Retinal pigment epithelial cell expression of active Rap 1a by scAAV2 inhibits choroidal neovascularization

dc.contributor.authorWang, Haibo
dc.contributor.authorHan, Xiaokun
dc.contributor.authorBretz, Colin A.
dc.contributor.authorBecker, Silke
dc.contributor.authorGambhir, Deeksha
dc.contributor.authorSmith, George W.
dc.contributor.authorSamulski, R. Jude
dc.contributor.authorWittchen, Erika S.
dc.contributor.authorQuilliam, Lawrence A.
dc.contributor.authorChrzanowska-Wodnicka, Magdalena
dc.contributor.authorHartnett, M. Elizabeth
dc.contributor.departmentDepartment of Biochemistry & Molecular Biology, IU School of Medicineen_US
dc.date.accessioned2017-05-17T17:31:59Z
dc.date.available2017-05-17T17:31:59Z
dc.date.issued2016-08-24
dc.description.abstractTo 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 × 10(8) 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.en_US
dc.identifier.citationWang, H., Han, X., Bretz, C. A., Becker, S., Gambhir, D., Smith, G. W., … Hartnett, M. E. (2016). Retinal pigment epithelial cell expression of active Rap 1a by scAAV2 inhibits choroidal neovascularization. Molecular Therapy. Methods & Clinical Development, 3, 16056–. http://doi.org/10.1038/mtm.2016.56en_US
dc.identifier.urihttps://hdl.handle.net/1805/12579
dc.language.isoen_USen_US
dc.publisherElsevieren_US
dc.relation.isversionof10.1038/mtm.2016.56en_US
dc.relation.journalMolecular Therapy -- Methods & Clinical Developmenten_US
dc.rightsAttribution-NonCommercial-NoDerivs 3.0 United States
dc.rights.urihttp://creativecommons.org/licenses/by-nc-nd/3.0/us/
dc.sourcePMCen_US
dc.subjectRap1aen_US
dc.subjectRetinaen_US
dc.subjectEpithelial cellsen_US
dc.subjectChoroidal neovascularizationen_US
dc.titleRetinal pigment epithelial cell expression of active Rap 1a by scAAV2 inhibits choroidal neovascularizationen_US
dc.typeArticleen_US
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