| Hypoxia-regulated retinal glial cell-specific promoter for potential gene therapy in disease. | |
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MedLine Citation:
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PMID: 21960554 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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PURPOSE: Retinal Müller cells span the retina and secrete several trophic factors and represent the functional link between blood vessels and neurons, making them attractive targets for gene therapy. Therefore, a hypoxia-regulated, retinal glial cell-specific vector was constructed and tested for its response to hypoxia. METHODS: A hybrid promoter containing domains of human glial fibrillary acidic protein (GFAP) and several hypoxia-responsive and aerobically silenced elements (HRSE) was incorporated separately into plasmid vectors for generation of self-complementary adeno-associated virus. Müller cells trasfected with plasmids or virus were compared with other cell lines using standard METHODS: The mouse model of oxygen-induced retinopathy (OIR) was used to analyze retinas from mice exposed to high oxygen or room air to evaluate the induction of the regulated promoter. RESULTS: The regulated promoter was silenced under aerobic conditions in comparison with unregulated promoter in Müller cells. Hypoxia induced a 12-fold and 16-fold increase in promoter activity in primary Müller cells and human Müller cell lines, respectively. In the OIR model, intravitreal injection of the regulated promoter at postnatal day 7 (P7) resulted in high levels of green fluorescent protein expression only in retinal Müller cells at P17. GFP expression was absent in retinas of mice only exposed to room air. In vivo studies confirm normoxia silencing, hypoxic induction, and cell specificity of the regulated promoter in the mouse retina. CONCLUSIONS: This hypoxia-regulated, retinal glial cell-specific AAV vector provides a platform for gene therapy within regions of retinal hypoxia which are found in diabetic retinopathy and age-related macular degeneration. |
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Authors:
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Howard M Prentice; Manas R Biswal; C Kathleen Dorey; Janet C Blanks |
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Publication Detail:
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Type: Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't Date: 2011-11-01 |
Journal Detail:
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Title: Investigative ophthalmology & visual science Volume: 52 ISSN: 1552-5783 ISO Abbreviation: Invest. Ophthalmol. Vis. Sci. Publication Date: 2011 |
Date Detail:
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Created Date: 2011-11-02 Completed Date: 2012-01-05 Revised Date: 2013-01-09 |
Medline Journal Info:
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Nlm Unique ID: 7703701 Medline TA: Invest Ophthalmol Vis Sci Country: United States |
Other Details:
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Languages: eng Pagination: 8562-70 Citation Subset: IM |
Affiliation:
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Center for Complex Systems and Brain Sciences, Charles E. Schmidt College of Science, Florida Atlantic University, Boca Raton, FL 33431, USA. |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Adenoviridae
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genetics Animals Anoxia / physiopathology, therapy* Cells, Cultured Epithelial Cells / cytology, physiology Gene Silencing / drug effects Genetic Therapy / methods* Genetic Vectors / genetics* Glial Fibrillary Acidic Protein / genetics* HEK293 Cells Humans Mice Neuroglia / cytology, physiology* Oxygen / pharmacology Plasmids / genetics Promoter Regions, Genetic / drug effects, genetics Retina / cytology, physiology Retinal Diseases / physiopathology, therapy* |
| Grant Support | |
ID/Acronym/Agency:
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EYO16119//PHS HHS |
| Chemical | |
Reg. No./Substance:
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0/Glial Fibrillary Acidic Protein; 7782-44-7/Oxygen |
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