Highly Efficient Delivery of Adeno-Associated Viral Vectors to the Primate Retina.

Adeno-associated virus (AAV) has emerged as the preferred vector for targeting gene expression to the retina. Subretinally injected AAV can efficiently transduce retinal pigment epithelium and photoreceptors in primate retina. Inner and middle primate retina can be transduced by intravitreally delivered AAV, but with low efficiency. This is due to dilution of vector, potential neutralization of capsid because it is not confined to the immune-privileged retinal compartment, and the presence of the inner limiting membrane (ILM), a barrier separating the vitreous from the neural retina. We here describe a novel "subILM" injection method that addresses all three issues. Specifically, vector is placed in a surgically induced, hydrodissected space between the ILM and neural retina. In an initial experiment, we injected viscoelastic (Healon(®)), a substance we confirmed was biocompatible with AAV, to create a subILM bleb and subsequently injected AAV2-GFP into the bleb after irrigation with basic salt solution. For later experiments, we used a Healon-AAV mixture to place single, subILM injections. In all cases, subILM delivery of AAV was well tolerated-no inflammation or gross structural changes were observed by ophthalmological examination or optical coherence tomography. In-life fluorescence imaging revealed profound transgene expression within the area of the subILM injection bleb that persisted for the study duration. Uniform and extensive transduction of retinal ganglion cells (RGCs) was achieved in the areas beneath the subILM bleb. Transduction of Müller glia, ON bipolar cells, and photoreceptors was also observed. Robust central labeling from green fluorescent protein-expressing RGCs confirmed their continued survival, and was observed in the lateral geniculate nucleus, the superior colliculus, and the pretectum. Our results confirm that the ILM is a major barrier to transduction by AAV in primate retina and that, when it is circumvented, the efficiency and depth to which AAV2 promotes transduction of multiple retinal cell classes are greatly enhanced.

Comparison of visual field severity classification systems for glaucoma.

PURPOSE: To compare categorical severity classification systems for glaucoma. METHODS: This cross-sectional study included 1,921 eyes (49.5% right eye) from 1,137 participants from the Diagnostic Innovations in Glaucoma Study and African Descent and Glaucoma Evaluation Study. Standard automated perimetry fields were classified using the: (1) Advanced Glaucoma Intervention Study scoring system (AGIS), (2) Glaucoma Severity Staging system (GSS), and (3) Enhanced Glaucoma Severity Staging system (eGSS). Systems were characterized using the following continuous measures of severity: mean deviation, pattern standard deviation, and visual field index. Classifications between systems and with optic disc stereophotograph assessment were compared (κ) and some stages were consolidated to evaluate severity classification across systems (Wilcoxon test). RESULTS: Mean deviation, pattern standard deviation, and visual field index were significantly different between GSS and AGIS, and GSS and eGSS in normal and abnormal fields (P<0.005). Agreement between AGIS and eGSS was substantial (κ=0.715±0.012); agreement between GSS and eGSS (κ=0.559±0.014) and AGIS (κ=0.519±0.016) was moderate. eGSS tended to stage abnormal fields most severely followed by GSS and then AGIS (P<0.001). CONCLUSIONS: The presence of glaucomatous optic neuropathy increases with staging severity for all systems. However, different systems led to different severity staging. Of the systems examined in this study, eGSS may be the better choice for its ease of use for both clinicians and researchers.