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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Effective gene expression in the rat dorsal root ganglia with a non-viral vector delivered via spinal nerve injection
Scientific Reports Open Access 17 October 2016
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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.
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.
The authors declare no conflict of interest.
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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
- integration-deficient lentiviral vector
- spinal cord
- motor neuron
- RNA interference
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