The ‘adenobody’ approach to viral targeting: specific and enhanced adenoviral gene delivery
The ‘adenobody’ approach to viral targeting: specific and enhanced adenoviral gene delivery
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病毒靶向的“腺体”方法：特异性和增强的腺病毒基因传递
作者：
S. Watkins;VV Mesyanzhinov;L. Kurochkina;R. Hawkins
重组腺病毒作为基因治疗的载体具有巨大的潜力。它们已经进化出一种有效的感染方法和广泛的宿主范围，但这导致了对基因递送特异性的担忧。为了靶向基于5型腺病毒的载体，我们研究了抗体方法。从噬菌体抗体库中筛选出抗病毒单链抗体片段，并构建了其与表皮生长因子（EGF）的C-末端融合蛋白。这种融合蛋白或“腺抗体”既与腺病毒的纤维蛋白结合，又与人细胞上的EGF受体（EGFR）结合，并且能够指导腺病毒与新受体结合。使用该系统，病毒感染的效率显著增强，并且靶向EGFR。腺病毒介导的感染与EGF受体的表达水平有关，并可被纯EGF的竞争所阻断。肽抑制实验表明，感染是直接介导的附着EGFR，不需要五邻体整合素相互作用。这项工作表明，“腺抗体”的方法可以提高效率，以及靶向腺病毒感染，并有许多潜在的应用基因治疗。
Recombinant adenoviruses have enormous potential as vectors for gene therapy. They have evolved an efficient method of infection and a wide host range but this leads to concerns about the specificity of gene delivery. In order to target an adenovirus type 5-based vector we have investigated an antibody approach. A virus neutralising scFv antibody fragment was isolated from a phage library and a C-terminal fusion protein with epidermal growth factor (EGF) constructed. This fusion protein, or ‘adenobody’, bound both to the fibre protein of the adenovirus and to the EGF receptor (EGFR) on human cells, and was able to direct adenoviral binding to the new receptor. Using this system the efficiency of viral infection was markedly enhanced and was targeted to the EGFR. The adenobody-directed infection correlated with the level of EGF receptor expressed on the cells and could be blocked by competition with pure EGF. Peptide inhibition experiments suggest that infection is mediated directly through attachment to the EGFR and does not require penton–integrin interactions. This work shows that the ‘adenobody’ approach can enhance the efficiency as well as target adenoviral infection and has numerous potential applications for gene therapy.