September 2016
Volume 57, Issue 12
Open Access
ARVO Annual Meeting Abstract  |   September 2016
Optogenetic engineering of retinal ganglion cells with AAV2.7m8-ChrimsonR-tdTomato (GS030) in combination with an event-based camera photostimulation device
Author Affiliations & Notes
  • Jose Sahel
    Division of Experimental Ophthalmology, Institut de la Vision, Paris, France
  • Deniz Dalkara
    Division of Experimental Ophthalmology, Institut de la Vision, Paris, France
  • Jens Duebel
    Division of Experimental Ophthalmology, Institut de la Vision, Paris, France
  • Ryad Benosman
    Division of Experimental Ophthalmology, Institut de la Vision, Paris, France
  • Guillaume Chenegros
    Division of Experimental Ophthalmology, Institut de la Vision, Paris, France
  • Anne Douar
    GenSight Biologics, Paris, France
  • Joël Chavas
    GenSight Biologics, Paris, France
  • Céline Bouquet
    GenSight Biologics, Paris, France
  • Didier Pruneau
    GenSight Biologics, Paris, France
  • Botond Roska
    Friedrich Miescher Institute, Basel, Switzerland
  • Serge A Picaud
    Division of Experimental Ophthalmology, Institut de la Vision, Paris, France
  • Footnotes
    Commercial Relationships   Jose Sahel, Chronocam (I), Chronolife (I), Genesignal (C), GenSight Biologics (C), GenSight Biologics (I), GenSight Biologics (F), Pixium Vision (I), Pixium Vision (C), Sanofi-Fovea (C), Vision Medicines (C); Deniz Dalkara, GenSight Biologics (C), GenSight Biologics (F); Jens Duebel, GenSight Biologics (F); Ryad Benosman, GenSight Biologics (F), GenSight Biologics (C); Guillaume Chenegros, GenSight Biologics (F), GenSight Biologics (C); Anne Douar, GenSight Biologics (E); Joël Chavas, GenSight Biologics (E); Céline Bouquet, GenSight Biologics (E); Didier Pruneau, GenSight Biologics (E); Botond Roska, GenSight Biologics (F); Serge Picaud, Chronocam (I), GenSight Biologics (F), Pixium Vision (C), Pixium Vision (I)
  • Footnotes
    Support  Banque publique d'Investissement, Foundation Fighting Blindness, LabEx LIFESENSES (ANR-10-LABX-65), GenSight Biologics
Investigative Ophthalmology & Visual Science September 2016, Vol.57, No Pagination Specified. doi:
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      Jose Sahel, Deniz Dalkara, Jens Duebel, Ryad Benosman, Guillaume Chenegros, Anne Douar, Joël Chavas, Céline Bouquet, Didier Pruneau, Botond Roska, Serge A Picaud; Optogenetic engineering of retinal ganglion cells with AAV2.7m8-ChrimsonR-tdTomato (GS030) in combination with an event-based camera photostimulation device. Invest. Ophthalmol. Vis. Sci. 201657(12):.

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

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Abstract

Purpose : Ectopic expression of a light-sensitive opsin in retinal ganglion cells (RGCs) is an attractive strategy to restore vision. Here, we evaluated the ability of ChrimsonR-tdTomato (ChrR-tdT), developed by Ed Boyden (MIT), to convert light insensitive RGCs into photoactivable cells both in rd1 mice and normal macaques. A photostimulation device inspired by natural vision has also been developed.

Methods : rd1 mice and Cynomolgus macaques were injected intravitreally with AAV2.7m8 vector encoding ChrR-tdT (GS030). Multielectrode array (MEA) measurements and expression of tdT were assessed in explanted retinas 1 and 2 months after injections in mice and macaques, respectively. Local tolerance was also evaluated in macaques by ophthalmic exams and histology.

Results : In both rd1 mice and normal macaques, ChrR-tdT was essentially expressed in RGCs. In macaques, its expression was restricted to the perifoveal area, and was not associated with any structural modifications, inflammation or necrosis; all ophthalmic exams were normal. Patch-clamp experiments, conducted on monkey retina by targeting ChrR-tdT-expressing RGCs, allowed biophysical characterization of ChrR-tdT-elicited currents. MEA recordings showed that light responses followed the action spectrum of ChrR-tdT. Activity was correlated with TdT fluorescence reaching very high spiking frequencies (above 300Hz). Required light levels were below safety radiation limits. A complete prototype of the photostimulation device was developed with the optical chain, a functional electronic, firmware and software architecture. It captures external scenes through an event-based camera over a high dynamic range to deliver visual stimulations onto the ChrR-tdT-transduced retina.

Conclusions : Our GS030 vector efficiently and safely photo-engineered RGCs in both blind mice and normal macaques. ChrR-tdT-based gene therapy combined with a photostimulation device appears to be a viable treatment option for vision restoration in RP patients.

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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