Gene analysis of a novel thrombotic risk factor; antithrombin-resistance.

2012 Fiscal Year Final Research Report

• Project/Area Number: 22590524
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• 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(保健), 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(保健), 助教 (30135371)
• Project Period (FY): 2010 – 2012
• Keywords: 臨床血液学 / 静脈血栓塞栓症 / 先天性血栓性素因 / 遺伝子変異 / アンチトロンビン抵抗性

Research Abstract

Venous thromboembolism is a multifactorial disease resulting from a complex interaction between circumstantial and genetic factors. In this study, we investigated possible causative gene defects in a large Japanese family with inherited thrombophilia, and found a novel missense mutation in the prothrombin gene resulting in a variant prothrombin. The mutation located at one of the antithrombin (AT) binding sites of thrombin molecule. We prepared wild-type and mutant recombinant prothrombins, and compared by prothrombin activation, AT-binding, and thrombin generation assays. In thrombin-antithrombin complex (TAT) formation, the mutant thrombin was impareded to form TAT. In thrombin generation assay of reconstituted prothrombin-deficient plasma with recombinant prothrombins, the mutant showed a decreased peak thrombin and an increased endogenous thrombin potential. We identified a novel F2 mutation leading to the mutant AT-resistant thrombin, which could be a cause of the inherited thrombophilia in this family.

Research Products

• [Journal Article] Ribavirin-induced intracellular GTP depletion activates transcription elongation in coagulation factor VII gene expression (2013)
  • Author(s): Suzuki A, Miyawaki Y, Okuyama E, Murata M, Ando Y, Kato I, Takagi Y, Takagi A, Murate T, Saito H, Kojima T
  • Journal Title: Biochemical J Volume: 449(1) Pages: 231-239
  • Peer Reviewed
• [Journal Article] A possible mechanism for Inv22-related F8 large deletions in severe hemophilia a patients with high responding factor VIII inhibitors (2012)
  • Author(s): Fujita J, Miyawaki Y, Suzuki A, Maki A, Okuyama E, Murata M, Takagi A, Murate T, Suzuki N, Matsushita T, Saito H, Kojima T
  • Journal Title: J Thromb Haemost Volume: 10(10) Pages: 2099-2107
  • Peer Reviewed
• [Journal Article] The author reply (2012)
  • Author(s): Matsushita T, Saito H, Kojima T
  • Journal Title: N Engl J Med Volume: 367 Pages: 1069-1070
  • Peer Reviewed
• [Journal Article] A survey of the clinical course and management of Japanese patients deficient in natural anticoagulants. (2012)
  • Author(s): Yokoyama k, Kojima T, Sakata Y, Kawasaki T, Tsuji H, Miyata T, Okamoto S, Murata M
  • Journal Title: Clin Appl Thromb Hemost Volume: 18(5) Pages: 506-513
  • Peer Reviewed
• [Journal Article] Thrombosis from a prothrombin mutation conveying antithrombin resistance (2012)
  • Author(s): Miyawaki Y, Suzuki A, Fujita J, Maki A, Okuyama E, Murata M, Takagi A, Murate T, Kunishima S, Sakai M, Okamoto K, Matsushita T, Naoe T, Saito H, Kojima K
  • Journal Title: N Engl J Med Volume: 366 Pages: 2390-2396
  • Peer Reviewed
• [Journal Article] A novel ENG mutation causing impaired co-translational processing of endoglin associated with hereditary hemorrhagic telangiectasia. (2012)
  • Author(s): Suzuki A, Nakashima D, Miyawaki Y, Fujita J, Maki A, Fujimori Y, Takagi A, Murate T, Teranishi M, Matsushita T, Saito H, Kojima T
