Novel mutation search of antithrombin resistance and its pathological analysis research by mouse model

• Project/Area Number: 16K09825
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Hematology
• Research Institution: Nagoya University
• Principal Investigator: Kojima Tetsuhito 名古屋大学, 医学系研究科(保健), 教授 (40161913)
• Co-Investigator(Kenkyū-buntansha): 高木 明 名古屋大学, 医学系研究科(保健), 准教授 (30135371) ; 松下 正 名古屋大学, 医学部附属病院, 教授 (30314008)
• Project Period (FY): 2016-04-01 – 2019-03-31
• Project Status: Completed (Fiscal Year 2018)
• Budget Amount: ¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000); Fiscal Year 2018: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2017: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000); Fiscal Year 2016: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
• Keywords: アンチトロンビン・レジスタンス（ATR） / プロトロンビン / 遺伝子異常 / ATR検出・臨床検査法 / 自動化機器 / ATR残存凝固活性 / Na+結合領域 / Arg593Leuノックインマウス / トロンビン / アンチトロンビン / ジーンタ一ゲティング / R593Lノックインマウス / アンチトロンビン・レシスタンス（ATR） / 人工授精 / 内科

Outline of Final Research Achievements

We have reported dis-prothrombinemia (R596L) showing a novel thrombotic tendency, antithrombin resistance (ATR), and further established the ATR detection test and identified a different mutation (R596Q). In addition, as a result of examining the coagulation activity characteristics of single base substitution mutants in the Na+ binding domain amino acids, K599R and E592Q showed ATR residual coagulation activity equivalent to the reported mutations. Furthermore, although R593L (corresponding to human R596L) knock-in mice showed normal birth and exhibited ATR, the antigen amount of prothrombin was as low as 70%, the one-step coagulation activity was 35%, and the ATR residual coagulation activity was relatively low compared to R596L mutant in human, R593L mutant homozygous mice seemed to have a low risk of thrombosis.

Academic Significance and Societal Importance of the Research Achievements

静脈血栓塞栓症の遺伝子異常判明例の約2/3は原因特定に至っていない中、申請者らが世界で初めて報告した新たな血栓性素因概念 アンチトロンビン・レジスタンス（ATR）の遺伝子異常を検索・解析は、従来にない独創的な発想の研究である。
本研究は、これまで原因特定に至ってない症例でのATR血栓性素因の同定とその詳細な病態解析、遺伝子組換体による新たなATR遺伝子異常の同定解析、ATRマウスの血栓症発症病態解析を行うもので、加齢とともに増加する深部静脈血栓・肺塞栓症の新しい診断法や予防法、治療法の開発に極めて有用な情報を提供することが予想され、超高齢化社会を迎えつつある日本において大きな社会的意義をもつ。

Research Products

• [Int'l Joint Research] University of Belgrade(Serbia)
• [Int'l Joint Research] University of Belgrade(Serbia)
• [Journal Article] Molecular basis of SERPINC1 mutations in Japanese patients with antithrombin deficiency. (2019)
  • Author(s): Tamura S, Hashimoto E, Suzuki N, Kakihara M, Odaira K, Hattori Y, Tokoro M, Suzuki S, Takagi A, Katsumi A, Hayakawa F, Suzuki A, Okamoto S, Kanematsu T, Matsushita T, Kojima T
  • Journal Title: Thrombosis Research Volume: -
  • Peer Reviewed
• [Journal Article] Apparent synonymous mutation F9 c.87A>G causes secretion failure by in-frame mutation with aberrant splicing. (2019)
  • Author(s): Odaira K, Tamura S, Suzuki N, Kakihara M, Hattori Y, Tokoro M, Suzuki S, Takagi A, Katsumi A, Hayakawa F, Okamoto S, Suzuki A, Kanematsu T, Matsushita T, Kojima T
  • Journal Title: Thrombosis Research Volume: -
  • Peer Reviewed
• [Journal Article] Familial pulmonary thromboembolism with a prothrombin mutation and antithrombin resistance. (2018)
  • Author(s): 10)Yoshida R, Seki S, Hasegawa J, Koyama T, Yamazaki K, Takagi A, Kojima T, Yoshimura M
  • Journal Title: J Cardiol Cases. Volume: in press Issue: 6 Pages: 197-199
  • DOI: 10.1016/j.jccase.2018.02.001
  • Peer Reviewed
• [Journal Article] Vwf K1362A resulted in failure of protein synthesis in mice． (2018)
  • Author(s): Sanda N, Suzuki N, Suzuki A, Kanematsu T, Kishimoto M, Hasuwa H, Takagi A, Kojima T, Matsushita T, Nakamura S
  • Journal Title: Int J Hematol. Volume: in press Issue: 4 Pages: 428-435
  • DOI: 10.1007/s12185-017-2394-y
  • NAID: 120006501193
  • Peer Reviewed / Open Access
• [Journal Article] Optimisation of antithrombin resistance assay as a practical clinical laboratory test: Development of prothrombin activator using factors Xa/Va and automation of assay (2018)
  • Author(s): Tamura S.、Suga Y.、Tanamura M.、Murata-Kawakami M.、Takagi Y.、Hottori Y.、Kakihara M.、Suzuki S.、Takagi A.、Kojima T.
