1994 年度実績報告書

血栓、止血障害の遺伝子治療を目指した分子生物学的解析

研究課題

研究課題/領域番号	05304036
研究機関	広島大学
研究代表者	蔵本淳広島大学, 原爆放射能医学研究所, 教授 (50034070)
研究分担者	斉藤英彦名古屋大学, 医学部, 教授 (20153819) 大熊稔京都大学, 医学部, 教授 (50026986) 岩永貞昭九州大学, 理学部, 教授 (90029942) 千谷晃一藤田保健衛生大学, 総合医科学研究所, 教授 (60179942) 鈴木宏治三重大学, 医学部, 教授 (70077808)
キーワード	血栓制御因子 / 遺伝子診断 / 遺伝子治療 / 血小板膜レセプター / 外因系凝固機構 / アポリポプロテイン(a)遺伝子 / NF-KB / アンチセンスオリゴマー導入療法
研究概要	血栓制御因子系、凝固因子及びアポ蛋白系、血小板膜レセプター系、血管内皮細胞系の分野についての研究結果は以下のごとくである。 1.先天性血栓制御因子異常症のうち血栓症を繰り返すProtein C,Protein S,α2-plasmin inhibitor(α-PI)異常症或いは低下症について、又血小板機能異常症については,GPIb/IX-Vの低下による粘着能障害を示すBernard -Soulier症候群(BSS)、Thromboxan A2レセプター異常症により出血傾向を起こす症例について遺伝子解析が行われた。その結果これら蛋白やレセプターの機能異常部位や或は分泌異常やレセプター蛋白発現低下の機序が明らかとなった。またフィブリノーゲンレセプター(GPIIb/IIIa)の特異的機能ドメインが組み替え技術を用いて明らかにされた。これらの研究成果は遺伝子治療に対するターゲットをより明確にし、また機能性の高い遺伝子組み替え蛋白を作成することを可能ならしめるとともに、抗血栓薬の開発に貢献する。 2.凝固因子VIIと組織因子(TF)との間の親和性に関係したドメインの研究が行われ、外因系凝固機構が明らかにされた。今後これらの機能異常の解析に有用である。血栓症と関連するアポリポプロテイン(a)遺伝子の転写調節機構が明らかにされつつあり、血栓症の発症予防の可能性が期待される。 3.エンドトキシン、トロンビン,TNFなどにより血管内皮細胞より分泌される炎症性サイトカインやトロンビンによる血管平滑筋細胞増殖は動脈硬化や血栓症を考える上で重要である。これら炎症性サイトカイン分泌や血管平滑筋細胞増殖は転写因子NF-KBによって誘導されることを明らかにした。しかし、アンチセンスNF-KBオリゴマーを内皮細胞に導入するとこれら一連の反応が抑制された。従って本方法は動脈硬化症や血栓症、DICの進展防止治療に有効な治療である事を示したものである。

研究成果
(40件)

すべてその他

すべて文献書誌 (40件)

