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 November 1, 2019; First 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

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 2

Expression of GFP throughout the posterior retina and RPE 2 weeks after suprachoroidal or subretinal injection of AAV8.GFP in rats.

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Expression of GFP throughout the posterior retina and RPE 2 weeks after ...
Two weeks after suprachoroidal injection of 2.85 × 1010 GCs of AAV8.GFP in rats, a transverse section showed more intense fluorescence in the retina on the side of the eye where the injection was performed (bottom), but was detectable around the entire circumference of the eye. The section was immunohistochemically stained with anti-GFP (red), and magnified views of the boxed regions on the side opposite the injection site (A and B) or on the same side (C and D) are shown. (A) In this region remote from the injection, there is strong GFP expression in some but not all RPE cells. Almost all photoreceptor inner segments (IS) and outer segments (OS) contain GFP, but few photoreceptor cell bodies in the outer nuclear layer (ONL). (B) In this second remote region, there are high levels of GFP in the RPE cells and photoreceptor IS and OS, but not cell bodies. Some cells in the inner nuclear layer (INL) and GCL contain GFP (C and D). In the 2 regions on the injected side of the eye, there are high levels of GFP in RPE cells and photoreceptor IS, OS, and cell bodies. There is a small artifactual retinal fold adjacent to the boxed area shown in D. Two weeks after subretinal injection of 2.85 × 1010 GCs of AAV8.GFP, there was strong GFP fluorescence and anti-GFP staining in RPE, photoreceptor IS, OS, and cell bodies in half of the eye where the retina had been detached by the vector (G and H), but not the other half (E and F). Scale bars: 100 μm for high power; 500 μm for low power.