生体への遺伝子導入法による難治性循環器疾患の病態解析と治療への応用・心血管用ベクターとしてのイムノリポソームの開発

• 研究課題/領域番号: 11470163
• 研究種目: 基盤研究(B)
• 配分区分: 補助金
• 応募区分: 一般
• 研究分野: 循環器内科学
• 研究機関: 大阪大学
• 研究代表者: 森下 竜一 大阪大学, 医学系研究科, 助教授 (40291439)
• 研究分担者: 金田 安史 大阪大学, 医学系研究科, 教授 (10177537) ; 檜垣 実男 大阪大学, 医学系研究科, 助教授 (70189744) ; 守口 篤 大阪大学, 医学系研究科, 助手 (10273666)
• 研究期間 (年度): 1999 – 2001
• 研究課題ステータス: 完了 (2001年度)
• 配分額: 14,600千円 (直接経費: 14,600千円); 2001年度: 3,500千円 (直接経費: 3,500千円); 2000年度: 3,500千円 (直接経費: 3,500千円); 1999年度: 7,600千円 (直接経費: 7,600千円)
• キーワード: 血管新生 / 遺伝子治療 / デコイ / NFKB / 再狭窄 / HGF / ASO / 再狭 / NFkB / 核酸医薬 / E2F / 閉塞性動脈硬化症 / 心筋梗塞

研究概要

本研究では、難治性循環器疾患の遺伝子治療ストラテジー開発とその臨床応用を目指している。HGF遺伝子によるヒト遺伝子治療臨床研究は、2001年6月より日本で始めての循環器疾患の遺伝子治療として開始された。現在までに6人の閉塞性動脈硬化症およびビュルガー病患者にヒトHGF遺伝子を投与した。前例で、血管造影で有意な血管陰影の増強が確認される一方、全例でO.1以上のABI改善もしくは疼痛の改善においてもVASスケールで1cm以上の改善を認めている(Efficacy rate=100%)、一方で、VEGF遺伝子治療で見られた投与部位の浮腫は認められなかった。現在までの成績では、VEGF遺伝子治療より有効性が高く、安全性に優れる可能性が示されつつある。また、同様のHGF遺伝子導入による血管新生をブタ心筋梗塞モデルにおいて明らかにし、重症虚血性心疾患の遺伝子治療として「HGF遺伝子プラスミドを用いたカテーテルデリバリーを基盤とした重症虚血性心疾患の治療のための遺伝子治療臨床研究」を大阪大遺伝子治療臨床研究審査委員会へ2001年申請した。また、申請者らはおとり型核酸医薬(デコイ)を開発し、細胞周期に必須な転写因子E2Fをターゲットにしたデコイによる再狭窄予防を目的とするヒト臨床研究の許可を大阪大学倫理委員会より得て、2000年4月より実施し、既に5人の患者に投与した。国内で初めての核酸医薬を用いた臨床研究であったが、全例で再狭窄は認められず、安全性にも問題ないことが明らかになった。今後に期待のもてる結果で、更に症例を増やしている。また、「血管拡張術後及びステント後再狭窄に対するNFKBデコイによる治療」計画を申請し、2001年8月に承認を得た。

研究成果

• [文献書誌] Taniyama Y, Morishita R, Aoki M, Nakagami H, Yamamoto K, Yamazaki K, Matsumoto K, Nakamura T, Kaneda Y, Ogihara T: "Therapeutic angioge nesis induced by human hepatocyte growth factor gene in rat and rabbit hindlimb ischemia models: preclinical study for treatment of peripheral arterial disease"Gene Therapy. 8. 181-189 (2001)
  • 説明: 「研究成果報告書概要(和文)」より
• [文献書誌] Hayashi K, Morishita R, Nakagami H, Yoshimura S, Hara A, Matsumoto K, Nakamura T, Kaneda Y, Ogihara T, Sakai N: "Gene therapy for preventing neuronal death using hepatocyte growth factor: in vivo gene transfer of HGF to subarachnoid space prevents delayed neuronal death in gerbil hippocampal CA1 neurons"Gene Therapy. 8. 1167-1173 (2001)
  • 説明: 「研究成果報告書概要(和文)」より
• [文献書誌] Taniyama Y, Morishita R, Hiraoka K, Aoki M, Nakagami H, Yamasaki K, Matsumoto K, Nakamura T, Kaneda Y, Ogihara T: "Therapeutic angiogenesis induced by human hepatocyte growth factor gene in rat diabetic hind limb ischemia model: molecular mechanisms of delayed angiogenesis in diabetes"Circulation. 104. 2344-2350 (2001)
  • 説明: 「研究成果報告書概要(和文)」より
• [文献書誌] Yoshimura S, Morishita R, Hayashi K, Yamamoto K, Nakagami H, Kaneda Y, Sakai N, Ogihara T: "Inhibition of intimal hyperplasia after balloon injury in rat carotid artery model using cis-element 'decoy' of nuclear factor-kB binding site as a novel molecular strategy"Gene Therapy. 8. 1635-1642 (2001)
  • 説明: 「研究成果報告書概要(和文)」より
