1996 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 Category	Grant-in-Aid for Scientific Research (B)
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	アデノウィルス / 血管平滑筋細胞 / 内皮細胞 / cGMP / 増殖 / ホメオボックス / ずり応力(shear stress) / 動脈硬化
Research Abstract	1.我々が明らかにした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た。このCNPアデノウィルスを培養血管平滑筋細胞(VSMC)に感染させ、内皮細胞の10000倍に及ぶCNP産生を確認した。その結果、VSMCのG1期増殖抑制を認めた。この際、cGMPによるp21の活性化に伴うcyclin E,cyclin Aの活性化の低下を認めた。 2.CNPによるVSMC増殖抑制の分子機構を明らかにするため、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(Grpwth Arrest-specific Homeobox)の発現を検討したところ、血管収縮因子であり血管増殖に対し促進的に働くアンジオテンシンIIによりGax発現は著しく抑制され、一方、血管増殖抑制因子であるCNPによりその発現が亢進することを明らかにした。更に、Gax mRNAに対するアンセチンスオリゴヌレクオチド投与によりGax発現を抑制すると、cGMPカスケードによるVSMCの増殖抑制作用はブロックされた。以上より、血管内皮由来ペプチドによるvascular growthにおける共通の転写因子Gaxの意義が明らかとなった。 3.血管内皮由来ペプチドであるCNP及び最近副腎より単離同定され血管でもその発現を認める血管弛緩物質であるアドレノメジュリン(AM)に関して、ずり応力(shear stress)によりその発現がup-regulationされることを明らかにした。またshear stressの低下部に動脈硬化病変が好発することが知られているが、実際ヒト冠動脈硬化病変において、内皮細胞でのCNPの発現が低下していることを明らかにした。 4.ナトリウム利尿ペプチド及び一酸化窒素NOの共通のシグナル伝達経路であるcGMPカスケードの主要なシグナル伝達物質であるヒトcGMP依存性プロティンキナーゼ(G-キナーゼ)タイプIαをクローニングした。更に、その持続活性化ミュータントを作成し、これを用い現在トランスジェニックマウスを開発中である。

Research Products

(20 results)

All Other

All Publications (20 results)

[Publications] T.Ikeda,et al.: "Natriuretic peptide receptors in human arterial and venous coronary bypass vessels and rabbit vein grafts." Hypertension. 27. 833-837 (1996)
[Publications] K.Doi,et al.: "Vascular cndothelial growth factor suppresses C-type natriurctic peptide secretion." Hypertension. 27. 811-815 (1996)
[Publications] N.Tamura,et al.: "cDNA cloning and gene expression of human type I alpha cGNP-dependent protein kinase." Hypertension. 27. 552-557 (1996)
[Publications] Y.Komatsu,et al.: "Regulation of endothelial production of C-type natriuretic peptide in coculture with vascular smooth muscle cells--The role of vascular natriuretic peptide system in vascular growht inhibition" Circ.Res.78. 606-614 (1996)
[Publications] Y.Komatsu,et al.: "Regulation of secretion and clearance of C-type natriuretic peptide in the interaction of vascular endothelial cells and smooth muscle cells." J.Hypertens.14. 585-592 (1996)
[Publications] Y.Miyamoto,et al.: "Alternative RNA splicing of human endothelin-A receptor generates multiple transcripts." Biochemical J.313. 795-801 (1996)
[Publications] T.Igaki,et al.: "C-type natriuretic peptide in chronic renal failure and its action in humans." Kidney Int.49 Suppl 55. S144-S147 (1996)
[Publications] T.Igaki,et al.: "Insulin suppresses endothelial sccretion of C-type natriuretic peptide, a novei endothelium-derived relaxing peptide." Diabetes. 45 Suppl 3. S62-S64 (1996)
[Publications] K.Takaya,et al.: "The RET proto-oncogene in sporadic pheochromicytomas." Internal Medicine. 35. 449-452 (1996)
[Publications] M.Suda,et al.: "C-type natriuretic peptide as an autocrine/paracrine regulator of osteoblast." Biochem.Biophys.Res.Commun.223. 1-6 (1996)
[Publications] R.Iwata,et al.: "Sensitive chemilunimescent enzyine immunoassay for deternination of endothelin-1." Jpn.J.Clin.Chem.25. 91-96 (1996)
[Publications] R.Iwata,et al.: "Direct determination of plasma endothelin-1 by chemiluminescence enzyme immunoassay." Clin.Chem.48. 1155-1158 (1996)
[Publications] K.Nakao,et al.: "The natriuretic peptide family." Curr.Opin.Niphrol.Hypertens.5. 4-11 (1996)
[Publications] N.Tamura,et al.: "Two cardiac natriuretic peptide genes (atrial natriuretic peptide and brain natriuretic peptide) are organized in tandem in the mouse and human genomes." J.Mol.Cell.Cardiol.28. 1811-1815 (1996)
[Publications] T.Matsumoto,et al.: "Type 2 angiotensin II receptor is expressed in human myometrium and uterine leiomyoma and is down-regulated during pregnancy." J.Clin.Endocrinol.Metab.81. 4366-4372 (1996)
[Publications] K.Takaya,et al.: "Expression of the RET proto-oncogene in normal human tissues,pheochromocytomas,and other tumors of neural crest origin." J.Mol.Med.74. 617-621 (1996)
[Publications] T.Naruko,et al.: "C-type natriuretic peptide in human coronary atherosclerotic lesions." Circulation. 94. 3103-3108 (1996)
[Publications] J.Yamashita,et al.: "Opposite regulation of Gax homeobox expression by angiotensin II and C-type natriuretic peptide." Hyertension. 29 part 2. 381-387 (1997)
[Publications] T-H.Chun,et al.: "Shear stress augments expression of C-type natriuretic peptide and adrenomedullin." Hyertension. (in press). (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." Amerecan Heart J.(in press). (1997)

