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

The Journal of biological chemistry

PubMedID: 24753246

Hislop JN, Islam TA, Eleftheriadou I, Carpentier DC, Trabalza A, Parkinson M, Schiavo G, Mazarakis ND. Rabies virus envelope glycoprotein targets lentiviral vectors to the axonal retrograde pathway in motor neurons. J Biol Chem. 2014;.
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 labelling techniques and confocal microscopy we demonstrate that pseudotyping with rabies virus glycoprotein (RV-G) enables the axonal retrograde transport of two distinct subtypes of lentiviral vector in motor neuron cultures. Analysis of this process reveals that these vectors traffic through Rab5-positive endosomes and accumulate within a non-acidic Rab7 compartment. RV-G pseudotyped vectors are co-transported with both the tetanus neurotoxin binding fragment as well as the membrane proteins thought to mediate rabies virus endocytosis (neural cell adhesion molecule, nicotinic acetylcholine receptor, 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 cultures in compartmentalised chambers, we demonstrate that axonal retrograde transport of this vectors is 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, and suggests a novel area for improvement of gene therapy vectors.