  • Journal Title: International Journal of Laboratory Hematology Volume: 40 Issue: 3 Pages: 312-319
  • DOI: 10.1111/ijlh.12786
  • Peer Reviewed
• [Journal Article] Combined deficiency of factors V and VIII by chance coinheritance of parahaemophilia and haemophilia A, but not by mutations of either LMAN1 or MCFD2, in a Japanese family. (2018)
  • Author(s): Suzuki S, Nakamura Y, Suzuki N, Yamazaki T, Takagi Y, Tamura S, Takagi A, Kanematsu T, Matsushita T, Kojima T
  • Journal Title: Haemophilia. Volume: 24 Issue: 1
  • DOI: 10.1111/hae.13360
  • Peer Reviewed / Open Access
• [Journal Article] Diagnosis and treatment guidelines for aberrant portal hemodynamics. (2017)
  • Author(s): Moriyasu F, Furuichi Y, Tanaka A, Takikawa H, Yoshida H, Sakaida I, Obara K, Hashizume M, Kage M, Ohfuji S, Kitano S, Kawasaki S, Kokubu S, Matsutani S, Eguchi S, Shiomi S, Kojima T, Maehara Y, Kuniyoshi Y.
  • Journal Title: Hepatol Res. Volume: 47 Issue: 5 Pages: 373-386
  • DOI: 10.1111/hepr.12862
  • Peer Reviewed / Open Access
• [Journal Article] Clinical and biochemical characterization of the prothrombin Belgrade mutation in a large Serbian pedigree: new insights into the antithrombin resistance mechanism. (2017)
  • Author(s): Miljic P, Gvozdenov M, Takagi Y, Takagi A, Pruner I, Dragojevic M, Tomic B, Bodrozic J, Kojima T, Radojkovic D, Djordjevic V.
  • Journal Title: J Thromb Haemost. Volume: 15 Issue: 4 Pages: 670-677
  • DOI: 10.1111/jth.13618
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] First case report of hemophilia B Leyden in Japan. (2017)
  • Author(s): Yamashita A, Nagae C, Mori M, Ashikaga T, Kojima T, Taki M
  • Journal Title: Int J Hematol. Volume: 106 Issue: 1 Pages: 135-137
  • DOI: 10.1007/s12185-017-2194-4
  • Peer Reviewed
• [Journal Article] In vitro exploration of latent prothrombin mutants conveying antithrombin resistance. (2017)
  • Author(s): Tamura S, Murata-Kawakami M, Takagi Y, Suzuki S, Katsumi A, Takagi A, Kojima T.
  • Journal Title: Thromb Res. Volume: 159 Pages: 33-38
  • DOI: 10.1016/j.thromres.2017.09.020
  • Peer Reviewed
• [Journal Article] Missense mutations in the gene encoding prothrombin corresponding to Arg596 cause antithrombin resistance and thrombomodulin resistance (2016)
  • Author(s): Takagi Y, Murata M, Kozuka T, Nakata Y, Hasebe R, Tamura S, Takagi A, Matsushita T, Saito H, Kojima T.
