Differentiation and time specific gene Identification by the mapping of transcriptional start site

2017 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: 転写産物 / 5'末端 / 細胞特異的転写物

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。
自らデザイン改良した乗り換え用の5’末端オリゴヌクレオチドを利用する際に，Switching による concatamer 形成をある程度抑制する（完全ではない）オリゴヌクレオチド改良や修飾法を試し，技術的な問題の克服を行った。本年度，本研究で進めている改良 SMART法と mRNA のCap構造を利用して転写５’末端部の濃縮法である CAGE 法 の比較検討を行った。改良 SMART法は，やはり concatemer 形成の欠点があり，False positive があるようだが，sensitivity は高い様である。改良 SMART法でモデル動物で特殊な5'末端のクローニングは可能であった。最終年度の細胞特異的な転写5'末端ライブラリー作成に向け，モデルマウス導入予定である。
本研究過程で，ヒト疾患モデル動物から，ある大きな遺伝子の３’末端側から新規の転写産物が転写され　目の発生に関わっていることがわかってきており，技術開発は遅れているものの新規遺伝子同定という面では，一つの成果を出せた。結果は，現在論文作成中である。

Current Status of Research Progress

3: Progress in research has been slightly delayed.

Reason

Switching オリゴヌクレオチドのconcatemer 形成が抑制できずに，進行が遅れている。細胞特異的な5'末端解析は，switchng 法と CAGE法を併用することで進められると考えられ，今後研究が追いついていけるように進める。

Strategy for Future Research Activity

細胞特異的な5'末端解析は，switchng 法と CAGE法を併用することで進める。細胞特異的 5'末端ライブラリー作成の為にモデルマウス導入を進めて，研究を進展させる。

