In vivo gene transfer into rat arterial walls with novel adeno-associated virus vectors.
OBJECTIVE
We studied the ability of recombinant adeno-associated virus (rAAV) vectors to achieve gene transfer in vivo to intact rat carotid arteries.


METHODS
Isolated segments of uninjured rat carotid arteries were incubated with (1) rAAV vectors that expressed a beta-galactosidase gene, (2) a related vector with no promoter, or (3) a normal saline solution. Gene transfer was evaluated with in situ polymerase chain reaction (PCR). Transgene expression was assessed at intervals that ranged from 24 hours to 2 months by measurement of beta-galactosidase activity and protein mass in tissue extracts with fluorometric and enzyme-linked immunosorbent assays, respectively. Dose dependence of expression was determined for virus concentrations that ranged from 5 x 10(4) to 5 x 10(5) infectious units (iu)/ml.


RESULTS
Light microscopic analysis of in situ PCR-stained histologic sections of transduced vessel walls showed approximately 90% of intimal and medial cell nuclei contained the beta-galactosidase gene, compared with none in control arteries. In vivo beta-galactosidase expression was (1) highest 24 hours after gene transfer, (2) elevated for 1 month, and (3) dose responsive.


CONCLUSIONS
rAAV vectors can mediate focal gene transfer into the intact rat carotid artery with detectable levels of transgene expression for 1 month and are potentially useful agents for in vivo gene transfer into intact arteries.
Arnold
T E
TE
Department of Surgery, Health Sciences Center, State University of New York at Stony Brook 11794-8191, USA.
Gnatenko
D
D
Bahou
W F
WF
D000818:Animals; D002339:Carotid Arteries / Q000201:enzymology*; D000229:Dependovirus; D004797:Enzyme-Linked Immunosorbent Assay; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D008297:Male; D016133:Polymerase Chain Reaction; D051381:Rats; D017207:Rats, Sprague-Dawley; D011994:Recombinant Proteins; D019076:Transgenes; D001616:beta-Galactosidase / Q000235:genetics*
D011994:Recombinant Proteins; D001616:beta-Galactosidase