Wong Liang-Fong, Azzouz Mimoun, Walmsley Lucy E, Askham Zoe, Wilkes Fraser J, Mitrophanous Kyriacos A, Kingsman Susan M, Mazarakis Nicholas D
Neurobiology Group, Oxford Biomedica (UK) Ltd., Medawar Centre, Oxford OX4 4GA, United Kingdom.
Mol Ther. 2004 Jan;9(1):101-11. doi: 10.1016/j.ymthe.2003.09.017.
We have developed a non-primate-based lentiviral vector based on the equine infectious anemia virus (EIAV) for efficient gene transfer to the central and peripheral nervous systems. Previously we have demonstrated that pseudotyping lentiviral vectors with the rabies virus glycoprotein confers retrograde axonal transport to these vectors. In the present study we have successfully produced high-titer EIAV vectors pseudotyped with envelope glycoproteins from Rhabdovirus vesicular stomatitis virus (VSV) serotypes (Indiana and Chandipura strains); rabies virus [various Evelyn-Rokitnicki-Abelseth ERA strains and challenge virus standard (CVS)]; Lyssavirus Mokola virus, a rabies-related virus; and Arenavirus lymphocytic choriomeningitis virus (LCMV). These vectors were delivered to the striatum or spinal cord of adult rats or muscle of neonatal mice by direct injection. We report that the lentiviral vectors pseudotyped with envelopes from the VSV Indiana strain, wild-type ERA, and CVS strains resulted in strong transduction in the striatum, while Mokola- and LCMV-pseudotyped vectors exhibited moderate and weak transduction, respectively. Furthermore ERA- and CVS-pseudotyped lentiviral vectors demonstrated retrograde transport and expression in distal neurons after injection in brain, spinal cord, and muscle. The differences in transduction efficiencies and retrograde transport conferred by these envelope glycoproteins present novel opportunities in designing therapeutic strategies for different neurological diseases.
我们基于马传染性贫血病毒(EIAV)开发了一种非灵长类慢病毒载体,用于将基因高效转移至中枢神经系统和外周神经系统。此前我们已证明,用狂犬病病毒糖蛋白对慢病毒载体进行假型化可使这些载体实现逆行轴突运输。在本研究中,我们成功制备了高滴度的EIAV载体,其假型化包膜糖蛋白来自弹状病毒水疱性口炎病毒(VSV)血清型(印第安纳株和钱迪普拉株)、狂犬病病毒[各种伊夫林 - 罗基特尼基 - 阿贝尔塞斯(ERA)株和攻击病毒标准株(CVS)]、狂犬病相关病毒莫科拉病毒以及沙粒病毒淋巴细胞性脉络丛脑膜炎病毒(LCMV)。通过直接注射将这些载体递送至成年大鼠的纹状体或脊髓或新生小鼠的肌肉。我们报告称,用VSV印第安纳株、野生型ERA和CVS株的包膜假型化的慢病毒载体在纹状体中产生了强烈的转导,而莫科拉病毒和LCMV假型化的载体分别表现出中等和较弱的转导。此外,ERA和CVS假型化的慢病毒载体在脑、脊髓和肌肉注射后,在远端神经元中显示出逆行运输和表达。这些包膜糖蛋白赋予的转导效率和逆行运输的差异为设计针对不同神经疾病的治疗策略提供了新的机会。