June 2017
Volume 58, Issue 8
Open Access
ARVO Annual Meeting Abstract  |   June 2017
Intravitreal injection of AAV7m8.hLCA5 restores photoreceptor function in Lca5-/- mice to nearly WT levels.
Author Affiliations & Notes
  • Puya Aravand
    Ophthalmology, University of Pennsylvania, Philadelphia, Pennsylvania, United States
  • Ji Yun Song
    Ophthalmology, University of Pennsylvania, Philadelphia, Pennsylvania, United States
  • Shangzhen Zhou
    Ophthalmology, University of Pennsylvania, Philadelphia, Pennsylvania, United States
  • Jieyan Pan
    Ophthalmology, University of Pennsylvania, Philadelphia, Pennsylvania, United States
  • Arkady Lyubarsky
    Ophthalmology, University of Pennsylvania, Philadelphia, Pennsylvania, United States
  • Jean Bennett
    Ophthalmology, University of Pennsylvania, Philadelphia, Pennsylvania, United States
  • Sergei S Nikonov
    Ophthalmology, University of Pennsylvania, Philadelphia, Pennsylvania, United States
    Vision Research Center, University of Pennsylvania, Philadelphia, Pennsylvania, United States
  • Footnotes
    Commercial Relationships   Puya Aravand, None; Ji Yun Song, None; Shangzhen Zhou, None; Jieyan Pan, None; Arkady Lyubarsky, None; Jean Bennett, Biogen (F), Gensight Biologics (S), Limelight (F), Sanofi (C), Spark Therapeutics (S); Sergei Nikonov, None
  • Footnotes
    Support  National Institutes of Health (5 P30 EY001583-42), Foundation Fighting Blindness, Research to Prevent Blindness, Center for Advanced Retina and Ocular Therapeutics, and the Paul and Evanina Bell Mackall Foundation Trust.
Investigative Ophthalmology & Visual Science June 2017, Vol.58, 4551. doi:
  • Views
  • Share
  • Tools
    • Alerts
      ×
      This feature is available to authenticated users only.
      Sign In or Create an Account ×
    • Get Citation

      Puya Aravand, Ji Yun Song, Shangzhen Zhou, Jieyan Pan, Arkady Lyubarsky, Jean Bennett, Sergei S Nikonov; Intravitreal injection of AAV7m8.hLCA5 restores photoreceptor function in Lca5-/- mice to nearly WT levels.. Invest. Ophthalmol. Vis. Sci. 2017;58(8):4551.

      Download citation file:


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

      ×
  • Supplements
Abstract

Purpose : Early onset vision loss results from mutations in LCA5, which encodes Lebercilin, a protein critical for photoreceptor health and function. We tested the possibility that gene augmentation therapy restores photoreceptor and retinal function using multi electrode array (MEA) in LCA5 mouse model (Lca5-/-).

Methods : Postnatal day 5 Lca5-/- mice received unilateral intravitreal injection with AAV7m8.CMV/CBA.hLCA5 (~9.87E+10 vg/eye) spiked with 5% (v/v) AAV7m8.CBA.GFP. Contralateral eyes were uninjected. 5 age-matched wildtype (WT) mice were positive controls. Retinas from light-adapted mice were dissected under red light and mounted ganglion cell side down in the MEA chamber. Calibrated full field flashes of 455 nm light (efficiency of pigment excitation ~40%:rhodopsin and M-opsin; ~0.2%:S-opsin) of different intensities were used for light stimulation (2 s flashes at 0.1 Hz or 50 ms flashes at 4 Hz). Data were analyzed using custom code in Matlab; spike sorting was performed in Plexon Offline Sorter.

Results : Presence of GFP confirmed exposure of photoreceptors to AAV. After 3-4 months, 3 out of 4 injected retinas had strong light responses and 1 showed minimal response in MEA testing. Responses became detectable at scotopic intensities (42-112 photons*s-1*μm-2, 455 nm) and strong responses were observed through the brightest photopic intensity of 2.00e9 photons*s-1*μm-2. Retinas from uninjected eyes showed minimal to abolished responses. As expected for rod/cone driven responses, after exposure to brightest light, scotopic responses disappeared but photopic responses were not significantly affected. Response kinetics were similar to those in WT retinas. Light sensitivity was slightly lower in treated Lca5-/- vs the most sensitive WT retinas where first responses were detected at 8-21 photons*s-1*μm-2. All ganglion cell types identifiable in the WT retinas under full field stimulation (ON-, OFF- and ON/OFF-types) were detected in the Lca5-/- treated retinas after spike sorting. WT and treated Lca5-/- retinas were responsive to 4Hz flicker stimulation at 3.53e2-2.00e9 photons*s-1*μm-2. All untreated Lca5-/- retinas showed slow melanopsin driven responses at brighter intensities which were absent in the light sensitive treated Lca5-/- and WT retinas.

Conclusions : Given successful transduction, gene therapy can restore degenerating retinal cells to a state close to the WT conditions.

This is an abstract that was submitted for the 2017 ARVO Annual Meeting, held in Baltimore, MD, May 7-11, 2017.

×
×

This PDF is available to Subscribers Only

Sign in or purchase a subscription to access this content. ×

You must be signed into an individual account to use this feature.

×