Uronic acids functionalized polyethyleneimine (PEI)-polyethyleneglycol (PEG)-graft-copolymers as novel synthetic gene carriers.
Uronic acids functionalized polyethyleneimine (PEI)-polyethyleneglycol (PEG)-graft-copolymers as novel synthetic gene carriers.
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DOI：
10.1016/j.biomaterials.2005.11.011
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发表时间：
2006-04
期刊：
影响因子：
14
通讯作者：
S. Weiss;Nathalie Sieverling;Maren Niclasen;C. Maucksch;A. Thünemann;H. Möhwald;D. Reinhardt;J. Rosenecker;C. Rudolph
中科院分区：
文献类型：
--
作者：
S. Weiss;Nathalie Sieverling;Maren Niclasen;C. Maucksch;A. Thünemann;H. Möhwald;D. Reinhardt;J. Rosenecker;C. Rudolph
在这项研究中，我们研究了半乳糖醛酸（GalAc）和甘露醛酸（ManAc）作为受体介导的基因传递的新型靶向配体。GalAc和ManAc分别偶联于聚乙烯亚胺（PEI）或PEI -聚乙二醇（PEG）。此外，乳酸菌酸（LacAc）包含一个与半乳糖酸相关的碳水化合物环，通过其开链葡萄糖酸部分与每个聚合物偶联。用分析超离心和粒径排除色谱对聚合物的摩尔质量分布进行了表征。pei - conjugated - pdna复合物转染HepG2-、HeLa-和16HBE14o -细胞。GalAc和lacac功能化PEI偶联物介导的基因表达量低于PEI。相比之下，manac功能化pei偶联物介导的基因表达比其他测试的pei偶联物高出3个数量级，特别是在低N/P比的负电荷基因载体上，与细胞系无关。转染前用过量的ManAc对细胞进行预孵育，仅对ManAc功能化的pei偶联物显著抑制转染率。甲基-α-d-甘露醛酸与PEI偶联导致转染率明显低于基于ManAc-PEI的配合物。结合荧光素标记的manac功能化右旋糖酐的荧光显微镜图像和细胞的FACS分析，这些结果表明，受体介导的manac - pei - pdna复合物的内吞作用通过manac特异性受体参与了基因转移。总之，pei聚合物的manac修饰代表了一种新的受体介导的基因传递策略，在体内应用前景广阔。
In this study, we investigated galacturonic (GalAc)- and mannuronic (ManAc) acids as novel targeting ligands for receptor-mediated gene delivery. GalAc and ManAc were coupled to either polyethyleneimine (PEI) or PEI–polyethyleneglycol (PEG). Furthermore, lactobionic acid (LacAc), which comprises a GalAc-related carbohydrate ring, was coupled to each of the polymers through its open-chain gluconic acid moiety. The molar mass distributions of the polymers were characterized by analytical ultracentrifugation and size exclusion chromatography. PEI-conjugate–pDNA complexes were transfected into HepG2-, HeLa-, and 16HBE14o−-cells. Gene expression mediated by GalAc- and LacAc-functionalized PEI-conjugates was lower than for PEI. In contrast, gene expression mediated by ManAc-functionalized PEI-conjugates was up to three orders of magnitude higher than for the other tested PEI-conjugates, in particular for negatively charged gene vectors at low N/P ratios, independent of the cell line. Pre-incubation of cells with an excess of ManAc before transfection significantly inhibited transfection rates only for ManAc-functionalized PEI-conjugates. Coupling of methyl-α-d-mannuronic acid to PEI resulted in significantly lower transfection rates than for ManAc-PEI based complexes. Together with fluorescence microscopy images of fluorescein-labelled ManAc-functionalized dextrans and FACS analyses of cells, these results demonstrate that receptor-mediated endocytosis of ManAc–PEI-conjugate–pDNA complexes via ManAc-specific receptors was involved in gene transfer. In conclusion, ManAc-modification of PEI-polymers represents a novel strategy for receptor-mediated gene delivery which could be promising for in vivo application.