Extracellular barriers to in Vivo PEI and PEGylated PEI polyplex-mediated gene delivery to the liver
Extracellular barriers to in Vivo PEI and PEGylated PEI polyplex-mediated gene delivery to the liver
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DOI：
10.1021/bc700388u
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发表时间：
2008-03-01
影响因子：
4.7
通讯作者：
Pun, Suzie H.
中科院分区：
文献类型：
--
作者：
Burke, Rob S.;Pun, Suzie H.
复合物介导的基因治疗是一种有前途的替代病毒基因治疗。这些合成载体的一个挑战是与体外实现的转染效率相比体内转染效率降低。许多基因传递的细胞内屏障已被阐明，但类似的定量细胞外屏障基因传递仍然是一个需要。在这项研究中，通过YOYO-I荧光猝灭试验证明了血清蛋白、可溶性糖胺聚糖和细胞外基质提取物对复合物的解包装。此外，在体外转染前将复合物暴露于血清或蛋白聚糖导致DNA的细胞摄取减少。最后，将PEI复合物和PEG化PEI复合物注射到小鼠门静脉中，并通过共聚焦显微镜评估标记的DNA和聚合物在肝内的分布。PEI复合物将DNA递送至肝脏，但观察到广泛的载体解包装，PEI主要与细胞外基质共定位。聚乙二醇化复合物介导较少的DNA递送到肝脏，可能是由于在血液中过早的载体解包装。通过这项工作，血液和细胞外基质已被确定为聚合物介导的体内基因递送到肝脏的重要细胞外屏障。
Polyplex-mediated gene therapy is a promising alternative to viral gene therapy. One challenge to these synthetic carriers is reduced transfection efficiencies in vivo compared to those achieved in vitro. Many of the intracellular barriers to gene delivery have been elucidated, but similar quantification of extracellular barriers to gene delivery remains a need. In this study, the unpackaging of polyplexes by serum proteins, soluble glycosaminoglycans, and an extracellular matrix extract was demonstrated by a YOYO-I fluorescence quenching assay. Additionally, exposing polyplexes to serum or proteoglycans before in vitro transfection caused decreased cellular uptake of DNA. Lastly, PEI polyplexes and PEGylated PEI polyplexes were injected into the portal vein of mice,and the intrahepatic distributions of labeled DNA and polymer were assessed by confocal microscopy. PEI polyplexes delivered DNA to the liver, but extensive vector unpackaging was observed, with PEI primarily colocalized with the extracellular matrix. PEGylated polyplexes mediated less DNA delivery to the liver, possibly due to premature vector unpackaging in the blood. Through this work, both the blood and the extracellular matrix have been determined to be significant extracellular barriers to polyplex-mediated in vivo gene delivery to the liver.