• [文献書誌] Taniyama Y, Tachibana K, Hiraoka K, Namba T, Yamasaki K, Hashiya N, Aoki M, Ogihara T, Yasufumi K, Morishita R: "Local delivery of plasmid DNA into rat carotid artery using ultrasound"Circulation. (in press). (2001)
  • 説明: 「研究成果報告書概要(和文)」より
• [文献書誌] Nakamura T, Morishita R, Asai T, Tsuboniwa N, Aoki M, Sakonjo H, Yamasaki K, Hashiya N, Kaneda Y, Ogihara T: "Molecular strategy using cis-element 'decoy' of E2F binding site inhibits neointimal formation in porcine balloon-injured coronary artery model"Gene Therapy. (in press).
  • 説明: 「研究成果報告書概要(和文)」より
• [文献書誌] Morishita R, Aoki M, Hashiya N, Yamasaki K, Makino H, Wakayama K, Azuma J, Ogihara T: Gene Therapy & Regulation. (in press).
  • 説明: 「研究成果報告書概要(和文)」より
• [文献書誌] 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 hepatocyte growth factor gene in rat and rabbit hind limb ischemia module: preclintcal study of treatment of peripheral arterial disease"Gene Therapy. (8). 181-189 (2001)
  • 説明: 「研究成果報告書概要(欧文)」より
• [文献書誌] Hayashi K, Morishita R, Nakagami H, Yoshimura S, Hara A, Matsumoto K, Nakamura T, Kaneda Y, Sakai N: "Gene therapy for preventing neuronal death using hepatocyte growth factor in vivo gene transfer of HGF to subarachnoid space prevents delayed neuronal death in gerbil hippocampal CAI neurons"Gene Therapy. (8). 1167-1173 (2001)
  • 説明: 「研究成果報告書概要(欧文)」より
• [文献書誌] Taniyama Y, Morishita R, Hiraoka K, Aoki M, Nakagami H, Yamasaki K, Matsumoto K, Nakamura T, Kaneda Y, Ogihara T: "Therapeutic angiogenesis induced by human hepatocyte growth factor gene in rat diabetic hind limb ischernia module: molecular of delayed angiogenesis in diabetes"Circulation. (104). 2344-2350 (2001)
  • 説明: 「研究成果報告書概要(欧文)」より
• [文献書誌] Yoshimura Y, Morishita R, Hayashi K, Yamamoto K, Nakagami H, Kaneda Y, Sakai N, Ogihara T: "Inhibition of intimal hyperplasic after balloon injury in rat carotid artery model using cis-elememt "decoy" of nuclear factor-kB binding afte a novel molecular strategy"Gene therapy. (8). 1635-1642 (2001)
  • 説明: 「研究成果報告書概要(欧文)」より
• [文献書誌] Taniyama Y, Tachibana K, Hiraoka K, Namba T, Yamasaki K, Hashiya N, Aoki M, Ogihara T, Kaneda Y, Morishita R: "Local delivery of plasmid DNA into rat carotid artery using ultrasound"Circulation. (in press). (2001)
  • 説明: 「研究成果報告書概要(欧文)」より
• [文献書誌] Nakamura T, Morishita R, Asai T, Tsuboniwa N, Aoki M, Sakonjo H, Yamasaki K, Hashiya N, Kaneda Y, Ogihara T: "Molecular strategy using cis-element "decoy" of E2F binding alto inhibits neointimal formation in porcine balloon-injured coronary asiesy model."Gene Therapy. (in press). (2001)
  • 説明: 「研究成果報告書概要(欧文)」より
• [文献書誌] Morishita R, Aoki M, Hashiya N, Yamasaki K, Makino H, Wakayama K, Azuma J, Ogihara T: Gene Therapy & Regulation. (in press).