[文献書誌] Kuramoto A.et al: "Amino terminal regions of the α-subunits of GPIIb/IIIa and the vitronectin receptor possess the epitopes for the complex-specific antibodies A2A9 and LM609,respectively." Blood. 82. 75a- (1993)
[文献書誌] Shimomura T,et al: "Cloning and characterization of the gene encoding the human platelet glycoprotein V." J Biol Chem. 268. 20801- (1993)
[文献書誌] 一瀬白帝: "アポリポプロテイン(a)." カレントテラピー. 11. 84-87 (1993)
[文献書誌] 一瀬白帝: "プラスミノゲンで電子ファミリー" 血液・腫瘍科. 26. 382-390 (1993)
[文献書誌] 一瀬白帝: "線維素溶解現象の開始と制御の分子機序" 山形医学. 11. 15-24 (1993)
[文献書誌] 一瀬白帝: "アポリポプロテイン(a):動脈硬化と血栓症" 日本血栓止血学会誌. 4. 363-371 (1993)
[文献書誌] Kuramoto A.et al: "Expression of platelet membrane glycoprotein V in human megakaryocytes and megakaryocytic cell lines:A study using a novel monoclonal antibody against GPV." Thromb Haemost.72. 762- (1994)
[文献書誌] Kuramoto A.et al: "Nonsanse mutation in the glycoprotein IX coding sequence associated with Bernard-Soulier syndrome." Blood. 82. 742a (1994)
[文献書誌] Kuramoto A.et al: "Precise diagnosis by gene analysis and successful management of delivery in three patients with type IIB von Willebrand disease." Int J Haemat. 60. 163- (1994)
[文献書誌] Hirosawa S.et al: "Secretion of α2-plasmin inhibitor is impaired by amino acid deletion in a small region of the molecule." J.Biochem.115. 293-297 (1994)
[文献書誌] Titani K.et al: "C-type galactoside-binding lectin from Bothrops jararaca venom:comparison of its structure and function with botrocetin." Arch.Biochem.Biophys.308. 306-310 (1994)
[文献書誌] Titani K.et al: "Purification and characterization of bothrombin,a fibrinogen-clotting serine protease from the venom of Bothrops jararaca." Biochemistry. 33. 1843-1849 (1994)
[文献書誌] Titani K.et al: "Immuno-enzymometric analysis for plasma von Willebrand factor degradation in diabetes mellitus using monoclonal antibodies recognizing protease-sensitive sites." Clinical Biochem.27. 123-131 (1994)
[文献書誌] Titai K.et al: "A 28kDa-protein with disintegrin-like structure (Jararhagin-C)purified from Bothrops jararaca venom inhibits collagen-and ADP-induced platelet aggregation." Biochem.Biophys.Res.Commun.201. 331-339 (1994)
[文献書誌] Titani K.et al: "Structural elements influencing von Willebrand factor(vWF)binding affinity for platelet glycoprotein Ib within a dispase-digested vWF fragment." Blood. 84. 1553-1558 (1994)
[文献書誌] Titani K.et al: "Alternatively-spliced isoform of P-selectin is present in vivo as a soluble molecule." J.Biol.Chem.269. 23708-23715 (1994)
[文献書誌] Titani K.et al: "Fibronectin and 130-kDa molecule complex mimics snake venom botrocetin-like structure." J.Biochem.(in press). (1994)
[文献書誌] Titani K.et al: "Isolation and characterization of Bothrops jararaca GPIb-BP,a snake venom antagonist specific to platelet glycoprotein Ib." Thromb.Haemostas.(in press). (1994)
[文献書誌] Titani K.et al: "ABO blood group genotype and plasma von Willebrand factor in normal individuals." Vox Sang.(in press). (1994)
[文献書誌] Okuma M,et al: "Arg^<60> to Leu mutation of the human thromboxane A_2 receptor in a dominantly inherited bleeding disorder." J.Clin.Invest.94. 1662-1667 (1994)
[文献書誌] 大熊稔: "血小板機能異常症" 日内会誌. 83. 259-264 (1994)
