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

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 4

Suprachoroidal versus subretinal injection of RGX-314.

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Suprachoroidal versus subretinal injection of RGX-314. Rats were given s...
Rats were given suprachoroidal (SC) or subretinal (SR) injection of 1.2 × 108 GCs of RGX-314 in one eye and SC or SR vehicle injection in the other eye. After 2 weeks, intravitreous VEGF (100 ng) was given and 24 hours later, fundus photographs showed normal retinas and retinal vessels in eyes given SC (A) or SR (C) injection of RGX-314, whereas SC (B) or SR (D) vehicle controls showed dilated, engorged vessels and hemorrhages (B, arrow). Fluorescein angiograms (FAs) showed normal caliber vessels with sharp margins in SC (E) or SR (G) RGX-314–injected eyes, whereas vehicle controls showed dilated vessels with blurred margins (F and H). Results were similar 7 weeks after vector injection. Twenty-four hours after VEGF (100 ng) injection, eyes that had been given SC (I and M) or SR (K and O) injection of RGX-314 showed normal retinal vessels with sharp margins in FAs, whereas vehicle controls showed dilated retinal vessels and hemorrhages (J and L, arrows), and hazy margins in FAs (N and P). (Q) Two or 7 weeks after SC or SR injection of RGX-314 or AAV8.GFP in the fellow eye, mean (± SEM) vitreous albumin measured by ELISA 24 hours after VEGF (100 ng) injection was significantly less in SC or SR RGX-314–injected eyes versus fellow eye controls (n = 10, *P < 0.03 by unpaired t test). (R) Mean (± SEM) anti–VEGF Fab in the retina and RPE/choroid were high 2 or 7 weeks after RGX-314 injection with no significant difference between the SC and SR routes of injection at each time point (n ≥ 5).