In utero delivery of rAAV2/9 induces neuronal expression of the transgene in the brain: towards new models of Parkinson’s disease

Abstract

Animal models are essential tools for basic pathophysiological research as well as validation of therapeutic strategies for curing human diseases. However, technical difficulties associated with classical transgenesis approaches in rodent species higher than Mus musculus have prevented this long-awaited development. The availability of viral-mediated gene delivery systems in the past few years has stimulated the production of viruses with unique characteristics. For example, the recombinant adeno-associated virus serotype 9 (rAAV2/9) crosses the blood-brain barrier, is capable of transducing developing cells and neurons after intravenous injection and mediates long-term transduction. Whilst post-natal delivery is technically straightforward, in utero delivery bears the potential of achieving gene transduction in neurons at embryonic stages during which the target area is undergoing development. To test this possibility, we injected rAAV2/9 carrying either A53T mutant human α-synuclein or green fluorescent protein, intracerebroventricularly in rats at embryonic day 16.5. We observed neuronal transgene expression in most regions of the brain at 1 and 3 months after birth. This proof-of-concept experiment introduces a new opportunity to model brain diseases in rats.

Pilot Study Assessing the Impact of Intrathecal Administration of Variants AAV-PHP.B and AAV-PHP.eB on Brain Transduction in Adult Rhesus Macaques

“…Since both serotypes share the ability to cross the blood-brain barrier (BBB) in newborns, they may be delivered through various administration routes ( Bourdenx et al, 2014 ; Bedbrook et al, 2018 ). They are associated with widespread, long-term transduction in nondividing cells in rodents ( Cearley and Wolfe, 2006 ; Duque et al, 2009 ; Foust et al, 2009 ; Zhang et al, 2011 ; Chansel-Debordeaux et al, 2017 ; Chansel-Debordeaux et al, 2018 ) and non-human primates (NHPs) ( Dehay et al, 2012 ; Gray et al, 2013 ; Samaranch et al, 2013 ; Hinderer et al, 2014 ; Yang et al, 2014 ; Bey et al, 2020 ), making them a powerful tool for delivering genetic material to the CNS. Recently, a novel AAV9-derived variant that far more efficiently crosses the mouse BBB (named AAV-PHP.…”

GRK2‐Targeted Knockdown as Therapy for Multiple System Atrophy

“…Degeneration and symptoms of MSA begin in adulthood, but the transgenic PLP‐SYN mouse model 10 involves constitutive oligodendroglial human α‐syn overexpression. We sought to determine whether the intrauterine (E14) intracerebroventricular delivery 11 of GRK2 miRNA could offer adequate protection for the late‐onset development of behavioral and pathological abnormalities in this mouse model (Fig. 1).…”

Advances in cellular resolution microscopy for brain imaging in rats

“…91,[107][108][109][110][111] In addition, in utero AAV injections can be used to achieve widespread cortical labeling in rats. 112 A strength of in utero gene delivery is that it can be implemented during different stages of development to yield spatially specific expression within the neocortex without the need for laminar specific promoters. Moreover, the method can be optimized to produce widespread infection from a single injection.…”