From the Cardiac Unit (S.J., N. v P.) and the Radiotherapy Department
(C.H.), University Hospital Gasthuisberg, University of Leuven, and the Center
for Transgene Technology and Gene Therapy, Flanders Interuniversity Institute
for Biotechnology (D.F., Z.N., O.V., P.Z., R.G., D.C.), Leuven, Belgium.
Correspondence to Stefan Janssens, MD, PhD, Cardiac Unit and Center for Transgene Technology and Gene Therapy, KU-Leuven, Campus Gasthuisberg, Herestraat 49, B-3000 Leuven, Belgium. E-mail stefan.janssens{at}med.kuleuven.ac.be
Background—Loss of
endothelial NO production after
arterial injury may contribute to restenosis,
characterized by neointima formation and elastic recoil.
Adenovirus-mediated transfer of the gene encoding NO synthase (NOS) in
balloon-injured arteries may restore NO production and inhibit
neointima formation.
Methods and Results—After balloon injury, rat carotid
arteries were transduced with 3x1010 pfu/mL recombinant
adenovirus carrying the human endothelial constitutive
NOS cDNA (AdCMVceNOS, n=8) or no cDNA (AdRR5, n=8). ceNOS expression
was confirmed by immunoblot analysis of vascular
extracts and was localized by immunostaining in 30% of
medial smooth muscle cells (SMCs) and in the adventitia of
AdCMVceNOS-transduced arteries. Vascular cGMP levels were reduced from
3.9 pmol/g wet wt in uninjured arteries to 0.7 pmol cGMP/g after AdRR5
but were restored after ceNOS gene transfer (3.8 pmol cGMP/g wet wt,
P<.05 versus AdRR5). Intima-to-media ratio 2 weeks
after injury was significantly reduced (0.19±0.02 in
AdCMVceNOS-infected versus 0.69±0.07 in AdRR5-infected arteries,
P<.05). In vitro, BrdU incorporation of
AdCMVceNOS-infected SMCs was reduced by 28% compared with
AdRR5-infected SMCs. Transduced cells from injured carotid arteries
subjected to FACS sorting showed a significantly lower BrdU labeling
index in ceNOS-infected rats (29±6% versus 43±5% and 45±4% in
control, injured, and AdRR5-infected rats, respectively,
P<.05).
Conclusions—AdCMVceNOS gene transfer to balloon-injured rat
carotid arteries restores vascular NO productionand reduces
neointima formation, at least in part because of an
antiproliferative effect on medial SMCs. Adenovirus-mediated
ceNOS gene transfer might reduce arterial
restenosis after balloon angioplasty.
© 1998 American Heart Association, Inc.
Basic Science Reports
Human Endothelial Nitric Oxide Synthase Gene Transfer Inhibits Vascular Smooth Muscle Cell Proliferation and Neointima Formation After Balloon Injury in Rats
Key Words: nitric oxide • muscle, smooth • stenosis • genes
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