Targeting of adenovirus via genetic modification of the viral capsid combined with a protein bridge
Targeting of adenovirus via genetic modification of the viral capsid combined with a protein bridge
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DOI：
10.1128/jvi.77.24.12931-12940.2003
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发表时间：
2003-12-01
影响因子：
5.4
通讯作者：
Krasnykh, V
中科院分区：
文献类型：
--
作者：
Korokhov, N;Mikheeva, G;Krasnykh, V
开发用于细胞特异性递送基因治疗剂的遗传靶向腺病毒（Ad）载体的潜在障碍在于以下事实：几种类型的靶向蛋白配体需要翻译后修饰，例如二硫键的形成，这对于Ad衣壳蛋白是不可用的，这是由于它们在病毒体组装期间的核定位。为了克服这一问题，我们开发了一种新的靶向策略，该策略将Ad衣壳的遗传修饰与蛋白质桥方法相结合，从而产生载体-配体靶向复合物。复合物的组分通过遗传修饰与Ad衣壳和靶向配体结合。这种结合机制的一个组成部分，金黄色葡萄球菌蛋白A的Fc结合结构域，被基因整合到Ad纤维蛋白中。配体由与免疫球蛋白的Fc结构域融合的靶向组分组成，其在Ad-配体复合物形成期间充当与这些遗传修饰的纤维结合的对接部分。配体的模块化设计解决了与Ad的结构和生物合成相容性的问题，从而促进载体靶向多种细胞受体。我们的研究表明，结合Fc结构域和抗CD 40单链抗体或CD 40 L的靶向配体与蛋白A修饰的Ad载体形成稳定的复合物，导致基因递送至CD 40阳性靶细胞的显著增强。由于这种基因转移不依赖于靶细胞对天然Ad 5受体的表达，因此这种策略导致了适用于组织特异性基因治疗的真正靶向的Ad载体的衍生。
A potential barrier to the development of genetically targeted adenovirus (Ad) vectors for cell-specific delivery of gene therapeutics lies in the fact that several types of targeting protein ligands require posttranslational modifications, such as the formation of disulfide bonds, which are not available to Ad capsid proteins due to their nuclear localization during assembly of the virion. To overcome this problem, we developed a new targeting strategy, which combines genetic modifications of the Ad capsid with a protein bridge approach, resulting in a vector-ligand targeting complex. The components of the complex associate by virtue of genetic modifications to both the Ad capsid and the targeting ligand. One component of this mechanism of association, the Fc-binding domain of Staphylococcus aureus protein A, is genetically incorporated into the Ad fiber protein. The ligand is comprised of a targeting component fused with the Fc domain of immunoglobulin, which serves as a docking moiety to bind to these genetically modified fibers during the formation of the Ad-ligand complex. The modular design of the ligand solves the problem of structural and biosynthetic compatibility with the Ad and thus facilitates targeting of the vector to a variety of cellular receptors. Our study shows that targeting ligands incorporating the Fc domain and either an anti-CD40 single-chain antibody or CD40L form stable complexes with protein A-modified Ad vectors, resulting in significant augmentation of gene delivery to CD40-positive target cells. Since this gene transfer is independent of the expression of the native Ad5 receptor by the target cells, this strategy results in the derivation of truly targeted Ad vectors suitable for tissue-specific gene therapy.