Degeneration of photoreceptors, retinal cells including retinal pigment epithelium, and retinal ganglion cells characterizes visual disorders involving retinal dysfunction. The efficient and targeted gene delivery into retinal cells is critical for treatment of a variety of visual disorders. Here, we report use of an OCT guided, spatially targeted, near-infrared laser microirradiation platform to successfully deliver genes encoding ambient-light activatable multi-characteristic opsin (MCO) to retinal cells in non-human primates. The efficacy of gene delivery was evaluated by fundus imaging of the reporter (mCherry) fluorescence, and immunohistochemistry. Functional improvement was measured by red-ERG which was attributed to MCO’s broadband activation spectrum.
Stargardt Disease is an inherited macular degeneration caused by mutation of genes, primarily ABCA4 in the photoreceptor cells leading to their dysfunction and degeneration in the macula. Here, we report results of ambient-light vision restoration in Stargardt mice (Abca4tm1Ght/J) upon intravitreal delivery of AAV-carried MCO (vMCO) as evaluated by electrophysiology, Optical Coherence Tomography (OCT) and visually guided behavior. Further, safety of MCO-enabled vision restoration therapy was evaluated by measurement of Intraocular Pressure, OCT and Immunohistochemistry. The vMCO based ambient light activatable optogenetic therapy has potential to be a disease-agnostic therapy to address the unmet need of inherited retinal degenerative diseases.
We hypothesized that PEDF gene transduction in retina can provide single-dose treatment to prevent ganglion cell damage. Here, we present OCT guided ultrafast laser based non-viral targeted delivery PEDF-encoding genes to retina for neuroprotection. The ultrafast laser gene delivery showed layer-specific reliable expression of PEDF gene in retina without any detectable damage. Monitoring of IOP and electroretinogram after ultrafast laser transfection showed no adverse changes. The ultrafast laser transfection of large PEDF genes in retina exhibited significant therapeutic benefit in an injury model. Absence of any immune response in retina subsequent to ultrafast-laser transfection provides unique opportunity for repeated dosing.
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