AAV2 mediated transduction of the mouse retina after optic nerve injury Journal Article

Author(s): Nickells, Robert W; Schmitt, Heather M; Maes, Margaret E; Schlamp, Cassandra L
Article Title: AAV2 mediated transduction of the mouse retina after optic nerve injury
Affiliation IST Austria
Abstract: PURPOSE. Gene therapy of retinal ganglion cells (RGCs) has promise as a powerful therapeutic for the rescue and regeneration of these cells after optic nerve damage. However, early after damage, RGCs undergo atrophic changes, including gene silencing. It is not known if these changes will deleteriously affect transduction and transgene expression, or if the therapeutic protein can influence reactivation of the endogenous genome. METHODS. Double-transgenic mice carrying a Rosa26-(LoxP)-tdTomato reporter, and a mutant allele for the proapoptotic Bax gene were reared. The Bax mutant blocks apoptosis, but RGCs still exhibit nuclear atrophy and gene silencing. At times ranging from 1 hour to 4 weeks after optic nerve crush (ONC), eyes received an intravitreal injection of AAV2 virus carrying the Cre recombinase. Successful transduction was monitored by expression of the tdTomato reporter. Immunostaining was used to localize tdTomato expression in select cell types. RESULTS. Successful transduction of RGCs was achieved at all time points after ONC using AAV2 expressing Cre from the phosphoglycerate kinase (Pgk) promoter, but not the CMV promoter. ONC promoted an increase in the transduction of cell types in the inner nuclear layer, including Müller cells and rod bipolar neurons. There was minimal evidence of transduction of amacrine cells and astrocytes in the inner retina or optic nerve. CONCLUSIONS. Damaged RGCs can be transduced and at least some endogenous genes can be subsequently activated. Optic nerve damage may change retinal architecture to allow greater penetration of an AAV2 virus to transduce several additional cell types in the inner nuclear layer.
Keywords: gene therapy; optic nerve damage; retinal ganglion cells
Journal Title: Investigative Ophthalmology and Visual Science
Volume: 58
Issue 14
ISSN: 01460404
Publisher: Association for Research in Vision and Ophthalmology Inc.  
Date Published: 2017-12-14
Start Page: 6091
End Page: 6104
Copyright Statement: CC BY
DOI: 10.1167/iovs.17-22634
Notes: The authors thank Kimberly Toops for helpful discussions on imaging of tdTomato fluorescence. Supported by the National Eye Institute Grant R01 EY012223 (RWN), Vision Science CORE Grant P30 EY016665 (Department of Ophthalmology and Visual Sciences, University of Wisconsin-Madison), National Institutes of Health Grant T32 GM081061 (Department of Pathology and Laboratory Medicine, University of Wisconsin-Madison), a Research to Prevent Blindness Senior Investigator Award (RWN), and an unrestricted research grant from the Research to Prevent Blindness.
Open access: yes (OA journal)