| Project/Area Number |
05044170
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| Research Category |
Grant-in-Aid for international Scientific Research
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| Allocation Type | Single-year Grants |
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| Section | Joint Research |
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| Research Institution | Osaka University |
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Principal Investigator |
KANEDA Yasufumi Institute for Molecular and Cellular Biology, Osaka University, 細胞生体工学センター, 助教授 (10177537)
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| Co-Investigator(Kenkyū-buntansha) |
DZAU Victor j. Stanford University, School of Medicine, 医学部, 教授
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| Project Period (FY) |
1993 – 1994
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| Project Status |
Completed (Fiscal Year 1994)
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| Budget Amount *help |
¥8,500,000 (Direct Cost: ¥8,500,000)
Fiscal Year 1994: ¥3,000,000 (Direct Cost: ¥3,000,000)
Fiscal Year 1993: ¥5,500,000 (Direct Cost: ¥5,500,000)
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| Keywords | HVJ-liposomes / Neointima formation / Antisense oligonucleotides / E2F decoys / NO synthase / Antistenosis / 遺伝子導入 / 遺伝子治療 / 循環器疾患 |
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| Research Abstract |
We developed efficient gene delivery system called HVJ-liposomes, Using this delivery system, we attempted to prevent restsnosis after angioplasty which is a barrier to the therapy of myocardial infarction. We examined the effect of AS-ODNs for the therapy of restenosis. First we introduced AS-PCNA and AS-cdc2 kinase into balloon injured vessels by HVJ-liposome. Cotransfer of both AS-ODNs resulted in the simultanuous inhibition of PCNA and cdc2 kinase mRNA expression to undetectable levels. Then HVJ-liposome containing 15 uM AS-PCNA and AS-cdc2 kinase was injected into balloon injured rat carotid artery. Neointima formation was completely inhibited by this treatment, whereas the sense-ODNs had no effect. The blockade of neointima formation by these AS continiued for 2 weeks by a single intraluminal administration and the inhibitory effect lasted for a period of 8 weeks after transfection.We further examined effect of AS-ODN against various cell-cycle regulators on neointima formation.
… More
The combination of AS-PCNA and cdc2 kinase or AS-cdc2 kinase and cdk2 kinase were most effective for the prevention of neointima formation. Then, we developed new strategy to inhibit cell proliferation by the introduction of a single molecule. The transcription of PCNA,cdc2 kinase and some protooncogene were activated by a common transcription factor, E2F,and the consensus DNA sequence recognized by E2F is known to be TTTCGCGC.We investigated the effect of double-stranded ODN designated as competitor of E2F on the prevention of neointima formation. We examined the effect of E2F decoy on the prevention of restenosis. E2F decoy was introduced into balloon injured rat carotid artery by HVJ-liposome. Our results demonstrated a marked suppression of neointima formation at 2 weeks sfter angioplasty by the decoy against E2F.At a dosage of 3 uM,decoy ODN inhibited neointima formation by approximately 80 % compared to vessels treated with HVJ-liposome alone or mismatched decoy ODN-treated vessels. Then, we employed NO synthase cDNA to inhibit neointima formation.NOS introduced into blood walls by HVJ-liposomes inhibited neointima formation for at least one month. These strategies will be able to be applied for the treatment of restenosis in humans. Less
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