  • Journal Title: THROMBOSIS AND HAEMOSTASIS Volume: 116 Issue: 12 Pages: 1022-31
  • DOI: 10.1160/th16-03-0223
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Progestin isoforms provide different levels of protein S expression in HepG2 cells. (2016)
  • Author(s): Kozuka, Toshihiro Tamura, Shogo Kawamura, Nami Nakata, Yukiko Hasebe, Ryo Makiyama, Ayumi Takagi, Yuki Murata, Moe Mizutani, Naoki Takagi, Akira Kojima, Tetsuhito
  • Journal Title: Thrombosis research Volume: 145 Pages: 40-5
  • DOI: 10.1016/j.thromres.2016.07.007
  • NAID: 120005895232
  • Peer Reviewed
• [Journal Article] Distinct X chromosomal rearrangements in four haemophilia B patients with entire F9 deletion. (2016)
  • Author(s): Nakamura Y, Ando Y, Takagi Y, Murata M, Kozuka T, Nakata Y, Hasebe R, Takagi A, Matsushita T, Shima M, Kojima T.
  • Journal Title: Haemophilia. Volume: 22 Issue: 3 Pages: 433-439
  • DOI: 10.1111/hae.12849
  • Peer Reviewed / Acknowledgement Compliant
• [Presentation] スプライシング異常を引き起こす血液凝固第IX遺伝子サイレント変異・血友病B分子病態 (2018)
  • Author(s): 大平晃也,田村彰吾,坂根寛人,所真昼,垣原美紗樹,服部有那,橋本恵梨華,鈴木幸子,高木夕希,高木明,兼松毅,岸本麻由子,鈴木伸明,松下正,小嶋哲人
  • Organizer: 第40回日本血栓止血学会学術集会
• [Presentation] F8 Inv22を含む複雑な変異を同定した重症血友病A症例の遺伝学的変異アレル由来の検索 (2018)
  • Author(s): 垣原美紗樹,田村彰吾,服部有那,高木夕希,鈴木幸子,橋本恵梨華,坂根寛人,大平晃也,所真昼,高木明,小川実加,兼松毅,鈴木伸明,松下正,小嶋哲人
  • Organizer: 第40回日本血栓止血学会学術集会
• [Presentation] 脳梗塞を発症したフィブリノケゲン異常症の分子病態機能解析 (2018)
  • Author(s): 服部有那,垣原美紗樹,高木夕希,鈴木幸子,橋本恵梨華,坂根寛人,大平晃也,所真昼,田村彰吾,高木明,鈴木伸明,松下正,小嶋哲人
  • Organizer: 第40回日本血栓止血学会学術集会
• [Presentation] 血液凝固第V因子欠乏症を合併する血友病 A 症例:第2報・凝血学的評価 (2018)
  • Author(s): 鈴木幸子,高木夕希,坂根寛人,橋本恵梨華,垣原美紗樹,服部有那,大平晃也,所真昼,田村彰吾,高木明,鈴木敦夫,鈴木伸明,松下正,山崎鶴夫,小嶋哲人
  • Organizer: 第40回日本血栓止血学会学術集会
• [Presentation] Antithrombin resistance: a unique inherited thrombophilia (2018)
  • Author(s): Tetsuhito Kojima
  • Organizer: The 10th Congress of the Asian-Pacific Society on Thrombosis and Hemostasis
  • Int'l Joint Research / Invited
• [Presentation] Functional analysis of a variant fibrinogen from dysfibrinogenemia patient with cerebral (2018)
  • Author(s): Yuna Hattori, Takagi Yuki, Misaki Kakihara, Sachiko Suzuki, Koya Odaira, Mahiru Tokoro, Shogo Tamura, Tsuyoshi Kanemetsu, Nobuaki Suzuki, Tadashi Matsushita, Tetsuhito Kojima
  • Organizer: The 10th Congress of the Asian-Pacific Society on Thrombosis and Hemostasis
  • Int'l Joint Research
• [Presentation] Haploid origin of unusual Inv22 X-chromosome carrying wild-type telomere region in severe hemophilia A patients (2018)
  • Author(s): Misaki Kakihara, Shogo Tamura, Yuna Hattori, Sachiko Suzuki, Koya Odaira, Mahiru Tokoro, Takeshi Kanematsu, Nobuaki Suzuki, Tadashi Matsushita, Tetsuhito Kojima
