マイクロアレイを利用した血管内皮細胞の分化・再生因子同定と診断・治療への応用

• Project/Area Number: 12877108
• Research Category: Grant-in-Aid for Exploratory Research
• Allocation Type: Single-year Grants
• Research Field: Circulatory organs internal medicine
• Research Institution: Osaka University
• Principal Investigator: 森下 竜一 大阪大学, 大学院・医学研究科, 助教授 (40291439)
• Project Period (FY): 2000 – 2001
• Project Status: Completed (Fiscal Year 2001)
• Budget Amount: ¥1,800,000 (Direct Cost: ¥1,800,000); Fiscal Year 2001: ¥600,000 (Direct Cost: ¥600,000); Fiscal Year 2000: ¥1,200,000 (Direct Cost: ¥1,200,000)
• Keywords: 内皮 / マクロアレイ / 血管新生 / HGF / VEGF / 血管 / マイクロアレイ / DNAチッブ

Research Abstract

本研究では、ヒト大動脈由来内皮細胞を用いて、増殖過程の内皮(成長期:growing stage)及び接触阻害(contact inhibition)後の内皮、更にスクラツチによる修復過程の内皮(regrowing stage)、を用いて、ESTデータベースを元にしたマイクロアレイを行った。その中には、増殖期及び静止期に特異的に発現している転写因子、増殖因子の受容体などの遺伝子を幾つか同定した。また、肝細胞増殖因子(HGF)添加により、ヒト内皮細胞増殖時に発現亢進している遺伝子をマイクロアレイを利用しで、明らかにした。現在、HGFによる内皮再生促進におけるこれら遺伝子の関連を検討している。
また、血管再生を増強する治療戦略として、HGFやVEGFなど血管新生因子の遺伝子導入により再生された血管は、血管拡張物質であるプロスタサイクリンを合成するPG12合成酵素遺伝子との共導入により、再生が促進されることをラット及びウサギ下肢虚血モデルで明らかにした。今後、第2世代の血管新生療法として期待される。

Research Products

• [Publications] Taniyama Y, Morishita R, Aoki M, Nakagami H, Yamamoto K, Yamazaki K, Matsumoto K, Nakamura T, Kaneda, Y, Ogihara T.: "Therapeutic angiogenesis induced by human nepatocyte growth factor gene in rat and rabbit hind limb ischerma models: precliniceal study for treatment of peripheral arterial disease"Gene Therapy. 8. 181-189 (2001)
• [Publications] Yamainoto K, Morishita R, Hayashi S, Matsushita H, Nakagami H, Moriguchi A, Matsumoto K, Nakamura T, Kaneda Y. Ogihara T.: "Contribution of Bcl-2. but not Bcl-xL and Bax, to anti-apoptotic actions of hepatocyte growth factor in hypoxic conditioned human endothelial cells."Hypertension. 37. 1341-1348 (2001)
• [Publications] Nakagaini H, Morishita, R, Yamamoto K, Yoshimura S, Taniyama Y, Aoki M, Matsubara H, Kim S, Kaneda Y, Ogihara T.: "Phosphorylation of p38MAP kinase downstream of bax-caspase-3 pathway leads to cell death induced by high D-glucosec in human endothelial cells"Diabetes. 50. 1472-1481 (2001)
• [Publications] Ahn JD, Morishita R, Kaneda Y, Lee KU, Park JY, Jeon YJ, Song HS Lee IK: "Transcription factor decoy DON for activator protcin-1 (AP-1 inhibits expression of type 1 plasmlnogen activator inhibitor (PAI-1) gene induced by high glucose and arigiotensin II in cultured human vascular smooth muscle cells."Diabetologia. 44. 713-720 (2001)
• [Publications] Aoki M, Nata T, Morishita R, Matsushita H, Nakagami H, Yamamoto K, Yamazaki K, Nakabayashi M, Ogihara T, Kaneda Y.: "Endothelial apoptosis induced by oxidative stress through activation of NFkB : anti-apoptotic effect of antioxidant agents on endothelial cells."Hypertension. 38. 48-55 (2001)
• [Publications] Aoki M, Morishita R, Hayashi SI, Jo N, Matsumoto K, Nakamaura T. Kaneda Y. Ogihara T.: "Inhibition of neointimal formation after balloon injury by cilostazol, accompanied by improvement of endothelial dysfunction and induction of hepatocyte growth factor in rat diabetes model."Diabetologia. 44. 1034-1042 (2001)
• [Publications] Morishita R, Aoki M, Kaneda Y, Ogihara T.: "Pharmacology & Therapeutics"Alan C. Sartorelli, William C, Bowman, David R, Sibley, Michael J, Curtis. 247 (2001)
• [Publications] Hayashi Si,Morlshita R,Matsushita H,Nakagami H,Taniyama Y,Nakamura T,Aoki,M,unamoto K,Higaki J,Ogihara T.: "Cyclic AMP inhibited proliferation of human aortic vascular smooth muscle cells, accompanied by induction of p53 and p21."Hypertension. 35. 237-243 (2000)
• [Publications] Matsushita H,Morishita R,Nata T,Aoki M,Nakagami H,Tanitama Y,Yamamoto K,Higaki J,Kaneda Y,Ogihara T.: "Hypoxia induced endothelial apoptosis through NFkB-mediated bcl-2 suppression : in vivo evidence of importance of NFkB in endothelial cell regulation."Circulation Research. 86. 974-981 (2000)
• [Publications] Taniyama Y,Morishita R,Nakagami H,Moriguchi A,Sakonjo H,Kim S,Matsumoto K,Nakamura T,Higaki J,Ogihara T.: "Potential contribution of a novel anti-fibrotic factor, hepatocyte growth factor, to prevention of myocardial fibrosis by angiotensin II blockade in cardiomyopathic hamster."Circulation. 102. 246-252 (2000)
• [Publications] Nakagami H,Morishita R.Yamamoto K,Taniyama Y,Aoki M,Kim S,Matsumoto K,Nakamura T,Higaki J,Ogihara T.: "Anti-apoptotic action of hepatocyte growth factor(HGF)through mitogen-activated protein kinase on human aortic endothelial cells."Journal of Hypertension. 18. 1411-1420 (2000)
• [Publications] Yamada S,Morishita R,Nakamura S,Ogihara T,Kusumi Y,Sakurai I,Kubo N,Sakurabayashi I.: "Development of antibody againgst epitope of lipoprotein (a) modified by oxidation : evaluation of new enzyme-linked immunoassay for oxidized Lp (a)."Circulation. 102. 1639-1644 (2000)
• [Publications] Nakagami H,Morishita R,Yamamoto K,Yoshimura S,Taniyama Y,Aoki M,Matsubara H,Kim S,Kaneda Y,Ogihara T.: "Phosphorylation of p38MAP kinase downstream of bax-caspase-3 pathway leads to cell death induced by high D-glucose in human endothelial cells."Diabetes. (in press).
• [Publications] Morishita R,Aoki M,Nakagami H,Taniyama Y,Yamamoto K,Matsushita H,Matsumoto K,Higaki J,Kaneda Y,Ogihara T.: "Heart Failure ; Frontiers in Cardiology"Kitabatake A,Sasayama S,Francis G.S.. 8 (2000)
• [Publications] Kaneda Y,Nakasima H,Tsuboniwa N,Morishita R.: "Progress of fusigenic viral-liposomes. Progress in Gene Therapy : basic and clinical frontiers."Bertolottis R et al.. 19 (2000)