Department of Gene Therapy, Centre for Neurosciences, Division of Experimental Medicine, Faculty of Medicine, Imperial College London, Hammersmith Hospital, UK.
Gene Ther. 2012 Jul;19(7):761-74. doi: 10.1038/gt.2011.124. Epub 2011 Sep 8.
Rabies virus glycoprotein (RVG) can pseudotype lentiviral vectors, although at a lower efficiency to that of vesicular stomatitis virus glycoprotein (VSVG). Transduction with VSVG-pseudotyped vectors of rodent central nervous system (CNS) leads to local neurotropic gene transfer, whereas with RVG-pseudotyped vectors additional disperse transduction of neurons located at distal efferent sites occurs via axonal retrograde transport. Attempts to produce high-titre RVG-pseudotyped lentiviral vectors for preclinical and clinical trials has to date been problematic. We have constructed several chimeric RVG/VSVG glycoproteins and found that a construct bearing the external/transmembrane domain of RVG and the cytoplasmic domain of VSVG shows increased incorporation onto HIV-1 lentiviral particles and has increased infectivity in vitro in 293T cells and in differentiated neuronal cell lines of human, rat and murine origin. Stereotactic application of vector pseudotyped with this RVG/VSVG chimera in the rat striatum resulted in efficient gene transfer at the site of injection showing both neuronal and glial tropism. Distal neuronal transduction in the substantia nigra, thalamus and olfactory bulb via retrograde axonal transport also occurs after intrastriatal administration of chimera-pseudotyped vectors at similar levels to that observed with a RVG-pseudotyped vector. This is the first report of distal transduction in the olfactory bulb. The enhanced pseudotyping with this envelope should enable easier production of higher-titre pseudotyped lentiviral vectors that exhibit efficient local and dispersed neuronal transduction in the CNS.
狂犬病病毒糖蛋白 (RVG) 可以假型慢病毒载体，尽管效率低于水疱性口炎病毒糖蛋白 (VSVG)。用 VSVG 假型载体转导啮齿动物中枢神经系统 (CNS) 会导致局部嗜神经基因转移，而用 RVG 假型载体则通过轴突逆行转运发生神经元的额外弥散转导。迄今为止，为临床前和临床试验生产高滴度 RVG 假型慢病毒载体的尝试一直存在问题。我们构建了几种嵌合 RVG/VSVG 糖蛋白，发现一种带有 RVG 的外膜/跨膜结构域和 VSVG 的细胞质结构域的构建体显示出对 HIV-1 慢病毒颗粒的更高整合，并在 293T 细胞和分化的神经元细胞系中人、大鼠和小鼠来源的细胞系中具有更高的体外感染性。用这种 RVG/VSVG 嵌合体假型的载体立体定向应用于大鼠纹状体导致注射部位的有效基因转移，表现出神经元和神经胶质嗜性。通过逆行轴突转运，在纹状体给药后，通过嗅球、丘脑和黑质的远端神经元转导也以与 RVG 假型载体观察到的相似水平发生。这是嗅球中发生远端转导的第一个报道。这种包膜的增强假型化应该能够更容易地生产更高滴度的假型慢病毒载体，从而在 CNS 中实现有效的局部和弥散神经元转导。
