AAV2/4-RS1 gene therapy in the retinoschisin knockout mouse model of X-linked retinoschisis
<h4>Objective</h4> To evaluate efficacy of a novel adeno-associated virus (AAV) vector, AAV2/4-RS1, for retinal rescue in the retinoschisin knockout (Rs1-KO) mouse model of X-linked retinoschisis (XLRS). Brinzolamide (Azopt®), a carbonic anhydrase inhibitor, was tested for its ability to...
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| Format: | Article |
| Language: | English |
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Public Library of Science (PLoS)
2022-01-01
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| Series: | PLoS ONE |
| Online Access: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9728878/?tool=EBI |
| _version_ | 1828269059021471744 |
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| author | Brittni A. Scruggs Sajag Bhattarai Megan Helms Ioana Cherascu Adisa Salesevic Elliot Stalter Joseph Laird Sheila A. Baker Arlene V. Drack |
| author_facet | Brittni A. Scruggs Sajag Bhattarai Megan Helms Ioana Cherascu Adisa Salesevic Elliot Stalter Joseph Laird Sheila A. Baker Arlene V. Drack |
| author_sort | Brittni A. Scruggs |
| collection | DOAJ |
| description | <h4>Objective</h4> To evaluate efficacy of a novel adeno-associated virus (AAV) vector, AAV2/4-RS1, for retinal rescue in the retinoschisin knockout (Rs1-KO) mouse model of X-linked retinoschisis (XLRS). Brinzolamide (Azopt®), a carbonic anhydrase inhibitor, was tested for its ability to potentiate the effects of AAV2/4-RS1. <h4>Methods</h4> AAV2/4-RS1 with a cytomegalovirus (CMV) promoter (2x1012 viral genomes/mL) was delivered to Rs1-KO mice via intravitreal (N = 5; 1μL) or subretinal (N = 21; 2μL) injections at postnatal day 60–90. Eleven mice treated with subretinal therapy also received topical Azopt® twice a day. Serial full field electroretinography (ERG) was performed starting at day 50–60 post-injection. Mice were evaluated using a visually guided swim assay (VGSA) in light and dark conditions. The experimental groups were compared to untreated Rs1-KO (N = 11), wild-type (N = 12), and Rs1-KO mice receiving only Azopt® (N = 5). Immunofluorescence staining was performed to assess RS1 protein expression following treatment. <h4>Results</h4> The ERG b/a ratio was significantly higher in the subretinal plus Azopt® (p<0.0001), subretinal without Azopt® (p = 0.0002), and intravitreal (p = 0.01) treated eyes compared to untreated eyes. There was a highly significant subretinal treatment effect on ERG amplitudes collectively at 7–9 months post-injection (p = 0.0003). Cones showed more effect than rods. The subretinal group showed improved time to platform in the dark VGSA compared to untreated mice (p<0.0001). RS1 protein expression was detected in the outer retina in subretinal treated mice and in the inner retina in intravitreal treated mice. <h4>Conclusions</h4> AAV2/4-RS1 shows promise for improving retinal phenotype in the Rs1-KO mouse model. Subretinal delivery was superior to intravitreal. Topical brinzolamide did not improve efficacy. AAV2/4-RS1 may be considered as a potential treatment for XLRS patients. |
| first_indexed | 2024-04-13T05:26:02Z |
| format | Article |
| id | doaj.art-b3a196fcb39c44e6bff52826d22be2a5 |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-04-13T05:26:02Z |
| publishDate | 2022-01-01 |
| publisher | Public Library of Science (PLoS) |
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| spelling | doaj.art-b3a196fcb39c44e6bff52826d22be2a52022-12-22T03:00:35ZengPublic Library of Science (PLoS)PLoS ONE1932-62032022-01-011712AAV2/4-RS1 gene therapy in the retinoschisin knockout mouse model of X-linked retinoschisisBrittni A. ScruggsSajag BhattaraiMegan HelmsIoana CherascuAdisa SalesevicElliot StalterJoseph LairdSheila A. BakerArlene V. Drack<h4>Objective</h4> To evaluate efficacy of a novel adeno-associated virus (AAV) vector, AAV2/4-RS1, for retinal rescue in the retinoschisin knockout (Rs1-KO) mouse model of X-linked retinoschisis (XLRS). Brinzolamide (Azopt®), a carbonic anhydrase inhibitor, was tested for its ability to potentiate the effects of AAV2/4-RS1. <h4>Methods</h4> AAV2/4-RS1 with a cytomegalovirus (CMV) promoter (2x1012 viral genomes/mL) was delivered to Rs1-KO mice via intravitreal (N = 5; 1μL) or subretinal (N = 21; 2μL) injections at postnatal day 60–90. Eleven mice treated with subretinal therapy also received topical Azopt® twice a day. Serial full field electroretinography (ERG) was performed starting at day 50–60 post-injection. Mice were evaluated using a visually guided swim assay (VGSA) in light and dark conditions. The experimental groups were compared to untreated Rs1-KO (N = 11), wild-type (N = 12), and Rs1-KO mice receiving only Azopt® (N = 5). Immunofluorescence staining was performed to assess RS1 protein expression following treatment. <h4>Results</h4> The ERG b/a ratio was significantly higher in the subretinal plus Azopt® (p<0.0001), subretinal without Azopt® (p = 0.0002), and intravitreal (p = 0.01) treated eyes compared to untreated eyes. There was a highly significant subretinal treatment effect on ERG amplitudes collectively at 7–9 months post-injection (p = 0.0003). Cones showed more effect than rods. The subretinal group showed improved time to platform in the dark VGSA compared to untreated mice (p<0.0001). RS1 protein expression was detected in the outer retina in subretinal treated mice and in the inner retina in intravitreal treated mice. <h4>Conclusions</h4> AAV2/4-RS1 shows promise for improving retinal phenotype in the Rs1-KO mouse model. Subretinal delivery was superior to intravitreal. Topical brinzolamide did not improve efficacy. AAV2/4-RS1 may be considered as a potential treatment for XLRS patients.https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9728878/?tool=EBI |
| spellingShingle | Brittni A. Scruggs Sajag Bhattarai Megan Helms Ioana Cherascu Adisa Salesevic Elliot Stalter Joseph Laird Sheila A. Baker Arlene V. Drack AAV2/4-RS1 gene therapy in the retinoschisin knockout mouse model of X-linked retinoschisis PLoS ONE |
| title | AAV2/4-RS1 gene therapy in the retinoschisin knockout mouse model of X-linked retinoschisis |
| title_full | AAV2/4-RS1 gene therapy in the retinoschisin knockout mouse model of X-linked retinoschisis |
| title_fullStr | AAV2/4-RS1 gene therapy in the retinoschisin knockout mouse model of X-linked retinoschisis |
| title_full_unstemmed | AAV2/4-RS1 gene therapy in the retinoschisin knockout mouse model of X-linked retinoschisis |
| title_short | AAV2/4-RS1 gene therapy in the retinoschisin knockout mouse model of X-linked retinoschisis |
| title_sort | aav2 4 rs1 gene therapy in the retinoschisin knockout mouse model of x linked retinoschisis |
| url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9728878/?tool=EBI |
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