  • Journal Title: Thromb Res Volume: 129(5) Pages: e200-208
  • Peer Reviewed
• [Journal Article] Venous thromboembolism and a race-specific genetic variation, protein S K196E, in Japanese. (2011)
  • Author(s): Miyata T, Hamasaki N, Wada H, Kojima T
  • Journal Title: J Thromb Haemost Volume: (Epub ahead of print)
  • Peer Reviewed
• [Journal Article] Historical perspective and future direction of coagulation research (2011)
  • Author(s): Saito H, Matsushita T, Kojima T
  • Journal Title: J Thromb Haemost Volume: Suppl 1 Pages: 352-63
  • Peer Reviewed
• [Journal Article] Life-threatening hemorrhage and prolonged wound healing are remarkable phenotypes manifested by complete plasminogen activator inhibitor-1 deficiency in humans (2011)
  • Author(s): Iwaki T, Tanaka A, Miyawaki Y, Suzuki A, Kobayashi T, Takamatsu J, Matsushita T, Umemura K, Urano T, Kojima T, Terao T, Kanayama N
  • Journal Title: J Thromb Haemost Volume: 9(6) Pages: 1200-6
  • Peer Reviewed
• [Journal Article] Syndecan-4 deficiency limits neointimal formation after vascular injury by regulating vascular smooth muscle cell proliferation and vascular progenitor cell mobilization. (2011)
  • Author(s): Ikesue M, Matsui Y, Ohta D, Danzaki K, Ito K, Kanayama M, Kurotaki D, Morimoto J, Kojima T, Tsutsui H, Uede T
  • Journal Title: Arterioscler Thromb Vasc Biol Volume: 31(5) Pages: 1066-74
  • Peer Reviewed
• [Journal Article] Syndecan-4 prevents cardiac rupture and dysfunction after myocardial infarction. (2011)
  • Author(s): Matsui Y, Ikesue M, Danzaki K, Morimoto J, Sato M, Tanaka S, Kojima T, Tsutsui H, Uede T
  • Journal Title: Circ Res Volume: 108(11) Pages: 1328-39
  • Peer Reviewed
• [Journal Article] Down-regulation of PROS1 gene expression by 17.-estradiol via estrogen receptor . (ER.)-Sp1 interaction recruiting receptor-interacting protein140 and the corepressor-HDAC3 complex. (2010)
  • Author(s): A Suzuki, N Sanda, Y Miyawaki, Y Fujimori, T Yamada, A Takagi, T Murate, H Saito, T Kojima
  • Journal Title: J. Biol. Chem Volume: 285(18) Pages: 13444-13453
  • Peer Reviewed
• [Journal Article] A novel splice site mutation in intron C of PROS1 leads to markedly reduced mutant mRNA level, absence of thrombin-sensitive region, and impaired secretion and cofactor activity of mutant protein S. (2010)
  • Author(s): H Okada, S Kunishima, M Hamaguchi, A Takagi, K Yamamoto, J Takamatsu, T Matsushita, H Saito, T Kojima, T Yamazaki
  • Journal Title: Thromb Res Volume: 125(5) Pages: e246-e250
  • Peer Reviewed
• [Journal Article] Activated protein C resistance in the Japanese population due to homozygosity for the factor V R2 haplotype (2010)
  • Author(s): H Okada, Y Toyoda, A Takagi, H Saito, T Kojima, T Yamazaki
  • Journal Title: Int J Hematol Volume: 91(3) Pages: 549-550
  • Peer Reviewed
• [Journal Article] Severe hemophilia A in a Japanese female caused by an F8-intron 22 inversion associated with skewed X chromosome inactivation (2010)
  • Author(s): Y Miyawaki, A Suzuki, Y Fujimori, A Takagi, T Murate, N Suzuki, A Katsumi, T Naoe, K Yamamoto, T Matsushita, J Takamatsu, T Kojima
  • Journal Title: Int J Hematol Volume: 92(2) Pages: 405-408
  • Peer Reviewed
• [Presentation] Up-regulation mechanisms of syndecan-4, a heparan sulfate proteoglycan, expression by TNF-a in human endothelial like EAhy926 cells (2012)