Research Products

• [Journal Article] Whole-exome sequencing and gene-based rare variant association tests suggest that PLA2G4E might be a risk gene for panic disorder (2018)
  • Author(s): Morimoto Y, Shimada-Sugimoto M, Otowa T, Yoshida S, Kinoshita A, Mishima H, Yamaguchi N, Mori T, Imamura A, Ozawa H, Kurotaki N, Ziegler C, Domschke K, Deckert J, Umekage T, Tochigi M, Kaiya H, Okazaki Y, Tokunaga K, Sasaki T, Yoshiura KI, Ono S.
  • Journal Title: Translational Psychiatry Volume: 8 Pages: 41～41
  • DOI: 10.1038/s41398-017-0088-0
  • Peer Reviewed / Open Access
• [Journal Article] A hot-spot mutation in CDC42 (p.Tyr64Cys) and novel phenotypes in the third patient with Takenouchi-Kosaki syndrome (2018)
  • Author(s): Motokawa Midori、Watanabe Satoshi、Nakatomi Akiko、Kondoh Tatsuro、Matsumoto Tadashi、Morifuji Kanako、Sawada Hirotake、Nishimura Toyoki、Nunoi Hiroyuki、Yoshiura Koh-ichiro、Moriuchi Hiroyuki、Dateki Sumito
  • Journal Title: Journal of Human Genetics Volume: 63 Pages: 387～390
  • DOI: 10.1038/s10038-017-0396-5
  • Peer Reviewed
• [Journal Article] Sick sinus syndrome with HCN4 mutations shows early onset and frequent association with atrial fibrillation and left ventricular noncompaction (2017)
  • Author(s): Ishikawa Taisuke、Ohno Seiko、Murakami Takashi、Yoshida Kentaro、Mishima Hiroyuki、Fukuoka Tetsuya、Kimoto Hiroki、Sakamoto Risa、Ohkusa Takafumi、Aiba Takeshi、Nogami Akihiko、Sumitomo Naokata、Shimizu Wataru、Yoshiura Koh-ichiro、Horigome Hitoshi、Horie Minoru、Makita Naomasa
  • Journal Title: Heart Rhythm. Volume: 14 Pages: 717～724
  • DOI: 10.1016/j.hrthm.2017.01.020
  • Peer Reviewed
• [Journal Article] Analysis of clinical symptoms and ABCC6 mutations in 76 Japanese patients with pseudoxanthoma elasticum (2017)
  • Author(s): Iwanaga Akira、Okubo Yumi、Yozaki Mariko、Koike Yuta、Kuwatsuka Yutaka、Tomimura Saori、Yamamoto Yosuke、Tamura Hiroshi、Ikeda Satoshi、Maemura Koji、Tsuiki Eiko、Kitaoka Takashi、Endo Yuichiro、Mishima Hiroyuki、Yoshiura Koh-Ichiro、Ogi Tomoo、Tanizaki Hideaki、Wataya-Kaneda Mari、Hattori Tomoyasu、Utani Atsushi
  • Journal Title: The Journal of Dermatology Volume: 44 Pages: 644～650
  • DOI: 10.1111/1346-8138.13727
  • Peer Reviewed
• [Journal Article] Deep sequencing reveals variations in somatic cell mosaic mutations between monozygotic twins with discordant psychiatric disease (2017)
  • Author(s): Morimoto Yoshiro、Ono Shinji、Imamura Akira、Okazaki Yuji、Kinoshita Akira、Mishima Hiroyuki、Nakane Hideyuki、Ozawa Hiroki、Yoshiura Koh-ichiro、Kurotaki Naohiro
  • Journal Title: Human Genome Variation Volume: 4 Pages: 17032～17032
  • DOI: 10.1038/hgv.2017.32
  • Peer Reviewed / Open Access
• [Journal Article] Identification of a novel heterozygous mutation of the Aggrecan gene in a family with idiopathic short stature and multiple intervertebral disc herniation (2017)
  • Author(s): Dateki Sumito、Nakatomi Akiko、Watanabe Satoshi、Shimizu Hitomi、Inoue Yukiko、Baba Hideo、Yoshiura Koh-ichiro、Moriuchi Hiroyuki
  • Journal Title: Journal of Human Genetics Volume: 62 Pages: 717～721
  • DOI: 10.1038/jhg.2017.33
  • Peer Reviewed
• [Journal Article] Progressive Atrial Conduction Defects Associated With Bone Malformation Caused by a Connexin-45 Mutation (2017)