  • 説明: 「研究成果報告書概要(欧文)」より
• [文献書誌] Taniyama Y, Morishita R, Aoki M, Nakagami H, Yamamoto K, Yamazaki K, Matsumoto K, Nakamura T, Kaneda Y, Ogihara T: "Therapeutic anglogenesis Induced by human hepatocyte growth factor gene in rat and rabbit hind limb ischemfa models: preclinical study for treatment of pertphera1 arterial disease"Gene Therapy. 8. 181-189 (2001)
• [文献書誌] Hayashi K, Morishita R, Nakagami H, Yoshimura S, Hara A, Matsumoto K, Nakamura T, Kaneda Y, Ogihara T, Sakai N: "Gene therapy for preventing neuronal death using hepatocyte growth factor: in vivo gene transfer of HGF to subarachnoid space prevents delayed neuronal death in gerbil hippocampal CA1 neurons"Gene Therapy. 8. 1167-1173 (2001)
• [文献書誌] Taniyama Y, Morishita R, Hiraoka K, Aoki M, Nakagami H, Yamasaki K, Matsumoto K, Nakamura T, Kaneda Y, Ogihara T: "Therapeutic angiogenesis induced by human hepatocyte growth factor gene in rat diabetic hind limb ischemia model: molecular mechanisms of delayed angiogenesis in diabetes"Circulation. 104. 2344-2350 (2001)
• [文献書誌] Yoshimura S, Morishita R, Hayashi K, Yamamoto K, Nakagami H, Kaneda Y, Sakai N, Ogihara T: "Inhibition of intimal hyperplasia after balloon injury in rat carotid artery model using cis-element "decoy" of unclear factor-kB binding site as a novel molecular strategy"Gene Therapy. 8. 1635-1642 (2001)
• [文献書誌] Taniyama Y, Tachibana K, Hiraoka K, Namba T, Yamasaki K, Hashiya N, Aoki M, Ogihara T, Kaneda Y, Morishita R: "Local delivery of plasmid DNA into rat carotid artery using ultrasound"Circulation. (in press). (2001)
• [文献書誌] Nakamura T, Morishita R, Asai T, Tsuboniwa N, Aoki M, Sakonjo H, Yamasaki K, Hashiya N, Kaneda Y, Ogihara T: "Molecular strategy using cis-element "decoy" of E2F binding site inhibits neointimal formation in porcine balloon-injured coronary artery model"Gene Therapy. (in press).
• [文献書誌] Morishita R, Aoki M, Hashiya N, Yamasaki K, Makino H, Wakayama K, Azuma J, Ogihara T: "Gene Therapy & Regulation"(in press).
