Molecular elucidation of antithrombin resistance mechanism and development of thrombophilia model mouse.

• Project/Area Number: 25293129
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Partial Multi-year Fund
• Section: 一般
• Research Field: Laboratory medicine
• Research Institution: Nagoya University
• Principal Investigator: Kojima Tetsuhito 名古屋大学, 医学(系)研究科(研究院), 教授 (40161913)
• Co-Investigator(Kenkyū-buntansha): MATSUSHITA Tadashi 名古屋大学, 医学部附属病院, 教授 (30314008) ; TAKAGI Akira 名古屋大学, 大学院医学系研究科, 准教授 (30135371)
• Project Period (FY): 2013-04-01 – 2016-03-31
• Project Status: Completed (Fiscal Year 2015)
• Budget Amount: ¥18,850,000 (Direct Cost: ¥14,500,000、Indirect Cost: ¥4,350,000); Fiscal Year 2015: ¥5,070,000 (Direct Cost: ¥3,900,000、Indirect Cost: ¥1,170,000); Fiscal Year 2014: ¥5,070,000 (Direct Cost: ¥3,900,000、Indirect Cost: ¥1,170,000); Fiscal Year 2013: ¥8,710,000 (Direct Cost: ¥6,700,000、Indirect Cost: ¥2,010,000)
• Keywords: アンチトロンビン-レジスタンス（ATR） / プロトロンビン異常症 / トロンビン / 遺伝性血栓症 / 一塩基置換 / トロンボモジュリン / R593Lプロトロンビン-ノックインマウス / 相同組換え / ｱﾝﾁﾄﾛﾝﾋﾞﾝ・ﾚｼﾞｽﾀﾝｽ(ATR) / R593Lﾌﾟﾛﾄﾛﾝﾋﾞﾝ-ﾏｳｽ / ﾄﾛﾝﾎﾞﾓｼﾞｭﾘﾝ/ﾌﾟﾛﾃｲﾝC経路 / アンチトロンビン・レジスタンス（ATR） / R593Lプロトロンビン-マウス / トロンボモジュリン／プロテインC経路

Outline of Final Research Achievements

We established a laboratoly test to detect antithromnbin resistance (ATR) in plasma, identified Serbia mutation (p.R596Q) using this method, and newly found 2 distinct Japanese families with ATR caused by Serbian mutation. We reported one of them as the first Japanese thrombophilia family associated with Serbian ATR mutation. In addition, we revealed that mutant thrombin derived from prothrombin Yukuhasi conveying ATR showed thrombomodulin resistance (TMR), in termas of fibrinogen clotting inhibition. Single-base substitution missense variants at 596R of the prothrombin induced ATR and TMR. Furthermore, we established a heterozygous R593L (correspond to R596L in human) knock-in mouse, and observed that 14 out of 40 new born mouse pups in the breeding of the heterozygous mice soon died, suggesting new born lethality of homozygous R593L knock-in mouse.

Research Products

• [Int'l Joint Research] University of Belgrade/Blood Transfusion Institute of Serbia/Clinical Center of Serbia(Serbia)
• [Journal Article] Syndecan 4 Mediates Nrf2-dependent Expansion of Bronchiolar Progenitors That Protect Against Lung Inflammation. (2016)
  • Author(s): Santoso A, Kikuchi T, Tode N, Hirano T, Komatsu R, Damayanti T, Motohashi H, Yamamoto M, Kojima T, Uede T, Nukiwa T, Ichinose M.
  • Journal Title: Mol Ther. Volume: 24 Issue: 1 Pages: 41-52
  • DOI: 10.1038/mt.2015.153
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] The first case of antithrombin-resistant prothrombin Belgrade mutation in Japanese. (2016)
  • Author(s): Kishimoto M, Suzuki N, Murata M, Ogawa M, Kanematsu T, Takagi A, Kiyoi H, Kojima T, Matsushita T
  • Journal Title: Ann Hematol. Volume: 95 Issue: 3 Pages: 541-542
  • DOI: 10.1007/s00277-015-2533-6
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Increased SPHK2 transcription of human colon cancer cells in serum-depleted culture: the involvement of CREB transcription factor. (2015)
  • Author(s): Mizutani N, Omori Y, Tanaka K, Ito H, Takagi A, Kojima T, Nakatochi M, Ogiso H, Kawamoto Y, Nakamura M, Suzuki M, Kyogashima M, Koizumi K, Nozawa Y, Murate T.
