The ciliary smooth muscle electrotransfer: basic principles and potential for sustained intraocular production of therapeutic proteins.

Details

Serval ID
serval:BIB_CD27BD4D6BC6
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
The ciliary smooth muscle electrotransfer: basic principles and potential for sustained intraocular production of therapeutic proteins.
Journal
Journal of Gene Medicine
Author(s)
Touchard E., Kowalczuk L., Bloquel C., Naud M.C., Bigey P., Behar-Cohen F.
ISSN
1521-2254 (Electronic)
ISSN-L
1099-498X
Publication state
Published
Issued date
2010
Peer-reviewed
Oui
Volume
12
Number
11
Pages
904-919
Language
english
Notes
Publication types: Comparative Study ; Journal Article ; Research Support, Non-U.S. Gov'tPublication Status: ppublish
Abstract
BACKGROUND: We have developed a nonviral gene therapy method based on the electrotransfer of plasmid in the ciliary muscle. These easily accessible smooth muscle cells could be turned into a biofactory for any therapeutic proteins to be secreted in a sustained manner in the ocular media.
METHODS: Electrical conditions, design of electrodes, plasmid formulation, method and number of injections were optimized in vivo in the rat by localizing β-galactosidase expression and quantifying reporter (luciferase) and therapeutic (anti-tumor necrosis factor) proteins secretion in the ocular media. Anatomical measurements were performed via human magnetic resonance imaging to design a human eye-sized prototype that was tested in the rabbit.
RESULTS: In the rat, transscleral injection of 30 µg of plasmid diluted in half saline (77 mM NaCl) followed by application of eight square-wave electrical pulses (15 V, 10 ms, 5.3 Hz) using two platinum/iridium electrodes, an internal wire and an external sheet, delivered plasmid efficiently to the ciliary muscle fibers. Gene transfer resulted in a long-lasting (at least 5 months) and plasmid dose-/injection number- dependent secretion of different molecular weight proteins mainly in the vitreous, without any systemic exposure. Because ciliary muscle anatomical measurements remained constant among ages in adult humans, an integrated device comprising needle-electrodes was designed and manufactured. Its usefulness was validated in the rabbit.
CONCLUSIONS: Plasmid electrotransfer to the ciliary muscle with a suitable medical device represents a promising local and sustained protein delivery system for treating posterior segment diseases, avoiding repeated intraocular injections.
Keywords
Adult, Aged, Aged, 80 and over, Animals, Aqueous Humor/metabolism, Child, Child, Preschool, Ciliary Body/metabolism, Electroporation/methods, Eye/anatomy & histology, Eye/metabolism, Female, Gene Transfer Techniques, Genetic Therapy, Humans, Luciferases/genetics, Luciferases/metabolism, Male, Middle Aged, Models, Animal, Muscle, Smooth/metabolism, Plasmids, Rabbits, Rats, Rats, Inbred Lew, beta-Galactosidase/genetics, beta-Galactosidase/metabolism
Pubmed
Web of science
Create date
23/08/2013 7:16
Last modification date
20/08/2019 15:47
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