2000 Fiscal Year Annual Research Report

血栓症の成因と病態、とくに血液凝固とその制御機構障害への分子生物学的アプローチ

Research Project

Project/Area Number	11470250
Research Institution	Jichi Medical School
Principal Investigator	松田道生自治医科大学, 医学部, 教授 (50048980)
Co-Investigator(Kenkyū-buntansha)	三室淳自治医科大学, 医学部, 講師 (10221607) 諏合輝子自治医科大学, 医学部, 講師 (60183844) 坂田洋一自治医科大学, 医学部, 助教授 (40129028) 窓岩清治自治医科大学, 医学部, 講師 (70296119)
Keywords	フィブリノゲン / 遺伝性異常フィブリノゲン / 構造異常 / 血栓症 / 出血傾向 / フィブリン超微細構造 / モノクロナル抗体 / 抗血管新生
Research Abstract	1.血栓症や出血など成因としての異常フィブリノゲン(fibrinogen,Fbg)の解析: 2度の出産に際して重篤な出血を来した36歳の女性に見出された異常Fbg Osaka VIでは、Bβ鎖遺伝子の停止コドン(TAG)がLysを規定する(AAG)に置換しており、C末に12アミノ酸残基断片が付加されていることが判明した。しかも、そのC末第2位のCys残基を介して異常分子間でが形成され、両側あるいは一側でヂスルフィド結合した異常分子のホモダイマー(end-linked homodimer)が電顕学的解析によって証明された。その結果、フィブリン線維は異常に細く且つ分岐に富む緻密な網目を作り、フィブリン網が緻密であるにも拘わらず機械的外力により網目構造は容易に圧縮損傷された。このためOsaka VI Fbn塊は堅固な止血血栓として機能出来ず、出血に結びついたものと推察した。 2.その他:遺伝性異常Fbg分子の研究に資するために、これまでも種々のモノクロナル抗体の作製に努めてきたが、本研究においても白血球由来のエラスターゼによるFbgの分解産物中、Aα(196-204)断片を特異的に認識する抗体を作製することが出来た。この抗体はいわゆる線維素溶解系以外の反応系を介したFbg/Fbnの分解経路の同定に役立つものと期待される。また、プラスミノゲンのクリングル1-4領域に抗血管新生作用があることが指摘されていることから、これを誘導するブタの膵臓エラスターゼの遺伝子を組み込んだマウス実験腫瘍細胞において、腫瘍の発育が著明に阻止されることが示された。血液凝固線維素溶解系の新しい側面が指摘され、今後の検討課題として示されたことになる。その他、接着分子としてのFbg/Fbn、および種々の血液細胞を用いた実験で興味ある成績が得られた。これらの成績はすべて関連する国際的学術誌に報告した(研究業績参照)。

Research Products

(18 results)

All Other

All Publications (18 results)

[Publications] Yang W: "Two-step spreading mode of human glioma cells on fibrin monomer : Interaction of aVb3 with the substratum followed by interaction of a5b1 with endogenous cellular fibronectin secreted in the extracellular matrix."Thromb Res. 93. 279-290 (1999)
[Publications] Mimuro J: "A new type of Ser substitution for g Arg-275 in fibrinogen Kamogawa I characterized by impaired fibrin assembly."Thromb Haemost. 81. 940-944 (1999)
[Publications] Madoiwa S: "Developmental expression of plasminogen activator inhibitor-1 associated with thrombopoietin-dependent megakaryocytic differentiation."Blood. 94. 475-482 (1999)
[Publications] Nakamura M,: "Simultaneous core 2b1→6N-acetylglucosaminyltransferase up-regulation and sialyl-Lex expression during activation of human tonsillar B lymphocytes."FEBS Lett. 463. 125-128 (1999)
[Publications] Matsuda M: "Structure and function of fibrinogen : Insights from dysfibrinogens."Thromb Haemost. 82. 283-290 (1999)
[Publications] DiOrio JP: "Fibrinogen Marburg fibrin network structure."Microsc Microanal. 5. 1120-1121 (1999)
[Publications] Sugo T: "Fibrinogen Niigata : An abnormal fibrinogen with a Bb Asn-160 to Ser substitution associated with extra glycosylation at Bb Asn-158."Blood. 93. 279-290 (1999)
[Publications] Kikuchi J: "Induction of ubiquitin-conjugationg enzyme by aggregated low density lipoprotein in human macrophages and its implications for"Arterioscler Thromb Basc Biol. 20. 128-134 (2000)
[Publications] Kohno I: "A monoclonal antibody specific to the granulocyte-derived elastase-fragment D species of human fibrinogen and fibrin : Its application to the measurement of granulocyte-derived elastase-digests in plasma."Blood. 95. 1721-1728 (2000)
[Publications] Matsuda KM: "A novel strategy for the tumor angiogenesis-targeted gene therapy : Generation of angiostatin from endogenous plasminogen by protease gene transfer."Cancer Gene Therapy. 7. 589-596 (2000)
[Publications] Matsuda M: "Structure and function of fibrinogen inferred from hereditary dysfibrinogens."Fibrinolysis & Proteolysis. 14. 187-197 (2000)
[Publications] Furukawa Y: "Phosphorylation of Bcl-2 protein by CDC2 Kinase during G2/M phases and its role in cell cycle regulation."J Biol Chem. 275. 21661-21667 (2000)
[Publications] Matsuda M: "Structure and function of fibrinogen inferred from hereditary dysfibrinogens."Intern J Haematol. 72. 436-447 (2000)
[Publications] Sugo T: "End-linked homodimers in fibrinogen Osaka VI with a Bb-chain extension lead to fragile clot structure."Blood. 96. 3779-3785 (2000)
[Publications] Kikuchi J: "A simple semisolid subtraction method using carbodiimide-coated microplates."Mol Biotechnol. 15. 193-200 (2000)
[Publications] Furukawa Y: "Lineage-specific regulation of cell cycle control gene expression during haematop oietic cell differentiation."Br J Haemat. 110. 663-673 (2000)
[Publications] Matsuda M: "Structur, function and physiological roles of human fibrinogen."The Japanese Journal of Obstetrical, Gynecologcal & Neonatal Hematology. 9(2). 1-10 (2000)
[Publications] Matsuda M: "Hereditary disorders of fibrinogen"Ann N Y Acad Sci. (in press). (2000)

