Characteriziation of regulatory molecules of platelet integrin function and their clinical application

• Project/Area Number: 18K08326
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 54010:Hematology and medical oncology-related
• Research Institution: Osaka University
• Principal Investigator: Tomiyama Yoshiaki 大阪大学, 医学部附属病院, 特任教授(常勤) (80252667)
• Co-Investigator(Kenkyū-buntansha): 柏木 浩和 大阪大学, 医学系研究科, 講師 (10432535) ; 加藤 恒 大阪大学, 医学系研究科, 助教 (20705214)
• Project Period (FY): 2018-04-01 – 2021-03-31
• Project Status: Completed (Fiscal Year 2020)
• Budget Amount: ¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000); Fiscal Year 2020: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2019: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2018: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
• Keywords: GPIIb-IIIa / 血小板 / inside-outシグナル / 活性化変異 / kindlin-3 / インテグリン / Kundlin3 / CalDAG-GEFI / 血小板減少 / 後天性血小板無力症 / 自己抗体 / CMK細胞 / Kindlin 3

Outline of Final Research Achievements

The objective of this project is to identify and characterize regulatory molecules that are involved in αIIbβ3 (GPIIb-IIIa) function, especially αIIbβ3 activation. We focused on investigating human disease or human platelets for this purpose. Firstly, we identified a patient with a unique type of acquired Glanzmann thrombasthenia whose platelets lacked GPIIb-IIIa expression on their platelet surface, but contained significant amounts of GPIIb-IIIa in their inside. We identified autoantibody against GPIIb-IIIa which did not inhibit GPIIb-IIIa function, but may induce GPIIb-IIIa deficiency. Secondarily, we characterized αIIb(R990W) knock-in mice to reveal the molecular defects of human GPIIb-IIIa related macrothrombocytopenia(in human αIIb R995W). Finally, we identified the first Japanese case of kindlin-3 deficiency with sever bleeding tendency from a new born. We demonstrated that kindkin-3 is a prerequisite molecule regarding not only αIIbβ3 but also αLβ2 activation.

Academic Significance and Societal Importance of the Research Achievements

血管病、特に動脈血栓性疾患の発症抑制およびその制御法の開発は現代医療に課せられた最重要課題の一つであると言える。血小板は動脈血栓発症の中核を形成するだけでなく、動脈硬化進展や血管新生、組織修復に大きく関与していることが明らかとなってきた。このように血小板機能の制御機構を解明し標的分子を同定することは、出血性疾患の解明のみならず、血管病発症を制御する上で必須のことと考えられる。
この観点より本研究では、申請者は稀で示唆に富むヒトの血小板インテグリンαIIbβ3(GPIIb-IIIa)異常症を同定しその分子異常を解析した。本研究の成果は、より良い血小板機能制御法の開発に寄与するものと確信する。

