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AAV8-vectored suprachoroidal gene transfer produces widespread ocular transgene expression
Kun Ding, … , Olivier Danos, Peter A. Campochiaro
Kun Ding, … , Olivier Danos, Peter A. Campochiaro
Published August 13, 2019
Citation Information: J Clin Invest. 2019;129(11):4901-4911. https://doi.org/10.1172/JCI129085.
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Research Article Ophthalmology

AAV8-vectored suprachoroidal gene transfer produces widespread ocular transgene expression

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Abstract

There has been great progress in ocular gene therapy, but delivery of viral vectors to the retinal pigmented epithelium (RPE) and retina can be challenging. Subretinal injection, the preferred route of delivery for most applications, requires a surgical procedure that has risks. Herein we report a novel gene therapy delivery approach, suprachoroidal injection of AAV8 vectors, which is less invasive and could be done in an outpatient setting. Two weeks after suprachoroidal injection of AAV8.GFP in rats, GFP fluorescence covered 18.9% of RPE flat mounts and extended entirely around sagittal and transverse sections in RPE and photoreceptors. After 2 suprachoroidal injections of AAV8.GFP, GFP fluorescence covered 30.5% of RPE flat mounts. Similarly, widespread expression of GFP occurred in nonhuman primate and pig eyes after suprachoroidal injection of AAV8.GFP. Compared with subretinal injection in rats of RGX-314, an AAV8 vector expressing an anti-VEGF Fab, suprachoroidal injection of the same dose of RGX-314 resulted in similar expression of anti-VEGF Fab and similar suppression of VEGF-induced vascular leakage. Suprachoroidal AAV8 vector injection provides a noninvasive outpatient procedure to obtain widespread transgene expression in retina and RPE.

Authors

Kun Ding, Jikui Shen, Zibran Hafiz, Sean F. Hackett, Raquel Lima e Silva, Mahmood Khan, Valeria E. Lorenc, Daiqin Chen, Rishi Chadha, Minie Zhang, Sherri Van Everen, Nicholas Buss, Michele Fiscella, Olivier Danos, Peter A. Campochiaro

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Figure 5

Widespread expression of GFP 3 weeks after suprachoroidal injection of AAV8.GFP in nonhuman primates or pigs.

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Widespread expression of GFP 3 weeks after suprachoroidal injection of A...
Rhesus monkeys were given a suprachoroidal injection of 50 μL containing 4.75 × 1011 GCs of AAV8.GFP and after 3 weeks flat mounts were examined by fluorescence microscopy. The flat mounts and sections from each of the eyes showed similar results and therefore representative images are shown. A collage was made from one eye by aligning areas of overlap. The collage shows strong expression of GFP throughout approximately one-third of the retinal flat mount (A; scale bar: 1000 μm). In a RPE flat mount, high magnification view of the mid-periphery in the quadrant of injection shows heterogeneity of GFP expression with intense fluorescence in some RPE cells and little or none in others (B; scale bar: 100 μm). A higher magnification view shows the hexagonal shape of RPE cells, some seen in negative relief (C; scale bar: 50 μm). Posteriorly there is less intense, but more uniform GFP expression in RPE cells extending almost to the border of the optic nerve (ON) which is outlined by fluorescence (D; scale bar: 250 μm). Higher magnification shows the hexagonally shaped RPE cells with GFP in the cytoplasm and the nuclei in negative relief (E; scale bar: 50 μm). Retinal flat mounts showed GFP expression in many cells of the multilayered retina extending posteriorly to the cut edge of retina where it had been severed from the optic nerve (F; scale bar: 50 μm). Two weeks after suprachoroidal injection of 50 μL containing 4.75 × 1011 GCs of AAV8.GFP in a pig, merged images from ocular sections showed colocalization of fluorescence and anti-GFP staining in RPE cells and photoreceptor inner and outer segments (G; scale bar: 50 μm). There is also some GFP expression in the inner retina.

Copyright © 2025 American Society for Clinical Investigation
ISSN: 0021-9738 (print), 1558-8238 (online)

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