Polyelectrolyte nanoparticles mediate vascular gene delivery.
OBJECTIVE
The purpose is to develop a non-viral gene delivery system that meets the requirements of colloidal stability of DNA complexes expressed in terms of no particle aggregation under physiologic conditions. The system should be used to transfect cardiovascular tissues.


METHODS
We used a strategy based on the formation of polyelectrolyte nanoparticles by deposition of alternatively charged polyelectrolytes onto a DNA core. Polyelectrolytes were transfer RNA as well as the synthetic polyanion, polyvinyl sulfate (PVS), and the polycation polyethylenimine (PEI). The PEI/DNA complex formed the DNA core.


RESULTS
We observed that the DNA is condensed by polycations and further packaged by association with a polyanion. These nanoparticles exhibited negative surface charge and low aggregation tendency. In vivo rat carotid artery experiments revealed high transfection efficiency, not only with the reporter gene but also with the gene encoding human urokinase plasminogen activator (Hu-uPA). Hu-uPA is one of the proteins involved in the recovery of the blood vessels after balloon catheter injury and therefore clinically relevant.


CONCLUSIONS
A strategy for in vivo gene transfer is proposed that uses the incorporation of polyanions as RNA or PVS into PEI/DNA complexes in order to overcome colloidal instability and to generate a negative surface charge. The particles proved to be transfectionally active in vascular gene transfer.
Zaitsev
Sergey
S
Franz Volhard Clinic, HELIOS Klinikum-Berlin at the Max Delbrück Center for Molecular Medicine, D-13125 Berlin-Buch, Germany.
Cartier
Régis
R
Vyborov
Oleg
O
Sukhorukov
Gleb
G
Paulke
Bernd-Reiner
BR
Haberland
Annekathrin
A
Parfyonova
Yelena
Y
Tkachuk
Vsevolod
V
Böttger
Michael
M
D000818:Animals; D020212:Carotid Artery Injuries / Q000378:metabolism / Q000628:therapy; D004573:Electrolytes / Q000737:chemistry*; D015870:Gene Expression; D015316:Genetic Therapy; D008297:Male; D049329:Nanostructures / Q000737:chemistry*; D010957:Plasmids / Q000235:genetics; D011094:Polyethyleneimine; D051381:Rats; D011921:Rats, Inbred WKY; D014162:Transfection / Q000379:methods*; D017539:Tunica Intima / Q000378:metabolism; D014568:Urokinase-Type Plasminogen Activator / Q000096:biosynthesis / Q000235:genetics; D014945:Wound Healing; D001616:beta-Galactosidase / Q000096:biosynthesis / Q000235:genetics
D004573:Electrolytes; D011094:Polyethyleneimine; D001616:beta-Galactosidase; D014568:Urokinase-Type Plasminogen Activator