Research Products

• [Journal Article] Curative potential of fludarabine, melphalan, and non-myeloablative dosage of busulfan in elderly patients with myeloid malignancy (2020)
  • Author(s): Tomoaki Ueda, Tomoyasu Jo, Kazuya Okada, Yasuyuki Arai, Takayuki Sato, Takeshi Maeda, Tatsuhito Onishi, Yasunori Ueda
  • Journal Title: Int J Hematol Volume: 111 Issue: 2 Pages: 247-255
  • DOI: 10.1007/s12185-019-02763-2
  • Peer Reviewed
• [Journal Article] . Efficacy and safety of romiplostim in refractory aplastic anaemia: a Phase II/III, multicentre, open-label study. (2020)
  • Author(s): Jang JH, Tomiyama Y, Miyazaki K, Nagafuji K, Usuki K, Uoshima N, Fujisaki T, Kosugi H, Matsumura I, Sasaki K, Kizaki M, Sawa M, Hidaka M, Kobayashi N, Ichikawa S, Yonemura Y, Enokitani K, Matsuda A, Ozawa K, Mitani K, Lee JW, Nakao S.
  • Journal Title: Br J Haematol. Volume: 192 Issue: 1 Pages: 190-199
  • DOI: 10.1111/bjh.17190
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Risk factors for skin, mucosal, and organ bleeding in adults with primary ITP: a nationwide study in Japan. (2020)
  • Author(s): Hato T, Shimada N, Kurata Y, Kuwana M, Fujimura K, Kashiwagi H, Takafuta T, Murata M, Tomiyama Y.
  • Journal Title: Blood Adv. Volume: 4 Issue: 8 Pages: 1648-1655
  • DOI: 10.1182/bloodadvances.2020001446
  • Peer Reviewed / Open Access
• [Journal Article] Reevaluation of platelet function in chronic immune thrombocytopenia: impacts of platelet size, platelet-associated anti-αIIbβ3 antibodies and thrombopoietin receptor agonists (2020)
  • Author(s): Nishiura N, Kashiwagi H, Akuta K, Hayashi S, Kato H, Kanakura Y, Tomiyama Y.
  • Journal Title: Br J Haematol . Volume: 189 Issue: 4 Pages: 760-771
  • DOI: 10.1111/bjh.16439
  • Peer Reviewed
• [Journal Article] Reference guide for management of adult immune thrombocytopenia in Japan: 2019 Revision. (2020)
  • Author(s): Kashiwagi H, Kuwana M, Hato T, Takafuta T, Fujimura K, Kurata Y, Murata M, Tomiyama Y
  • Journal Title: International Journal of Hematology Volume: 111 Issue: 3 Pages: 329-351
  • DOI: 10.1007/s12185-019-02790-z
  • Peer Reviewed
• [Journal Article] Additional validation of Osaka method (0.01 mol/L dithiothreitol) for negating the daratumumab interference. (2019)
  • Author(s): Hosokawa M, Kashiwagi H, Nakayama K, Sakuragi M, Nakao M, Morikawa T, Kiyokawa T, Aochi H, Nagamine K, Shibayama H, Tomiyama Y.
  • Journal Title: Transfusion. Volume: 59 Issue: 7 Pages: 2479-2480
  • DOI: 10.1111/trf.15305
  • Peer Reviewed / Open Access
• [Journal Article] 血小板β3インテグリンと血小板血栓形成. (2019)
  • Author(s): 冨山佳昭
  • Journal Title: 日本血栓止血学会誌 Volume: 30 Pages: 579-585
  • NAID: 130007689121
  • Peer Reviewed
• [Journal Article] A unique phenotype of acquired Glanzmann thrombasthenia due to non-function-blocking anti-αIIbβ3 autoantibodies (2018)
  • Author(s): Akuta K.、Kashiwagi H.、Yujiri T.、Nishiura N.、Morikawa Y.、Kato H.、Honda S.、Kanakura Y.、Tomiyama Y.
  • Journal Title: Journal of Thrombosis and Haemostasis Volume: 17 Issue: 1 Pages: 206-219
  • DOI: 10.1111/jth.14323
  • Peer Reviewed
• [Journal Article] Distinct effects of daratumumab on indirect and direct antiglobulin tests: a new method employing 0.01 mol/L dithiothreitol for negating the daratumumab interference with preserving K antigenicity (Osaka method). (2018)