  • Organizer: The 10th Congress of the Asian-Pacific Society on Thrombosis and Hemostasis
  • Int'l Joint Research
• [Presentation] 特発生血栓 : アンチトロンビン抵抗性の診断－凝検査法を中心に－ (2018)
  • Author(s): 高木明,田村彰吾,小嶋哲人
  • Organizer: 第19回日本検査血液学会学術集会
  • Invited
• [Presentation] 血漿検体測定を目指したアンチトロンビン抵抗性凝固第X因子検出法の構築 (2018)
  • Author(s): 田村彰吾,高木明,藤岡亮也,服部有那,垣原美紗樹,高木夕希,鈴木幸子,高木夕希,小嶋哲人
  • Organizer: 第19回日本検査血液学会学術集会
• [Presentation] Peri-arteriolar megakaryopoieticmicroenviroment via reciprocal CLEC-2/PDPN axis mouse bone marrow (2018)
  • Author(s): Shogo Tamura, Katsue Suzuki-Inoue, Yukio Ozaki, Tetsuhito Kojima
  • Organizer: 第80回日本血液学会学術集会
  • Invited
• [Presentation] Molecular genetic analysis of dysfibrinogenemia including novel mutations with thrombosis (2018)
  • Author(s): Yuna Hattori, Shogo Tamura, Misaki Kakihara, Sachiko Suzuki, Koya Odaira, Mahiru Tokoro, Humihiko Hayakawa, Tsuyoshi Kanemetsu, Nobuaki Suzuki, Tadashi Matsushita, Tetsuhito Kojima
  • Organizer: 第80回日本血液学会学術集会
• [Presentation] 新しい遺伝性血栓性素因：アンチトロンビン・レジスタンス (2017)
  • Author(s): 小嶋哲人
  • Organizer: 第59回日本小児血液・がん学会学術集会
  • Invited
• [Presentation] Assessment of F9 mutation associated with inhibitor development in Japanese hemophilia B patients (2017)
  • Author(s): Hiroto Sakane, Shogo Tamura, Erika Hashimoto, Akiya Fujioka, Ayumi Makiyama, Nami Kawamura, Sachiko Suzuki, Yuki Takagi, Tsuyoshi Kanematsu, Mayuko Kishimoto, Mika Ogawa, Nobuaki Suzuki, Tadashi Matsushita, Hiroshi Yada, Midori Shima, Tetsuhito Kojima
  • Organizer: XXVI Congress of International Society on Thrombosis and Haemostasis
  • Int'l Joint Research
• [Presentation] Thrombin generation assay CANNOT identify antithrombin resistance during anticoagulant therapy (2017)
  • Author(s): Yuki Takagi, Sachiko Suzuki, Nami Kawamura, Ayumi Makiyama, Hiroto Sakane, Erika Hashimoto, Akiya Fujioka, Shogo Tamura, Akira Takagi, Valentina Djordjevic, Tetsuhito Kojima
  • Organizer: XXVI Congress of International Society on Thrombosis and Haemostasis
  • Int'l Joint Research
• [Presentation] Compound heterozygosity for mutations in ITGA2B including a novel p.Cys198Ser in Glanzmann Thrombasthenia (2017)
  • Author(s): Erika Hashimoto, Shinji Kunishima, Yuki Takagi, Sachiko Suzuki, Ayumi Makiyama, Hiroto Sakane, Akiya Fujioka, Takahiro Uehara, Shogo Tamura, Akira Takagi, Tetsuhito kojima
  • Organizer: XXVI Congress of International Society on Thrombosis and Haemostasis
  • Int'l Joint Research
• [Presentation] Unusual Genomic Rearrangement combined with Inv22 and wild-type X-chromosome in severe hemophilia A patients (2017)
  • Author(s): Shogo Tamura, Misaki Kakihara, Yuna Hattori, Erika Hashimoto, Hiroto Sakane, Yuki Takagi, Mika Ogawa, Takeshi Kanematsu, Nobuaki Suzuki, Akira Takagi, Tadashi Matsushita, Tetsuhito kojima
