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Venezuelan equine encephalitis virus glycoprotein pseudotyping confers neurotropism to lentiviral vectors

Gene Therapy volume 20, pages 723–732 (2013)Cite this article

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Abstract

We have produced high-titre HIV-1 green fluorescent protein-expressing lentiviral (LV) vectors pseudotyped with strain 3908 Venezuelan equine encephalitis virus glycoprotein (VEEV-G) and used them to study transduction of: (1) rat embryonic motor neuron (MN) and striatal neuron primary cultures, (2) differentiated MN cell line NSC-34 and (3) adult rat striatum. In primary neuronal cultures, transduction with VEEV-G-pseudotyped LV was more efficient and more neuronal than with vesicular stomatitis virus glycoprotein (VSV-G)-pseudotyped LV. In NSC-34 cells clear retrograde transport of VEEV-G vector particles was observed. In the striatum at the injection site, transduction with the VEEV-G vectors driven by cytomegalovirus or phosphoglycerate kinase promoters exhibited a distinct neuronal tropism with no microglial and only a minor astroglial component, superior to that obtained with VSV-G-pseudotyped LV, irrespective of the promoter used. Neuronal transduction efficiency increased over time. Distal to the injection site transduction of mitral cells in the olfactory bulb, thalamic neurons and dopaminergic neurons in the substantia nigra pars compacta was detected. This, together with observations of retrograde axonal trafficking in vitro indicates that these vectors also possess low level of retrograde neuronal transduction capability in vivo. In this study, we demonstrate both strong neurotropism as well as sustainability of expression and minimal host immune response in vivo, making the VEEV-G-pseudotyped LV vectors potentially useful for gene therapy of neurodegenerative diseases.

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Acknowledgements

We would like to thank our ex-MSc students Petros Patsali, Daniel M Lipiski and Joanne Crowe for technical contributions to this study. Dr Robert A. Davey, University of Texas for giving us the VEEV-G TRD and 3908 plasmids. Professor Richard Reynolds and Dr Owain Howell for the use of and help with fluorescence microscopy to document histological staining of brain sections. We would also like to thank Dr Egle Solito and Enrico Cristante for letting us use several antibodies. This work was partly supported by Imperial College London starting funds to NDM and a Seventh Framework Programme European Research Council Advanced Grant, no: 23314 to NDM supporting AT, IE, JH and SME.

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  1. C Georgiadis

    Present address: 2Current address: Institute of Child Health & Great Ormond Street Hospital, 30 Guilford Street, London WC1N 1EH, UK.,

Authors and Affiliations

  1. Division of Brain Sciences, Faculty of Medicine, Gene Therapy, Centre for Neuroinflammation & Neurodegeneration, Imperial College London, Hammersmith Hospital Campus, London, UK

    A Trabalza, C Georgiadis, I Eleftheriadou, J N Hislop, S M Ellison, M E Karavassilis & N D Mazarakis

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  1. A Trabalza
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Correspondence to N D Mazarakis.

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Trabalza, A., Georgiadis, C., Eleftheriadou, I. et al. Venezuelan equine encephalitis virus glycoprotein pseudotyping confers neurotropism to lentiviral vectors. Gene Ther 20, 723–732 (2013). https://doi.org/10.1038/gt.2012.85

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