  • Author(s): Hosokawa M, Kashiwagi H, Nakayama K, Sakuragi M, Nakao M, Morikawa T, Kiyokawa T, Aochi H, Nagamine K, Shibayama H, Tomiyama Y.
  • Journal Title: Transfusion Volume: 58 Issue: 12 Pages: 3003-3013
  • DOI: 10.1111/trf.14900
  • Peer Reviewed
• [Journal Article] アテローム血栓症発症の分子機構の解明：Patient-Orianted Researchからの解析. (2018)
  • Author(s): 冨山佳昭
  • Journal Title: 日本血栓止血学会誌 Volume: 29 Pages: 765-774
  • Peer Reviewed
• [Journal Article] 抗血小板抗体の検出とその臨床的意義 (2018)
  • Author(s): 冨山佳昭
  • Journal Title: 日本輸血細胞治療学会誌 Volume: 64 Pages: 681-687
  • NAID: 130007556256
• [Presentation] ガイドラインの「科学的根拠」に関する考察. (2020)
  • Author(s): 冨山佳昭
  • Organizer: 第68回日本輸血・細胞治療学会総会（2020.5.29, 紙上開催, 紀野修一）
• [Presentation] ITPと血小板輸血 (2020)
  • Author(s): 柏木浩和, 冨山佳昭
  • Organizer: 第68回日本輸血・細胞治療学会総会（2020.5.29, 紙上開催, 紀野修一）
• [Presentation] 新規のインテグリンαIIbβ3非活性化変異β3(R734C)を認めた一家系 (2020)
  • Author(s): 芥田敬吾, 柏木浩和, 西浦伸子, 加藤恒, 保仙直毅, 金倉譲, 冨山佳昭
  • Organizer: 第42回日本血栓止血学会学術集会（2020.7.1-31, 紙上開催・オンデマンド, 冨山佳昭）
• [Presentation] 血小板機能異常症解析からのメッセージ：血小板血栓制御機構（会長シンポジウム） (2020)
  • Author(s): 加藤 恒
  • Organizer: 第42回日本血栓止血学会学術集会（2020.7.1-31, 紙上開催・オンデマンド, 冨山佳昭）
• [Presentation] Platelet function of chronic immune thrombocytopenia: Impact of platelet size, platelet-associated anti-αIIbβ3 antibodies and thrombopoietin receptor agonists. (2019)
  • Author(s): Nshiura N, Kashiwagi H, Akuta K, Kato H, Kanakura Y, Tomiyama Y
  • Organizer: The XXVII Congress of the International Society of Thrombosis and Haemostasis
  • Int'l Joint Research
• [Presentation] The critical role of Kindlin-3 in initiation of physiological thrombus formation; analysis of Kindlin-3 deficient patient. (2019)
  • Author(s): Kato H, Honda S,Kashiwagi H, Nishiura N,Akuta K, Kokame K, Miyata T, Tomiyama Y
  • Organizer: he American Society of Hematology 61th Annual Meeting
  • Int'l Joint Research
• [Presentation] 新規のインテグリンαIIbβ3非活性化変異β3(R734C)を認めた一家系 (2019)
  • Author(s): 芥田敬吾, 柏木浩和, 西浦伸子, 加藤 恒, 金倉 譲, 冨山佳昭
  • Organizer: 第81回日本血液学会学術集会
• [Presentation] Acquired Glanzmann thrombasthenia due to marked reduction of surface αIIbβ3 expression with non function-blocking anti-αIIbβ3 antibodies. (2018)
  • Author(s): Akuta K, Kashiwagi H, Yujiri T, Nishiura N, Morikawa Y, Kato H, Honda S, Kanakura Y, Tomiyama Y
  • Organizer: The 10th Congress of the Asian-Pacific Society on Thrombosis and Hemostasis
  • Int'l Joint Research
• [Presentation] Platelet integrin αIIbβ3 activation kinetics in inherited platelet functional disorders: the role of ADP receptor P2Y12, CalDAG-GEFI and kindlin-3 in αIIbβ3 activation by inside-out signaling. (2018)
  • Author(s): Kato H, Nishiura N, Akura K, Kashiwagi H, Kokame K, Miyata T, Kanakura Y, Tomiyama Y
  • Organizer: The American Society of Hematology 60th Annual Meeting
  • Int'l Joint Research
• [Presentation] αIIb(R990W), a Gain-of Function Mutation of αIIbβ3, Knock-in Mice Show Moderately Impaired Thrombopoiesis. (2018)
  • Author(s): Akuta K, Kiyomizu K, Kashiwagi H, Kunishima S, Banno F, Kokame K, Nishiura N, Kato H, Honda S, Kanakura Y, Miyata T, Tomiyama Y
  • Organizer: The American Society of Hematology 60th Annual Meeting
  • Int'l Joint Research
• [Presentation] 血小板抗体の検出とその臨床的意義 (2018)
  • Author(s): 冨山佳昭
  • Organizer: 第66回日本輸血・細胞治療学会総会
  • Invited
• [Presentation] 「Shosuke Award」Molecular mechanism of atherothrombosis: lessons from patient-oriented research (2018)
  • Author(s): 冨山佳昭
  • Organizer: 第40回日本血栓止血学会学術集会
  • Invited