Rabies virus envelope glycoprotein targets lentiviral vectors to the axonal retrograde pathway in motor neurons

James N. Hislop, Tarin A. Islam, Ioanna Eleftheriadou, David C. J. Carpentier, Antonio Trabalza, Michael Parkinson, Giampietro Schiavo, Nicholas D. Mazarakis

Research output: Contribution to journalArticle

21 Citations (Scopus)

Abstract

Rabies pseudotyped lentiviral vectors have great potential in gene therapy, not least because of their ability to transduce neurons following their distal axonal application. However, very little is known about the molecular processes that underlie their retrograde transport and cell transduction. Using multiple labeling techniques and confocal microscopy, we demonstrated that pseudotyping with rabies virus envelope glycoprotein (RV-G) enabled the axonal retrograde transport of two distinct subtypes of lentiviral vector in motor neuron cultures. Analysis of this process revealed that these vectors trafficked through Rab5-positive endosomes and accumulated within a non-acidic Rab7 compartment. RV-G pseudotyped vectors were co-transported with both the tetanus neurotoxin-binding fragment and the membrane proteins thought to mediate rabies virus endocytosis (neural cell adhesion molecule, nicotinic acetylcholine receptor, and p75 neurotrophin receptor), thus demonstrating that pseudotyping with RV-G targets lentiviral vectors for transport along the same pathway exploited by several toxins and viruses. Using motor neurons cultured in compartmentalized chambers, we demonstrated that axonal retrograde transport of these vectors was rapid and efficient; however, it was not able to transduce the targeted neurons efficiently, suggesting that impairment in processes occurring after arrival of the viral vector in the soma is responsible for the low transduction efficiency seen in vivo, which suggests a novel area for improvement of gene therapy vectors.

Original languageEnglish
Pages (from-to)16148-16163
Number of pages16
JournalThe Journal of Biological Chemistry
Volume289
Issue number23
Early online date21 Apr 2014
DOIs
Publication statusPublished - 6 Jun 2014

Fingerprint

Rabies virus
Motor Neurons
Viruses
Neurons
Glycoproteins
Axonal Transport
Genetic Therapy
Nerve Growth Factor Receptor
Neural Cell Adhesion Molecules
Gene therapy
Rabies
Endosomes
Nicotinic Receptors
Carisoprodol
Endocytosis
Confocal Microscopy
Membrane Proteins
Confocal microscopy
Labeling

Keywords

  • Axon
  • Endosome
  • Glycoprotein
  • Lentivirus
  • Rab proteins
  • Trafficking

Cite this

Hislop, J. N., Islam, T. A., Eleftheriadou, I., Carpentier, D. C. J., Trabalza, A., Parkinson, M., ... Mazarakis, N. D. (2014). Rabies virus envelope glycoprotein targets lentiviral vectors to the axonal retrograde pathway in motor neurons. The Journal of Biological Chemistry, 289(23), 16148-16163. https://doi.org/10.1074/jbc.M114.549980

Rabies virus envelope glycoprotein targets lentiviral vectors to the axonal retrograde pathway in motor neurons. / Hislop, James N.; Islam, Tarin A.; Eleftheriadou, Ioanna; Carpentier, David C. J.; Trabalza, Antonio; Parkinson, Michael; Schiavo, Giampietro; Mazarakis, Nicholas D.

In: The Journal of Biological Chemistry, Vol. 289, No. 23, 06.06.2014, p. 16148-16163.

Research output: Contribution to journalArticle

Hislop, JN, Islam, TA, Eleftheriadou, I, Carpentier, DCJ, Trabalza, A, Parkinson, M, Schiavo, G & Mazarakis, ND 2014, 'Rabies virus envelope glycoprotein targets lentiviral vectors to the axonal retrograde pathway in motor neurons', The Journal of Biological Chemistry, vol. 289, no. 23, pp. 16148-16163. https://doi.org/10.1074/jbc.M114.549980
Hislop, James N. ; Islam, Tarin A. ; Eleftheriadou, Ioanna ; Carpentier, David C. J. ; Trabalza, Antonio ; Parkinson, Michael ; Schiavo, Giampietro ; Mazarakis, Nicholas D. / Rabies virus envelope glycoprotein targets lentiviral vectors to the axonal retrograde pathway in motor neurons. In: The Journal of Biological Chemistry. 2014 ; Vol. 289, No. 23. pp. 16148-16163.
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N1 - Acknowledgments We thank Kinga Bercsenyi, Maciej J. M. Szukszto, and Sultana Mahjabeen Miah for contributing to this project. We also thank Professor Ronald Montelaro (University of Pittsburgh School of Medicine) for providing the p26 EIAV reference serum. This work was supported by Seventh Framework Programme European Research Council Advanced Investigators Grant 23314 (to N. D. M. supporting J. N. H., T. A. I., I. E., A. T., and D. C. J. C.), Imperial College funds (to N. D. M.), and Cancer Research UK (to M. P., K. B., and G. S.).

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N2 - Rabies pseudotyped lentiviral vectors have great potential in gene therapy, not least because of their ability to transduce neurons following their distal axonal application. However, very little is known about the molecular processes that underlie their retrograde transport and cell transduction. Using multiple labeling techniques and confocal microscopy, we demonstrated that pseudotyping with rabies virus envelope glycoprotein (RV-G) enabled the axonal retrograde transport of two distinct subtypes of lentiviral vector in motor neuron cultures. Analysis of this process revealed that these vectors trafficked through Rab5-positive endosomes and accumulated within a non-acidic Rab7 compartment. RV-G pseudotyped vectors were co-transported with both the tetanus neurotoxin-binding fragment and the membrane proteins thought to mediate rabies virus endocytosis (neural cell adhesion molecule, nicotinic acetylcholine receptor, and p75 neurotrophin receptor), thus demonstrating that pseudotyping with RV-G targets lentiviral vectors for transport along the same pathway exploited by several toxins and viruses. Using motor neurons cultured in compartmentalized chambers, we demonstrated that axonal retrograde transport of these vectors was rapid and efficient; however, it was not able to transduce the targeted neurons efficiently, suggesting that impairment in processes occurring after arrival of the viral vector in the soma is responsible for the low transduction efficiency seen in vivo, which suggests a novel area for improvement of gene therapy vectors.

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