Department of Molecular Genetics, Institute of Biomedical Sciences, Fukushima Medical University School of Medicine, Fukushima, Japan.
Hum Gene Ther. 2011 Feb;22(2):197-206. doi: 10.1089/hum.2009.179. Epub 2011 Jan 27.
The lentiviral vector system based on human immunodeficiency virus type 1 (HIV-1) is used extensively in gene therapy trials of neurological and neurodegenerative diseases. Retrograde axonal transport of viral vectors offers a great advantage to the delivery of genes into neuronal cell bodies that are situated in regions distant from the injection site. Pseudotyping of HIV-1-based vectors with selective variants of rabies virus glycoprotein (RV-G) increases gene transfer via retrograde transport into the central nervous system. Because large-scale application for gene therapy trials requires high titer stocks of the vector, pseudotyping of a lentiviral vector that produces more efficient retrograde transport is needed. In the present study, we developed a novel vector system for highly efficient retrograde gene transfer by pseudotyping an HIV-1 vector with a fusion envelope glycoprotein (termed FuG-B) in which the cytoplasmic domain of RV-G was substituted by the corresponding part of vesicular stomatitis virus glycoprotein. The FuG-B pseudotype shifted the transducing property of the lentiviral vector and enhanced the retrograde transport-mediated gene transfer into different brain regions innervating the striatum with greater efficiency than that of the RV-G pseudotype in mice. In addition, injection of the FuG-B-pseudotyped vector into monkey striatum (caudate and putamen) allowed for highly efficient gene delivery into the nigrostriatal dopamine system, which is a major target for gene therapy of Parkinson's disease. Our strategy provides a powerful tool for the treatment of certain neurological and neurodegenerative diseases by promoting retrograde gene delivery via a lentiviral vector.
基于人类免疫缺陷病毒 1 型 (HIV-1) 的慢病毒载体系统广泛应用于神经和神经退行性疾病的基因治疗试验。病毒载体的逆行轴突运输为将基因递送到远离注射部位的神经元细胞体提供了巨大优势。用狂犬病病毒糖蛋白 (RV-G) 的选择性变体对基于 HIV-1 的载体进行假型化可增加通过逆行运输进入中枢神经系统的基因转移。因为基因治疗试验的大规模应用需要载体的高滴度库存，所以需要假型化产生更有效的逆行运输的慢病毒载体。在本研究中，我们通过用融合包膜糖蛋白 (称为 FuG-B) 假型化 HIV-1 载体开发了一种新型高效逆行基因转移载体系统，其中 RV-G 的细胞质结构域被水疱性口炎病毒糖蛋白的相应部分取代。FuG-B 假型改变了慢病毒载体的转导特性，并增强了逆行运输介导的基因转移到不同的脑区，与 RV-G 假型相比，向纹状体支配的脑区的转移效率更高在小鼠中。此外，将 FuG-B 假型化载体注射到猴纹状体 (尾状核和壳核) 中允许高效地将基因递送到黑质纹状体多巴胺系统，这是帕金森病基因治疗的主要靶点。我们的策略通过促进慢病毒载体的逆行基因传递，为某些神经和神经退行性疾病的治疗提供了一种强大的工具。
