Targeting of Adenovirus Vectors to the LRP Receptor Family With the High-affinity Ligand RAP via Combined Genetic and Chemical Modification of the pIX Capsomere
Targeting of Adenovirus Vectors to the LRP Receptor Family With the High-affinity Ligand RAP via Combined Genetic and Chemical Modification of the pIX Capsomere
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DOI：
10.1038/mt.2008.174
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发表时间：
2008-11-01
影响因子：
12.4
通讯作者：
Kreppel, Florian
中科院分区：
文献类型：
--
作者：
Corjon, Stephanie;Wortmann, Andreas;Kreppel, Florian
腺病毒（Ad）载体靶向需要在病毒体表面呈现特定的配体。遗传化学靶向是基于将携带活性巯基的半胱氨酸残基遗传引入到病毒粒子的溶剂可及的衣壳中，并随后进行配体的化学偶联。在这里，我们利用这种技术用高亲和力配体修饰pIX衣体。将c端半胱氨酸遗传引入pIX，允许全长蛋白与病毒粒子特异性偶联，同时不影响载体的产生。对两种高亲和力配体受体相关蛋白（RAP）和转铁蛋白（Tf）的直接比较表明，高亲和力配体与pIX偶联后的靶向可能需要配体在进入细胞后从受体释放。此外，通过标记载体颗粒的活细胞成像获得的数据表明，将非常大的蛋白质偶联到pIX可以损害细胞内载体颗粒的运输。最后，我们证明了基因化学靶向技术适用于静脉注射后肝脏的体内靶向。我们的数据为成功定位经pix修饰的广告载体的基本要求提供了重要的见解。
Adenovirus (Ad) vector targeting requires presentation of specific ligands on the virion's surface. Geneti-chemical targeting is based on the genetic introduction of cysteine residues bearing reactive thiol groups into solvent-accessible capsomeres of the virion and subsequent chemical coupling of ligands. Here, we exploited this technology to modify the pIX capsomere with high-affinity ligands. Genetic introduction of C-terminal cysteines to pIX allowed for specific coupling of full-length proteins to the virion, while not affecting vector production. Direct comparison of the two high-affinity ligands receptor-associated protein (RAP) and transferrin (Tf) revealed that targeting after coupling of a high-affinity ligand to pIX presumably requires release of the ligand from its receptor after cell entry. In addition, data obtained by live cell imaging of labeled vector particles demonstrated that coupling of very large proteins to pIX can impair intracellular vector particle trafficking. Finally, we demonstrate that the geneti-chemical targeting technology is suitable for in vivo targeting to liver after intravenous injection. Our data provide significant insight into basic requirements for successful targeting of pIX-modified Ad vectors.