Chimeric rabies SADB19-VSVg-pseudotyped lentiviral vectors mediate long-range retrograde transduction from the mouse spinal cord

Abstract

Lentiviral vectors have proved an effective method to deliver transgenes into the brain; however, they are often hampered by a lack of spread from the site of injection. Modifying the viral envelope with a portion of a rabies envelope glycoprotein can enhance spread in the brain by using long-range axon projections to facilitate retrograde transport. In this study, we generated two chimeric envelopes containing the extra-virion and transmembrane domain of rabies SADB19 or CVS-N2c with the intra-virion domain of vesicular stomatitis virus. Viral particles were packaged containing a green fluorescent protein reporter construct under the control of the phosphoglycerokinase promoter. Both vectors produced high-titer particles with successful integration of the glycoproteins into the particle envelope and significant transduction of neurons in vitro. Injection of the SADB19 chimeric viral vector into the lumbar spinal cord of adult mice mediated a strong preference for gene transfer to local neurons and axonal terminals, with retrograde transport to neurons in the brainstem, hypothalamus and cerebral cortex. Development of this vector provides a useful means to reliably target select populations of neurons by retrograde targeting.

Lentiviral vectors as tools to understand central nervous system biology in mammalian model organisms

“…The spread of one microliter of VSVg pseudotyped vector in the brain is restricted to 1–2 mm from the injection site ( Desmaris et al, 2001 ; Linterman et al, 2011 ), with no retrograde transport. By exchanging the VSVg for a rabies glycoprotein (Rbg; Mazarakis et al, 2001 ), or a chimera of these two ( Kato et al, 2011a , b , 2014 ; Carpentier et al, 2012 ; Schoderboeck et al, 2015 ) retrograde transport is enabled so that after transduction of axonal terminals, viral contents (minus envelope) are transported to the cell body ( Figure 1 ).…”

“…Scale bar = 50 μm. Reproduced with permission from Schoderboeck et al (2015) . (B) An electron micrograph of a lentivirus transduced spine in the monkey striatum expressing eYFP.…”

“…Here, lentiviral vectors offer improved specificity because most AAV serotypes would label both afferent and efferent neurons in the region ( Klaw et al, 2013 ; Figure 1 ). In addition, the flexibility to have the vectors endocytosed at the terminals or somata of target neurons means that the investigator can select the vector so that injection and uptake is in an area that will not be visualized ( Schoderboeck et al, 2015 ; Figure 4A ). The advantage for experiments with an anatomical focus is that the physical damage or artifact caused by the injection does not interfere with the interpretation or analysis of images taken at high-resolution, such as electron microscopy.…”

Tropism, intracerebral distribution, and transduction efficiency of HIV- and SIV-based lentiviral vectors after injection into the mouse brain: a qualitative and quantitative in vivo study

“…No EGFP could be detected in the olfactory tract, and it has to be taken into account that a minimum of two synapses has to be bridged on the way from piriform lobe to olfactory bulb. The mechanism of the retrograde transport of viral particles within the brain is not known, but glycoproteins of the viral envelope seem to play a role during the interaction with host cells and endosomal vector transport (Desmaris et al 2001 ; Kato et al 2011 ; Carpentier et al 2012 ; Schoderboeck et al 2015 ). Our results confirm that transduction of olfactory bulb neurons is possible for simple VSVG-pseudotyped lentiviruses, even if rabies or chimeric rabies/VSVG pseudotyping might enhance retrograde axonal transport of vectors (Kato et al 2011 ; Carpentier et al 2012 ; Schoderboeck et al 2015 ).…”

Viral vector-based tools advance knowledge of basal ganglia anatomy and physiology

“…However, using glycoproteins from other viruses or VSVg chimerics with rabies glycoproteins can allow strong retrograde uptake (Kato et al 2011(Kato et al , 2014Schoderboeck et al 2015). Additional envelopes have been used to select specific neural types, e.g., the lymphocytic choriomeningitis virus (LCMV) glycoprotein results in preferential transduction of astrocytes in the rat substantia nigra (Cannon et al 2011), or rabies to confer retrograde transport (Kato et al 2011(Kato et al , 2014Schoderboeck et al 2015). The packaging capacity of lentiviruses is ϳ10 kb, allowing relatively large and complex constructs to be introduced.…”