A novel formulation based on 2,3-di(tetradecyloxy)propan-1-amine cationic lipid combined with polysorbate 80 for efficient gene delivery to the retina.
OBJECTIVE
The aim of the present study was to evaluate the potential application of a novel formulation based on a synthesized cationic lipid 2,3-di(tetradecyloxy)propan-1-amine, combined with polysorbate 80 to deliver the pCMS-EGFP plasmid into the rat retina.


METHODS
We elaborated lipoplexes by mixing the formulation containing the cationic lipid and the polysorbate 80 with the plasmid at different cationic lipid/DNA ratios (w/w). Resulted lipoplexes were characterized in terms of size, charge, and capacity to condense, protect and release the DNA. In vitro transfection studies were performed in HEK-293 and ARPE-19 cells. Formulations were also tested in vivo by monitoring the expression of the EGFP after intravitreal and subretinal injections in rat eyes.


RESULTS
At 2/1 cationic lipid/DNA mass ratio, the resulted lipoplexes had 200 nm of hydrodynamic diameter; were positive charged, spherical, protected DNA against enzymatic digestion and transfected efficiently HEK-293 and ARPE-19 cultured cells exhibiting lower cytotoxicity than LipofectamineTM 2000. Subretinal administrations transfected mainly photoreceptors and retinal pigment epithelial cells; whereas intravitreal injections produced a more uniform distribution of transfection through the inner part of the retina.


CONCLUSIONS
These results hold great expectations for other gene delivery formulations based on this cationic lipid for retinal gene therapy purposes.
University of the Basque Country, Vitoria, Spain.
Gustavo Puras
GP
Ochoa
Jon Zárate
JZ
Sesma
Mireia Agirre
MA
Díez
Ariadna
A
Díaz-Tahoces
Marcelino
M
Avilés-Trigeros
Santiago
S
Grijalvo
Ramón
R
Eritja
Eduardo
E
Fernández
Jose Luis
JL
Pedraz
D000818:Animals; D002412:Cations / Q000737:chemistry; D002460:Cell Line; D004247:DNA / Q000008:administration & dosage* / Q000235:genetics; D004075:Diglycerides / Q000737:chemistry*; D057809:HEK293 Cells; D006801:Humans; D008297:Male; D011136:Polysorbates / Q000737:chemistry*; D051381:Rats; D017207:Rats, Sprague-Dawley; D012160:Retina / Q000378:metabolism*; D014162:Transfection / Q000379:methods*
D002412:Cations; D004075:Diglycerides; D011136:Polysorbates; C057475:1,2-ditetradecylglycerol; D004247:DNA