  • Author(s): Seki A, Ishikawa T, Daumy X, Mishima H, Barc J, Sasaki R, Nishii K, Saito K, Urano M, Ohno S, Otsuki S, Kimoto H, Baruteau AE, Thollet A, Fouchard S, Bonnaud S, Parent P, Shibata Y, Perrin JP, Le Marec H, Hagiwara N, Mercier S, Horie M, Probst V, Yoshiura KI, Redon R, Schott JJ, Makita N.
  • Journal Title: Journal of the American College of Cardiology Volume: 70 Pages: 358～370
  • DOI: 10.1016/j.jacc.2017.05.039
  • Peer Reviewed
• [Journal Article] Fetiform teratoma was a parthenogenetic tumor arising from a mature ovum (2017)
  • Author(s): Miura Kiyonori、Kurabayashi Takumi、Satoh Chisei、Sasaki Kensaku、Ishiguro Tatsuya、Yoshiura Koh-ichiro、Masuzaki Hideaki
  • Journal Title: Journal of Human Genetics Volume: 62 Pages: 803～808
  • DOI: 10.1038/jhg.2017.45
  • Peer Reviewed
• [Journal Article] Effects of HLA-DRB1 alleles on susceptibility and clinical manifestations in Japanese patients with adult onset Still’s disease (2017)
  • Author(s): Asano T, Furukawa H, Sato S, Yashiro M, Kobayashi H, Watanabe H, Suzuki E, Ito T, Ubara Y, Kobayashi D, Iwanaga N, Izumi Y, Fujikawa K, Yamasaki S, Nakamura T, Koga T, Shimizu T, Umeda M, Nonaka F, Yasunami M, Ueki Y, Eguchi K, Tsuchiya N, Tohma S, Yoshiura KI, Ohira H, Kawakami A, Migita K.
  • Journal Title: Arthritis Research & Therapy Volume: 19 Pages: 199～199
  • DOI: 10.1186/s13075-017-1406-x
  • Peer Reviewed
• [Journal Article] Two consecutive successful live birth in woman with 17α hydroxylase deficiency by frozen?thaw embryo transfer under hormone replacement endometrium preparation (2017)
  • Author(s): Kitajima Michio、Miura Kiyonori、Inoue Tsuneo、Murakami Yuko、Kitajima Yuriko、Murakami Naoko、Taniguchi Ken、Yoshiura Ko-Ichiro、Masuzaki Hideaki
  • Journal Title: Gynecological Endocrinology Volume: 34 Pages: 381～384
  • DOI: 10.1080/09513590.2017.1393512
  • Peer Reviewed
• [Journal Article] Postmortem genetic analysis of sudden unexpected death in infancy: neonatal genetic screening may enable the prevention of sudden infant death (2017)
  • Author(s): Oshima Yuki、Yamamoto Takuma、Ishikawa Taisuke、Mishima Hiroyuki、Matsusue Aya、Umehara Takahiro、Murase Takehiko、Abe Yuki、Kubo Shin-ichi、Yoshiura Koh-ichiro、Makita Naomasa、Ikematsu Kazuya
  • Journal Title: Journal of Human Genetics Volume: 62 Pages: 989～995
  • DOI: 10.1038/jhg.2017.79
  • Peer Reviewed
• [Journal Article] Chromosomal analysis of myelodysplastic syndromes among atomic bomb survivors in Nagasaki (2017)
  • Author(s): Horai M, Satoh S, Matsuo M, Iwanaga M, Horio K, Jo T, Takasaki Y, Kawaguchi Y, Tsushima H, Yoshida S, Taguchi M, Itonaga H, Sawayama Y, Taguchi J, Imaizumi Y, Hata T, Moriuchi Y, Haase D, Yoshiura KI, Miyazaki Y.
  • Journal Title: British Journal of Haematology Volume: 180 Pages: 381～390
  • DOI: 10.1111/bjh.15050
  • Peer Reviewed
• [Journal Article] Detection of de novo single nucleotide variants in offspring of atomic-bomb survivors close to the hypocenter by whole-genome sequencing (2017)
  • Author(s): Horai Makiko、Mishima Hiroyuki、Hayashida Chisa、Kinoshita Akira、Nakane Yoshibumi、Matsuo Tatsuki、Tsuruda Kazuto、Yanagihara Katsunori、Sato Shinya、Imanishi Daisuke、Imaizumi Yoshitaka、Hata Tomoko、Miyazaki Yasushi、Yoshiura Koh-ichiro
  • Journal Title: Journal of Human Genetics Volume: 63 Pages: 357～363
  • DOI: 10.1038/s10038-017-0392-9
  • Peer Reviewed
• [Presentation] Next-generation sequencing targeting disease-associated candidate genes unveils novel missense mutations for long QT syndrome in Japanese patients. (2018)
