Retinal pigment epithelial cell expression of active Rap 1a by scAAV2 inhibits choroidal neovascularization
To test the hypothesis that increased Rap1a activity specifically in retinal pigment epithelial cells resists choroidal neovascularization (CNV), self-complementary adeno-associated virus 2 (scAAV2) with RPE65-promoter-driven GFP vectors were generated and introduced subretinally into Rap1b-deficien...
| Main Authors: | , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2016-01-01
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| Series: | Molecular Therapy: Methods & Clinical Development |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2329050117300281 |
| _version_ | 1818193065007906816 |
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| author | Haibo Wang Xiaokun Han Colin A Bretz Silke Becker Deeksha Gambhir George W Smith R Jude Samulski Erika S Wittchen Lawrence A Quilliam Magdalena Chrzanowska-Wodnicka M Elizabeth Hartnett |
| author_facet | Haibo Wang Xiaokun Han Colin A Bretz Silke Becker Deeksha Gambhir George W Smith R Jude Samulski Erika S Wittchen Lawrence A Quilliam Magdalena Chrzanowska-Wodnicka M Elizabeth Hartnett |
| author_sort | Haibo Wang |
| collection | DOAJ |
| description | To test the hypothesis that increased Rap1a activity specifically in retinal pigment epithelial cells resists choroidal neovascularization (CNV), self-complementary adeno-associated virus 2 (scAAV2) with RPE65-promoter-driven GFP vectors were generated and introduced subretinally into Rap1b-deficient mice. Six-week-old mice that received subretinal control (scAAV2-Con) or constitutively active Rap1a (scAAV2-CARap1a) showed strong GFP at the 5 × 108 viral particle/μl dose 5 weeks later without altering retinal morphology or function. Compared to scAAV2-Con- or phosphate-buffered saline (PBS)-injected, eyes injected with scAAV2-CARap1a had increased Rap1 in retinal pigment epithelial (RPE)/choroidal lysates and a significant reduction in CNV volume 7 days after laser, comparable to eyes that received intravitreal anti-VEGF versus IgG control. scAAV2-CARap1a-, but not anti-VEGF-, injected eyes had increased pan-cadherin in RPE/choroids. In cultured RPE cells, increased active Rap1a inhibited TNFα-induced disassociation of junctional pan-cadherin/β-catenin complexes, increased transepithelial electrical resistance through an interaction of β-catenin with phosphorylated scaffold protein, IQGAP1, and inhibited choroidal endothelial cell (CEC) transmigration of an RPE monolayer. This evidence shows that increased Rap1a activity specifically in RPE cells is sufficient to reduce CEC transmigration and CNV and involves IQGAP1-mediated protection of RPE junctional complexes. |
| first_indexed | 2024-12-12T00:40:28Z |
| format | Article |
| id | doaj.art-cd8e2b0f679940ba84f1a0b4c6314f90 |
| institution | Directory Open Access Journal |
| issn | 2329-0501 |
| language | English |
| last_indexed | 2024-12-12T00:40:28Z |
| publishDate | 2016-01-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Molecular Therapy: Methods & Clinical Development |
| spelling | doaj.art-cd8e2b0f679940ba84f1a0b4c6314f902022-12-22T00:44:16ZengElsevierMolecular Therapy: Methods & Clinical Development2329-05012016-01-013C10.1038/mtm.2016.56Retinal pigment epithelial cell expression of active Rap 1a by scAAV2 inhibits choroidal neovascularizationHaibo Wang0Xiaokun Han1Colin A Bretz2Silke Becker3Deeksha Gambhir4George W Smith5R Jude Samulski6Erika S Wittchen7Lawrence A Quilliam8Magdalena Chrzanowska-Wodnicka9M Elizabeth Hartnett10The John Moran Eye Center, University of Utah, Salt Lake City, Utah, USAThe John Moran Eye Center, University of Utah, Salt Lake City, Utah, USAThe John Moran Eye Center, University of Utah, Salt Lake City, Utah, USAThe John Moran Eye Center, University of Utah, Salt Lake City, Utah, USAThe John Moran Eye Center, University of Utah, Salt Lake City, Utah, USAThe John Moran Eye Center, University of Utah, Salt Lake City, Utah, USAUNC Vector Core, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, USADepartment of Cell Biology and Physiology, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, USADepartment of Biochemistry and Molecular Biology, Indiana University School of Medicine, Indianapolis, Indiana, USABlood Research Institute, Blood Center of Wisconsin, Milwaukee, Wisconsin, USAThe John Moran Eye Center, University of Utah, Salt Lake City, Utah, USATo test the hypothesis that increased Rap1a activity specifically in retinal pigment epithelial cells resists choroidal neovascularization (CNV), self-complementary adeno-associated virus 2 (scAAV2) with RPE65-promoter-driven GFP vectors were generated and introduced subretinally into Rap1b-deficient mice. Six-week-old mice that received subretinal control (scAAV2-Con) or constitutively active Rap1a (scAAV2-CARap1a) showed strong GFP at the 5 × 108 viral particle/μl dose 5 weeks later without altering retinal morphology or function. Compared to scAAV2-Con- or phosphate-buffered saline (PBS)-injected, eyes injected with scAAV2-CARap1a had increased Rap1 in retinal pigment epithelial (RPE)/choroidal lysates and a significant reduction in CNV volume 7 days after laser, comparable to eyes that received intravitreal anti-VEGF versus IgG control. scAAV2-CARap1a-, but not anti-VEGF-, injected eyes had increased pan-cadherin in RPE/choroids. In cultured RPE cells, increased active Rap1a inhibited TNFα-induced disassociation of junctional pan-cadherin/β-catenin complexes, increased transepithelial electrical resistance through an interaction of β-catenin with phosphorylated scaffold protein, IQGAP1, and inhibited choroidal endothelial cell (CEC) transmigration of an RPE monolayer. This evidence shows that increased Rap1a activity specifically in RPE cells is sufficient to reduce CEC transmigration and CNV and involves IQGAP1-mediated protection of RPE junctional complexes.http://www.sciencedirect.com/science/article/pii/S2329050117300281 |
| spellingShingle | Haibo Wang Xiaokun Han Colin A Bretz Silke Becker Deeksha Gambhir George W Smith R Jude Samulski Erika S Wittchen Lawrence A Quilliam Magdalena Chrzanowska-Wodnicka M Elizabeth Hartnett Retinal pigment epithelial cell expression of active Rap 1a by scAAV2 inhibits choroidal neovascularization Molecular Therapy: Methods & Clinical Development |
| title | Retinal pigment epithelial cell expression of active Rap 1a by scAAV2 inhibits choroidal neovascularization |
| title_full | Retinal pigment epithelial cell expression of active Rap 1a by scAAV2 inhibits choroidal neovascularization |
| title_fullStr | Retinal pigment epithelial cell expression of active Rap 1a by scAAV2 inhibits choroidal neovascularization |
| title_full_unstemmed | Retinal pigment epithelial cell expression of active Rap 1a by scAAV2 inhibits choroidal neovascularization |
| title_short | Retinal pigment epithelial cell expression of active Rap 1a by scAAV2 inhibits choroidal neovascularization |
| title_sort | retinal pigment epithelial cell expression of active rap 1a by scaav2 inhibits choroidal neovascularization |
| url | http://www.sciencedirect.com/science/article/pii/S2329050117300281 |
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