September 2016
Volume 57, Issue 12
Open Access
ARVO Annual Meeting Abstract  |   September 2016
Optogenetic visual restoration using ChrimsonR: Validation in degenerative rodent models, rd1 and P23H.
Author Affiliations & Notes
  • Romain Caplette
    Institut de la Vision, Paris, France
  • Elisabeth Dubus
    Institut de la Vision, Paris, France
  • Gregory Gauvain
    Institut de la Vision, Paris, France
  • Olivier Marre
    Institut de la Vision, Paris, France
  • Melissa Desrosiers
    Institut de la Vision, Paris, France
  • Deniz Dalkara
    Institut de la Vision, Paris, France
  • José-Alain Sahel
    Institut de la Vision, Paris, France
  • Anne Douar
    Gensight Biologics, Paris, France
  • Didier Pruneau
    Gensight Biologics, Paris, France
  • Serge A Picaud
    Institut de la Vision, Paris, France
  • Footnotes
    Commercial Relationships   Romain Caplette, None; Elisabeth Dubus, None; Gregory Gauvain, None; Olivier Marre, None; Melissa Desrosiers, None; Deniz Dalkara, Gensight Biologics (C), Gensight Biologics (P); José-Alain Sahel, Chronocam (I), Chronocam (C), Chronocam (P), Gensight Biologics (C), Gensight Biologics (P), Gensight Biologics (I), Gensight Biologics (F), Pixium (I), Pixium (C), Pixium (P); Anne Douar, Gensight Biologics (E), Gensight Biologics (I); Didier Pruneau, Gensight Biologics (E), Gensight Biologics (I); Serge Picaud, Chronocam (I), Chronocam (C), Gensight Biologics (C), Gensight Biologics (F), Pixium (C)
  • Footnotes
    Support  Funding Sources: Banque publique d'Investissement, Foundation Fighting Blindness, LabEx LIFESENSES (ANR-10-LABX-65), Fondation de la recherche médicale, Gensight Biologics.
Investigative Ophthalmology & Visual Science September 2016, Vol.57, 600. doi:
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      Romain Caplette, Elisabeth Dubus, Gregory Gauvain, Olivier Marre, Melissa Desrosiers, Deniz Dalkara, José-Alain Sahel, Anne Douar, Didier Pruneau, Serge A Picaud; Optogenetic visual restoration using ChrimsonR: Validation in degenerative rodent models, rd1 and P23H.. Invest. Ophthalmol. Vis. Sci. 2016;57(12):600.

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

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Abstract

Purpose : Ectopic expression of ChannelRhodopsin2 in retinal ganglion cells was shown to restore vision in blind rd1 mice. However, concerns on phototoxicity were raised by the required high excitation threshold in the blue wavelength range. In this study, we investigated the use of ChrimsonR (ChrR), a red shifted opsin, as radiation safety limits are much higher in the red light range. We compared further the functional efficacy of ChrR and the construct ChrimsonR-TdTomato (ChrR-TdT) in two degenerative models: rd1 and P23H.

Methods : rd1 mice and P23H rats were injected intravitreally with AAV2.7m8 vector encoding ChrR-Tdt and ChrR under the CAG promoter. Expression of TdT was examined by its fluorescence under classic and confocal microscopy. Multielectrode array (MEA) recordings were performed ex vivo on flat-mounted retina in presence of synaptic blockers to assess photoactivation of ChrR-expressing retinal ganglion cells at least a month after injection.

Results : ChrR-tdT expression was widely distributed across the RGC layer in the rd1 retina. The fluorescence extended in RGC cell bodies and their dendritic arbors. For MEA recordings, retinal pieces were selected close to the optic nerve where transfection was maximal. For rd1 experiments, retinal ganglion cell responses were measured in 6 out of 8 retinas with ChrR-TdT whereas only 4 retinas out of 8 were active with ChrR. The activity spectrum matched that of ChrR. In the red wavelength range, light level to elicit spiking was below radiation safety limits. Light thresholds were similar with ChrR and ChrRTdT but spiking frequencies were greater in ChrR-TdT expressing cells. Recording in P23H rats confirmed retinal ganglion cell photoactivity following ChR-TdT delivery.

Conclusions : This study demonstrated the potential of ChrR for the reactivation of retinal ganglion cells in a blind retina. The data suggested that ChrR-TdT was more potent than ChrR and paved the way for further investigation of ChrR-TdT expression and function in the primate retina (see posters Gauvain et al. and Chaffiol et al.).

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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