| Project/Area Number |
18590144
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Medical pharmacy
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| Research Institution | Kumamoto University |
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Principal Investigator |
ARIMA Hidetoshi Kumamoto University, Graduate School of Pharmaceutical Sciences, Professor (50260964)
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| Co-Investigator(Kenkyū-buntansha) |
KAI Hirofumi Kumamoto University, Graduate School of Pharmaceutical Sciences, Professor (30194658)
HIRAYAMA Fumitoshi Sojo University, Faculty of Pharmaceutical Sciences, Professor (90094036)
UEKAMA Kaneto Sojo University, Faculty of Pharmaceutical Sciences, Professor (90040328)
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| Project Period (FY) |
2006 – 2007
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| Project Status |
Completed (Fiscal Year 2007)
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| Budget Amount *help |
¥4,080,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥480,000)
Fiscal Year 2007: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
Fiscal Year 2006: ¥2,000,000 (Direct Cost: ¥2,000,000)
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| Keywords | siRNA / carrier / Dendrimer / Cyclodextrin / cell specific delivery / シクロテキストリン / 結合体 / デリバリー / 糖修飾体 |
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| Research Abstract |
We previously reported the potential use of starburst polyamidoamine dendrimer (dendrimer, generation 3, G3)conjugate with a-cyclodextrin (α-CDE) as a novel gene delivery carrier. In the present study, the potential of α-CDE and their sugar-appended α-CDEs as novel and cell-specific carriers of small interfering RNA(siRNA) and short hairpin RNA-expressing vector(shpDNA)was evaluated in vitro and in vivo. The RNAi effects of the siRNA complex with cc-CDE were preferable to those of the complexes with commercially-available siRNA carriers in NIH3T3 cells stably expressing pGL3 luciferase. In addition, the siRNA complex with a-CDE showed negligible cytotoxicity up to higher charge ratios. However, we found that a-CDE did not have the potential as shpDNA carriers because of low RNAi effect. When the solution containing the binary complex of siRNA with α-CDE was injected into tumor tissues of mice bearing Colon-26 cells stably expressing the GL3 luciferase gene, the sufficient RNAi effects were provided, although the complex of Lipofectamine^TM2000 with siRNA showed the off-target effect. Therefore, α-CDE was found to provide the sequence-specific gene silencing effect by the complexation with siRNA. On the other hand, we prepared mannosylated α-CDE(M-α-CDE)and lactosylated α-CDE(L-α-CDE)to delivery siRNA and shpDNA specifically to antigen-presenting cells and hepatocytes, respectively. However, we found that L-α-CDE was able to bind to asialoglycoprotein receptor expressing on hepatocytes, although M-a-CDE could not bind to mannose receptor expressing on antigen-presenting cells. Both L-α-CDE and M-α-CDE had negligible cytotoxicity. These results suggest that L-α-CDE has the potential for hepatocyte-specific siRNA and shpDNA carriers.
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