[文献書誌] Saito H.et al: "Impaired human tissue factor-mediated activity in blood clotting factor VII Nagoya(Arg304 to Trp)" J.Biol.Chem.269. 7355-7363 (1994)
[文献書誌] Saito H.ey al: "Humoral and cellular immunity to an encode protein induced by direct DNA injection." Human gene therapy. 5. 1335-1339 (1994)
[文献書誌] Suzuki K.et al: "Factor VII Mie:Homozygous asymptomatic type I deficiency caused by an amino acid substitution of His(CAC)for Arg(247)(CGC)in the catalytic domain." Thromb.Haemostas.71. 773-777 (1994)
[文献書誌] Suzuki K.et al: "The role of human factor X activation peptide in activation of factor X by factor IXa." J.Biochem.116. 335-340 (1994)
[文献書誌] Suzuki K.et al: "Receptor-mediated endocytosis of thrombin-antithrombin III complex by the human monocvtoid cell line U937." Biochem.Biphys.Res.Commun.200. 1334-1340 (1994)
[文献書誌] Suzuki K.et al: "The thrombin receptor transmits siggnals through a 330Kda protein kinase in human platelets." Biochem.Biophys.Res.Commun.302. 907-913 (1994)
[文献書誌] Suzuki K.et al: "Blood coagulation factor Xa interacts with a linear sequence of the kringle 2 domain of prothrombin." J. Biochem.116. 589-597 (1994)
[文献書誌] Suzuki K.et al: "Increased vascular endothelial cell markers in patients with chronic renal failure on maintenance hemodialvsis." Blood Coag.Fibrinol.5. 731-717 (1994)
[文献書誌] Suzuki K.et al: "Protein S-Tokushima:Abnormal molecule with a substitution of Glu for Lys-155 in the second epidermal growth factor-like domain of protein S." Blood. 83. 683-690 (1994)
[文献書誌] Maruyama I.et al: "Intravenous extended infusion of recombinant human soluble thrombomodulin prevented tissue factor-induced disseminated intravascular coagulation in rats." Am.J.Hematol.45. 293-303 (1994)
[文献書誌] Maruyama I.et al: "Effect of recombinant human soluble thrombomodulin (rhs-TM)on a rat model of disseminated intravascular coagulation with decreased level of plasma antithrombin III." Thromb.Haemost.71. 452-455 (1994)
[文献書誌] Maruvama I.et al: "Thrombin receptor agonist peptide decreases thrombomodulin activity in cultured human umbilical vein endothelial cells." Biocherm.Biophys.Res.Comm.199. 1262-1269 (1994)
[文献書誌] Kitajima I.et al: "Transcriptional suppression of the HTLV-1 LTR occurs by an unconvention interaction of a CREB with R region." Mol.Cell.Biol.14. 5371-5383 (1994)
[文献書誌] Kitajima I.et al: "E5510 antagonizes thrombin receptor singles by inhibiting NF-kB activation." Biocherm.Biphys.Res.Comm.203. 1181-1187 (1994)
[文献書誌] Kitajima I.et al: "Nitric oxide-mediated apoptosis in murine mastocytoma." Biocherm.Biophys.Res.Comm.204. 244-251 (1994)
[文献書誌] Kitajima I.et al: "Involvement of NF-kB activation in thrombin-induced human smooth muscle cell proliferation." Biocherm.Biophys. Res.Comm.204. 950-955 (1994)
[文献書誌] Kitajima I.et al: "Genomic footprinting of the HTLV-1 LTR in a transgenic mouse tumorigenesis model." Transgene. (in press). (1994)
[文献書誌] Sakata Y.et al: "Effect of glycosidase inhibitors on the biosynthesis,of α2-plasmin inhibitor and antithrombin III." Biochim.Biophys.Acta.1226. 300-306 (1994)
[文献書誌] 一瀬白帝: "血小板・凝固・線溶系疾患と遺伝子異常" バイオクリニカ. 9. 636-666 (1994)