  • Journal Title: J Cell Biochem Volume: 116 Issue: 10 Pages: 2227-2238
  • DOI: 10.1002/jcb.25173
  • Peer Reviewed
• [Journal Article] Increased Acid Ceramidase Expression depends on Upregulation of Androgen-dependent Deubiquitinases, USP2, in a Human Prostate Cancer Cell Line, LNCaP. (2015)
  • Author(s): Naoki Mizutani, Minami Inoue, Yukari Omori, Hiromi Ito, Keiko Tamiya-Koizumi, Akira Takagi, Tetsuhito Kojima, Mitsuhiro Nakamura, Soichiro Iwaki, Masahiro Nakatochi, Motoshi Suzuki, Yoshinori Nozawa, Takashi Murate
  • Journal Title: J. Biochem. Volume: 印刷中 Issue: 4 Pages: 309-319
  • DOI: 10.1093/jb/mvv039
  • Peer Reviewed
• [Journal Article] Serum syndecan-4 as a possible biomarker in patients with acute pneumonia (2015)
  • Author(s): Nikaido T, Tanino Y, Wang X, Sato S, Misa K, Fukuhara N, Sato Y, Fukuhara A, Uematsu M, Suzuki Y, Kojima T, Tanino M, Endo Y, Tsuchiya K, Kawamura I, Frevert C, Munakata M
  • Journal Title: J Infect Dis Volume: 212 Issue: 9 Pages: 1500-1508
  • DOI: 10.1093/infdis/jiv234
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Antithrombin resistance: a new mechanism of inherited thrombophilia (2015)
  • Author(s): 小嶋哲人, 高木明, 村田萌, 高木夕希
  • Journal Title: Rinsho Ketsueki Volume: 56 Issue: 6 Pages: 632-638
  • DOI: 10.11406/rinketsu.56.632
  • NAID: 130005093096
  • ISSN: 0485-1439, 1882-0824
  • Peer Reviewed
• [Journal Article] A complex genomic abnormality found in a patient with antithrombin deficiency and autoimmune disease-like symptoms. (2014)
  • Author(s): Kato I, Takagi Y, Ando Y, Nakamura Y, Murata M, Takagi A, Murate T, Matsushita T, Nakashima T, Kojima T.
  • Journal Title: Int J Hematol Volume: 100 Issue: 2 Pages: 200-205
  • DOI: 10.1007/s12185-014-1596-9
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] RhoF Promotes Murine Marginal Zone B Cell Development. (2014)
  • Author(s): Kishimoto M, Matsuda T, Yanase S, Katsumi A, Sezuki N, Ikejiri M, Takagi A, Ikawa M, Kojima T, Kunishima S, Kiyoi H, Naoe T, Matsushita T, Maruyama M
  • Journal Title: Nagoya J Med Sci. Volume: 76 Pages: 293-305
  • NAID: 120005468127
  • Peer Reviewed / Open Access
• [Journal Article] Antithrombin- resistant prothrombin Yukuhashi mutation also causes thrombomodulin resistance in fibrinogen clotting but not in protein C activation. (2014)
  • Author(s): Takagi Y, Kato I, Ando Y, NakamuraY, Murata M, Takagi A, Murate T, Kojima T
  • Journal Title: Thromb Res. Volume: 134 Issue: 4 Pages: 914-917
  • DOI: 10.1016/j.thromres.2014.07.040
  • Peer Reviewed
• [Journal Article] High prophylactic LMWH dose successfully suppressed hemostatic activation in pregnant woman with a new prpthrombin c.1787G>A mutation. (2014)
  • Author(s): Kovac M, Elezovic I, Mikovic V, Djordjevic V, Radojkovic D, Lalic-Cosic S, Murata M, Takagi A, Kojima T
  • Journal Title: Thromb Res. Volume: 135 Issue: 2 Pages: 420-422
  • DOI: 10.1016/j.thromres.2014.12.008
  • Peer Reviewed
• [Journal Article] 新たな血栓性素因：アンチトロンビンレジスタンス (2014)
  • Author(s): 小嶋哲人
  • Journal Title: 日本検査血液学会雑誌 Volume: 15 Pages: 289-296
• [Journal Article] Development of a new laboratory test to evaluate antithrombin resistance in plasma. (2014)
  • Author(s): Murata M, Takagi A, Suzuki A, Okuyama E, Takagi Y, Ando Y, Kato I, Nakamura Y, Murate T, Matsushita T, Saito H, Kojima T.
