Dynamic changes in the characteristics of cationic lipidic vectors after exposure to mouse serum: implications for intravenous lipofection
Dynamic changes in the characteristics of cationic lipidic vectors after exposure to mouse serum: implications for intravenous lipofection
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DOI：
10.1038/sj.gt.3300865
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发表时间：
1999-04-01
期刊：
影响因子：
5.1
通讯作者：
Huang, L
中科院分区：
文献类型：
--
作者：
Li, S;Tseng, WC;Huang, L
通过阳离子介导载体的静脉内基因递送提供全身性基因表达，特别是在肺中。按顺序;为了理解静脉内脂质转染的机制，进行了系统的研究，以研究脂质载体与小鼠血清的相互作用，强调血清如何影响不同脂质组成的载体的生物物理和生物学性质。本研究结果表明，在暴露于血清后，介体的特性发生了动态变化。向血清中加入无菌载体导致载体立即聚集。如浊度研究、蔗糖梯度离心分析和荧光共振能量转移（FRET）研究所示，延长血清与载体的孵育时间导致载体崩解。载体崩解与DNA释放和降解相关，如EtBr嵌入测定和DNA消化研究所示。血清诱导的载体崩解是本研究中测试的所有阳离子脂质载体的普遍现象。然而，不同脂质组成的载体在崩解速率方面差异很大。体外转染载体的崩解速率与其体内转染效率呈负相关。具有快速崩解速率的载体，例如含有二油酰磷脂酰乙醇胺（DOPE）的那些载体在肺中停留较差，并且在崩解细胞中几乎没有活性。相反，具有快速聚集和缓慢崩解的含胆固醇载体在体内转染细胞中是高效的。本研究的结果解释了为什么不同脂质组合物的阳离子介导载体在其体内转染效率方面具有显著差异。这些结果还表明，研究的相互作用的载体与血清可以作为一个预测模型的体内效率的载体的进一步研究的许多相互作用的载体与血清可能导致的发展，可以提供一个基因的靶细胞在组织特异性的方式。
Intravenous gene delivery via cationic lipidic vectors gives systemic gene expression particularly in the lung. in order;to understand the mechanism of intravenous lipofection, a systematic study was performed to investigate the interactions of lipidic vectors with mouse serum emphasizing how serum affects the biophysical and biological properties of vectors of different lipid compositions. Results from this study showed that lipidic vectors underwent dynamic changes in their characteristics after exposure to serum. Addition of lipidic vectors into serum resulted in an immediate aggregation of vectors. Prolonged incubation of lipidic vectors with serum led to vector disintegration as shown in turbidity study, sucrose-gradient centrifugation analysis and fluorescence resonance energy transfer (FRET) study. Vector disintegration was associated with DNA release and degradation as shown in EtBr intercalation assay and DNA digestion study. Serum-induced disintegration of vectors is a general phenomenon for all cationic lipidic Vectors tested in this study Yet, vectors of different lipid compositions vary greatly in the rate of disintegration. There is an inverse correlation between the disintegration rate of lipidic vectors and their in vivo transfection efficiency. Vectors with a rapid rate of disintegration such as those containing dioleoylphosphatidylethanolamine (DOPE) poorly stayed in the lung and were barely active in transfecting cells. In contrast, cholesterol-containing vectors that had a rapid aggregation and a slow disintegration were highly efficient in transfecting cells in vivo. The results of this study explain why cationic lipidic vectors of different lipid compositions have a dramatic difference in their in vivo transfection efficiency. These results also suggest that the study of the interactions of lipidic vectors with serum may serve as a predictive model for the in vivo efficiency of a lipidic vector Further study of the numerous interactions of lipidic vectors with serum might lead to the development of a vector which can deliver a gene to target cells in a tissue-specific manner.