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Innocuous full-length botulinum neurotoxin targets and promotes the expression of lentiviral vectors in central and autonomic neurons

Gene Therapy volume 18, pages 656–665 (2011)Cite this article

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Abstract

Fragments of botulinum neurotoxin (BoNT) have been explored as potential targeting moieties and carriers of biomolecules into neurons, although with lower binding and translocation efficiency compared with intact proteins. This study exploits a detoxified recombinant form of full-length BoNT/B (BoTIM/B) fused with core streptavidin (CS-BoTIM/B) for lentiviral targeting to central and autonomic neurons. CS-BoTIM/B underwent an activity-dependent entry into cultured spinal cord neurons. Coupling CS-BoTIM/B to biotinylated lentivirus-encoding green fluorescent protein (GFP) endowed considerable neuron selectivity to the vector as evident from the preferential expression of the reporter in neurons co-cultured with skeletal muscle cells. CS-BoTIM/B-guided lentiviral transduction with the expression of a SNARE protein, SNAP-25 (S25), rendered non-susceptible to proteolysis by three BoNT serotypes, yielded a sizable decrease in cleaved S25 upon exposure of spinal cord neurons to these toxins. This was accompanied by synaptic transmission being spared from blockade by BoNT/A or BoNT/E, reflecting adequate translation and functional competence of recombinant multi-toxin-resistant S25. The augmented neurotropism conveyed on the lentivirus by CS-BoTIM/B was also demonstrated in vivo through enhanced expression of a reporter in intramural ganglionic neurons in the rat trachea, after injection of the targeted GFP-encoding lentivirus. Thus, a novel and realistic prospect for gene therapy of peripheral neuropathies is offered in this study through lentiviral targeting to neurons by CS-BoTIM/B.

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Acknowledgements

This study is supported by USAMRIID (grant number HDTRA1-07-C-0034) and a Research Professorship plus Principal Investigator award (to JOD) from the Science Foundation Ireland. We are grateful to Dr Jiafu Wang for providing the pET-29a clone of BoNT/B and Liam Ryan for technical assistance.

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Authors and Affiliations

  1. International Centre for Neurotherapeutics, Dublin City University, Dublin, Ireland

    V B O'Leary, S V Ovsepian, A Raghunath, Q Huo, G W Lawrence & J O Dolly

  2. Office of the Chief Scientist, United States Army Medical Research Institute of Infectious Diseases, Frederick, MD, USA

    L Smith

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  1. V B O'Leary
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  2. S V Ovsepian
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  3. A Raghunath
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Correspondence to J O Dolly.

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O'Leary, V., Ovsepian, S., Raghunath, A. et al. Innocuous full-length botulinum neurotoxin targets and promotes the expression of lentiviral vectors in central and autonomic neurons. Gene Ther 18, 656–665 (2011). https://doi.org/10.1038/gt.2011.8

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