  • Journal Title: Thromb Res Volume: 133 Issue: 2 Pages: 293-298
  • DOI: 10.1016/j.thromres.2013.11.021
  • Peer Reviewed
• [Journal Article] 1Current venous thromboembolism management and its outcomes in Japan: the nationwide JAVA observational study. (2014)
  • Author(s): Nakamura M, Miyata T, Ozeki Y, Takayama M, Komori K, Yamada N, Origasa H, Satokawa H, Maeda H, Tanabe N, Unno N, Shibuya T, Tanemoto K, Kondo K, Kojima T.
  • Journal Title: Circ J. Volume: 78 Pages: 708-717
  • Peer Reviewed
• [Journal Article] Impact of antithrombin deficiency on efficacy of edoxaban and antithrombin-dependent anticoagulants, fondaparinux, enoxaparin, and heparin. (2013)
  • Author(s): Fukuda T, Kamisato C, Honda Y, Matsushita T, Kojima T, Furugohri T, Morishima Y, Shibano T
  • Journal Title: Thromb Res. Volume: 131 Issue: 6 Pages: 540-546
  • DOI: 10.1016/j.thromres.2013.04.016
  • Peer Reviewed
• [Journal Article] Establishment of mouse model of MYH9 disorders: heterozygous R702C mutation provokes macrothrombocytopenia with leukocyte inclusion bodies, renal glomerulosclerosis and hearing disability. (2013)
  • Author(s): Suzuki N, Kunishima S, Ikejiri M, Maruyama S, Sone M, Takagi A, Ikawa M, Okabe M, Kojima T, Saito H, Naoe T, Matsushita T.
  • Journal Title: PLoS One Volume: 8 Issue: 8 Pages: 41-50
  • DOI: 10.1371/journal.pone.0071187
  • NAID: 120006957397
  • Peer Reviewed
• [Presentation] Practical laboratory assay to detect abnormal prothrombin conveying thrombomodulin resistance. (2015)
  • Author(s): 高木夕希、水谷直貴、村田萌、小塚敏弘、中田悠紀子、長谷部瞭、河村奈美、槇山愛弓、橋本恵梨華、高木明、小嶋哲人
  • Organizer: 第77回日本血液学会学術集会
  • Place of Presentation: 金沢
  • Year and Date: 2015-10-16
• [Presentation] Investigation of molecular mechanisms of PS mRNA up-regulation by progestin isoforms in HepG2 cells. (2015)
  • Author(s): 小塚敏弘、長谷部瞭、中田悠紀子、槇山愛弓、河村奈美、中村友紀、高木夕希、村田萌、水谷直貴、高木明、小嶋哲人
  • Organizer: 第77回日本血液学会学術集会
  • Place of Presentation: 金沢
  • Year and Date: 2015-10-16
• [Presentation] アンチトロンビン抵抗性検出検査法の自動凝固検査機器への最適化 Part 2 (2015)
  • Author(s): 村田 萌、水谷直貴、高木夕希、長谷部瞭、小塚敏弘、中田悠紀子、高木 明、松下 正、小嶋哲人
  • Organizer: 第16回日本検査血液検査血液学会学術集会
  • Place of Presentation: 名古屋
  • Year and Date: 2015-07-11
• [Presentation] 深部静脈血栓症で発症したアンチトロンビンレジスタンス症例の抗凝固療法 (2015)
  • Author(s): 岸本磨由子、鈴木伸明、村田 萌、小川実加、兼松 毅、高木 明、小嶋哲人、松下 正
  • Organizer: 第16回日本検査血液検査血液学会学術集会
  • Place of Presentation: 名古屋
  • Year and Date: 2015-07-11
