Palmitic acid substitution on cationic polymers for effective delivery of plasmid DNA to bone marrow stromal cells
Palmitic acid substitution on cationic polymers for effective delivery of plasmid DNA to bone marrow stromal cells
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DOI：
10.1002/jbm.a.31249
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发表时间：
2007-05-01
影响因子：
4.9
通讯作者：
Uludag, Hasan
中科院分区：
文献类型：
--
作者：
Incani, Vanessa;Tunis, Emily;Uludag, Hasan
由于病毒载体的安全性问题，非病毒基因载体在基因治疗中被积极探索。为了设计有效的基因载体用于骨髓基质细胞（BMSC）的修饰，将阳离子聚合物聚乙烯亚胺（PEI）和聚-L-赖氨酸（PLL）通过酰胺键与棕榈酸（PA）连接。根据反应条件，PEI和PLL被每个聚合物链2.2-5.2和13.4-16.2 PA取代。在琼脂糖凝胶结合试验中，PA取代的聚合物对含有增强型绿色荧光蛋白（pEGFP）的质粒显示出略低的结合效率。PLL-PA而非PEI-PA的细胞结合特别增强，导致更高百分比的细胞显示出显著的聚合物摄取。PLL-PA（vs. PLL）也显著增加了pEGFP向BMSC的递送，但PEI-PA（vs. PEI）没有显著增加。PLL-PA的转染效率比未修饰的聚合物显著更高（接近5倍）。我们的结论是，PA取代PLL提供了一个有效的载体，转染的原代细胞来自骨髓。(c)2007 Wiley Periodicals，Inc.
Nonviral gene carriers are actively explored in gene therapy due to safety concerns of the viral carriers. To design effective gene carriers for modification of bone marrow stromal cells (BMSC), an important cell phenotype for clinical application of gene therapy, cationic polymers polyethyleneimine (PEI), and poly-L-Lysine (PLL) were substituted with palmitic acid (PA) via amide linkages. Depending on the reaction conditions, PEI and PLL was substituted with 2.2-5.2 and 13.4-16.2 PA per polymer chain. The PA substituted polymers displayed slightly lower binding efficiency towards a plasmid containing Enhanced Green Fluorescent Protein (pEGFP) in an agarose gel binding assay. The cell binding of PLL-PA, but not PEI-PA, was particularly enhanced, resulting in higher percentage of the cells displaying a significant polymer uptake. pEGFP delivery into the BMSC was also significantly increased with the PLL-PA (vs. PLL), but not PEI-PA (vs. PEI). The transfection efficiency of PLL-PA was significantly higher (similar to fivefold) than the unmodified polymer. We conclude that PA substitution on PLL provides an effective carrier for transfection of primary cells derived from the bone marrow. (c) 2007 Wiley Periodicals, Inc.