  • Author(s): Okuyama E, Murata M, Suzuki A, Ando Y, Kato I, Takagi Y, Takagi A, Murate T, Saito T, Kojima T
  • Organizer: 第35回日本分子生物学会年
  • Place of Presentation: 福岡
  • Year and Date: 20121211-14
• [Presentation] A Possible Mechanism for Inv22-related F8 Large Deletions in Severe Hemophilia A Patients with High Responding Factor VIII Inhibitors (2012)
  • Author(s): Fujita J, Miyawaki Y, Suzuki A, Okuyama E, Murata M, Ando Y, Kato I, Takagi Y, Hosaka M, Takagi A, Murate T, Kajiura Y, Suzuki N, Matsushita T, Saito T, Kojima T
  • Organizer: VIIth Congress of the Asian-Pacific Society on Thrombosis and Hemostasis
  • Place of Presentation: Melbourne, Australia
  • Year and Date: 20121028-31
• [Presentation] Molecular Basis of Coagulation Factor IX Gene Abnormalities in 16 Japanese Hemophilia B Patients (2012)
  • Author(s): Murata M, Okuyama E, Suzuki A, Miyawaki Y, Ando Y, Kato I, Takagi Y, Hosaka M, Takagi A, Murate T, Kishimoto, M, Suzuki N, Matsushita T, Saito T, Kojima T
  • Organizer: VIIth Congress of the Asian-Pacific Society on Thrombosis and Hemostasis
  • Place of Presentation: Melbourne, Australia
  • Year and Date: 20121028-31
• [Presentation] NF-kappaB is essential for Syndecan-4 up-regulation by TNF-alpha in Vascular Endothelial-like EAhy926 Cells (2012)
  • Author(s): Okuyama E, Murata M, Suzuki A, Miyawaki Y, Ando Y, Kato I, Takagi Y, Takagi A, Murate T, Saito T, Kojima T
  • Organizer: VIIth Congress of the Asian-Pacific Society on Thrombosis and Hemostasis
  • Place of Presentation: Melbourne, Australia
  • Year and Date: 20121028-31
• [Presentation] Immunosuppressant IMP dehydrogenase inhibitors up-regulated coagulation factor VII mRNA (2012)
  • Author(s): Suzuki A, Miyawaki Y, Okuyama E, Murata M, Ando Y, Kato I, Takagi Y, Takagi A, Murate T, Saito T, Kojima T
  • Organizer: 第74回日本血液学会学術集会
  • Place of Presentation: 京都
  • Year and Date: 20121019-21
• [Presentation] Molecular mechanisms of syndecan-4 upregulation by TNF-. in the endothelial-like EAhy926 cells (2012)
  • Author(s): Okuyama E, Murata M, Ando Y, Kato I, Takagi Y, Miyawaki Y, Suzuki A, Takagi A, Murate T, Saito T, Kojima T
  • Organizer: 第74回日本血液学会学術集会
  • Place of Presentation: 京都
  • Year and Date: 20121019-21
• [Presentation] 教育講演:基本シリーズ:血栓・止血 血栓性素因の病因と病態 (2011)
  • Author(s): 小嶋哲人
  • Organizer: 第73回日本血液学会学術集会
  • Place of Presentation: 名古屋
  • Year and Date: 20111014-16
• [Presentation] OS-3-120: Molecular basis of an AT-resistant variant thrombin found in a Japanese family with thrombophilia (2011)
  • Author(s): Miyawaki Y, Suzuki A, Fujita J, Maki A, Okuyama E, Murata M, Takagi A, Murate T, Kunishima S, Sakai M, Okamoto K, Matsushita T, Kojima T
  • Organizer: 第73回日本血液学会学術集会
  • Place of Presentation: 名古屋
  • Year and Date: 20111014-16
• [Presentation] PS-1-63: Thrombophilia and JAK2 V617F mutations in Japanese patients with Budd-Chiari syndrome (2011)
  • Author(s): Maki A, Fujita J, Okuyama E, Murata M, Miyawaki Y, Suzuki A, Takagi A, Murate T, Kojima T
  • Organizer: 第73回日本血液学会学術集会
  • Place of Presentation: 名古屋
  • Year and Date: 20111014-16
• [Presentation] PS-1-72: Unusual X-chromosomal rearrangements with F8-int22 inversion in three severe hemophilia A patients (2011)
  • Author(s): Fujita J, Miyawaki Y, Suzuki A, Maki A, Okuyama E, Murata M, Takagi A, Murate T, Matsushita T, Suzuki N, Saito H, Kojima T
  • Organizer: 第73回日本血液学会学術集会
