Hexon-specific PEGylated adenovirus vectors utilizing avidin-biotin interaction.
Hexon-specific PEGylated adenovirus vectors utilizing avidin-biotin interaction.
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DOI：
10.1016/j.biomaterials.2010.10.060
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发表时间：
2011-02
期刊：
影响因子：
14
通讯作者：
Emi Suzuki-Kouyama;K. Katayama;F. Sakurai;Tomoko Yamaguchi;S. Kurachi;K. Kawabata;S. Nakagawa;H. Mizuguchi
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文献类型：
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作者：
Emi Suzuki-Kouyama;K. Katayama;F. Sakurai;Tomoko Yamaguchi;S. Kurachi;K. Kawabata;S. Nakagawa;H. Mizuguchi
聚乙二醇修饰的重组腺病毒（Ad）载体是一种很有前途的方法，不仅可以避免中和抗Ad抗体和被吞噬细胞摄取，而且可以延长Ad载体全身给药后的血液滞留时间。然而，常规PEG化导致Ad载体的转导活性显著降低，这可能是因为PEG非特异性地缀合至Ad衣壳蛋白并抑制Ad载体与主要受体柯萨奇病毒-腺病毒受体（CAR）的结合。为了PEG化Ad载体而不显著降低转导活性，将生物素结合肽（BAP）插入六邻体的高变区（HVR）5中，其不参与与CAR的结合，然后PEG通过抗生物素蛋白-生物素相互作用特异性缀合至六邻体HVR 5。在体外转导实验表明，六邻体特异性聚乙二醇化并没有引起明显的降低，在腺病毒载体的转导效率，虽然插入的BAP到HVR 5本身的转导效率降低了50倍，与传统的腺病毒载体相比，在没有抗腺病毒血清。在抗Ad血清的存在下，六邻体HVR 5中具有BAP的Ad载体的转导被显著阻断;然而，抗Ad血清仅轻微抑制六邻体特异性PEG化Ad载体（Ad-BAP/Bio/Avi/Bio-PEG-L2）的转导。静脉注射Ad-BAP/Bio/Avi/Bio-PEG-L2导致血液滞留时间延长，肝脏中的转导显著减少，并在肿瘤中蓄积;然而，出乎意料的是，Ad-BAP/Bio/Avi/Bio-PEG-L2在肿瘤中的转导效率几乎处于背景水平。
PEGylation of recombinant adenovirus (Ad) vectors is a promising approach for not only evasion from neutralizing anti-Ad antibodies and uptake by phagocytic cells, but also prolongation of the blood retention time of Ad vectors after systemic administration. However, the conventional PEGylation leads to significant reduction in the transduction activity of Ad vectors, probably because PEG is nonspecifically conjugated to the Ad capsid protein and inhibits the binding of Ad vectors to the primary receptor, coxsackievirus-adenovirus receptor (CAR). In order to PEGylate an Ad vector without significant reduction in the transduction activity, the biotin-binding peptide (BAP) was inserted into the hypervariable region (HVR) 5 of the hexon, which is not involved in the binding to CAR, and PEG was then specifically conjugated to the hexon HVR5 via avidin-biotin interaction. In vitro transduction experiments demonstrated that the hexon-specific PEGylation did not cause an apparent reduction in the transduction efficiency of the Ad vector, although the insertion of the BAP into the HVR5 itself reduced the transduction efficiency by 50-fold, compared with the conventional Ad vector, in the absence of anti-Ad serum. In the presence of anti-Ad serum, the transduction with the Ad vector with the BAP in the hexon HVR5 was significantly blocked; however, anti-Ad serum only slightly inhibited the transduction with the hexon-specifically PEGylated Ad vector (Ad-BAP/Bio/Avi/Bio-PEG-L2). Intravenous administration of Ad-BAP/Bio/Avi/Bio-PEG-L2 resulted in prolonged blood retention, significant reduction in the transduction in the liver, and accumulation in the tumor; however, unexpectedly, the transduction efficiency of Ad-BAP/Bio/Avi/Bio-PEG-L2 in the tumor was almost at the background level.