  • Author(s): Ryo Watanabe, Naomasa Makita, Koh-ichiro Yoshiura, Toshihiro Tanaka for Japanese LQTS Deep Sequencing Consortium.
  • Organizer: 第1回日本循環器学会基礎研究フォーラム
• [Presentation] 顕著な睡眠障害，発達障害を有するCREBBP遺伝子のエクソン31の新規突然変異例． (2018)
  • Author(s): 福井香織，渡邊順子，吉浦孝一郎．
  • Organizer: 第40回日本小児遺伝学会学術集会
• [Presentation] Evans症候群および低ガンマグロブリン血症を発症した歌舞伎症候群の一例． (2018)
  • Author(s): 蒸野寿紀，南 弘一，鈴木啓之，森尾友宏，今井耕輔，三嶋博之，吉浦孝一郎，田村志宣，山下友佑，園木 孝志．
  • Organizer: 第1回日本免疫不全・自己炎症学会総会・学術集会
• [Presentation] Novel transcript of IPTR1 contributes to the development of anterior eye segment. (2018)
  • Author(s): Kinoshita A., Mishima H., Saitoh S., Yoshiura K.
  • Organizer: The 2nd International Symposium of the Network-type Joint Usage/Research Center for Radiation Disaster Medical Science,
  • Int'l Joint Research
• [Presentation] Identification of novel long QT syndrome-associated mutations by targeted sequencing analyses. (2017)
  • Author(s): Watanabe R., Ohno S., Aiba T., Ishikawa T., Nakano Y., Mishima H., Minamino N., Makiyama T., Tanaka Y., Murata H., Hayashi M., Iwasaki Y., Watanabe A., Tachibana M., Morita H., Miyamoto Y., Yoshiura K., Tsunoda T., Watanabe H., Kurabayashi M., Nogami A., Kihara Y., Makita N., Shimizu W., Horie M., Tanaka T.
  • Organizer: European Human Genetics Conference
  • Int'l Joint Research
• [Presentation] De novo gain-of-function mutations in the epigenetic regulator SMCHD1 cause Bosma arhinia microphthalmia syndrome. (2017)
  • Author(s): Gordon C. T., Xue S., Yigit G., Filali H., Chen K., Rosin N., Yoshiura K., Oufadem M., Beck T., Dion C., Sefiani A., Kayserili H., Murphy J., Chatdokmaiprai C., Hillmer A., Wattanasirichaigoon D., Lyonnet S., Magdinier F., Javed A., Blewitt M., Amiel J., Wollnik B., Reversade B.
  • Organizer: European Human Genetics Conference
  • Int'l Joint Research
• [Presentation] A case of epileptic encephalopathy combined with various involuntary movement, sever psychomotor retardation with KCNA2 gene mutation. (2017)
  • Author(s): 里龍晴，原口康平，森山薫，渡辺聡，神村直久，吉浦孝一郎，森内浩幸．
  • Organizer: 第59回日本小児神経学会学術集会
• [Presentation] 全エクソーム解析で判明したosteogenesis imperfectaの１例． (2017)
  • Author(s): 三春範夫，吉浦孝一郎，佐々木晃，正路貴代．
  • Organizer: 第２4回遺伝性疾患に関する出生前診断研究会
• [Presentation] A Heterozygous FLNB Mutation in a Japanese Boy with Spondylocarpotarsal Synostosis Syndrome and his Mother with Short Stature. (2017)
  • Author(s): Hitomi Shimizu, Satoshi Watanabe, Hiroyuki Moriuchi, Koh-ichiro Yoshiura, Sumito Dateki.
  • Organizer: 10th International Joint Meeting of Pediatric Endocrinology
  • Int'l Joint Research
• [Presentation] 低身長を契機に診断に至ったSpondylocarpotarsal synostosisの1幼児例． (2017)
  • Author(s): 清水日智，渡辺聡，中富明子，吉浦孝一郎，森内浩幸，伊達木澄人．
  • Organizer: 第51回日本小児内分泌学会学術集会
• [Presentation] Identification of I287S homozygous mutation in the MLX gene in an infant with non-alcoholic steatohepatitis: A case report. (2017)
  • Author(s): Yoriko Watanabe, Satoshi Watanabe, Chisei Sato, Sachiko Ogasawara, Jun Akiba, Masayoshi Kage, Hirohisa Yano, Kaori Fukui, Tatsuki Mizuochi, Shoji Yano, Kiyoshi Hayasaka, Yushiro Yamashita, Takumi Kawaguchi, Takuji Torimura, Koh-ichiro Yoshiura.
  • Organizer: American Society of Human Genetics Annual Meeting 2017
  • Int'l Joint Research
• [Presentation] BCORナンセンス変異にもとづくOFCD症候群の一例． (2017)
