Overexpression of a constitutively active protein kinase G mutant reduces neointima formation and in-stent restenosis.
BACKGROUND
Neointima formation after arterial injury is associated with reduced vascular cyclic guanosine monophosphate (cGMP) and cGMP-dependent protein kinase (PKG), a major cGMP effector in vascular smooth muscle. We tested the effect of PKG overexpression on the neointimal response to vascular injury. Methods and Results- Infection of cultured rat aortic smooth muscle cells (RASMCs) with an adenoviral vector specifying a cGMP-independent, constitutively active PKG mutant (AdPKGcat) reduced serum-induced migration by 33% and increased serum-deprivation-induced apoptosis 2-fold (P<0.05 for both). Infection with wild-type PKG (AdPKG), in the absence of cGMP, did not affect migration or apoptosis. Two weeks after balloon-injured rat carotid arteries were infected with 1x 10(10) pfu AdPKGcat (n=12), AdPKG (n=8), or a control adenovirus (n=8), intima-to-media ratio was less in AdPKGcat-infected arteries than in AdPKG- or control adenovirus-infected vessels (0.26+/-0.06 versus 0.61+/-0.12 and 0.70+/-0.12, respectively, P<0.05 for both). Two weeks after intramural administration of 1.75x10(10) pfu AdPKGcat (n=8) or a control adenovirus (n=8) into porcine coronary arteries with in-stent restenosis, luminal diameter was greater in AdPKGcat-infected arteries than in control adenovirus-infected vessels (2.32+/-0.16 versus 1.81+/-0.13 mm, P=0.028), associated with reduced neointimal area (3.30+/-0.24 versus 4.15+/-0.13 mm(2), P=0.008), neointima-to-vessel area ratio (0.42+/-0.05 versus 0.58+/-0.04, P<0.05), and percent stenosis (45+/-6% versus 70+/-4%, P<0.05).


CONCLUSIONS
Expression of a constitutively active PKG reduces neointima formation after balloon injury in rats and reduces coronary in-stent restenosis in pigs. PKGcat gene transfer may be a promising strategy for vasculoproliferative disorders.
Sinnaeve
Peter
P
Center for Transgene Technology and Gene Therapy, Flanders Interuniversity Institute for Biotechnology, and the Cardiac Unit, University Hospital Gasthuisberg, University of Leuven, Belgium.
Chiche
Jean-Daniel
JD
Gillijns
Hilde
H
Van Pelt
Natascha
N
Wirthlin
Douglas
D
Van De Werf
Frans
F
Collen
Desire
D
Bloch
Kenneth D
KD
Janssens
Stefan
S
D000256:Adenoviridae / Q000235:genetics; D000800:Angioplasty, Balloon / Q000009:adverse effects; D000818:Animals; D017209:Apoptosis; D016893:Carotid Stenosis / Q000201:enzymology / Q000209:etiology / Q000473:pathology / Q000628:therapy; D002465:Cell Movement; D002478:Cells, Cultured; D023903:Coronary Restenosis / Q000209:etiology / Q000473:pathology / Q000628:therapy*; D017869:Cyclic GMP-Dependent Protein Kinases / Q000235:genetics* / Q000378:metabolism / Q000502:physiology; D004789:Enzyme Activation; D005822:Genetic Vectors; D006083:Graft Occlusion, Vascular / Q000201:enzymology / Q000209:etiology / Q000473:pathology / Q000628:therapy*; D007700:Kinetics; D008297:Male; D009131:Muscle, Smooth, Vascular / Q000166:cytology / Q000502:physiology; D051381:Rats; D017208:Rats, Wistar; D015607:Stents / Q000009:adverse effects*; D013552:Swine; D014161:Transduction, Genetic
D017869:Cyclic GMP-Dependent Protein Kinases