  • Place of Presentation: 名古屋
  • Year and Date: 20111014-16
• [Presentation] A NOVEL PROTHROMBIN GENE MUTATION LEADS TO AN AT-RESISTANT THROMBIN IN A FAMILY WITH INHERITED THROMBOPHILIA (2011)
  • Author(s): Miyawaki Y, Suzuki A, Fujimori Y, Fujita J, Maki A, Takagi A, Murate T, Sakai M, Okamoto K, Matsushita T, Kojima T
  • Organizer: XXIIIrd Congress of International Society on Thrombosis and Haemostasis
  • Place of Presentation: Kyoto, Japan
  • Year and Date: 20110723-28
• [Presentation] DISTINCT X-CHROMOSOMAL REARRANGEMENTS WITH F8-INTRON22 INVERSION IN THREE UNRELATED JAPANESE SEVERE HEMOPHILIA A PATIENTS WITH HIGH TITER INHIBITOR (2011)
  • Author(s): Fujita J, Suzuki A, Miyawaki Y, Maki A, Fujimori Y, Takagi A, Murate T, Matsushita T, Suzuki N, Saito H, Kojima T
  • Organizer: XXIIIrd Congress of International Society on Thrombosis and Haemostasis
  • Place of Presentation: Kyoto, Japan
  • Year and Date: 20110723-28
• [Presentation] Historical perspective and future direction of coagulation research (2011)
  • Author(s): Saito H, Matsushita T, Kojima T
  • Organizer: XXIIIrd Congress of International Society on Thrombosis and Haemostasis
  • Place of Presentation: XXIIIrd Congress of International Society on Thrombosis and Haemostasis
  • Year and Date: 20110723-28
• [Presentation] Receptor interacting protein 140 (RIP140) mediated ERα-Sp1 and corepressor-HDAC3 complex association in 17β-estradiol dependent down-regulation of protein S (2010)
  • Author(s): A Suzuki, Y Miyawaki, J Fujita, A Maki, Y Fujimori, A Takagi, T Murate, H Saito, T Kojima
  • Organizer: 第33回日本分子生物学会
  • Place of Presentation: 神戸
  • Year and Date: 20101207-10
• [Presentation] Genetic Disorders Related to Coagulopathy Prevalent in the Far East (2010)
  • Author(s): T Kojima
  • Organizer: 6th Congress of the Asia Pacific Society on Thrombosis and Haemostasis (APSTH Bali 2010)
  • Place of Presentation: Bali
  • Year and Date: 20101013-16
• [Presentation] A survey of venous thromboembolism in Japanese patients with inherited anticoagulant deficiency (2010)
  • Author(s): K Yokoyama, T Kojima, Y Sakata, T Kawasaki, H Tsuji, T Miyata, S Okamoto, M Murata
  • Organizer: 第72回日本血液学会総会
  • Place of Presentation: 横浜
  • Year and Date: 20100924-26
• [Presentation] A novel endoglin gene mutation associated with hereditary hemorrhagic telangiectasia in a Japanese (2010)
  • Author(s): A Suzuki, Y Miyawaki, J Fujita, A Maki, Y Fujimori, A Takagi, T Murate, M Teranishi, H Saito, T Kojima
  • Organizer: 第72回日本血液学会総会
  • Place of Presentation: 横浜
  • Year and Date: 20100924-26
• [Book] あなたも名医!新しい経口抗凝固薬、どう使う (2012)
  • Author(s): 小嶋哲人
  • Total Pages: 149-152
  • Publisher: 日本医事新報社
• [Book] フォンダパリヌクスの基礎循環器内科医のためのXa阻害薬のすべて (2012)
  • Author(s): 小嶋哲人
  • Total Pages: 147-151
  • Publisher: メディカルレビュー社
• [Book] わかりやすい血栓と止血の臨床 (2011)
  • Author(s): 小嶋哲人
  • Total Pages: 107-109
  • Publisher: 南江堂
• [Book] 抗血栓薬の最前線 -基礎と臨床 (2011)
  • Author(s): 小嶋哲人
  • Total Pages: 75-81
  • Publisher: 医薬ジャーナル社
• [Book] 血液疾患 最新の治療 2011-2013 (2010)
  • Author(s): 小嶋哲人
  • Total Pages: 271-274
  • Publisher: 南江堂
• [Remarks]
  • URL: http://www.met.nagoya-u.ac.jp/KENSAWEB/labo/blood/index.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及び試験方法、並びに、ポリヌクレオチド (2010)
  • 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 Number: 特許 2010-289686
  • Filing Date: 2010-12-27