title: Significant changes in endogenous retinal gene expression assessed 1 year after a single intraocular injection of AAV-CNTF or AAV-BDNF.
abstract: 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.
authors.affiliation: School of Anatomy, Physiology, and Human Biology, The University of Western Australia , Western Australia, Australia.
authors.forename: Chrisna J
authors.initials: CJ
authors.lastname: LeVaillant
authors.affiliation: School of Anatomy, Physiology, and Human Biology, The University of Western Australia , Western Australia, Australia.
authors.forename: Anil
authors.initials: A
authors.lastname: Sharma
authors.affiliation: School of Anatomy, Physiology, and Human Biology, The University of Western Australia , Western Australia, Australia.
authors.forename: Jill
authors.initials: J
authors.lastname: Muhling
authors.affiliation: School of Anatomy, Physiology, and Human Biology, The University of Western Australia , Western Australia, Australia.
authors.forename: Lachlan Pg
authors.initials: LP
authors.lastname: Wheeler
authors.affiliation: School of Anatomy, Physiology, and Human Biology, The University of Western Australia , Western Australia, Australia.
authors.forename: Greg S
authors.initials: GS
authors.lastname: Cozens
authors.affiliation: School of Anatomy, Physiology, and Human Biology, The University of Western Australia, Western Australia, Australia; Department of Obstetrics and Gynecology, Laboratory for Transplantation and Regenerative Medicine, Sahlgrenska Academy, University of Gothenburg, Gothenburg, Sweden.
authors.forename: Mats
authors.initials: M
authors.lastname: Hellström
authors.affiliation: School of Animal Biology, The University of Western Australia, Western Australia, Australia; Western Australian Neuroscience Research Institute, Western Australia, Australia.
authors.forename: Jennifer
authors.initials: J
authors.lastname: Rodger
authors.affiliation: School of Anatomy, Physiology, and Human Biology, The University of Western Australia, Western Australia, Australia; Western Australian Neuroscience Research Institute, Western Australia, Australia.
authors.forename: Alan R
authors.initials: AR
authors.lastname: Harvey