Genetics (ICHG2016)
  • Place of Presentation: Kyoto International Conference Center, Japan.（京都府，京都市）
  • Year and Date: 2016-04-03 – 2016-04-07
  • Int'l Joint Research
• [Presentation] Mutaions in the patients with Nakajo Nishimura Syndrome-like autoinflammatory diseases. (2016)
  • Author(s): Akira Kinoshita, Nobuo Kanazawa, Noriko Kinjo, Hiroyuki Mishima, Koh-ichiro Yoshiura.
  • Organizer: The international Congress of Human Genetics (ICHG2016)
  • Place of Presentation: Kyoto International Conference Center, Japan.（京都府，京都市）
  • Year and Date: 2016-04-03 – 2016-04-07
  • Int'l Joint Research
• [Presentation] Aberrant methylation at imprinted DMRs is associated with placental mesenchymal dysplasia. (2016)
  • Author(s): Saori Aoki, Ken Higashimoto, Hidenori Hidaka, Hidetaka Watanabe, Yasufumi Ohtauka, Mishima Hiroyuki, Koh-ichiro Yoshiura, Hitomi Yatsuki, Kenichi Nishioka, Keiichiro Joh, Takashi Ohba, Hidetaka Katabuchi, Hidenobu Soejima.
  • Organizer: The international Congress of Human Genetics (ICHG2016)
  • Place of Presentation: Kyoto International Conference Center, Japan.（京都府，京都市）
  • Year and Date: 2016-04-03 – 2016-04-07
  • Int'l Joint Research
• [Presentation] Circulation levels of C19Mc-cluster microRNAs in pregnant women with abruptio placenta. (2016)
  • Author(s): Yuri Hasegawa, Kiyonori Miura, Ai Higashijima, Yuko Murakami, Ozora Tsukamoto, Shuhei Abe, Naoki Fuchi, Shoko Miura, Atsushi Yoshida, Hiroyuki Mishima, Akira Kinoshita, Koh-ichiro Yoshiura, Hideaki Masuzaki.
  • Organizer: The international Congress of Human Genetics (ICHG2016)
  • Place of Presentation: Kyoto International Conference Center, Japan.（京都府，京都市）
  • Year and Date: 2016-04-03 – 2016-04-07
  • Int'l Joint Research
• [Presentation] Normal ranges of plasma concentrations of pregnancy-associated microRNAs during pregnancy. (2016)
  • Author(s): Yuko Murakami, Kiyonori Miura, Ai Higashijima, Naoki Fuchi, Shuhei Abe, Yuri Hasegawa, Atsushi Yoshida, Masanori Kaneuchi, Yuko Murakami, Ozora Tsukamoto, Shoko Miura, Hiroyuki Mishima, Akira Kinoshita, Koh-ichiro Yoshiura, Hideaki Masuzaki.
  • Organizer: The international Congress of Human Genetics (ICHG2016)
  • Place of Presentation: Kyoto International Conference Center, Japan.（京都府，京都市）
  • Year and Date: 2016-04-03 – 2016-04-07
  • Int'l Joint Research