AAV2 mediated transduction of the mouse retina after optic nerve injury

R. Nickells, H. Schmitt, M.E. Maes, C. Schlamp, Investigative Ophthalmology and Visual Science 58 (2017) 6091–6104.

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Journal Article | Published | English

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Author
Nickells, Robert; Schmitt, Heather; Maes, Margaret EIST Austria ; Schlamp, Cassandra
Department
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.
Publishing Year
Date Published
2017-12-14
Journal Title
Investigative Ophthalmology and Visual Science
Volume
58
Issue
14
Page
6091 - 6104
ISSN
IST-REx-ID
557

Cite this

Nickells R, Schmitt H, Maes ME, Schlamp C. AAV2 mediated transduction of the mouse retina after optic nerve injury. Investigative Ophthalmology and Visual Science. 2017;58(14):6091-6104. doi:10.1167/iovs.17-22634
Nickells, R., Schmitt, H., Maes, M. E., & Schlamp, C. (2017). AAV2 mediated transduction of the mouse retina after optic nerve injury. Investigative Ophthalmology and Visual Science, 58(14), 6091–6104. https://doi.org/10.1167/iovs.17-22634
Nickells, Robert, Heather Schmitt, Margaret E Maes, and Cassandra Schlamp. “AAV2 Mediated Transduction of the Mouse Retina after Optic Nerve Injury.” Investigative Ophthalmology and Visual Science 58, no. 14 (2017): 6091–6104. https://doi.org/10.1167/iovs.17-22634.
R. Nickells, H. Schmitt, M. E. Maes, and C. Schlamp, “AAV2 mediated transduction of the mouse retina after optic nerve injury,” Investigative Ophthalmology and Visual Science, vol. 58, no. 14, pp. 6091–6104, 2017.
Nickells R, Schmitt H, Maes ME, Schlamp C. 2017. AAV2 mediated transduction of the mouse retina after optic nerve injury. Investigative Ophthalmology and Visual Science. 58(14), 6091–6104.
Nickells, Robert, et al. “AAV2 Mediated Transduction of the Mouse Retina after Optic Nerve Injury.” Investigative Ophthalmology and Visual Science, vol. 58, no. 14, Association for Research in Vision and Ophthalmology Inc., 2017, pp. 6091–104, doi:10.1167/iovs.17-22634.
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2018-12-12
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