title: Human endothelial nitric oxide synthase gene transfer inhibits vascular smooth muscle cell proliferation and neointima formation after balloon injury in rats.
abstract: 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.
  
  
  RESULTS
  After balloon injury, rat carotid arteries were transduced with 3x10(10) 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 production and 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.
authors.affiliation: Cardiac Unit, University Hospital Gasthuisberg, University of Leuven, Belgium. stefan.janssens@med.kuleuven.ac.be
authors.forename: S
authors.initials: S
authors.lastname: Janssens
authors.forename: D
authors.initials: D
authors.lastname: Flaherty
authors.forename: Z
authors.initials: Z
authors.lastname: Nong
authors.forename: O
authors.initials: O
authors.lastname: Varenne
authors.forename: N
authors.initials: N
authors.lastname: van Pelt
authors.forename: C
authors.initials: C
authors.lastname: Haustermans
authors.forename: P
authors.initials: P
authors.lastname: Zoldhelyi
authors.forename: R
authors.initials: R
authors.lastname: Gerard
authors.forename: D
authors.initials: D
authors.lastname: Collen
mesh_terms: D000256:Adenoviridae / Q000235:genetics; D000800:Angioplasty, Balloon / Q000009:adverse effects*; D000818:Animals; D001011:Aorta / Q000293:injuries / Q000473:pathology; D002455:Cell Division / Q000502:physiology; D004730:Endothelium, Vascular / Q000201:enzymology*; D018014:Gene Transfer Techniques; D006801:Humans; D009131:Muscle, Smooth, Vascular / Q000166:cytology*; D019001:Nitric Oxide Synthase / Q000235:genetics*; D052250:Nitric Oxide Synthase Type III; D051381:Rats; D017539:Tunica Intima / Q000293:injuries* / Q000473:pathology
chemical_list: C496314:NOS3 protein, human; D019001:Nitric Oxide Synthase; D052250:Nitric Oxide Synthase Type III; C496316:Nos3 protein, rat