1996 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] T.Ikeda,et al.: "Natriuretic peptide receptors in human arterial and venous coronary bypass vessels and rabbit vein grafts." Hypertension. 27. 833-837 (1996)

[Publications] K.Doi,et al.: "Vascular cndothelial growth factor suppresses C-type natriurctic peptide secretion." Hypertension. 27. 811-815 (1996)

[Publications] N.Tamura,et al.: "cDNA cloning and gene expression of human type I alpha cGNP-dependent protein kinase." Hypertension. 27. 552-557 (1996)

[Publications] Y.Komatsu,et al.: "Regulation of endothelial production of C-type natriuretic peptide in coculture with vascular smooth muscle cells--The role of vascular natriuretic peptide system in vascular growht inhibition" Circ.Res.78. 606-614 (1996)

[Publications] Y.Komatsu,et al.: "Regulation of secretion and clearance of C-type natriuretic peptide in the interaction of vascular endothelial cells and smooth muscle cells." J.Hypertens.14. 585-592 (1996)

[Publications] Y.Miyamoto,et al.: "Alternative RNA splicing of human endothelin-A receptor generates multiple transcripts." Biochemical J.313. 795-801 (1996)

[Publications] T.Igaki,et al.: "C-type natriuretic peptide in chronic renal failure and its action in humans." Kidney Int.49 Suppl 55. S144-S147 (1996)

[Publications] T.Igaki,et al.: "Insulin suppresses endothelial sccretion of C-type natriuretic peptide, a novei endothelium-derived relaxing peptide." Diabetes. 45 Suppl 3. S62-S64 (1996)

[Publications] K.Takaya,et al.: "The RET proto-oncogene in sporadic pheochromicytomas." Internal Medicine. 35. 449-452 (1996)

[Publications] M.Suda,et al.: "C-type natriuretic peptide as an autocrine/paracrine regulator of osteoblast." Biochem.Biophys.Res.Commun.223. 1-6 (1996)

[Publications] R.Iwata,et al.: "Sensitive chemilunimescent enzyine immunoassay for deternination of endothelin-1." Jpn.J.Clin.Chem.25. 91-96 (1996)

[Publications] R.Iwata,et al.: "Direct determination of plasma endothelin-1 by chemiluminescence enzyme immunoassay." Clin.Chem.48. 1155-1158 (1996)

[Publications] K.Nakao,et al.: "The natriuretic peptide family." Curr.Opin.Niphrol.Hypertens.5. 4-11 (1996)

[Publications] N.Tamura,et al.: "Two cardiac natriuretic peptide genes (atrial natriuretic peptide and brain natriuretic peptide) are organized in tandem in the mouse and human genomes." J.Mol.Cell.Cardiol.28. 1811-1815 (1996)

[Publications] T.Matsumoto,et al.: "Type 2 angiotensin II receptor is expressed in human myometrium and uterine leiomyoma and is down-regulated during pregnancy." J.Clin.Endocrinol.Metab.81. 4366-4372 (1996)

[Publications] K.Takaya,et al.: "Expression of the RET proto-oncogene in normal human tissues,pheochromocytomas,and other tumors of neural crest origin." J.Mol.Med.74. 617-621 (1996)

[Publications] T.Naruko,et al.: "C-type natriuretic peptide in human coronary atherosclerotic lesions." Circulation. 94. 3103-3108 (1996)

[Publications] J.Yamashita,et al.: "Opposite regulation of Gax homeobox expression by angiotensin II and C-type natriuretic peptide." Hyertension. 29 part 2. 381-387 (1997)

[Publications] T-H.Chun,et al.: "Shear stress augments expression of C-type natriuretic peptide and adrenomedullin." Hyertension. (in press). (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." Amerecan Heart J.(in press). (1997)

伊藤裕京都大学, 医学研究科, 助手 (40252457)