September 2016
Volume 57, Issue 12
Open Access
ARVO Annual Meeting Abstract  |   September 2016
Vitreous body: A barrier for AAV-mediated gene transfer to inner retinal neurons of macaque
Author Affiliations & Notes
  • Tshitoko Kizito Tshilenge
    Atlantic Gene Therapies INSERM UMR 1089, Nantes, France
  • Baptiste Ameline
    Atlantic Gene Therapies INSERM UMR 1089, Nantes, France
  • Michel Weber
    Service d'Ophtalmologie, CHU-Hôtel Dieu, Nantes, France
  • Guylène Le Meur
    Service d'Ophtalmologie, CHU-Hôtel Dieu, Nantes, France
  • Jack-Yves Deschamps
    ONIRIS, Nantes-Atlantic College of Veterinary Medicine, Food Science and Engineering, Emergency and Critical Care Unit, Nantes, France
  • Alexandra Mendes-Madeira
    Atlantic Gene Therapies INSERM UMR 1089, Nantes, France
  • Steven Nedellec
    Cellular and Tissular Imaging Core Facility of Nantes University, SFR Santé François Bonamy INSERM UMS016 / CNRS UMS3556, Nantes, France
  • Véronique Blouin
    Atlantic Gene Therapies INSERM UMR 1089, Nantes, France
  • Virginie Pichard
    Atlantic Gene Therapies INSERM UMR 1089, Nantes, France
  • Philippe Moullier
    Atlantic Gene Therapies INSERM UMR 1089, Nantes, France
    Department of Molecular Genetics and Microbiology, University of Florida, College of Medicine, Gainesville, Florida, United States
  • Fabienne Rolling
    Atlantic Gene Therapies INSERM UMR 1089, Nantes, France
  • Footnotes
    Commercial Relationships   Tshitoko Kizito Tshilenge, None; Baptiste Ameline, None; Michel Weber, None; Guylène Le Meur, None; Jack-Yves Deschamps, None; Alexandra Mendes-Madeira, None; Steven Nedellec, None; Véronique Blouin, None; Virginie Pichard, None; Philippe Moullier, None; Fabienne Rolling, None
  • Footnotes
    Support  None
Investigative Ophthalmology & Visual Science September 2016, Vol.57, 100. doi:
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      Tshitoko Kizito Tshilenge, Baptiste Ameline, Michel Weber, Guylène Le Meur, Jack-Yves Deschamps, Alexandra Mendes-Madeira, Steven Nedellec, Véronique Blouin, Virginie Pichard, Philippe Moullier, Fabienne Rolling; Vitreous body: A barrier for AAV-mediated gene transfer to inner retinal neurons of macaque. Invest. Ophthalmol. Vis. Sci. 2016;57(12):100.

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

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Abstract

Purpose : For retinal gene delivery, several studies have illustrated that AAV2/2 injected intravitreally transduces retinal ganglion cells (RGCs) efficiently in rodent models. However, considerable anatomical differences between eyes of rodent models and human subjects make difficult the translation of these results. Indeed, contrary to rodent eyes, the vitreous body fills a significantly larger space in human eyes, which consequently might be a physical barrier for vector diffusion and transduction. Because macaque eyes recapitulate the anatomical characteristics of the human eye, we evaluated the effect of the vitreous body on AAV2/2 transduction in macaque retinas following intravitreal injection.

Methods : We generated AAV2/2 vectors carrying the enhanced green fluorescence protein (eGFP) cDNA under the control of the cytomegalovirus promoter. AAV2/2 vector was administered intravitreally in macaque eyes either directly in the vitreous chamber without removing the vitreous body (n=2) or after a complete vitrectomy, a surgical procedure aiming to remove the vitreous body (n=2). Native eGFP expression was monitored by funduscopic imaging and by confocal microscopy in retinal flat mounts.

Results : In macaque eyes where vitrectomy was performed, extensive eGFP expression was detectable in RGCs axons projecting toward the optic nerve head. The transduction efficiency was mainly localized in peripheral retina while in central retina low eGFP expression was observed except for RGCs transduction around the macular region. On the contrary, eGFP expression was not detected in central and peripheral retinas in macaque eyes in which the vitreous body was present.

Conclusions : This observation suggests that the vitreous body interferes in AAV2/2-mediated transduction to inner retinal neurons of macaque retinas. Our hypothesis is that the vector injected intravitreally without prior vitrectomy is obstructed by the vitreous -a gel-like structure-, which may trap the vector and in turn reduces its diffusion towards inner retinal neurons. We illustrated that vitrectomy prior vector injection is an effective procedure to overcome this physical barrier and consequently improves the transduction of RGCs in macaque retinas. Overall, these results set the stage for the translation of vitrectomy in human subjects as relevant approach to promote gene delivery to inner retinal neurons after AAV vector intravitreal administration.

This is an abstract that was submitted for the 2016 ARVO Annual Meeting, held in Seattle, Wash., May 1-5, 2016.

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