• [文献書誌] Aoki M,Morishita R,Taniyama Y,Kida I,Moriguchi A,Matsumoto K,Nakamura T,Kaneda Y,Higaki J,Ogihara T.: "Angiogenesis induced by hepatocyte growth factor in non-infarcted myocardium and infarcted myocardium : up-regulation of essential transcription factor for angiogenesis,ets."Gene Therapy. 7. 417-427 (2000)
• [文献書誌] Matsushita H,Morishita R,Aoki M,Tomita N,Taniyama Y,Nakagami H,Shimozato T,Higaki J,Kaneda Y,Ogihara T.: "Transfection of antisense p53 tumor suppressor gene oligodeoxynucleotides into rat carotid artery resulted in abnormal growth of vascular smooth muscle cells."Circulation. 101. 1447-1452 (2000)
• [文献書誌] Matsushita H,Morishita R,Nata T,Aoki M,Nakagami H,Taniyama Y,Yamamoto K,Higaki J,Kaneda Y,Ogihara T.: "Hypoxis 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)
• [文献書誌] Tomita N,Morishita R,Lan HY,Yamamoto K,Hashizume M,Notakae M,Toyosawa K.Fulitani B,Mu W,Nikolic-Paterson DJ,Atkins RC,Kaneda Y,Higaki J,Ogihara T.: "In vivo administration of a nlenr transcription factor-kappaB decoy suppresses experimental crescentic glomcrulonophritis."Journal of American Society of Nephrology. 11. 1244-1252 (2000)
• [文献書誌] Yamamoto K,Morishita R,Tomita N,Shimozato T,Nakagami H,Kikuchi A,Aoki M,Higaki J,Kaneda Y,Ogihara T.: "Ribozyme oligonucleotides against transforming growth factor-β inhibited neointimal formation after vascular injury in rat model : potentisl application of ribozyme strategy to treat cardiovascular diseage."Circulation. 102. 1308-1314 (2000)
• [文献書誌] Hayashi K,Nakamura S,Morishita R,Moriguchi A,Aoki M,Matsumoto K,Nakamura T,Kaneda Y,Sakai N,Ogihara T.: "In vivo transfer of human hepatocyte growth factor gene accelcrates re-endothelialization and inhibits neointimal formation after balloon injury in rat model."Gene Therapy. 7. 1664-1671 (2000)
• [文献書誌] Morishita R,Aoki M,Nakamura S,Higaki J,Kaneda Y,Ogihara T.: "Gene therapy for cardiovascular disease using hepatocyte growth factor.Atherosclerosis V.The Fifth Saratoga International Conference."Edited by Numano F & Gimbrone MA.Ann New York Acad. 369-376 (2000)
• [文献書誌] Kaneda Y,Nakasima H,Tsuboniwa N,Morishita R.: "Progress of fusigenic viral-liposomes.Progress in Gene Therapy : basic and clinical frontiers."edited by Bertolottis R et al.VSP,. 225-243 (2000)
• [文献書誌] Kawamura I, Morishita R, Tomita N, Aketa M, Tsujimoto S, Manda T, Tomoi M, Kida I, Hifakai J, Kaneda Y, Shimomura K, Ogihara T.: "Intratumoral injection of oligonucleotides to the NFkB binding site inhibits cachexia in a mouse tumor model"Gene Therapy. 6. 91-97 (1999)
• [文献書誌] Takahashi H, Oakamoto E, Fujimoto J, Ueki T, Kaneda Y, Tsutusi H, Nakanishi K,Sawa Y, Morishita R, Matusmoto K, Nakamura T,: "Curative gene therapy of liver cirrhosis by hepatocyte growth factor in rats"Nature Medicine. 5. 226-230 (1999)
• [文献書誌] Morishita R, Nakamura S, Hayashi S, Taniyama Y, Moriguchi A, Nagano T, Taiji M, Noguchi H, Takeshita S, Matsumoto K, Nakamura T, Higaki J, Ogihara T.: "Therapeutic angiogenesis induced by human recombinant hepatocyte growth factor in rabbit hind limb ischemia model as "cytokine supplement therapy""Hypertension. 33. 1379-1384 (1999)
• [文献書誌] Hayashi S, Morishita R, Nakamura S, Yamamoto K, Moriguchi A, Nagano T, Taizi M, Noguchi H, Matsumoto K, Nakamura T, Higaki J, Ogihara T.: "Potential role of hepatocyte growth factor,a novel angiogenic growth factor, in peripheral arterial disease : down-regulation of HGF in response to hypoxia in vascular in vascular cells"Circulation. 100. II301-II308 (1999)
• [文献書誌] Morishita R, Gibbons GH, Tomita N, Zhang L, Kaneda Y, Ohihara T, Dzau VJ.: "Anitisense oligodeozynucleotide inhibition of vascular angiotensin converting enzyme expression attenuates neointimal formation : evidence for tissue ACE function"Arteriosclerosis,Thrombosis & Vascular Biology. (in press).
• [文献書誌] Tomika N, Morishita R, Lan HY, Yamamoto K, Hashizume M, Notakae M, Toyosawa K, Fujitani B,Mu W, Nikolic-Paterson DJ, Atkins RC, Kaneda Y, Higaki J, Ogihara T.: "In vivo administration of a clear transcription factor-kappaB decoy suppresses experimental crescentic glomerulonephritis"Journal of American Society of Nephrology. (in press).