title: Gene Delivery of Calreticulin Anti-Angiogenic Domain Attenuates the Development of Choroidal Neovascularization in Rats.
abstract: Choroidal neovascularization (CNV) is a common pathological feature in neovascular age-related macular degeneration, which is the leading cause of vision loss among elderly populations in developed countries. This study evaluated the effect of a novel endogenous inhibitor of angiogenesis, calreticulin anti-angiogenic domain (CAD), subconjunctivally delivered by an adenoviral vector (Ad-CAD) in a rat model of laser-induced CNV. CAD was expressed in Ad-CAD-infected cells and inhibited the angiogenic activity in human umbilical vein endothelial cells in vitro. CAD expression was also found in various ocular tissues after in vivo subconjunctival Ad-CAD injection. Via bioluminescence imaging it is shown that a single subconjunctival injection of Ad-luciferase induced the expression of the transgene in the injected eyes within 24 h, which lasted for at least 112 days. Forty-two days after subconjunctival injection of Ad-CAD, retinal structure and function were unaffected, as measured using optical coherence tomography and electroretinography, respectively. After laser injury, subconjunctival Ad-CAD gene delivery significantly inhibited CNV lesions as measured via choroid flat-mounts (51% reduction at 21 days; p < 0.001), as well as by fundus fluorescein angiography (19.3%, 28.2%, 31%, and 27.5% reductions at days 21, 28, 35, and 42, respectively; p < 0.05) in rats. The data suggest that subconjunctival Ad-CAD gene therapy could effectively inhibit laser-induced CNV and might be an attractive therapeutic approach for the management of choroidal neovascularization.
authors.affiliation: 1 Department of Ophthalmology, Kaohsiung Veterans General Hospital , Kaohsiung, Taiwan .|2 Yuh-Ing Junior College of Health Care and Management, Kaohsiung, Taiwan .|3 National Defense Medical Center , Taipei, Taiwan .
authors.forename: Youn-Shen
authors.initials: YS
authors.lastname: Bee
authors.affiliation: 4 Centre for Eye Research Australia, Royal Victorian Eye and Ear Hospital , East Melbourne, Australia .|5 Department of Ophthalmology, the First Affiliated Hospital of Jinan University, Guangzhou, China .
authors.forename: Leilei
authors.initials: L
authors.lastname: Tu
authors.affiliation: 1 Department of Ophthalmology, Kaohsiung Veterans General Hospital , Kaohsiung, Taiwan .|6 School of Medicine, National Yang-Ming University , Taipei, Taiwan .
authors.forename: Shwu-Jiuan
authors.initials: SJ
authors.lastname: Sheu
authors.affiliation: 1 Department of Ophthalmology, Kaohsiung Veterans General Hospital , Kaohsiung, Taiwan .
authors.forename: Hsiu-Chen
authors.initials: HC
authors.lastname: Lin
authors.affiliation: 7 Department of Medical Education and Research, Kaohsiung Veterans General Hospital , Kaohsiung, Taiwan .
authors.forename: Jia-Hua
authors.initials: JH
authors.lastname: Tang
authors.affiliation: 4 Centre for Eye Research Australia, Royal Victorian Eye and Ear Hospital , East Melbourne, Australia .|8 Ophthalmology, Department of Surgery, University of Melbourne , East Melbourne, Australia .
authors.forename: Jiang-Hui
authors.initials: JH
authors.lastname: Wang
authors.affiliation: 9 Department of Optometry and Vision Sciences, University of Melbourne , East Melbourne, Australia .
authors.forename: Selwyn M
authors.initials: SM
authors.lastname: Prea
authors.affiliation: 4 Centre for Eye Research Australia, Royal Victorian Eye and Ear Hospital , East Melbourne, Australia .|8 Ophthalmology, Department of Surgery, University of Melbourne , East Melbourne, Australia .
authors.forename: Gregory J
authors.initials: GJ
authors.lastname: Dusting
authors.affiliation: 10 Center for Stem Cell Research, Kaohsiung Medical University , Kaohsiung, Taiwan .|11 Division of Gastroenterology, Department of Internal Medicine, Kaohsiung Medical University , Kaohsiung, Taiwan .
authors.forename: Deng-Chyang
authors.initials: DC
authors.lastname: Wu
authors.affiliation: 5 Department of Ophthalmology, the First Affiliated Hospital of Jinan University, Guangzhou, China .
authors.forename: Jingxiang
authors.initials: J
authors.lastname: Zhong
authors.affiliation: 9 Department of Optometry and Vision Sciences, University of Melbourne , East Melbourne, Australia .
authors.forename: Bang V
authors.initials: BV
authors.lastname: Bui
authors.affiliation: 10 Center for Stem Cell Research, Kaohsiung Medical University , Kaohsiung, Taiwan .|12 Institute of Biomedical Sciences, National Sun Yat-Sen University , Kaohsiung, Taiwan .|13 Center for Neuroscience, National Sun Yat-Sen University , Kaohsiung, Taiwan .
authors.forename: Ming-Hong
authors.initials: MH
authors.lastname: Tai
authors.affiliation: 4 Centre for Eye Research Australia, Royal Victorian Eye and Ear Hospital , East Melbourne, Australia .|8 Ophthalmology, Department of Surgery, University of Melbourne , East Melbourne, Australia .
authors.forename: Guei-Sheung
authors.initials: GS
authors.lastname: Liu
mesh_terms: D000818:Animals; D037282:Calreticulin / Q000235:genetics* / Q000627:therapeutic use; D020256:Choroidal Neovascularization / Q000235:genetics / Q000628:therapy*; D004195:Disease Models, Animal; D018014:Gene Transfer Techniques; D015316:Genetic Therapy; D005822:Genetic Vectors / Q000627:therapeutic use; D061307:Human Umbilical Vein Endothelial Cells; D006801:Humans; D051381:Rats; D012160:Retina / Q000187:drug effects / Q000473:pathology; D057135:Wet Macular Degeneration / Q000235:genetics / Q000473:pathology / Q000628:therapy*
chemical_list: C576483:CALR protein, human; D037282:Calreticulin
keywords: CAD; adenovirus; choroidal neovascularization; gene delivery