Combined genetic and chemical capsid modifications enable flexible and efficient de- and retargeting of adenovirus vectors
Combined genetic and chemical capsid modifications enable flexible and efficient de- and retargeting of adenovirus vectors
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DOI：
10.1016/j.ymthe.2005.03.006
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发表时间：
2005-07-01
影响因子：
12.4
通讯作者：
Kochanek, S
中科院分区：
文献类型：
--
作者：
Kreppel, F;Gackowski, J;Kochanek, S
将病毒基因治疗载体靶向特定细胞的许多尝试都取得了有限的成功。在这里，我们描述了一种新的病毒载体靶向平台的基础上，遗传和化学载体颗粒修饰的独特组合，以克服在病毒载体靶向的典型限制。我们在腺病毒衣壳的溶剂暴露位置遗传地引入半胱氨酸。相应的硫醇基团是高度反应性的，我们建立了控制共价偶联到它们的蛋白质和非蛋白质配体的程序。在转铁蛋白的偶联后，颗粒被有效地靶向转铁蛋白受体途径。根据所使用的化学，配体可以在硫醚或二硫键的形成下偶联，后者允许在细胞进入内体后分离配体和颗粒。此外，该技术可以有效地与载体屏蔽结合以实现真正的重靶向：在载体颗粒的氨基-PEG化之后，遗传引入的硫醇仍然可用于配体偶联，并且颗粒可以重靶向转铁蛋白受体。由于该平台是稳健的，可以扩展，与工业标准兼容，并且可以整合化学上不同的分子作为配体，因此它可以用于临床基因治疗，并且还可能用于疫苗接种。
Numerous attempts to target viral gene therapy vectors to specific cells have met with limited success. Here we describe a novel virus vector-targeting platform based on a unique combination of genetic and chemical vector particle modifications to overcome typical restrictions in virus vector targeting. We genetically introduced cysteines at solvent-exposed positions of the adenovirus capsid. The corresponding thiol groups were highly reactive, and we established procedures for controlled covalent coupling to them of protein and nonprotein ligands. After the coupling of transferrin, the particles were efficiently targeted to the transferrin receptor pathway. Depending on the chemistry used, ligands could be coupled under the formation of thioether or disulfide bonds, the latter allowing for separation of ligand and particle after cell entry in the endosome. Furthermore, this technology could be efficiently combined with vector shielding for true retargeting: after amino-PEGylation of the vector particles the genetically introduced thiols were still accessible for ligand coupling, and particles could be retargeted to the transferrin receptor. Since this platform is robust, can be scaled, is compatible with industrial standards, and can integrate chemically diverse molecules as ligands, it may be used for clinical gene therapy and, potentially, also for vaccination.