PEGylated adenovirus vectors containing RGD peptides on the tip of PEG show high transduction efficiency and antibody evasion ability
PEGylated adenovirus vectors containing RGD peptides on the tip of PEG show high transduction efficiency and antibody evasion ability
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DOI：
10.1002/jgm.699
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发表时间：
2005-05-01
影响因子：
3.5
通讯作者：
Nakagawa, S
中科院分区：
文献类型：
--
作者：
Eto, Y;Gao, JQ;Nakagawa, S
背景腺病毒载体（Ads）的聚乙二醇化是基因治疗中一种有吸引力的策略。尽管已经开发了许多类型的表现出抗体逃避活性和长血浆半衰期的聚乙二醇化Ad（PEG-Ad），但是聚乙二醇（PEG）链的空间位阻阻止了它们进入细胞。同样，足够的基因表达的医疗treatment.Methods，这有不同的PEG修饰率，一组的PEG-广告，构建，并使用A549细胞的基因表达进行了评估。本文报道了一种新型的聚乙二醇化腺病毒（RGD-PEG-Ad），它在PEG的末端含有RGD（Arg-Gly-Asp）肽段。我们评估了柯萨奇-腺病毒受体（CAR）阳性和阴性细胞中的基因表达，并测定了体内基因表达。结果PEG-Ads能降低CAR阳性细胞的基因表达，而RGD-PEG-Ad能显著增强CAR阳性细胞的基因表达，其表达水平是PEG-Ads的200倍。此外，RGD-PEG-Ad的基因表达几乎等于Ad-RGD的基因表达，Ad-RGD在纤维中含有RGD基序，并且在CAR阳性和阴性细胞中表现出最高的基因表达。此外，尽管在抗Ad抗血清存在下Ad-RG-D基因表达显著降低，但RGD-PEG-Ad保持其抗抗体活性。体内实验也表明，修饰的广告与RGD-PEG诱导有效的基因express.Conclusions在本研究中，我们证明了一种新的策略，结合整合素靶向的RGD肽的PEG尖端和修饰的广告使用这种材料，可以提高基因表达在CAR阳性和阴性细胞。同时，这种新型PEG化Ad保持了对抗体的强保护活性。这种策略也可以很容易地修改，以开发使用其他靶向分子的其他载体。版权所有（c）2004年约翰威利父子有限公司。
Background PEGylation of adenovirus vectors (Ads) is an attractive strategy in gene therapy. Although many types of PEGylated Ad (PEG-Ads), which exhibit antibody evasion activity and long plasma half-life, have been developed, their entry into cells has been prevented by steric hindrance by polyethylene glycol (PEG) chains. Likewise, sufficient gene expression for medical treatment could not be achieved.Methods A set of PEG-Ads, which have different PEG modification rates, was constructed, and gene expression was evaluated using A549 cells. A novel PEGylated Ad (RGD-PEG-Ad), which contained RGD (Arg-Gly-Asp) peptides on the tip of PEG, was developed. We evaluated gene expression both in Coxsackie-adenovirus receptor (CAR)-positive as well as -negative cells, and in vivo gene expression was also determined. Furthermore, the antibody evasion ability and the specificity of infection exhibited by this RGD-PEG-Ad were also evaluated.Results Whereas PEG-Ads decreased gene expression in CAR-positive cells, RGD-PEG-Ad enhanced gene expression notably, to a level about 200-fold higher than that of PEG-Ads. Moreover, gene expression of RGD-PEG-Ad was almost equal to that of Ad-RGD, which contains an RGD-motif in the fiber and exhibits among the highest gene expression of CAR-positive and -negative cells. Furthermore, although Ad-RG-D gene expression decreased remarkably in the presence of anti-Ad antiserum, RGD-PEG-Ad maintained its activity against antibodies. In vivo experiments also demonstrated that the modification of Ads with RGD-PEG induced efficient gene expression.Conclusions In the present study, we demonstrated that a new strategy, which combined integrin-targeting the RGD peptide on the tip of PEG and modified the Ad using this material, could enhance gene expression in both CAR-positive and -negative cells. At the same time, this novel PEGylated Ad maintained strong protective activity against antibodies. This strategy could also be easily modified for developing other vectors using other targeting molecules. Copyright (c) 2004 John Wiley & Sons, Ltd.