1994 年度 実績報告書

血栓、止血障害の遺伝子治療を目指した分子生物学的解析

研究代表者

蔵本 淳 広島大学, 原爆放射能医学研究所, 教授 (50034070)

研究成果

[文献書誌] Kuramoto A.et al: "Amino terminal regions of the α-subunits of GPIIb/IIIa and the vitronectin receptor possess the epitopes for the complex-specific antibodies A2A9 and LM609,respectively." Blood. 82. 75a- (1993)

[文献書誌] Shimomura T,et al: "Cloning and characterization of the gene encoding the human platelet glycoprotein V." J Biol Chem. 268. 20801- (1993)

[文献書誌] 一瀬白帝: "アポリポプロテイン(a)." カレントテラピー. 11. 84-87 (1993)

[文献書誌] 一瀬白帝: "プラスミノゲンで電子ファミリー" 血液・腫瘍科. 26. 382-390 (1993)

[文献書誌] 一瀬白帝: "線維素溶解現象の開始と制御の分子機序" 山形医学. 11. 15-24 (1993)

[文献書誌] 一瀬白帝: "アポリポプロテイン(a):動脈硬化と血栓症" 日本血栓止血学会誌. 4. 363-371 (1993)

[文献書誌] Kuramoto A.et al: "Expression of platelet membrane glycoprotein V in human megakaryocytes and megakaryocytic cell lines:A study using a novel monoclonal antibody against GPV." Thromb Haemost.72. 762- (1994)

[文献書誌] Kuramoto A.et al: "Nonsanse mutation in the glycoprotein IX coding sequence associated with Bernard-Soulier syndrome." Blood. 82. 742a (1994)

[文献書誌] Kuramoto A.et al: "Precise diagnosis by gene analysis and successful management of delivery in three patients with type IIB von Willebrand disease." Int J Haemat. 60. 163- (1994)

[文献書誌] Hirosawa S.et al: "Secretion of α2-plasmin inhibitor is impaired by amino acid deletion in a small region of the molecule." J.Biochem.115. 293-297 (1994)

[文献書誌] Titani K.et al: "C-type galactoside-binding lectin from Bothrops jararaca venom:comparison of its structure and function with botrocetin." Arch.Biochem.Biophys.308. 306-310 (1994)

[文献書誌] Titani K.et al: "Purification and characterization of bothrombin,a fibrinogen-clotting serine protease from the venom of Bothrops jararaca." Biochemistry. 33. 1843-1849 (1994)

[文献書誌] Titani K.et al: "Immuno-enzymometric analysis for plasma von Willebrand factor degradation in diabetes mellitus using monoclonal antibodies recognizing protease-sensitive sites." Clinical Biochem.27. 123-131 (1994)

[文献書誌] Titai K.et al: "A 28kDa-protein with disintegrin-like structure (Jararhagin-C)purified from Bothrops jararaca venom inhibits collagen-and ADP-induced platelet aggregation." Biochem.Biophys.Res.Commun.201. 331-339 (1994)

[文献書誌] Titani K.et al: "Structural elements influencing von Willebrand factor(vWF)binding affinity for platelet glycoprotein Ib within a dispase-digested vWF fragment." Blood. 84. 1553-1558 (1994)

[文献書誌] Titani K.et al: "Alternatively-spliced isoform of P-selectin is present in vivo as a soluble molecule." J.Biol.Chem.269. 23708-23715 (1994)

[文献書誌] Titani K.et al: "Fibronectin and 130-kDa molecule complex mimics snake venom botrocetin-like structure." J.Biochem.(in press). (1994)

[文献書誌] Titani K.et al: "Isolation and characterization of Bothrops jararaca GPIb-BP,a snake venom antagonist specific to platelet glycoprotein Ib." Thromb.Haemostas.(in press). (1994)

[文献書誌] Titani K.et al: "ABO blood group genotype and plasma von Willebrand factor in normal individuals." Vox Sang.(in press). (1994)

[文献書誌] Okuma M,et al: "Arg^<60> to Leu mutation of the human thromboxane A_2 receptor in a dominantly inherited bleeding disorder." J.Clin.Invest.94. 1662-1667 (1994)

[文献書誌] 大熊 稔: "血小板機能異常症" 日内会誌. 83. 259-264 (1994)

[文献書誌] Saito H.et al: "Impaired human tissue factor-mediated activity in blood clotting factor VII Nagoya(Arg304 to Trp)" J.Biol.Chem.269. 7355-7363 (1994)

[文献書誌] Saito H.ey al: "Humoral and cellular immunity to an encode protein induced by direct DNA injection." Human gene therapy. 5. 1335-1339 (1994)

