July 2019
Volume 60, Issue 9
Open Access
ARVO Annual Meeting Abstract  |   July 2019
Novel recombinant Adeno-Associated Virus transfects photoreceptor cells following intravitreal injection in sheep.
Author Affiliations & Notes
  • Maya Ross
    Koret School of Veterinary Medicine, Hebrew University of Jerusalem Israel, Rehovot, Israel
  • Eyal Banin
    Department of Ophthalmology, Hadassah-Hebrew University Medical Center, Jerusalem, Israel
  • Edward Averbukh
    Department of Ophthalmology, Hadassah-Hebrew University Medical Center, Jerusalem, Israel
  • Melissa Desrosiers
    Institute de la Vision, Paris, France
  • Alexey Obolensky
    Department of Ophthalmology, Hadassah-Hebrew University Medical Center, Jerusalem, Israel
  • Raaya Ezra-Elia
    Koret School of Veterinary Medicine, Hebrew University of Jerusalem Israel, Rehovot, Israel
  • Hen Honig
    ARO, The Volcani Center, Rishon LeZion, Israel
  • Esther Yamin
    Department of Ophthalmology, Hadassah-Hebrew University Medical Center, Jerusalem, Israel
  • Alexasnder Rosov
    ARO, The Volcani Center, Rishon LeZion, Israel
  • Hay Dvir
    ARO, The Volcani Center, Rishon LeZion, Israel
  • Elisha Gootwine
    ARO, The Volcani Center, Rishon LeZion, Israel
  • Deniz Dalkara
    Institute de la Vision, Paris, France
  • Ron Ofri
    Koret School of Veterinary Medicine, Hebrew University of Jerusalem Israel, Rehovot, Israel
  • Footnotes
    Commercial Relationships   Maya Ross, None; Eyal Banin, AGTC (F); Edward Averbukh, None; Melissa Desrosiers, None; Alexey Obolensky, None; Raaya Ezra-Elia, None; Hen Honig, None; Esther Yamin, None; Alexasnder Rosov, None; Hay Dvir, None; Elisha Gootwine, AGTC (F); Deniz Dalkara, None; Ron Ofri, AGTC (F)
  • Footnotes
    Support  ISF Grant 1257/15, Israel Ministry oh Health Grant 300011892
Investigative Ophthalmology & Visual Science July 2019, Vol.60, 2897. doi:https://doi.org/
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      Maya Ross, Eyal Banin, Edward Averbukh, Melissa Desrosiers, Alexey Obolensky, Raaya Ezra-Elia, Hen Honig, Esther Yamin, Alexasnder Rosov, Hay Dvir, Elisha Gootwine, Deniz Dalkara, Ron Ofri; Novel recombinant Adeno-Associated Virus transfects photoreceptor cells following intravitreal injection in sheep.. Invest. Ophthalmol. Vis. Sci. 2019;60(9):2897. doi: https://doi.org/.

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

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Abstract

Purpose : Subretinal (SR) delivery of recombinant Adeno-Associated Viruses (AAV) resulted in successful treatment of several inherited retinal degenerations in humans and animal models. However, due to potential complications, drawbacks and challenges of SR injections, there is an impetus to develop less invasive delivery methods such as intravitreal (IVT) injections. In the current study we evaluated whether an engineered AAV2-7m8 vector, with improved retinal penetration properties, can transduce cones following IVT injection in sheep.

Methods : Surgeries were performed on seven AAV2 seronegative, ophthalmologically normal five-month-old Afec-Assaf sheep. We used an engineered AAV2-7m8 vector that differs from its parental serotype AAV2 by a peptide insertion. The vector, carrying GFP under the control of a ubiquitous (CAG) promoter, was injected IVT in five eyes. The same vector, carrying GFP under the control of a cone specific (pr1.7) promoter, was injected IVT in 6 eyes. SR injections of AAV2-7m8 with both promoters served as positive controls. GFP expression was evaluated in vivo every four weeks by fundus photography using a fluorescein angiography lens. Four months post-treatment sheep were sacrificed and GFP expression confirmed by immunohistochemistry (IHC).

Results : Four weeks post-treatment, in vivo imaging of SR-injected eyes demonstrated substantial fluorescence indicating high GFP expression. Eyes injected IVT with AAV2-7m8-CAG-GFP began showing fluorescence on in vivo imaging eight weeks post-treatment, while in eyes treated IVT with AAV2-7m8-pr1.7-GFP first signs of GFP expression were evident 12 weeks post-treatment. IHC confirmed retinal GFP expression in SR positive controls and IVT-treated eyes. IVT injection of AAV2-7m8-CAG-GFP resulted in GFP expression in the ganglion cell layer, while its SR injection resulted in expression in RPE cells and photoreceptors. Both SR and IVT injections of AAV2-7m8-pr1.7-GFP resulted in GFP expression in cones. In vivo imaging and IHC demonstrated a more robust GFP expression in SR than in IVT injected eyes.

Conclusions : IVT-injected AAV2-7m8 successfully penetrated the ovine retina, leading to photoreceptor transduction. The efficacy of IVT delivery of AAV2-7m8 as a potential treatment of achromatopsia will be further investigated in our day blind sheep model.

This abstract was presented at the 2019 ARVO Annual Meeting, held in Vancouver, Canada, April 28 - May 2, 2019.

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