Cyclic RGD peptide-conjugated polyplex micelles as a targetable gene delivery system directed to cells possessing alphavbeta3 and alphavbeta5 integrins.
Cyclic RGD peptide-conjugated polyplex micelles as a targetable gene delivery system directed to cells possessing alphavbeta3 and alphavbeta5 integrins.
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发表时间：
2007
影响因子：
4.7
通讯作者：
M. Oba;S. Fukushima;N. Kanayama;K. Aoyagi;N. Nishiyama;H. Koyama;K. Kataoka
中科院分区：
文献类型：
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作者：
M. Oba;S. Fukushima;N. Kanayama;K. Aoyagi;N. Nishiyama;H. Koyama;K. Kataoka
在温和的酸性条件下，以缩醛-聚乙二醇-聚赖氨酸（c（RGDfK）-PEG-PLys）为原料，合成了一种含有环状RGD肽的嵌段共聚物--环[RGDfK（CX-）]-聚乙二醇-聚赖氨酸（c（RGDfK）-PEG-PLys）。环状RGD肽识别α v β 3和α v β 5整联蛋白受体，其在血管生成、血管内膜增厚和恶性肿瘤增殖中起关键作用。c（RGDfK）-PEG-PLys/pDNA复合物胶束与PEG-PLys/pDNA复合物胶束相比，对具有α v β 3和α v β 5整合素的培养的HeLa细胞显示出显著增加的转染效率（TE）。另一方面，在针对不具有α v β 3和少量α v β 5整联蛋白的293 T细胞的转染中，与PEG-PLys/pDNA胶束的TE相比，c（RGDfK）-PEG-PLys/pDNA胶束的TE没有显示出增加。流式细胞术分析显示，c（RGDfK）-PEG-PLys/pDNA胶束对HeLa细胞的摄取高于PEG-PLys/pDNA胶束，这与转染结果一致。此外，共聚焦激光扫描显微镜观察显示，在c（RGDfK）-PEG-PLys胶束中的pDNA优先积累在HeLa细胞的核周区域内孵育3小时。对于不含c（RGDfK）配体的胶束，没有观察到pDNA如此快速且定向地积累到核周区域。这些结果表明，在TE的增加所诱导的引入的c（RGDfK）肽配体是由于细胞摄取的增加，以及促进细胞内运输胶束向核周区域通过α v β 3和α v β 5整合素受体介导的内吞作用，这表明，环RGD肽共轭的复合物胶束作为一个位点特异性靶向基因递送系统具有很好的可行性。
A cyclic RGD peptide-conjugated block copolymer, cyclo[RGDfK(CX-)]-poly(ethylene glycol)-polylysine (c(RGDfK)-PEG-PLys), was synthesized from acetal-PEG-PLys under mild acidic conditions and spontaneously associated with plasmid DNA (pDNA) to form a polyplex micelle in aqueous solution. The cyclic RGD peptide recognizes alphavbeta3 and alphavbeta5 integrin receptors, which play a pivotal role in angiogenesis, vascular intima thickening, and the proliferation of malignant tumors. The c(RGDfK)-PEG-PLys/pDNA polyplex micelle showed a remarkably increased transfection efficiency (TE) compared to the PEG-PLys/pDNA polyplex micelle for the cultured HeLa cells possessing alphavbeta3 and alphavbeta5 integrins. On the other hand, in the transfection against the 293T cells possessing no alphavbeta3 and a few alphavbeta5 integrins, the TE of the c(RGDfK)-PEG-PLys/pDNA micelle showed no increase compared to the TE of the PEG-PLys/pDNA micelle. Flow cytometric analysis revealed a higher uptake of the c(RGDfK)-PEG-PLys/pDNA micelle than the PEG-PLys/pDNA micelle against HeLa cells, consistent with the transfection results. Furthermore, a confocal laser scanning microscopic observation revealed that the pDNA in the c(RGDfK)-PEG-PLys micelle preferentially accumulated in the perinuclear region of the HeLa cells within 3 h of incubation. No such fast and directed accumulation of pDNA to the perinuclear region was observed for the micelles without c(RGDfK) ligands. These results indicate that the increase in the TE induced by the introduction of the c(RGDfK) peptide ligand was due to an increase in cellular uptake as well as facilitated intracellular trafficking of micelles toward the perinuclear region via alphavbeta3 and alphavbeta5 integrin receptor-mediated endocytosis, suggesting that the cyclic RGD peptide-conjugated polyplex micelle has promising feasibility as a site-specifically targetable gene delivery system.