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Lara Carroll, Hironori Uehara, Xiaohui Zhang, Balamurali K Ambati; Subretinal Injection of AAVBR1 Targets the RPE and Outer Retina. Invest. Ophthalmol. Vis. Sci. 2020;61(7):2901.
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AAVBR1 is a novel AAV vector with unique tropism for brain vascular endothelium. We tested this adeno-associated virus in comparison with AAV2 and AAV8 for its ability to target the retina and RPE.
C57Bl6 mice were injected with AAVBR1.GFP via systemic delivery (tail vein or retro-orbital sinus: 5 x 1010 VG in 100μl DPBS), or ocular delivery (intravitreal or subretinal: 5 x 108 VG in 1 μl DPBS) to assess its ocular tropism. A dosage study was subsequently performed to compare AAV2.GFP, AAV8.GFP and AAVBR1.GFP for targeting the RPE and retina with subretinal injection. In vivo fundus fluorescence was assessed using either a Heidelberg Spectralis or Phoenix Micron IV imaging system. Ex vivo analysis was performed on flat-mounted or sectioned retinas and RPE 1-2 months after injections for fluorescent quantification and detailed examination of cell tropism.
Subretinal injection was the only method of AAVBR1.GFP delivery resulting in ocular signal, showing strong targeting of RPE and outer nuclear layer with minimal to no targeting of the inner retina. Furthermore, subretinal delivery of AAVBR1.GFP showed GFP signal as early as 48 hours, with broad targeting of the RPE and outer retina. At peak fluorescence (1 month after subretinal injection), AAVBR1.GFP signal was >2x greater than that of either AAV2.GFP or AAV8.GFP.
The specificity and efficiency of AAVBR1 for RPE and outer retina following subretinal injection suggests this vector may be preferable to AAV2 and AAV8 for cell-specific targeting using therapeutics to treat retinitis pigmentosa, AMD, and other diseases that affect the outer retina. However, unlike its ability to target brain vasculature following systemic delivery, AAVBR1 does not target retinal vascular endothelium or choriocapillaris at clinically relevant dosages using any delivery route.
This is a 2020 ARVO Annual Meeting abstract.
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