[文献書誌] Suzuki K.et al: "Factor VII Mie:Homozygous asymptomatic type I deficiency caused by an amino acid substitution of His(CAC)for Arg(247)(CGC)in the catalytic domain." Thromb.Haemostas.71. 773-777 (1994)

[文献書誌] Suzuki K.et al: "The role of human factor X activation peptide in activation of factor X by factor IXa." J.Biochem.116. 335-340 (1994)

[文献書誌] Suzuki K.et al: "Receptor-mediated endocytosis of thrombin-antithrombin III complex by the human monocvtoid cell line U937." Biochem.Biphys.Res.Commun.200. 1334-1340 (1994)

[文献書誌] Suzuki K.et al: "The thrombin receptor transmits siggnals through a 330Kda protein kinase in human platelets." Biochem.Biophys.Res.Commun.302. 907-913 (1994)

[文献書誌] Suzuki K.et al: "Blood coagulation factor Xa interacts with a linear sequence of the kringle 2 domain of prothrombin." J. Biochem.116. 589-597 (1994)

[文献書誌] Suzuki K.et al: "Increased vascular endothelial cell markers in patients with chronic renal failure on maintenance hemodialvsis." Blood Coag.Fibrinol.5. 731-717 (1994)

[文献書誌] Suzuki K.et al: "Protein S-Tokushima:Abnormal molecule with a substitution of Glu for Lys-155 in the second epidermal growth factor-like domain of protein S." Blood. 83. 683-690 (1994)

[文献書誌] Maruyama I.et al: "Intravenous extended infusion of recombinant human soluble thrombomodulin prevented tissue factor-induced disseminated intravascular coagulation in rats." Am.J.Hematol.45. 293-303 (1994)

[文献書誌] Maruyama I.et al: "Effect of recombinant human soluble thrombomodulin (rhs-TM)on a rat model of disseminated intravascular coagulation with decreased level of plasma antithrombin III." Thromb.Haemost.71. 452-455 (1994)

[文献書誌] Maruvama I.et al: "Thrombin receptor agonist peptide decreases thrombomodulin activity in cultured human umbilical vein endothelial cells." Biocherm.Biophys.Res.Comm.199. 1262-1269 (1994)

[文献書誌] Kitajima I.et al: "Transcriptional suppression of the HTLV-1 LTR occurs by an unconvention interaction of a CREB with R region." Mol.Cell.Biol.14. 5371-5383 (1994)

[文献書誌] Kitajima I.et al: "E5510 antagonizes thrombin receptor singles by inhibiting NF-kB activation." Biocherm.Biphys.Res.Comm.203. 1181-1187 (1994)

[文献書誌] Kitajima I.et al: "Nitric oxide-mediated apoptosis in murine mastocytoma." Biocherm.Biophys.Res.Comm.204. 244-251 (1994)

[文献書誌] Kitajima I.et al: "Involvement of NF-kB activation in thrombin-induced human smooth muscle cell proliferation." Biocherm.Biophys. Res.Comm.204. 950-955 (1994)

[文献書誌] Kitajima I.et al: "Genomic footprinting of the HTLV-1 LTR in a transgenic mouse tumorigenesis model." Transgene. (in press). (1994)

[文献書誌] Sakata Y.et al: "Effect of glycosidase inhibitors on the biosynthesis,of α2-plasmin inhibitor and antithrombin III." Biochim.Biophys.Acta.1226. 300-306 (1994)

[文献書誌] 一瀬白帝: "血小板・凝固・線溶系疾患と遺伝子異常" バイオクリニカ. 9. 636-666 (1994)

1994 年度実績報告書

蔵本淳広島大学, 原爆放射能医学研究所, 教授 (50034070)

[文献書誌] 大熊稔: "血小板機能異常症" 日内会誌. 83. 259-264 (1994)