• [Presentation] 先天性アンチトロンビン欠乏症８症例SERPINC1遺伝子解析 (2015)
  • Author(s): 中田悠紀子、小塚敏弘、長谷部瞭、高木夕希、村田 萌、上牧 務、松尾真稔、高木 明、松下 正、小嶋哲人
  • Organizer: 第16回日本検査血液検査血液学会学術集会
  • Place of Presentation: 名古屋
  • Year and Date: 2015-07-11
• [Presentation] Analysis of prothrombin mutants in NA+ binding domain as a potential candidate conveying antithrombin resistance. (2015)
  • Author(s): Moe Murata, Naoki Mizutani, Yuki Takagi, Ryo Hasebe, Toshihiro Kozuka, Yukiko Nakata, Akira Takagi, Tetsuhito Kojima
  • Organizer: XXV Congress of International Society on Thrombosis and Haemostasis
  • Place of Presentation: Toronto, Canada
  • Year and Date: 2015-06-20
  • Int'l Joint Research
• [Presentation] Analysis of prothrombin missense mutants at 596ARG by single nucleotide substitution for anticoagulant system. (2015)
  • Author(s): Yuki Takagi, Moe Murata, Yuki Nakamura, Toshihiro Kozuka, Yukiko Nakata, Ryo Hasebe, Akira Takagi, Tetsuhito Kojima
  • Organizer: XXV Congress of International Society on Thrombosis and Haemostasis
  • Place of Presentation: Toronto, Canada
  • Year and Date: 2015-06-20
  • Int'l Joint Research
• [Presentation] A case of severe hemophilia B associated with a large insertion of SVA RETROTRANSPOSON in the coagulation factor IX gene. (2015)
  • Author(s): Naoki Mizutani, Yuki Nakamura, Moe Murata, Yuki Takagi, Ryo Hasebe, Toshihiro Kozuka, Yukiko Nakata, Akira Takagi, Jun-ichi Kitazawa, Midori Shima, Tetsuhito Kojima
  • Organizer: XXV Congress of International Society on Thrombosis and Haemostasis
  • Place of Presentation: Toronto, Canada
  • Year and Date: 2015-06-20
  • Int'l Joint Research
• [Presentation] 新たな血栓性素因：アンチトロンビン･レジスタンス (2015)
  • Author(s): 小嶋哲人
  • Organizer: 第25回日本産婦人科新生児血液学会
  • Place of Presentation: 東京
  • Year and Date: 2015-06-05
  • Invited
• [Presentation] Antidotes to dabigatran and anti-Xa anticoagulants (2015)
  • Author(s): 小嶋哲人
  • Organizer: 第37回日本血栓止血学会学術集会
  • Place of Presentation: 甲府
  • Year and Date: 2015-05-21
  • Invited
• [Presentation] プロトロンビン変異による新たなアンチトロンビン抵抗性変異の候補解析 (2015)
  • Author(s): 村田 萌、水谷直貴、高木夕希、長谷部瞭、小塚敏弘、中田悠紀子、槇山愛弓、河村奈美、橋本恵梨華、高木 明、松下 正、小嶋哲人
  • Organizer: 第37回日本血栓止血学会学術集会
  • Place of Presentation: 甲府
  • Year and Date: 2015-05-21
• [Presentation] 深部静脈血栓症で発症しアンチトロンビン抵抗性を示した本邦初 Prothrombin Belgrade 変異の一家系 (2015)
  • Author(s): 岸本磨由子、鈴木伸明、村田萌、小川実加、兼松毅、高木明、小嶋哲人、松下正
  • Organizer: 第37回日本血栓止血学会学術集会
  • Place of Presentation: 甲府
  • Year and Date: 2015-05-21
