Enhanced pseudotyping efficiency of HIV-1 lentiviral vectors by a rabies/vesicular stomatitis virus chimeric envelope glycoprotein
Enhanced pseudotyping efficiency of HIV-1 lentiviral vectors by a rabies/vesicular stomatitis virus chimeric envelope glycoprotein
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DOI：
10.1038/gt.2011.124
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发表时间：
2012-07-01
期刊：
影响因子：
5.1
通讯作者：
Mazarakis, N. D.
中科院分区：
文献类型：
--
作者：
Carpentier, D. C. J.;Vevis, K.;Mazarakis, N. D.
狂犬病病毒糖蛋白（RVG）可以假型化慢病毒载体，尽管其效率低于水泡性口炎病毒糖蛋白（VSVG）。啮齿动物中枢神经系统（CNS）的VSVG-假型载体转导导致局部亲神经基因转移，而RVG-假型载体位于远端传出部位的神经元的额外分散转导通过轴突逆行运输发生。迄今为止，生产用于临床前和临床试验的高滴度RVG假型慢病毒载体的尝试是有问题的。我们已经构建了几种嵌合RVG/VSVG糖蛋白，并发现带有RVG的外部/跨膜结构域和VSVG的胞质结构域的构建体显示出增加的掺入到HIV-1慢病毒颗粒上，并在体外在293 T细胞和人、大鼠和小鼠来源的分化的神经元细胞系中具有增加的感染性。在大鼠纹状体中用该RVG/VSVG嵌合体假型化的载体的立体定向应用导致在注射部位的有效基因转移，显示神经元和胶质细胞向性。通过逆行轴突运输的黑质，丘脑和嗅球中的远端神经元转导也发生在纹状体内施用嵌合体假型载体后，其水平与用RVG假型载体观察到的水平相似。这是第一个报告的远端转导嗅球。用这种包膜增强的假型化应该能够更容易地生产更高滴度的假型化慢病毒载体，其在CNS中表现出有效的局部和分散的神经元转导。Gene Therapy（2012）19，761-774; doi：10.1038/gt.2011.124; 2011年9月8日在线发表
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. Gene Therapy (2012) 19, 761-774; doi:10.1038/gt.2011.124; published online 8 September 2011