Polyplex Micelles with Cyclic RGD Peptide Ligands and Disulfide Cross-Links Directing to the Enhanced Transfection via Controlled Intracellular Trafficking
Polyplex Micelles with Cyclic RGD Peptide Ligands and Disulfide Cross-Links Directing to the Enhanced Transfection via Controlled Intracellular Trafficking
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DOI：
10.1021/mp800070s
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发表时间：
2008-11-01
影响因子：
4.9
通讯作者：
Kataoka, Kazunori
中科院分区：
文献类型：
--
作者：
Oba, Makoto;Aoyagi, Kazuhiro;Kataoka, Kazunori
制备了巯基化c（RGDfK）-聚乙二醇-聚赖氨酸（PEG-PLys）嵌段聚合物，并将其与质粒DNA（pDNA）离子络合，制备了靶向二硫键交联的聚复合物胶束。所获得的复合物胶束针对具有α（v）β（3）整联蛋白受体的培养的HeLa细胞实现了显著增强的转染效率，所述α（v）β（3）整联蛋白受体被环状RGD肽选择性识别，证明了环状RGD肽配体在胶束表面上和核心中的二硫键交联的协同作用，以通过还原切割在细胞内环境中发挥pDNA的平稳释放。这种增强是不是由于增加的摄取量的复合物胶束，但在其细胞内的运输路线的变化。详细的激光共聚焦显微镜观察显示，与环RGD肽配体的复合物胶束分布在核周区域的早期阶段，优先通过小窝介导的内吞作用，这可能是一个理想的途径，以避免溶酶体降解的交付基因。因此，这种将配体和交联引入复合物胶束中的方法对于构建通过控制细胞内分布来增强转染的非病毒基因载体是有希望的。
Thiolated c(RGDfK)-poly(ethylene glycol)-block-poly(lysine) (PEG-PLys), a novel block polymer that has a cyclic RGD peptide in the PEG terminus and thiol groups in the PLys side chain, was prepared and applied to the preparation of targetable disulfide cross-linked polyplex micelles through ion complexation with plasmid DNA (pDNA). The obtained polyplex micelles achieved remarkably enhanced transfection efficiency against cultured HeLa cells possessing alpha(v)beta(3) integrin receptors, which are selectively recognized by cyclic RGD peptides, demonstrating the synergistic effect of cyclic RGD peptide ligands on the micelle surface and disulfide cross-links in the core to exert the smooth release of pDNA in the intracellular environment via reductive cleavage. This enhancement was not due to an increase in the uptake amount of polyplex micelles but to a change in their intracellular trafficking route. Detailed confocal laser scanning microscopic observation revealed that polyplex micelles with cyclic RGD peptide ligands were distributed in the perinuclear region in the early stages preferentially through caveolae-mediated endocytosis, which may be a desirable pathway for avoiding the lysosomal degradation of delivered genes. Hence, this approach to introducing ligands and cross-links into the polyplex micelles is promising for the construction of nonviral gene vectors that enhance transfection by controlling intracellular distribution.