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Limbal Approach-Subretinal Injection of Viral Vectors for Gene Therapy in Mice Retinal Pigment Epithelium
Published on: August 7, 2015
Solid lipid nanoparticle-based vectors intended for the treatment of X-linked juvenile retinoschisis by gene therapy:
P S Apaolaza1, A Del Pozo-Rodríguez1, J Torrecilla1
1Pharmacokinetic, Nanotechnology and Gene Therapy Group (PharmaNanoGene), Faculty of Pharmacy, Centro de investigación Lascaray ikergunea, University of the Basque Country UPV/EHU, Paseo de la Universidad 7, 01006 Vitoria-Gasteiz, Spain.
Abstract:
X-linked juvenile retinoschisis (XLRS), which results from mutations in the gene RS1 that encodes the protein retinoschisin, is a retinal degenerative disease affecting between 1/5000 and 1/25,000 people worldwide. Currently, there is no cure for this disease and the treatment is based on the application of low-vision aids. The aim of the present work was the in vitro and in vivo evaluation of two different non-viral vectors based on solid lipid nanoparticles (SLNs), protamine and two anionic polysaccharides, hyaluronic acid (HA) or dextran (DX), for the treatment of XLRS. First, the vectors containing a plasmid which encodes both the reporter green fluorescent protein (GFP) and the therapeutic protein retinoschisin, under the control of CMV promoters, were characterized in vitro. Then, the vectors were subretinally or intravitreally administrated to C57BL/6 wild type mice. One week later, GFP was detected in all treated mice and in all retinal layers except in the Outer Nuclear Layer (ONL) and the Inner Nuclear Layer (INL), regardless of the administration route and the vector employed. Finally, two weeks after subretinal or intravitreal injection to Rs1h-deficient mice, GFP and retinoschisin expression was detected in all retinal layers, except in the ONL, which was maintained for at least two months after subretinal administration. The structural analysis of the treated Rs1h-deficient eyes showed a partial recovery of the retina related to the production of retinoschisin. This work shows for the first time a successful RS1 gene transfer to Rs1h-deficient animals using non-viral nanocarriers, with promising results that point to non-viral gene therapy as a feasible future therapeutic tool for retinal disorders.
Insights
New non-viral nanocarriers show promise for treating X-linked juvenile retinoschisis (XLRS). Solid lipid nanoparticles successfully delivered the RS1 gene in preclinical models, restoring retinoschisin protein and partially recovering retinal structure.
Area of Science:
- Ophthalmology
- Gene Therapy
- Nanomedicine
Background:
- X-linked juvenile retinoschisis (XLRS) is a genetic retinal disease caused by RS1 gene mutations, leading to vision loss.
- Current treatments for XLRS are limited to low-vision aids, with no cure available.
- Gene therapy offers a potential therapeutic strategy for XLRS by restoring functional retinoschisin protein.
Purpose of the Study:
- To evaluate the efficacy of novel non-viral vectors for gene therapy in X-linked juvenile retinoschisis (XLRS).
- To assess in vitro and in vivo gene delivery and expression of retinoschisin using solid lipid nanoparticles (SLNs).
Main Methods:
- Development and in vitro characterization of two non-viral vectors: SLNs with protamine and either hyaluronic acid (HA) or dextran (DX).
- Vectors carried a plasmid encoding green fluorescent protein (GFP) and the therapeutic protein retinoschisin.
- Subretinal and intravitreal administration in C57BL/6 wild-type and Rs1h-deficient mice, followed by GFP and retinoschisin expression analysis.
Main Results:
- GFP expression was detected in retinal layers of wild-type mice, excluding the Outer and Inner Nuclear Layers.
- Rs1h-deficient mice showed GFP and retinoschisin expression in most retinal layers after vector administration.
- Subretinal administration in Rs1h-deficient mice maintained expression for at least two months and showed partial retinal structural recovery.
Conclusions:
- Non-viral nanocarriers, specifically SLNs, are effective for RS1 gene transfer in a preclinical model of XLRS.
- This study demonstrates the potential of non-viral gene therapy as a future treatment for retinal disorders like XLRS.
- Successful gene delivery and partial functional recovery highlight the promise of this approach for inherited retinal diseases.
