Trans-Ocular Electric Current In Vivo Enhances AAV-Mediated Retinal Transduction in Large Animal Eye After Intravitreal Vector Administration
| Autor: | Sheik Pran Babu Sardar Pasha, Yong Zeng, Camasamudram Vijayasarathy, Haohua Qian, Paul A. Sieving, Lisa Wei, Hongman Song, Henry E. Wiley, Ronald A. Bush, Zhijian Wu |
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| Rok vydání: | 2020 |
| Předmět: |
0301 basic medicine
gene delivery and expression in the retina Genetic Vectors Basic fibroblast growth factor Biomedical Engineering Gene Expression Biology Retina Article 03 medical and health sciences chemistry.chemical_compound Transduction (genetics) 0302 clinical medicine Transduction Genetic medicine Animals electric current Glial fibrillary acidic protein large-animal models intravitreal administration Inner limiting membrane Retinal Intravitreal administration Dependovirus Cell biology AAV vector Ophthalmology 030104 developmental biology medicine.anatomical_structure chemistry Inner nuclear layer 030221 ophthalmology & optometry biology.protein Rabbits sense organs |
| Zdroj: | Translational Vision Science & Technology |
| ISSN: | 2164-2591 |
| DOI: | 10.1167/tvst.9.7.28 |
| Popis: | Author(s): Song, Hongman; Zeng, Yong; Sardar Pasha, Sheik Pran Babu; Bush, Ronald A; Vijayasarathy, Camasamudram; Qian, Haohua; Wei, Lisa; Wiley, Henry E; Wu, Zhijian; Sieving, Paul A | Abstract: PurposeElectric micro-current has been shown to enhance penetration and transduction of adeno-associated viral (AAV) vectors in mouse retina after intravitreal administration. We termed this: "electric-current vector mobility (ECVM)." The present study considered whether ECVM could augment retinal transduction efficiency of intravitreal AAV8-CMV-EGFP in normal rabbit and nonhuman primate (NHP) macaque. Potential mechanisms underlying enhanced retinal transduction by ECVM were also studied.MethodsWe applied an electric micro-current across the intact eye of normal rabbit and monkey in vivo for a brief period immediately after intravitreal injection of AAV8-CMV-EGFP. Retinal GFP expression was evaluated by fundus imaging in vivo. Retinal immunohistochemistry was performed to assess the distribution of retinal cells transduced by the AAV8-EGFP. Basic fibroblast growth factor (bFGF) was analyzed by quantitative RT-polymerase chain reaction (PCR). Muller glial reactivity and inner limiting membrane (ILM) were examined by the glial fibrillary acidic protein (GFAP) and vimentin staining in mouse retina, respectively.ResultsECVM significantly increased the efficiency of AAV reaching and transducing the rabbit retina following intravitreal injection, with gene expression in inner nuclear layer, ganglion cells, and Muller cells. Similar trend of improvement was observed in the ECVM-treated monkey eye. The electric micro-current upregulated bFGF expression in Muller cells and vimentin showed ILM structural changes in mouse retina.ConclusionsECVM promotes the transduction efficiency of AAV8-CMV-GFP in normal rabbit and monkey retinas following intravitreal injection.Translational relevanceThis work has potential translational relevance to human ocular gene therapy by increasing retinal expression of therapeutic vectors given by intravitreal administration. |
| Databáze: | OpenAIRE |
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