  • Organizer: XXVI Congress of International Society on Thrombosis and Haemostasis
  • Int'l Joint Research
• [Presentation] A NOVEL LARGE DELETION FOUND IN A JAPANESE FAMILY WITH ANTITHROMBIN DEFICIENCY (2016)
  • Author(s): Erika HASHIMOTO, Yuki TAKAGI, Nami KAWAMURA, Ayumi MAKIYAMA, Hiroto SAKANE, Akiya FUJIOKA, Shogo TAMURA, Akira TAKAGI, Yoshifumi FUKUSHIMA, Makoto KANEKO, Tetsuhito KOJIMA
  • Organizer: The 9th Congress of Asia Pacific Society of Thrombosis and Hemostasis (APSTH)
  • Place of Presentation: Taipei
  • Year and Date: 2016-10-06
  • Int'l Joint Research
• [Presentation] Molecular basis of F8 gene abnormality in hemophilia A patients in Nagoya (2016)
  • Author(s): Nami KAWAMURA, Ayumi MAKIYAMA, Yuki TAKAGI, Erika HASHIMOTO, Hiroto SAKANE, Akiya FUJIOKA, Shogo TAMURA, Akira TAKAGI, Nobuaki SUZUKI, Tadashi MATSUSHITA, Tetsuhito KOJIMA
  • Organizer: The 9th Congress of Asia Pacific Society of Thrombosis and Hemostasis (APSTH)
  • Place of Presentation: Taipei
  • Year and Date: 2016-10-06
  • Int'l Joint Research
• [Presentation] プロテインC欠乏症51家系のPROC遺伝子解析 (2016)
  • Author(s): 槇山愛弓、高木夕希、河村奈美、橋本恵梨華、田村彰吾、高木明、岸本磨由子、鈴木伸明、松下正、小嶋哲人
  • Organizer: 第17回日本検査血液検査血液学会学術集会
  • Place of Presentation: 博多
  • Year and Date: 2016-08-06
• [Presentation] 低フィブリノゲン血症3症例の遺伝子解析 (2016)
  • Author(s): 高木夕希、河村奈美、槇山愛弓、橋本恵梨華、安藤裕実、加藤衣央、田村彰吾、高木明、小嶋哲人
  • Organizer: 第38回日本血栓止血学会学術集会
  • Place of Presentation: 奈良
  • Year and Date: 2016-06-16
• [Presentation] Prothrombin missense mutations at 596Arg reduced the affinity of mutant thrombin to thrombomodulin controlled by Na+ concentration. (2016)
  • Author(s): Yuki Takagi, Nami Kawamura, Ayumi Makiyama, Erika Hashimoto, Shogo Tamura, Akira Takagi, Tetsuhito Kojima
  • Organizer: XXIX International Symposium on Technical Innovations in Laboratory Hematology (ISLH)
  • Place of Presentation: Milano, Italy
  • Year and Date: 2016-05-12
  • Int'l Joint Research
• [Book] 臨床に直結する血栓止血学 (2018)
  • Author(s): 朝倉英策 編著
  • Total Pages: 688
  • Publisher: 中外医学社
  • ISBN: 9784498125797
• [Book] 徹底ガイド DICのすべて 2019-20 (2018)
  • Author(s): 丸藤 哲 特集編集
  • Total Pages: 450
  • Publisher: 総合医学社
  • ISBN: 9784883785599
• [Book] 血液疾患 最新の治療 2017-2019 (2017)
  • Author(s): 兼松毅、小嶋哲人
  • Total Pages: 372
  • Publisher: 南江堂
• [Book] みんなに役立つ 血友病の基礎と臨床 改訂3版 (2016)
  • Author(s): 田村彰吾、小嶋哲人
  • Total Pages: 463
  • Publisher: 医歯ジャーナル社
• [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] Hematology & Gene research Lab
  • 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 病態形跡学講座 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