1997 Fiscal Year Annual Research Report

血管内皮由来ペプチドの血管再構築における臨床的意義の検討と遺伝子治療への応用

Research Project

Project/Area Number	08457264
Research Institution	KYOTO UNIVERSITY
Principal Investigator	伊藤裕京都大学, 医学研究科, 助手 (40252457)
Co-Investigator(Kenkyū-buntansha)	細田公則京都大学, 人間環境学研究科, 助手 (40271598)
Keywords	血管内皮細胞 / 血管平滑筋細胞 / 増殖 / ナトリウム利尿ペプチド / cGMP / トランスジェニックマウス
Research Abstract	我々は、これまで新しい内皮由来血管弛緩ペプチドであるCNPcDNAを組み込んだ非増殖性アデノウィルスを構築し、培養血管平滑筋細胞(VSMC)に感染させることで、VSMCのG1期増殖抑制が生じ、更にこのCNPによるVSMC増殖抑制において、最近単離同定された心血管系特異的ホメオボックス遺伝子Gax(Growth Arrest-specific Homeobox)が関与することを証明した。本年度は、我々が既にクローニングしたcGMPカスケードの主要なシグナル伝達分子と考えられるヒトcGMP依存性プロティンキナーゼ(Gキナーゼ)I_αに注目し、cGMPカスケードの血管再構築における意義を発生工学的手法を用い検討した。ヒトGキナーゼI_α完全長cDNAを、βアクチンプロモーター、サイトメガロウィルスエンハンサーを有する発現ベクターに組み込み、培養血管平滑筋細胞にtransfectionしたところ、cGMP添加により血管平滑筋細胞の遊走、増殖が有意に抑制された。次に、この発現ベクターを用いGキナーゼ過剰発現トランスジェニックマウスの開発を試みた。その結果、少なくとも10コピー以上のGキナーゼトランスジーンを有する2系統のF1マウスを得た。GキナーゼmRNAのNorthern blot解析及びGキナーゼ特異的kination assayを用い、このトランスジェニックマウスにおいて著名なGハナーゼ発現が心血管系において認められることを明らかにした。今後、このGキナーゼ過剰発現マウスを用いCNPを始めとする血管内皮由来ペプチドとcGMPカスケードの血管再構築における意義をin vivoで検討する予定である。

Research Products
(17 results)

All Other

All Publications (17 results)

[Publications] J.Yamashita, et al.: "Opposite regulation of Gax homeobox expression by angiotensin II and C-type natriuretic peptide." Hypertension. 29. 381-387 (1997)
[Publications] T-H.Chun, et al.: "Shear stress augments expression of C-type natriuretic peptide and adrenomedullin." Hypertension. 29. 1296-1302 (1997)
[Publications] T.Ikeda, et al.: "The plasma levels of brain and atrial natriuretic peptides elevate in proportion to the left ventricular end-systolic wall stress in patients with aortic stenosis." Am.Heart J.133. 307-314 (1997)
[Publications] H.Itoh, et al.: "Impaired guanosine 3',5'-cyclin phosphate production in severe pregnancy-induced hypertension with high plasma levels of atrial and brain natriuretic pepitdes." Endocrine J.44. 389-393 (1997)
[Publications] K.Yoshimine, et al.: "Altered expression of atrial natriuretic peptide and contractile protein genes in hypertrophied ventricle of JVS mice with systemic carnitine deficiency." J.Mol.Cell Cardiol.29. 571-578 (1997)
[Publications] H.Itoh, et al.: "Significance of vascular natriuretic peptide system in vascular remodeling in humans and its application to gene therapy." Annals New York Academy of Sciences. 811. 533-541 (1997)
[Publications] M.Harada, et al.: "Significance of ventricular myocytes and nonmyocytes interaction during cardiocyte hypertrophy-Evidence for endothelin-1 as a paracrine hypertrophic factor from cardiac nonmyocytes." Circulation. 96. 3737-3744 (1997)
[Publications] K.Doi, et al.: "Adenovirus-mediated gene transfer of C-type natriuretic peptide causes G1 growth inhibition of cultured vascular smooth muscle cells." Biochem.Biophys.Res.Commun.239. 889-894 (1997)
[Publications] J.Matsuda, et al.: "Cloning of rat uncoupling protein-3 and uncoupling protein-2 cDNAs : their gene expression in rats fed high-fat diet." FEBS Letters. 418. 200-204 (1997)
[Publications] K.Doi, et al.: "Expression of natriuretic peptide system during embryonic stem cell vasculogenesis." Heart and Vessels. Suppl.12. 18-22 (1997)
[Publications] M.Harada, et al.: "Role of cardiac nonmyocytes in cyclic mechanical stretch-induced myocyte hypertrophy." Heart and Vessels. Suppl.12. 198-200 (1997)
[Publications] K.Masatsugu, et al.: "Physiologic shear stress suppresses endothelin-converting enzyme-1 expression in vascular endothelial cells." J.Cardiovasc.Pharmacol.31 Suppl.1. S42-S45 (1998)
[Publications] T.Saito, et al.: "Oxidative stress suppresses the endothelial secretion of endothelin." J.Cardiovasc.Pharmacol.31 Suppl.1. S45-S47 (1998)
[Publications] S.Suga, et al.: "Regulation of endothelial production of C-type natriuretic peptide by interaction between endothelial cells and macrophages." Endocrinology. (in press). (1998)
[Publications] M.Suda, et al.: "Skeletal overgrowth in transgenic mice that overexpress brain natriuretic peptide." Proc.Natl.Acad.Sci,U.S.A.(in press). (1998)
[Publications] Y.Numata, et al.: "An immunoradiometric assay kit for N-terminal fragment of proatrial natriuretic peptide (ProANP N-peptide) in human plasma as a useful clinical test for heart failure." Endocrinol.Metabol.(in press). (1998)
[Publications] H.Itho, et al.ed by W.K.Samson and E.R.Levin: ""Contemporary Endocrinology : Natriuretic Peptides in Health and Disease"" Humana Press Inc.,Totowa,NJ, (1997)