  • Author(s): 鳥尾倫子，古賀のりこ，赤峰哲，吉浦孝一郎，酒井康成，大賀正一．
  • Organizer: 日本人類遺伝学会 第62回大会
• [Presentation] De novo mutations in the C-terminal region of ITPR1 cause aniridia, cerebellar ataxia and intellectual disability, Gillespie syndrome. (2017)
  • Author(s): Shinji Saitoh, Akira Kinoshita, Kana Hosoki, Hiroyuki Mishima, Kiyotaka Tomiwa, Naoko Ishihara, Yutaka Negishi, Naoko Asahina, Hideaki Shiraishi, Koh-ichiro Yoshiura.
  • Organizer: 日本人類遺伝学会 第62回大会
• [Presentation] Targeted deep sequencing reveals novel mutations in Japanese long QT syndrome patients. (2017)
  • Author(s): Watanabe R, Ohno S, Aiba T, Ishikawa T, Nakano Y, Mishima H, Sudo T, Higuchi C, Minamino N, Makiyama T, Tanaka Y, Murata H, Hayashi M, Iwasaki Y, Watanabe A, Tachibana M, Morita H, Miyamoto Y, Yoshiura K, sunoda T, Watanabe H, Kurabayashi M, Nogami A, Kihara Y, Makita N, Shimuzu W, Horie M, Tanaka T.
  • Organizer: 日本人類遺伝学会 第62回大会
• [Presentation] A case suspected of mitochondrial complex III deficiency carrying BCS1L mutation presented with Fanconi syndrome． (2017)
  • Author(s): Kanako Ishii, Vlad Tocan, Noriko Oyama, Naoko Toda, Kazuhiro Okubo, Yoshito Ishizaki, Koh-ichiro Yoshiura, Shouichi Ohga.
  • Organizer: 日本人類遺伝学会 第62回大会
• [Presentation] Identification of I285S homozygous mutation on the MLX gene in an infant with non-alcoholic steatohepatitis． (2017)
  • Author(s): Yoriko Watanabe, Satoshi Watanabe, Chisei Satoh, Sachiko Ogasawara, Jun Akiba, Masayoshi Kage, Hirohisa Yano, Kaori Fukui, Tatsuki Mizuochi, Shoji Yano, Kiyoshi Hayasaka, Yushiro Yamashita, Takumi Kawaguchi, Takuji Torimura, Koh-ichiro Yoshiura.
  • Organizer: 日本人類遺伝学会 第62回大会
• [Presentation] Genetic and clinical analyses in Japanese Patients with Sitosterolemia. (2017)
  • Author(s): Hitomi Shimizu, Hideki Motomura, Satoshi Watanabe, Hiroyuki Mishima, Hiroyuki Moriuchi, Koh-ichiro Yoshiura, Sumito Dateki.
  • Organizer: 日本人類遺伝学会 第62回大会
• [Presentation] Gene-based rare variants association test implicates PLA2G4E as a risk gene for panic disorder. (2017)
  • Author(s): Yoshiro Morimoto, Shintaro Yoshida, Akira Kinoshita, Hiroyuki Mishima, Naohiro Yamaguchi, Akira Imamura, Naohiro Kurotaki, Hiroki Ozawa, Koh-ichiro Yoshiura, Shinji Ono.
  • Organizer: 日本人類遺伝学会 第62回大会
• [Presentation] MicroRNA-204-3p inhibits TLR4-related cytokines in family Mediterranean fever by targeting the PI3K signaling. (2017)
  • Author(s): Tomohiro Koga, Kiyoshi Migita, Shuntaro Sato, Kiyonori Miura, Hideki Masuzaki, Koh-ichiro Yoshiura, Atsushi Kawakami.
  • Organizer: 日本人類遺伝学会 第62回大会
• [Presentation] Novel transcript of ITPR1 contributes to the development of anterior eye segment. (2017)
  • Author(s): Akira Kinoshita, Kaname Ohyama, Susumu Tanimura, Katsuya Matsuda, Miroyuki Mishima, Masahiro Nakajima, Shinji Saitoh, Koh-ichiro Yoshiura.
  • Organizer: 日本人類遺伝学会 第62回大会
• [Presentation] 胆道閉鎖症I cyst型を合併した歌舞伎症候群の1例． (2017)
  • Author(s): 石原 潤，水落建輝，高木祐吾，江田慶輔，進藤亮太，弓削康太郎，福井香織，渡邊順子，橋詰直樹，石井信二，深堀 優，浅桐公男，八木 実，吉浦孝一郎．
  • Organizer: 第44回日本胆道閉鎖症研究会
• [Presentation] 脂肪萎縮性糖尿病に先天性難聴を合併し、PIK3R1遺伝子に新生突然変異を認めSHORT症候群と診断した一例． (2017)
  • Author(s): 大久保一宏，石井加奈子，大山紀子，トカン ヴラッド，戸田尚子，海老原 健，吉浦孝一郎，井原健二，大賀正一．
  • Organizer: 第51回日本小児内分泌学会
• [Presentation] PIK3R1遺伝子に新生突然変異を認めSHORT症候群と診断した脂肪萎縮性糖尿病の一例． (2017)
  • Author(s): 大久保一宏，石井加奈子，大山紀子，トカン ヴラッド，戸田尚子，海老原健，吉浦孝一郎、井原健二，大賀正一．
  • Organizer: 第27回臨床内分泌代謝Update