title: Targeting the tight junction protein, zonula occludens-1, with the connexin43 mimetic peptide, αCT1, reduces VEGF-dependent RPE pathophysiology.
abstract: A critical target tissue in age-related macular degeneration (AMD) is the retinal pigment epithelium (RPE), which forms the outer blood-retina barrier (BRB). RPE-barrier dysfunction might result from attenuation/disruption of intercellular tight junctions. Zonula occludens-1 (ZO-1) is a major structural protein of intercellular junctions. A connexin43-based peptide mimetic, αCT1, was developed to competitively block interactions at the PDZ2 domain of ZO-1, thereby inhibiting ligands that selectively bind to this domain. We hypothesized that targeting ZO-1 signaling using αCT1 would maintain BRB integrity and reduce RPE pathophysiology by stabilizing gap- and/or tight-junctions. RPE-cell barrier dysfunction was generated in mice using laser photocoagulation triggering choroidal neovascularization (CNV) or bright light exposure leading to morphological damage. αCT1 was delivered via eye drops. αCT1 treatment reduced CNV development and fluid leakage as determined by optical coherence tomography, and damage was correlated with disruption in cellular integrity of surrounding RPE cells. Light damage significantly disrupted RPE cell morphology as determined by ZO-1 and occludin staining and tiling pattern analysis, which was prevented by αCT1 pre-treatment. In vitro experiments using RPE and MDCK monolayers indicated that αCT1 stabilizes tight junctions, independent of its effects on Cx43. Taken together, stabilization of intercellular junctions by αCT1 was effective in ameliorating RPE dysfunction in models of AMD-like pathology.
  
  
  
  The connexin43 mimetic αCT1 accumulates in the mouse retinal pigment epithelium following topical delivery via eye drops. αCT1 eye drops prevented RPE-cell barrier dysfunction in two mouse models. αCT1 stabilizes intercellular tight junctions. Stabilization of cellular junctions via αCT1 may serve as a novel therapeutic approach for both wet and dry age-related macular degeneration.
authors.affiliation: Department of Neuroscience, Medical University of South Carolina, Charleston, SC, 29425, USA.
authors.forename: Elisabeth
authors.initials: E
authors.lastname: Obert
authors.affiliation: Center for Heart and Regenerative Medicine, Virginia Tech Carilion Research Institute, Roanoke, VA, 24016, USA.
authors.forename: Randy
authors.initials: R
authors.lastname: Strauss
authors.affiliation: Department of Ophthalmology, Medical University of South Carolina, Charleston, 167 Ashley Avenue, Charleston, SC, 29425, USA.
authors.forename: Carlene
authors.initials: C
authors.lastname: Brandon
authors.affiliation: FirstString Research Inc., Mount Pleasant, SC, 29464, USA.
authors.forename: Christina
authors.initials: C
authors.lastname: Grek
authors.affiliation: FirstString Research Inc., Mount Pleasant, SC, 29464, USA.
authors.forename: Gautam
authors.initials: G
authors.lastname: Ghatnekar
authors.affiliation: Center for Heart and Regenerative Medicine, Virginia Tech Carilion Research Institute, Roanoke, VA, 24016, USA.
authors.forename: Robert
authors.initials: R
authors.lastname: Gourdie
authors.affiliation: Department of Neuroscience, Medical University of South Carolina, Charleston, SC, 29425, USA. rohrer@musc.edu.|Department of Ophthalmology, Medical University of South Carolina, Charleston, 167 Ashley Avenue, Charleston, SC, 29425, USA. rohrer@musc.edu.|Ralph H. Johnson VA Medical Center, Charleston, SC, 29401, USA. rohrer@musc.edu.
authors.forename: Bärbel
authors.identifier: 0000-0003-2758-3771
authors.initials: B
authors.lastname: Rohrer
mesh_terms: D000818:Animals; D002460:Cell Line; D018031:Connexin 43 / Q000737:chemistry*; D006801:Humans; D051379:Mice; D008807:Mice, Inbred BALB C; D010455:Peptides / Q000737:chemistry* / Q000494:pharmacology*; D055213:Retinal Pigment Epithelium / Q000187:drug effects / Q000378:metabolism*; D019108:Tight Junctions / Q000187:drug effects / Q000378:metabolism*; D042461:Vascular Endothelial Growth Factor A / Q000378:metabolism*; D062826:Zonula Occludens-1 Protein / Q000378:metabolism*
chemical_list: D018031:Connexin 43; D010455:Peptides; D042461:Vascular Endothelial Growth Factor A; D062826:Zonula Occludens-1 Protein
keywords: Age-related macular degeneration; Choroidal neovascularization; Connexin43; Light damage; Retinal pigment epithelium; Tight junctions; Vascular endothelial growth factor