Significant changes in endogenous retinal gene expression assessed 1 year after a single intraocular injection of AAV-CNTF or AAV-BDNF
| Autor: | Chrisna J. LeVaillant, Greg Cozens, Jennifer Rodger, Anil Sharma, Lachlan P.G. Wheeler, Alan R. Harvey, Jill Muhling, Mats Hellström |
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| Jazyk: | angličtina |
| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
lcsh:QH426-470 viruses Biology Ciliary neurotrophic factor Retinal ganglion Article 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine Neurotrophic factors Gene expression Genetics medicine lcsh:QH573-671 Molecular Biology Ganglion cell layer lcsh:Cytology Retinal Molecular biology lcsh:Genetics 030104 developmental biology Real-time polymerase chain reaction medicine.anatomical_structure Retinal ganglion cell chemistry biology.protein Molecular Medicine 030217 neurology & neurosurgery |
| Zdroj: | Molecular Therapy: Methods & Clinical Development, Vol 3, Iss C (2016) Molecular Therapy. Methods & Clinical Development |
| ISSN: | 2329-0501 |
| Popis: | Use of viral vectors to deliver therapeutic genes to the central nervous system holds promise for the treatment of neurodegenerative diseases and neurotrauma. Adeno-associated viral (AAV) vectors encoding brain-derived neurotrophic factor (BDNF) or ciliary derived neurotrophic factor (CNTF) promote the viability and regeneration of injured adult rat retinal ganglion cells. However, these growth-inducing transgenes are driven by a constitutively active promoter, thus we examined whether long-term AAV-mediated secretion of BDNF or CNTF affected endogenous retinal gene expression. One year after the intravitreal injection of AAV-green fluorescent protein (GFP), bi-cistronic AAV-BDNF-GFP or AAV-CNTF-GFP, mRNA was extracted and analyzed using custom 96 well polymerase chain reaction arrays. Of 93 test genes, 56% showed significantly altered expression in AAV-BDNF-GFP and/or AAV-CNTF-GFP retinas compared with AAV-GFP controls. Of these genes, 73% showed differential expression in AAV-BDNF versus AAV-CNTF injected eyes. To focus on retinal ganglion cell changes, quantitative polymerase chain reaction was undertaken on mRNA (16 genes) obtained from fixed retinal sections in which the ganglion cell layer was enriched. The sign and extent of fold changes in ganglion cell layer gene expression differed markedly from whole retinal samples. Sustained and global alteration in endogenous mRNA expression after gene therapy should be factored into any interpretation of experimental/clinical outcomes, particularly when introducing factors into the central nervous system that require secretion to evoke functionality. |
| Databáze: | OpenAIRE |
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