May 2007
Volume 48, Issue 13
ARVO Annual Meeting Abstract  |   May 2007
Ciliary Muscle Electrotransfer Allows for Controlled and Sustained Production of Therapeutic Proteins in Ocular Media
Author Affiliations & Notes
  • E. Touchard
    INSERM U598, Paris, France
  • C. Bloquel
    INSERM U598, Paris, France
  • P. Bigey
    INSERM U640, Paris, France
  • C. Gandolphe
    INSERM U598, Paris, France
    INSERM U640, Paris, France
  • D. BenEzra
    INSERM U598, Paris, France
    Hadassah Hebrew University Hospital, Jerusalem, Israel
  • D. Scherman
    INSERM U640, Paris, France
  • F. Behar-Cohen
    INSERM U598, Paris, France
    Hotel-Dieu, Department of Ophthalmology / Rothschild Ophthalmologic Foundation, Paris, France
  • Footnotes
    Commercial Relationships E. Touchard, None; C. Bloquel, None; P. Bigey, None; C. Gandolphe, None; D. BenEzra, None; D. Scherman, None; F. Behar-Cohen, None.
  • Footnotes
    Support None.
Investigative Ophthalmology & Visual Science May 2007, Vol.48, 5811. doi:
  • Views
  • Share
  • Tools
    • Alerts
      This feature is available to authenticated users only.
      Sign In or Create an Account ×
    • Get Citation

      E. Touchard, C. Bloquel, P. Bigey, C. Gandolphe, D. BenEzra, D. Scherman, F. Behar-Cohen; Ciliary Muscle Electrotransfer Allows for Controlled and Sustained Production of Therapeutic Proteins in Ocular Media. Invest. Ophthalmol. Vis. Sci. 2007;48(13):5811.

      Download citation file:

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

  • Supplements

Purpose:: Our aim was to define the optimal conditions for plasmid transfection into the ciliary muscle in order to achieve a sustained and controlled secretion of therapeutic proteins in the vitreous cavity.

Methods:: 8-10 weeks old Lewis rats were used in these experiments. Injections of 10µl to 30µl containing 10 to 30µg of pVAX2-gLuc (secreted Gaussia luciferase) or pVAX1-LacZ plasmids (driven by a CMV promoter) were performed in the ciliary muscle of the rat eyes. Several parameters were evaluated regarding the variability and efficacy of transfection and the amount of secreted protein: route of injection (corneal tunnel or transscleral), formulation of DNA plasmids, number of injection sites, amount and volume of injected plasmids, electrode shape and electrical parameters. A kinetic of expression was performed with the gLuc encoding plasmid. Finally other therapeutic proteins were dosed in the vitreous cavity (Epo, sTNFR1-Ig).

Results:: Transscleral injections of naked plasmids (versus PEI) in saline (versus distilled water) were more efficient to transfect reproducibly the ciliary muscle. The optimal electrical conditions were 200V/cm, 8 pulses, 5Hz. Multiple injection sites significantly improved the reproducibility of transfection. The amount of transfected plasmid correlated well with the levels of proteins detected in the vitreous. When no current was applied, a pick of secretion was observed in the vitreous but no sustained protein production. On the contrary, using optimized conditions of electrotransfer, a sustained secretion of gLuc was achieved for at least 5 months. All tested proteins were dosed in the vitreous without any detectable levels into the serum. No major side effects resulted from this smooth muscle transfection technique.

Conclusions:: Electrotransfer of plasmids into the ciliary muscle is an easy, reproducible and simple technique to achieve a sustained secretion of any therapeutic proteins into the ocular media. Applications to neurotrophic proteins or anti-angiogenic approaches are currently evaluated.

Keywords: gene transfer/gene therapy • ciliary muscle • vitreous 

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.