Overexpression of CD39/nucleoside triphosphate diphosphohydrolase‐1 decreases smooth muscle cell proliferation and prevents neointima formation after angioplasty

Abstract

Summary. Background: Growing evidence implicates the involvement of extracellular nucleotides in the regulation of platelet, leukocyte, endothelial cell (EC) and vascular smooth muscle cell (VSMC) phenotype and function. Within the quiescent vasculature, extracellular nucleotides are rapidly hydrolyzed by CD39, the dominant endothelial nucleoside triphosphate diphosphohydrolase (NTPDase-1). However, vascular CD39/NTPDase-1 activity is lost in EC activated by oxidative stress or proinflammatory mediators, and upon denudation of the endothelium following balloon injury. The consequent increase in extracellular nucleotide concentrations triggers signaling events leading to prothrombotic responses and increased VSMC proliferation. Objectives: To investigate the effect of overexpressed CD39/NTPDase-1 in injured aorta. Methods: Using adenoviral-mediated gene transfer we expressed CD39/NTPDase-1 in mechanically denudated rat aortas. We measured intima formation by morphometry and VSMC proliferation by the [ 3 H]-thymidine incorporation assay. Results: Targeted expression of CD39 in injured vessels increased NTPDase activity (from 2.91 ± 0.31 to 22.07 ± 6.7 nmols Pi mg )1 protein, 4 days after exposure to the adenovirus) and prevented the formation of neointima. The thickness of the intimal layer in injured aortas exposed to Ad-CD39 was 26.2 ± 3.9 lm vs. 51.8 ± 6.1 lm and 64.4 ± 22.2 lm (P < 0.001) in vessels treated with Ad-b-gal and saline, respectively. Moreover, targeted expression of CD39/NTPDase-1 caused a 70% (P < 0.01) decrease in proliferation of VSMC isolated from transduced rat aortas as compared with VSMC derived from control vessels. Conclusions: The presented data suggest that increasing CD39/ NTPDase-1 activity in VSMC could represent a novel therapeutic approach for the prevention of stenosis associated with angioplasty and other occlusive vascular diseases.

Vascular smooth muscle cell expression of ectonucleotidase CD39 (ENTPD1) is required for neointimal formation in mice

“…Furukoji et al, have shown that expression of placental derived CD39 in rat carotid arteries suppresses thrombus formation and also suggest that subsequent neointimal formation may be limited by this manipulation [30]. Our own studies with rat aortic intimal injury suggest that human vascular CD39 up regulation with adenoviral vectors has comparable protective effects [31].…”

“…These somewhat puzzling effects may be explained by the desensitization of select P2Y receptors that in the absence of NTPDase activity has the same effect on receptor-signaling events, as would excessive scavenging of nucleotide, when CD39 is upregulated [30,31]. Hence, both knock-out mice null for Cd39 and transgenic mice over expressing CD39 have prolonged bleeding times secondary to platelet dysfunction, predominantly at the P2Y1 receptor level albeit at different pericellular nucleotide levels [21,32].…”

Suppression of endothelial CD39/ENTPD1 is associated with pulmonary vascular remodeling in pulmonary arterial hypertension

“…This suggests that sustained CD39 expression could be protective against vascular remodeling in PAH. This is supported by a study on angioplasty-induced vascular injury, where overexpression of CD39 reduced the development of neointima (31). However, CD39 enzymatic activity can also be regulated at posttranslational level via formation of multimers, which further increases the CD39 activity, or oxidative inactivation (43,46).…”

Purinergic Signaling and Blood Vessels in Health and Disease

“…Overexpression of E-NTPDase 1/CD39 decreases vascular smooth muscle cell proliferation and prevents neointima formation after angioplasty, consistent with an involvement of nucleotides in these processes (Koziak et al, 2008). E-NTPDase 1/CD39 is expressed on mouse carotid artery smooth muscle cells and is required for neointimal formation (Behdad et al, 2009).…”