Perivascular gene transfer of dominant-negative N19RhoA attenuates neointimal formation via inhibition of TGF-beta1-Smad2 signaling in rats after carotid artery balloon injury.
Phenotypic differentiation of adventitial fibroblasts to myofibroblasts is an essential feature of vascular remodeling. Here, we carried out perivascular gene transfer of dominant-negative N19RhoA to investigate whether antagonism of RhoA signaling attenuates neointimal formation following rat carotid artery balloon injury and alters TGF-beta1-Smad2-induced differentiation of adventitial fibroblasts to myofibroblasts. Perivascular delivery of an adenovirus coexpressing dominant-negative N19RhoA and humanized Renilla green fluorescent protein (hrGFP) (Ad-N19RhoA-hrGFP), as demonstrated by hrGFP staining, suppressed neointimal formation at 7 and 14days post-injury. Ad-N19RhoA-hrGFP administration inhibited neointimal alpha-smooth muscle-actin and Calponin expression, as markers of myofibroblast differentiation and perivascular collagen deposition, at 14days after balloon injury. Ad-N19RhoA-hrGFP administration also inhibited adventitial Smad2 phosphorylation, but did not alter local TGF-beta1 and total-Smad2 expression after injury. Our results provide evidence that perivascular gene transfer of dominant-negative N19RhoA blocks TGF-beta1-Smad2-induced differentiation of adventitial fibroblasts to myofibroblasts, which contributes to intimal hyperplasia after balloon injury.
Chen
WenDong
W
Shanghai Key Laboratory of Vascular Biology at Ruijin Hospital and Shanghai Institute of Hypertension, Shanghai Jiaotong University School of Medicine, Shanghai 200025, China.
Chu
YuFeng
Y
Zhu
DingLiang
D
Yan
Chen
C
Liu
JianJun
J
Ji
KaiDa
K
Gao
PingJin
P
D000199:Actins / Q000096:biosynthesis; D000256:Adenoviridae; D000800:Angioplasty, Balloon / Q000009:adverse effects; D000818:Animals; D002135:Calcium-Binding Proteins / Q000096:biosynthesis; D002339:Carotid Arteries / Q000601:surgery; D020212:Carotid Artery Injuries / Q000473:pathology*; D002454:Cell Differentiation; D005347:Fibroblasts / Q000473:pathology*; D018014:Gene Transfer Techniques; D049452:Green Fluorescent Proteins / Q000235:genetics; D006965:Hyperplasia / Q000473:pathology; D008297:Male; D008840:Microfilament Proteins / Q000096:biosynthesis; D051381:Rats; D017207:Rats, Sprague-Dawley; D051899:Smad2 Protein / Q000037:antagonists & inhibitors*; D053773:Transforming Growth Factor beta1 / Q000037:antagonists & inhibitors*; D020742:rhoA GTP-Binding Protein / Q000235:genetics*; D000096985:Calponins
D000199:Actins; D002135:Calcium-Binding Proteins; D049452:Green Fluorescent Proteins; D008840:Microfilament Proteins; D051899:Smad2 Protein; D053773:Transforming Growth Factor beta1; D020742:rhoA GTP-Binding Protein; D000096985:Calponins; C498862:Acta2 protein, rat