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Yassin Djalali-Talab, Olga Kazanskaya, Sandra Johnen, Antonis Koutsonas, Narendra Armogan, Peter Walter, Gabriele Thumann; Gene Therapy of a CNV Rodent Model Using Non-virally Stably Transfected RPE Cells with the PEDF Gene. Invest. Ophthalmol. Vis. Sci. 2011;52(14):4380.
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Choroidal neovascularization (CNV) is the leading cause of vision loss in AMD. Since PEDF is a potent inhibitor of angiogenesis as well as a neuroprotective agent, it would be expected that transplantation of pigment cells genetically modified to secrete high levels of PEDF could be used to treat neovascular AMD. This study evaluates the effect of transplantation of PEDF-transfected homologous RPE cells into the subretinal space of rats after laser-induced CNV.
Inhibition of migration and tube formation by human umbilical vein endothelial cells (HUVECs) was employed to quantify anti-angiogenic activity of rPEDF produced by PEDF-transfected homologous RPE cells. The effect of rPEDF and transplanted non-virally mediated PEDF-transfected cells in vivo was examined on CNV lesions generated by laser photocoagulation of the fundus in 24 Long Evans rats. Immediately after laser treatment, 1 µg of rPEDF or 2x104cells expressing human PEDF at 100 ng/day or control protein (Venus, yellow fluorescent mutant of GFP) were injected into the subretinal space. Extent of CNV was evaluated by isolectin B4 staining of choroid-RPE-flat mounts at 8 weeks and by fluorescein angiography (FAG) on day 7 and at 8 weeks. Distribution and state of injected cells was evaluated by immunostaining with anti-5 His antibody.
At a concentration of 100 ng/ml rPEDF inhibited HUVEC migration by 76% and tube formation by 66%. In this series of rats 1 µg of rPEDF reduced cumulative CNV area by 60% and 2x104RPE cells overexpressing PEDF reduced cumulative CNV area by 50%. FAG analysis indicated that 1 µg rPEDF and PEDF-overexpressing RPE cells reduced the CNV leakage area by 42% and by 50%, respectively.
The results of this study show that as little as 1 µg of rPEDF and as few as 20,000 RPE cells transfected with the PEDF gene and secreting PEDF at a concentration of 100 ng/day are able to control the development of neovscularization in a rat model of CNV. These studies indicate that pigment cells transfected with the PEDF gene would be effective to inhibit the neovascularisation in neovascular AMD and other neovascular diseases of the retina.
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