2000 Fiscal Year Annual Research Report

血栓症の成因と病態、とくに血液凝固とその制御機構障害への分子生物学的アプローチ

Principal Investigator

松田 道生 自治医科大学, 医学部, 教授 (50048980)

Research Products

[Publications] Yang W: "Two-step spreading mode of human glioma cells on fibrin monomer : Interaction of aVb3 with the substratum followed by interaction of a5b1 with endogenous cellular fibronectin secreted in the extracellular matrix."Thromb Res. 93. 279-290 (1999)

[Publications] Mimuro J: "A new type of Ser substitution for g Arg-275 in fibrinogen Kamogawa I characterized by impaired fibrin assembly."Thromb Haemost. 81. 940-944 (1999)

[Publications] Madoiwa S: "Developmental expression of plasminogen activator inhibitor-1 associated with thrombopoietin-dependent megakaryocytic differentiation."Blood. 94. 475-482 (1999)

[Publications] Nakamura M,: "Simultaneous core 2b1→6N-acetylglucosaminyltransferase up-regulation and sialyl-Lex expression during activation of human tonsillar B lymphocytes."FEBS Lett. 463. 125-128 (1999)

[Publications] Matsuda M: "Structure and function of fibrinogen : Insights from dysfibrinogens."Thromb Haemost. 82. 283-290 (1999)

[Publications] DiOrio JP: "Fibrinogen Marburg fibrin network structure."Microsc Microanal. 5. 1120-1121 (1999)

[Publications] Sugo T: "Fibrinogen Niigata : An abnormal fibrinogen with a Bb Asn-160 to Ser substitution associated with extra glycosylation at Bb Asn-158."Blood. 93. 279-290 (1999)

[Publications] Kikuchi J: "Induction of ubiquitin-conjugationg enzyme by aggregated low density lipoprotein in human macrophages and its implications for"Arterioscler Thromb Basc Biol. 20. 128-134 (2000)

[Publications] Kohno I: "A monoclonal antibody specific to the granulocyte-derived elastase-fragment D species of human fibrinogen and fibrin : Its application to the measurement of granulocyte-derived elastase-digests in plasma."Blood. 95. 1721-1728 (2000)

[Publications] Matsuda KM: "A novel strategy for the tumor angiogenesis-targeted gene therapy : Generation of angiostatin from endogenous plasminogen by protease gene transfer."Cancer Gene Therapy. 7. 589-596 (2000)

[Publications] Matsuda M: "Structure and function of fibrinogen inferred from hereditary dysfibrinogens."Fibrinolysis & Proteolysis. 14. 187-197 (2000)

[Publications] Furukawa Y: "Phosphorylation of Bcl-2 protein by CDC2 Kinase during G2/M phases and its role in cell cycle regulation."J Biol Chem. 275. 21661-21667 (2000)

[Publications] Matsuda M: "Structure and function of fibrinogen inferred from hereditary dysfibrinogens."Intern J Haematol. 72. 436-447 (2000)

[Publications] Sugo T: "End-linked homodimers in fibrinogen Osaka VI with a Bb-chain extension lead to fragile clot structure."Blood. 96. 3779-3785 (2000)

[Publications] Kikuchi J: "A simple semisolid subtraction method using carbodiimide-coated microplates."Mol Biotechnol. 15. 193-200 (2000)

[Publications] Furukawa Y: "Lineage-specific regulation of cell cycle control gene expression during haematop oietic cell differentiation."Br J Haemat. 110. 663-673 (2000)

[Publications] Matsuda M: "Structur, function and physiological roles of human fibrinogen."The Japanese Journal of Obstetrical, Gynecologcal & Neonatal Hematology. 9(2). 1-10 (2000)

[Publications] Matsuda M: "Hereditary disorders of fibrinogen"Ann N Y Acad Sci. (in press). (2000)

松田道生自治医科大学, 医学部, 教授 (50048980)