Gene therapy knockdown of VEGFR2 in retinal endothelial cells to treat retinopathy

Abstract

Inhibition of vascular endothelial growth factor (VEGF) in retinopathy of prematurity (ROP) raises concerns for premature infants because VEGF is essential for retinovascular development as well as neuronal and glial health. This study tested the hypothesis that endothelial cell-specific knockdown of VEGF receptor 2 (VEGFR2), or downstream STAT3, would inhibit VEGF-induced retinopathy without delaying physiologic retinal vascular development. We developed an endothelial cell-specific lentiviral vector that delivered shRNAs to VEGFR2 or STAT3 and a green fluorescent protein reporter under control of the VE-cadherin promoter. The specificity and efficacy of the lentiviral vector-driven shRNAs were validated in vitro and in vivo. In the rat oxygen-induced retinopathy model highly representative of human ROP, the effects of endothelial cell knockdown of VEGFR2 or STAT3 were determined on intravitreal neovascularization (IVNV), physiologic retinal vascular development [assessed as area of peripheral avascular/total retina (AVA)], retinal structure, and retinal function. Targeted knockdown of VEGFR2 or STAT3 specifically in retinal endothelial cells by subretinal injection of lentiviral vectors into postnatal day 8 rat pup eyes efficiently inhibited IVNV, and knockdown of VEGFR2 also reduced AVA and increased retinal thickness without altering retinal function. Taken together, our results support specific knockdown of VEGFR2 in retinal endothelial cells as a novel therapeutic method to treat retinopathy.Electronic supplementary materialThe online version of this article (10.1007/s10456-018-9618-5) contains supplementary material, which is available to authorized users.

Relationship between Retinal Thickness Profiles and Visual Outcomes in Young Adults Born Extremely Preterm

“…Analyzing the parafoveal and perifoveal changes in premature infants could be applicable to rodent models, where a fovea is absent, to explore the pathophysiology and potential therapeutic options for ROP. 29,30 Of note, preterm children are also at an increased risk of developing cortical or cerebral visual impairment (CVI). 31 The CVI is impaired vision without primary ocular pathology and it is one of the leading causes of poor visual acuity among EP M A N U S C R I P T A C C E P T E D births.…”

Three-dimensional pattern of extraretinal neovascular development in retinopathy of prematurity

“…[2] To-date, 3D visualization in these animals are lacking. As suggested in the animal model studies, signaling pathways triggered by variable oxygen levels and oxidative stress lead to VEGF receptor 2-induced disordered angiogenesis [31], [32] and intravitreal neovascularization. [5,29] [31] OCT imaging with 3D visualization provides a more distinct appearance of the inter-connections between vascular structures at V-AV junction in human infants compared to indirect ophthalmoscopy or imaging with a contact camera.…”

“…Longitudinal studies show similar findings by 3D SDOCT as that seen in experimental models in which VEGF signaling is inhibited. [31,33,34] Similarities in features between the experimental rat OIR model and human disease lend greater ability to translate findings experimentally to the human disease.…”

Signaling Through the Erythropoietin Receptor Affects Angiogenesis in Retinovascular Disease

“…5C). 29 At p8, Epo was greater in hWtEPOR retinas compared with WT retinas from mice raised in RA (Fig. 5A).…”