Potocytosis and cellular exit of complexes as cellular pathways for gene delivery by polycations
Potocytosis and cellular exit of complexes as cellular pathways for gene delivery by polycations
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DOI：
10.1002/jgm.772
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发表时间：
2005-10-01
影响因子：
3.5
通讯作者：
Fajac, I
中科院分区：
文献类型：
--
作者：
Grosse, S;Aron, Y;Fajac, I
背景虽然聚阳离子是基因转移最有效的非病毒载体之一，但它们所允许的基因表达对于体内应用来说仍然太低。为了设计更有效的聚阳离子载体，需要确定与当前聚阳离子复合的质粒的胞内运输的控制因素。方法和结果。用电子显微镜观察人气道上皮细胞中质粒DNA与糖基化多聚赖氨酸或聚乙烯亚胺（PEI）衍生物复合物的转运。复合物的细胞加工随它们的大小和所用的聚阳离子衍生物而变化：用所研究的所有聚阳离子载体制成的大复合物（> 200 nm）通过巨胞饮内化。相反，中间（100-200 nm）配体偶联的聚赖氨酸和PEI复合物主要通过网格蛋白包被的凹坑进入。然后在核内体囊泡中发现复合物，在靠近核的溶酶体或囊泡中积累，并且它们的核进入受到限制。对于用PEI衍生物获得的小复合物（< 100 nm）的群体，它们通过小窝内化，并遵循胞吞作用的运输模式到达内质网，其中它们的命运仍不清楚。最后，一些复合物退出细胞回流时使用PEI衍生物或通过外泌体样途径糖基化聚赖氨酸complexes.Conclusions观察到的不同途径的复杂贩运与复杂的大小意味着开发和研究载体形成复合物具有一定的大小。此外，我们描述的复杂出口可能有助于基于合成载体的基因转移的良好建立的短期效率。它有利于允许重复治疗的载体的工程化。版权所有（c）2005年约翰威利父子有限公司。
Background Although polycations are among the most efficient nonviral vectors for gene transfer, the gene expression they allow is still too low for in vivo applications. To engineer more potent polycationic vectors, the factors governing the intracellular trafficking of a plasmid complexed with current polycations need to be identified.Methods and results. The trafficking of plasmid DNA complexed to glycosylated polylysines or polyethylenimine (PEI) derivatives was studied by electron microscopy of human airway epithelial cells. The cellular processing of complexes varied with their size and the polycation derivative used: large complexes ( > 200 nm) made with all polycationic vectors studied were internalized by macropinocytosis. In contrast, intermediate (100-200 nm) ligand-coupled polylysine and PEI complexes primarily entered through clathrin-coated pits. Complexes were then found in endosomal vesicles, accumulated in lysosomes or vesicles near the nucleus and their nuclear entry was limited. For the population of small complexes ( < 100 nm) obtained with PEI derivatives, they were internalized through caveolae and pursued a traffic pattern of potocytosis to the endoplasmic reticulum where their fate remains unclear. Finally, some complexes exited the cells either by regurgitation when PEI derivatives were used or through an exosome-like pathway for glycosylated-polylysine complexes.Conclusions The different pathways of complex trafficking observed in relation with complex size imply the development and study of vectors forming complexes with definite size. Moreover, the complex exit we describe may contribute to the well-established short-term efficiency of gene transfer based on synthetic vectors. it favors the engineering of vectors allowing repeated treatment. Copyright (c) 2005 John Wiley & Sons, Ltd.