• [Presentation] プロトロンビン Arg596 における一塩基置換ミスセンス変異体のトロンボモジュリン 抵抗性評価 (2015)
  • Author(s): 高木夕希、村田 萌、中村友紀、小塚敏弘、中田悠紀子、長谷部瞭、橋本恵梨華、高木 明、小嶋哲人
  • Organizer: 第37回日本血栓止血学会学術集会
  • Place of Presentation: 甲府
  • Year and Date: 2015-05-21
• [Presentation] 血小板無力症に同定された GPIIb 遺伝子のミスセンス変異とスプライス変異の複合ヘテロ変異解析 (2015)
  • Author(s): 橋本恵梨華、村田萌、槇山愛弓、河村奈美、小塚敏弘、中田悠紀子、長谷部瞭、高木夕希、水谷直貴、高木明、國島 伸治、松下正、小嶋哲人
  • Organizer: 第37回日本血栓止血学会学術集会
  • Place of Presentation: 甲府
  • Year and Date: 2015-05-21
• [Presentation] Antithrombin resistance: a new mechanism of inherited thrombophilia (2014)
  • Author(s): 小嶋哲人
  • Organizer: 第76回日本血液学会学術集会
  • Place of Presentation: 大阪国際会議場
  • Year and Date: 2014-10-31 – 2014-11-02
  • Invited
• [Presentation] Optimization of the antithrombin resistance assay for the automated analyzer (2014)
  • Author(s): Moe Murata, Yuki Takagi, Yuki Nakamura, Ryo Hasebe, Toshihiro Kozuka, Yukiko Nakata, Akira Takagi, Tetsuhito Kojima
  • Organizer: The 8th Congress of Asia Pacific Society of Thrombosis and Hemostasis (APSTH)
  • Place of Presentation: Hanoi, Vietnam
  • Year and Date: 2014-10-09 – 2014-10-11
• [Presentation] Precise genetic abnormalities in four hemophilia B patients with large deletions of X-chromosome including entire F9 (2014)
  • Author(s): Yuki Nakamura, Moe Murata, Yuki Takagi, Toshihiro Kozuka, Yukiko Nakata, Ryo Hasebe, Akira Takagi, Tadashi Matsushita, Tetsuhito Kojima
  • Organizer: The 8th Congress of Asia Pacific Society of Thrombosis and Hemostasis (APSTH)
  • Place of Presentation: Hanoi, Vietnam
  • Year and Date: 2014-10-09 – 2014-10-11
• [Presentation] 原因不明であった静脈血栓塞栓症にみられたアンチトロンビン抵抗性を示す本邦２家系目のプロトロンビン異常症 (2014)
  • Author(s): 村田 萌、高木夕希、中村友紀、長谷部 瞭、小塚敏弘、中田 悠紀子、高木 明、村手 隆、松下 正、小嶋哲人
  • Organizer: 第33回日本臨床検査医学会東海・北陸支部例会
  • Place of Presentation: 名古屋大学鶴舞キャンパス
  • Year and Date: 2014-08-02
• [Presentation] アンチトロンビン抵抗性検出検査法の自動凝固検査機器への最適化 (2014)
  • Author(s): 村田 萌、高木夕希、中村友紀、長谷部 瞭、小塚敏弘、中田 悠紀子、高木 明、村手 隆、松下 正、小嶋哲人
  • Organizer: 第15回日本検査血液検査血液学会学術集会
  • Place of Presentation: 仙台国際センター
  • Year and Date: 2014-07-20 – 2014-07-21
• [Presentation] A novel mechanism of hereditary thrombosis by antithrombin resistance. (2013)
  • Author(s): Tetsuhito Kojima
  • Organizer: XXIV Congress of nternational Society on Thrombosis and Haemostasis
  • Place of Presentation: Amsterdam, The Neterlands
  • Invited
• [Presentation] Genetic risk factors for thrombophilia in Japanese (2013)