1997 Fiscal Year Annual Research Report

血管内皮由来ペプチドの血管再構築における臨床的意義の検討と遺伝子治療への応用

Principal Investigator

伊藤 裕 京都大学, 医学研究科, 助手 (40252457)

Research Products

[Publications] J.Yamashita, et al.: "Opposite regulation of Gax homeobox expression by angiotensin II and C-type natriuretic peptide." Hypertension. 29. 381-387 (1997)

[Publications] T-H.Chun, et al.: "Shear stress augments expression of C-type natriuretic peptide and adrenomedullin." Hypertension. 29. 1296-1302 (1997)

[Publications] T.Ikeda, et al.: "The plasma levels of brain and atrial natriuretic peptides elevate in proportion to the left ventricular end-systolic wall stress in patients with aortic stenosis." Am.Heart J.133. 307-314 (1997)

[Publications] H.Itoh, et al.: "Impaired guanosine 3',5'-cyclin phosphate production in severe pregnancy-induced hypertension with high plasma levels of atrial and brain natriuretic pepitdes." Endocrine J.44. 389-393 (1997)

[Publications] K.Yoshimine, et al.: "Altered expression of atrial natriuretic peptide and contractile protein genes in hypertrophied ventricle of JVS mice with systemic carnitine deficiency." J.Mol.Cell Cardiol.29. 571-578 (1997)

[Publications] H.Itoh, et al.: "Significance of vascular natriuretic peptide system in vascular remodeling in humans and its application to gene therapy." Annals New York Academy of Sciences. 811. 533-541 (1997)

[Publications] M.Harada, et al.: "Significance of ventricular myocytes and nonmyocytes interaction during cardiocyte hypertrophy-Evidence for endothelin-1 as a paracrine hypertrophic factor from cardiac nonmyocytes." Circulation. 96. 3737-3744 (1997)

[Publications] K.Doi, et al.: "Adenovirus-mediated gene transfer of C-type natriuretic peptide causes G1 growth inhibition of cultured vascular smooth muscle cells." Biochem.Biophys.Res.Commun.239. 889-894 (1997)

[Publications] J.Matsuda, et al.: "Cloning of rat uncoupling protein-3 and uncoupling protein-2 cDNAs : their gene expression in rats fed high-fat diet." FEBS Letters. 418. 200-204 (1997)

[Publications] K.Doi, et al.: "Expression of natriuretic peptide system during embryonic stem cell vasculogenesis." Heart and Vessels. Suppl.12. 18-22 (1997)

[Publications] M.Harada, et al.: "Role of cardiac nonmyocytes in cyclic mechanical stretch-induced myocyte hypertrophy." Heart and Vessels. Suppl.12. 198-200 (1997)

[Publications] K.Masatsugu, et al.: "Physiologic shear stress suppresses endothelin-converting enzyme-1 expression in vascular endothelial cells." J.Cardiovasc.Pharmacol.31 Suppl.1. S42-S45 (1998)

[Publications] T.Saito, et al.: "Oxidative stress suppresses the endothelial secretion of endothelin." J.Cardiovasc.Pharmacol.31 Suppl.1. S45-S47 (1998)

[Publications] S.Suga, et al.: "Regulation of endothelial production of C-type natriuretic peptide by interaction between endothelial cells and macrophages." Endocrinology. (in press). (1998)

[Publications] M.Suda, et al.: "Skeletal overgrowth in transgenic mice that overexpress brain natriuretic peptide." Proc.Natl.Acad.Sci,U.S.A.(in press). (1998)

[Publications] Y.Numata, et al.: "An immunoradiometric assay kit for N-terminal fragment of proatrial natriuretic peptide (ProANP N-peptide) in human plasma as a useful clinical test for heart failure." Endocrinol.Metabol.(in press). (1998)

[Publications] H.Itho, et al.ed by W.K.Samson and E.R.Levin: ""Contemporary Endocrinology : Natriuretic Peptides in Health and Disease"" Humana Press Inc.,Totowa,NJ, (1997)

伊藤裕京都大学, 医学研究科, 助手 (40252457)