A novel bispecific molecule delivered by recombinant AAV2 suppresses ocular inflammation and choroidal neovascularization.
Elevated vascular endothelial growth factor (VEGF) and complement activation are implicated in the pathogenesis of different ocular diseases. The objective of this study was to investigate the hypothesis that dual inhibition of both VEGF and complement activation would confer better protection against ocular inflammation and neovascularization. In this study, we engineered a secreted chimeric VEGF inhibitor domain (VID), a complement inhibitor domain (CID) and a dual inhibitor (ACVP1). Vectors expressing these three inhibitors were constructed and packaged into AAV2 (sextY-F) particles. The expression and secretion of the proteins were validated by Western blot. The effects of these inhibitors expressed from AAV2 vectors were examined in endotoxin-induced uveitis (EIU), experimental autoimmune uveoretinitis (EAU) and choroidal neovascularization (CNV) mouse models. The AAV2 vectors expressing the CID- and ACVP1-attenuated inflammation in EIU and EAU model, whereas the vector expressing VID showed improved retinal structure damaged by EAU, but not affect the infiltration of inflammatory cells in EAU or EIU eyes. Both VID and CID vectors improved laser-induced retinal and choroid/RPE injuries and CNV, whereas ACVP1 vector provided significantly better protection. Our results suggest that gene therapy targeting VEGF and complement components could provide an innovative and long-term strategy for ocular inflammatory and neovascular diseases.
State Key Laboratory of Biotherapy and Cancer Center, West China Hospital, Sichuan University, and Collaborative Innovation Center for Biotherapy, Chengdu, Sichuan, China.|Department of Ophthalmology, College of Medicine, University of Florida, Gainesville, FL, USA.
Yiming
Y
Li
Department of Ophthalmology, College of Medicine, University of Florida, Gainesville, FL, USA.
Ping
P
Zhu
Department of Ophthalmology, College of Medicine, University of Florida, Gainesville, FL, USA.
Amrisha
A
Verma
Department of Ophthalmology, College of Medicine, University of Florida, Gainesville, FL, USA.
Tuhina
T
Prasad
State Key Laboratory of Biotherapy and Cancer Center, West China Hospital, Sichuan University, and Collaborative Innovation Center for Biotherapy, Chengdu, Sichuan, China.
Hongxin
H
Deng
State Key Laboratory of Biotherapy and Cancer Center, West China Hospital, Sichuan University, and Collaborative Innovation Center for Biotherapy, Chengdu, Sichuan, China.
Dechao
D
Yu
Department of Ophthalmology, College of Medicine, University of Florida, Gainesville, FL, USA.
Qiuhong
Q
Li
D000818:Animals; D001327:Autoimmune Diseases; D002829:Choroid / Q000276:immunology / Q000473:pathology; D020256:Choroidal Neovascularization / Q000235:genetics / Q000276:immunology / Q000473:pathology / Q000628:therapy*; D000229:Dependovirus / Q000235:genetics* / Q000378:metabolism; D004731:Endotoxins; D015870:Gene Expression; D017930:Genes, Reporter; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000737:chemistry / Q000378:metabolism*; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D007141:Immunoglobulin Fc Fragments / Q000235:genetics / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D000072417:Protein Domains; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology; D012160:Retina / Q000276:immunology / Q000473:pathology; D012173:Retinitis / Q000235:genetics / Q000276:immunology / Q000473:pathology / Q000628:therapy*; D014605:Uveitis / Q000139:chemically induced / Q000235:genetics / Q000276:immunology / Q000628:therapy*; D042461:Vascular Endothelial Growth Factor A / Q000235:genetics / Q000276:immunology; D040281:Vascular Endothelial Growth Factor Receptor-1 / Q000235:genetics / Q000276:immunology; D040301:Vascular Endothelial Growth Factor Receptor-2 / Q000235:genetics / Q000276:immunology
D004731:Endotoxins; D007141:Immunoglobulin Fc Fragments; D011993:Recombinant Fusion Proteins; C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A; D049452:Green Fluorescent Proteins; D040281:Vascular Endothelial Growth Factor Receptor-1; D040301:Vascular Endothelial Growth Factor Receptor-2

AAV
; age-related macular degeneration; angiogenesis; autoimmune uveitis; choroidal neovascularization; complement; gene therapy; ocular inflammation; uveitis; vascular endothelial growth factor