  • Author(s): Tetsuhito Kojima
  • Organizer: Annual Meeting of Korean Society on Thrombosis and Haemostasis
  • Place of Presentation: Seoul, Korea
  • Invited
• [Book] 臨床検査法提要（改訂第34版） (2015)
  • Author(s): 小嶋哲人、高木明
  • Total Pages: 1970
  • Publisher: 金原出版株式会社
• [Book] 新・血栓止血血管学 凝固と炎症 (2015)
  • Author(s): 小嶋哲人
  • Total Pages: 189
  • Publisher: 金芳堂
• [Book] 新・血栓止血血管学 抗凝固と線溶 (2015)
  • Author(s): 小嶋哲人
  • Total Pages: 133
  • Publisher: 金芳堂
• [Book] 血液疾患診療ハンドブック・改訂3版 (2015)
  • Author(s): 鈴木伸明、小嶋哲人
  • Total Pages: 591
  • Publisher: 医薬ジャーナル社
• [Book] Annual Review 2016血液 (2015)
  • Author(s): 高木夕希、小嶋哲人
  • Total Pages: 234
  • Publisher: 中外医学社
• [Book] ファーマナビゲーター抗凝固療法編 (2015)
  • Author(s): 高木夕希、小嶋哲人
  • Total Pages: 415
  • Publisher: 株式会社メディカルレビュー社
• [Book] 徹底ガイド DICのすべて2014-15 (2014)
  • Author(s): 小嶋哲人
  • Total Pages: 392
  • Publisher: 総合医学社
• [Book] 「EBM血液疾患の治療 2015-2016」 (2014)
  • Author(s): 村田萌、小嶋哲人
  • Total Pages: 555
  • Publisher: 中外医学社
• [Book] 改訂第8版 内科学書 Vol.６ 血液・造血器疾患、神経疾患（分担） (2013)
  • Author(s): 小嶋哲人
  • Total Pages: 493
  • Publisher: 中山書店
• [Remarks] 名古屋大学大学院医学系研究科医療技術学専攻病態解析学講座分子病態解析学部門血液・遺伝子研究室
  • URL: http://square.umin.ac.jp/bloodlab/Hematol_%26_Gene_Res_Lab/Welcome.html
• [Remarks] 名古屋大学大学院医学系研究科 医療技術学専攻 病態解析学講座 分子病態解析学部門 血液・遺伝子研究室
  • URL: http://square.umin.ac.jp/bloodlab/Hematol_%26_Gene_Res_Lab/Welcome.html
• [Remarks] JAPANESE THROMBOPHILIA MUTATION DATABASE
  • URL: http://square.umin.ac.jp/bloodlab/Hematol_%26_Gene_Res_Lab/Japanese_Thrombophilia_mutation_database_%28JTMD%29.html
• [Remarks] 名古屋大学大学院医学系研究科 医療技術学専攻 病態解析学講座 分子病態解析学部門 血液・遺伝子研究室
  • URL: http://square.umin.ac.jp/bloodlab/Hematol_%26_Gene_Res_Lab/Welcome.html
• [Remarks] JAPANESE THROMBOPHILIA MUTATION DATABASE
  • URL: http://square.umin.ac.jp/bloodlab/Hematol_%26_Gene_Res_Lab/Japanese_Thrombophilia_mutation_database_%28JTMD%29.html
• [Patent(Industrial Property Rights)] 凝固因子として作用する異常トロンビンのためのトロンビン不活化動態測定方法 (2013)
  • Inventor(s): 高木 明、小嶋 哲人、松下 正
  • Industrial Property Rights Holder: 国立大学法人名古屋大学
  • Industrial Property Rights Type: 特許
  • Filing Date: 2013-04-01
  • Acquisition Date: 2015-10-09
• [Patent(Industrial Property Rights)] 血栓症のリスク検査法 (2013)
  • Inventor(s): 高木明、小嶋哲人、高木夕希、村田萌
  • Industrial Property Rights Holder: 国立大学法人名古屋大学
  • Industrial Property Rights Type: 特許
  • Industrial Property Number: 2013-182541
  • Filing Date: 2013-09-03