Efficient transduction of vascular endothelial cells with recombinant adeno-associated virus serotype 1 and 5 vectors.
Recombinant adeno-associated virus (rAAV) has become an attractive tool for gene therapy because of its ability to transduce both dividing and nondividing cells, elicit a limited immune response, and the capacity for imparting long-term transgene expression. Previous studies have utilized rAAV serotype 2 predominantly and found that transduction of vascular cells is relatively inefficient. The purpose of the present study was to evaluate the transduction efficiency of rAAV serotypes 1 through 5 in human and rat aortic endothelial cells (HAEC and RAEC). rAAV vectors with AAV2 inverted terminal repeats containing the human alpha1-antitrypsin (hAAT) gene were transcapsidated using helper plasmids to provide viral capsids for the AAV1 through 5 serotypes. True type rAAV2 and 5 vectors encoding beta-galactosidase or green fluorescence protein were also studied. Infection with rAAV1 resulted in the most efficient transduction in both HAEC and RAEC compared to other serotypes (p < 0.001) at 7 days posttransduction. Interestingly, expression was increased in cells transduced with rAAV5 to levels surpassing rAAV1 by day 14 and 21. Transduction with rAAV1 was completely inhibited by removal of sialic acid with sialidase, while heparin had no effect. These studies are the first demonstration that sialic acid residues are required for rAAV1 transduction in endothelial cells. Transduction of rat aortic segments ex vivo and in vivo demonstrated significant transgene expression in endothelial and smooth muscle cells with rAAV1 and 5 serotype vectors, in comparison to rAAV2. These results suggest the unique potential of rAAV1 and rAAV5-based vectors for vascular-targeted gene-based therapeutic strategies.
Chen
Sifeng
S
Department of Medicine, University of Alabama at Birmingham, Birmingham, AL 35294, USA.
Kapturczak
Matthias
M
Loiler
Scott A
SA
Zolotukhin
Sergei
S
Glushakova
Olena Y
OY
Madsen
Kirsten M
KM
Samulski
Richard J
RJ
Hauswirth
William W
WW
Campbell-Thompson
Martha
M
Berns
Kenneth I
KI
Flotte
Terence R
TR
Atkinson
Mark A
MA
Tisher
C Craig
CC
Agarwal
Anupam
A
D000818:Animals; D001011:Aorta / Q000378:metabolism; D002213:Capsid / Q000378:metabolism; D002478:Cells, Cultured; D004274:DNA, Recombinant / Q000235:genetics*; D000229:Dependovirus / Q000235:genetics*; D004730:Endothelium, Vascular / Q000737:chemistry / Q000378:metabolism* / Q000821:virology; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006493:Heparin / Q000378:metabolism; D006801:Humans; D008297:Male; D032389:Myocytes, Smooth Muscle / Q000737:chemistry / Q000378:metabolism* / Q000821:virology; D019158:N-Acetylneuraminic Acid / Q000378:metabolism; D009439:Neuraminidase / Q000494:pharmacology; D051381:Rats; D011917:Rats, Inbred Lew; D014161:Transduction, Genetic; D000515:alpha 1-Antitrypsin / Q000235:genetics; D001616:beta-Galactosidase / Q000378:metabolism
D004274:DNA, Recombinant; D000515:alpha 1-Antitrypsin; D049452:Green Fluorescent Proteins; D006493:Heparin; D009439:Neuraminidase; D001616:beta-Galactosidase; D019158:N-Acetylneuraminic Acid