Venezuelan equine encephalitis virus glycoprotein pseudotyping confers neurotropism to lentiviral vectors

Abstract

We have produced high-titre HIV-1 green fluorescent protein-expressing lentiviral (LV) vectors pseudotyped with strain 3908 Venezuelan equine encephalitis virus glycoprotein (VEEV-G) and used them to study transduction of: (1) rat embryonic motor neuron (MN) and striatal neuron primary cultures, (2) differentiated MN cell line NSC-34 and (3) adult rat striatum. In primary neuronal cultures, transduction with VEEV-G-pseudotyped LV was more efficient and more neuronal than with vesicular stomatitis virus glycoprotein (VSV-G)-pseudotyped LV. In NSC-34 cells clear retrograde transport of VEEV-G vector particles was observed. In the striatum at the injection site, transduction with the VEEV-G vectors driven by cytomegalovirus or phosphoglycerate kinase promoters exhibited a distinct neuronal tropism with no microglial and only a minor astroglial component, superior to that obtained with VSV-G-pseudotyped LV, irrespective of the promoter used. Neuronal transduction efficiency increased over time. Distal to the injection site transduction of mitral cells in the olfactory bulb, thalamic neurons and dopaminergic neurons in the substantia nigra pars compacta was detected. This, together with observations of retrograde axonal trafficking in vitro indicates that these vectors also possess low level of retrograde neuronal transduction capability in vivo. In this study, we demonstrate both strong neurotropism as well as sustainability of expression and minimal host immune response in vivo, making the VEEV-G-pseudotyped LV vectors potentially useful for gene therapy of neurodegenerative diseases.

Selective transduction of astrocytic and neuronal CNS subpopulations by lentiviral vectors pseudotyped with Chikungunya virus envelope

“…This was rather seen as the result of more efficient particle concentration as unconcentrated titers were lower than VSV-G. We previously observed this with the VEEV-G pseudotyped vectors [22]. By contrast SINV-G pseudotyped vectors failed to concentrate well and the titres remained in the 10 7 TU/ml range comparable to those reported by others previously [27].…”

“…No increased neurotropism was observed as we had reported for the VEEV-G pseudotype [22] but instead a strong astrocytic tropism was apparent. NHAs are serum selected primary astrocytes that express high levels of reactive astrocyte genes [47].…”

“…without causing significant tissue damage [18][19][20][21][22]. This, coupled with the fact that some target the nervous system [23] and the idea that most humans do not have pre-existing antibodies against them due to their restricted geography [23] The alphaviral genome encodes several non-structural and structural proteins that are…”

“…No evidence of retrograde transport and transduction of distal neurons was apparent, in a manner similar to VSV-G pseudotyped vectors and contrasting to RVG or VEEV-G pseudotyped ones [22]. This is suggestive that the interactive receptor is not being internalised or does not engage the axonal retrograde pathway in neurons [63].…”

“…So far the only LVs efficiently pseudotyped with alphaviral glycoproteins include RRV [18,39] and VEEV [22], which have been studied in vivo in the CNS. We set out to produce and study SINV-G and CHIKV-G pseudotyped LVs in vitro and compare them to VSV-G pseudotyped vectors.…”