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Efficient gene expression from integration-deficient lentiviral vectors in the spinal cord

Gene Therapy volume 20, pages 645–657 (2013)Cite this article

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Abstract

Gene transfer to spinal cord cells may be crucial for therapy in spinal muscular atrophy, amyotrophic lateral sclerosis and spinal cord injury. Lentiviral vectors are efficient for transduction of a variety of cells, but like all integrating vectors they pose a risk of insertional mutagenesis. Integration-deficient lentiviral vectors (IDLVs) remain episomal but retain the transduction efficiency of standard integrating lentiviral vectors, particularly when the episomes are not diluted out through repeated cell division. We have now applied IDLVs for transduction of spinal cord in vitro, in explants and in vivo. Our results demonstrate similar efficiency of eGFP expression from integrating lentiviral vectors and IDLVs in most cell types analyzed, including motor neurons, interneurons, dorsal root ganglia (DRG) neurons and astroglia. IDLV-mediated expression of pro-glial-cell-derived neurotrophic factor (Gdnf) rescues motor neuron cultures from death caused by removal of exogenous trophic support. IDLVs also mediate efficient RNA interference in DRG neuron cultures. After intraparenchymal injection in the rat and mouse cervical and lumbar regions in vivo, transduction is mainly neuronal, with both motor neurons and interneurons being efficiently targeted. These results suggest that IDLVs could be efficient and safer tools for spinal cord transduction in future therapeutic strategies.

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Acknowledgements

We thank Luigi Naldini, Miguel Sena-Esteves and Rosario Osta for lentiviral, rabies envelope and Gdnf plasmids, respectively. We acknowledge Genoma España (GENAME project), the 7th EU Framework Program (PERSIST project, Grant agreement 222878), The Friends of Guy’s Hospital and Comisión de Investigación Científica (CSIC) de la UDELAR for financial support. SGA and VC were supported by PhD Studentships from the Egyptian Government and the ‘Govern Balear, Conselleria d’Innovació’, respectively.

Author contributions

RJY-M conceived the project; HP, EF, SBM, LM, GO, JL and RJY-M contributed to experimental design; HP, EF, SGA, NL, THH, PY, KW and VC-M conducted the experimental work; HP, EF and RJY-M wrote the manuscript, and all authors contributed to data analysis and commented on the manuscript.

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Author notes

  1. H Peluffo and E Foster: These authors contributed equally to this work.

Authors and Affiliations

  1. School of Biological Sciences, Royal Holloway-University of London, London, UK

    H Peluffo, S G Ahmed, K Wanisch, V Caraballo-Miralles & R J Yáñez-Muñoz

  2. Department of Histology and Embryology, Faculty of Medicine, UDELAR, Montevideo, Uruguay

    H Peluffo

  3. Neurodegeneration Laboratory, Institut Pasteur de Montevideo, Montevideo, Uruguay

    H Peluffo

  4. Neurorestoration Group, Wolfson Centre for Age-Related Diseases, King’s College London, London, UK

    E Foster, N Lago, T H Hutson, L Moon, P Yip & S B McMahon

  5. Department of Biology/IUNICS, Universitat de les Illes Balears, Palma, Spain

    V Caraballo-Miralles, G Olmos & J Lladó

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  1. H Peluffo
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  3. S G Ahmed
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Correspondence to R J Yáñez-Muñoz.

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Peluffo, H., Foster, E., Ahmed, S. et al. Efficient gene expression from integration-deficient lentiviral vectors in the spinal cord. Gene Ther 20, 645–657 (2013). https://doi.org/10.1038/gt.2012.78

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