June 2023
Volume 64, Issue 8
Open Access
ARVO Annual Meeting Abstract  |   June 2023
Intravitreal AAV2 gene delivery to feline retinal ganglion cells (RGCs)
Author Affiliations & Notes
  • Kazuya Oikawa
    Ophthalmology and Visual Sciences, University of Wisconsin-Madison, Madison, Wisconsin, United States
    Surgical Sciences, University of Wisconsin-Madison, Madison, Wisconsin, United States
  • Julie Kiland
    Ophthalmology and Visual Sciences, University of Wisconsin-Madison, Madison, Wisconsin, United States
  • Seth Eaton
    Surgical Sciences, University of Wisconsin-Madison, Madison, Wisconsin, United States
  • Odalys Torne
    Ophthalmology and Visual Sciences, University of Wisconsin-Madison, Madison, Wisconsin, United States
    Surgical Sciences, University of Wisconsin-Madison, Madison, Wisconsin, United States
  • Virginia Mathu
    Ophthalmology and Visual Sciences, University of Wisconsin-Madison, Madison, Wisconsin, United States
    Surgical Sciences, University of Wisconsin-Madison, Madison, Wisconsin, United States
  • Robert W Nickells
    Ophthalmology and Visual Sciences, University of Wisconsin-Madison, Madison, Wisconsin, United States
  • Gillian J McLellan
    Ophthalmology and Visual Sciences, University of Wisconsin-Madison, Madison, Wisconsin, United States
    Surgical Sciences, University of Wisconsin-Madison, Madison, Wisconsin, United States
  • Footnotes
    Commercial Relationships   Kazuya Oikawa None; Julie Kiland None; Seth Eaton None; Odalys Torne None; Virginia Mathu None; Robert Nickells None; Gillian McLellan None
  • Footnotes
    Support  NIH grants R01 EY027396, R01 EY030123, P30 EY016665 and S10 OD026957;UW-Madison ICTR CTSA grants supported by NIH UL1TR002373; an unrestricted award to the UW-Madison Department of Ophthalmology and Visual Sciences from Research to Prevent Blindness
Investigative Ophthalmology & Visual Science June 2023, Vol.64, 3848. doi:
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    • Get Citation

      Kazuya Oikawa, Julie Kiland, Seth Eaton, Odalys Torne, Virginia Mathu, Robert W Nickells, Gillian J McLellan; Intravitreal AAV2 gene delivery to feline retinal ganglion cells (RGCs). Invest. Ophthalmol. Vis. Sci. 2023;64(8):3848.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose : Studies in rodent models suggest that adeno-associated virus serotype 2 (AAV2) is a promising gene therapy platform for RGC diseases like glaucoma, but inter-species anatomic differences mean that mouse data may not be predictive of success in humans. We assessed feasibility of intravitreal gene delivery with AAV2 to feline RGCs, in a species with eye size similar to humans.

Methods : Posterior vitreal injection of AAV 2/2-CMV-GFP, was performed overlying the area centralis in 5 adult cats. Cats received a single injection of 50µL of either self-complementary or traditional single-stranded AAV 2/2 vector (~2 x1012 viral genomes/mL, UNC Vector Core) in one eye. Peri-operative, immunosuppressive prednisolone (2mg/kg/day, PO) was gradually tapered over 6-10wks. Ophthalmic examination (slit-lamp biomicroscopy, indirect fundoscopy and tonometry) were performed pre- and post-injection. GFP reporter expression and morphological effects of viral transduction on the retina were monitored in vivo using confocal scanning laser ophthalmoscopy (cSLO) and optical coherence tomography (OCT), respectively (Spectralis, Heidelberg), at 1-2wk intervals over the 6-10wk study period. Full-field electroretinogram (ERG) and visual evoked potential (VEP) were conducted at baseline and post-injection. Retinas were examined by histology and immunolabeling for the RGC marker RBPMS and Müller cell/astrocyte marker SOX9, and GFP expression was examined in retina, distal optic nerve and brain, by epifluorescence and confocal microscopy. GFP+ cells were quantified in ImageJ.

Results : GFP+ retinal cells and RGC axons were visualized by cSLO at 1-2 weeks post-injection with highest GFP expression in the inferior-nasal retina. No retinal morphological changes were observed by OCT in vivo or by histology but 2/5 eyes exhibited mild perivascular inflammation. Retinal and ON function were preserved in injected eyes compared to baseline and untreated eyes. GFP+ signals in the injected eyes were predominantly identified in RBPMS+ RGC cells and SOX9+ Müller cells. GFP fluorescence was observed throughout the visual pathway. Peak transduction in RGCs (~50%) was observed in the regions with high GFP expression, whereas <1% of RGCs expressed GFP across the whole retina.

Conclusions : These data provide proof of concept that pre-retinal injection of AAV2/2 may represent a promising platform for gene delivery to feline RGCs in vivo.

This abstract was presented at the 2023 ARVO Annual Meeting, held in New Orleans, LA, April 23-27, 2023.

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