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J.-J. Pang, X. Dai, D. Everhart, B. Lei, S. L. Boye, A. Dinculescu, Y. Umino, B. Chang, R. Barlow, W. W. Hauswirth; Long Term Rescue Following Gene Therapy With Capsid Mutant AAV8 in the rd10 Mouse, a Model of Recessive Retinitis Pigmentosa. Invest. Ophthalmol. Vis. Sci. 2010;51(13):2527.
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© ARVO (1962-2015); The Authors (2016-present)
To test capsid mutant AAV8-mediated gene therapy in the rd10 mouse, a natural model of recessive RP with a PDEβ mutation.
A wild type PDEβ cDNA with small CBA promoter was packaged into AAV serotype 8 capsids containing a point mutation in surface-exposed tyrosine residues, AAV8 (Y733F)-smCBA-PDEβ. One µl of this vector was subretinally injected into one eye of rd10 mice at postnatal day 14 under red light. The partner eye remained uninjected. Animals were maintained for 2 weeks in darkness before being moved to 12hr/12hr cycling light. Six months later, treated mice were examined by ERG and optomotor-mediated behavioral tests. Mice were then sacrificed for biochemical and morphological examinations.
Scotopic and photopic ERGs were seen in the treated eyes only. Rescue was most evident in eyes that experienced the largest retinal detachment during vector administration and the least surgical complications. Restored scotopic ERGs had typical a-waves; up to 46% of normal b-wave amplitudes were maintained in treated eyes. About 71% of normal photopic ERG amplitudes were observed in treated eyes. Optomotor-mediated behavioral tests showed nearly normal rod and cone-driven visual acuity and contrast sensitivity in the treated eyes of the rd10 mice, but very poor performance in untreated eyes. Strong PDEβ expression was observed in photoreceptor outer segments of treated rd10 eyes by immunostaining and this was confirmed by Western blot analysis. PDEβ expression was undetectable in untreated eyes. Light microscopy showed that outer nuclear layer of treated eyes contained 4-6 rows. Up to 50% of normal outer segment length was preserved in treated eyes. In contrast, photoreceptor nuclei and outer segments were nearly undetectable in untreated eyes.
These data demonstrate that P14 administration of AAV8 (Y733F)-smCBA-PDEβ can arrest retinal degeneration in rd10 mice for at least 6 months. This was shown by significant structural, biochemical, behavioral and electrophysiological preservation/restoration. Our data suggest that mutant AAV8 (Y733F) vector provides a better choice to rescue early-onset retinal degeneration in rd10 mice.
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