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Local lentiviral short hairpin RNA silencing of CCR2 inhibits vein graft thickening in hypercholesterolemic apolipoprotein E3-Leiden mice

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Author: Eefting, D. · Bot, I. · Vries, M.R. de · Schepers, A. · Bockel, J.H. van · Berkel, T.J.C. van · Biessen, E.A.L. · Quax, P.H.A.
Type:article
Date:2009
Institution: TNO Kwaliteit van Leven
Source:Journal of Vascular Surgery, 1, 50, 152-160
Identifier: 241609
doi: doi:10.1016/j.jvs.2009.03.027
Keywords: Biology · Biomedical Research · apolipoprotein E3 · chemokine receptor CCR2 · lentiviral short hairpin RNA · lentivirus vector · messenger RNA · monocyte chemotactic protein 1 · unclassified drug · animal cell · animal model · article · atherosclerosis · carotid artery · cell migration · cell proliferation · clinical feature · controlled study · disease course · drug delivery system · drug efficacy · drug targeting · gene silencing · gene therapy · hypercholesterolemia · in vitro study · in vivo study · infection · Lentivirinae · male · morphometrics · mouse · nonhuman · priority journal · protein expression · receptor upregulation · smooth muscle fiber · treatment outcome · vein graft · vein graft disease · vein surgery · viral gene delivery system · Animals · Apolipoprotein E3 · Atherosclerosis · Chemokine CCL2 · Disease Models, Animal · Graft Occlusion, Vascular · Hypercholesterolemia · Lentivirus · Male · Mice · Receptors, CCR2 · RNA Interference · Veins

Abstract

Objective: Inflammatory responses to vascular injury are key events in vein graft disease and accelerated atherosclerosis, which may result in bypass failure. The monocyte chemoattractant protein-1 (MCP-1)/CC-chemokine receptor (CCR)-2 pathway is hypothesized to play a central role. A murine model for vein graft disease was used to study the effect of local application of lentiviral short hairpin RNA (shRNA) targeted against CCR2. Methods: A venous interposition was placed into the carotid artery of hypercholesterolemic apolipoprotein E3-Leiden (APOE*3-Leiden) mice to induce vein graft thickening with features of accelerated atherosclerosis. To demonstrate the efficacy of the lentiviral shRNA targeting murine CCR2 (shCCR2) in blocking vein graft disease in vivo, lentiviral shCCR2 or a control lentivirus was used to infect the vein graft locally (n = 8). Results: Vascular CCR2 and MCP-1 messenger RNA expression levels were significantly upregulated during lesion progression in the vein graft. Infection of smooth muscle cells (SMCs) with a lentiviral shRNA targeting shCCR2 completely abolished MCP-1-induced SMC migration and inhibited SMC proliferation in vitro (n = 3 per group). Morphometric analysis of sections of grafts showed a significant 38% reduction in vein graft thickening in the shCCR2-treated mice 4 weeks after surgery (control, 0.42 ± 0.05 mm2; shCCR2, 0.26 ± 0.03 mm2; P = .007). Conclusion: Vascular CCR2 contributes to vein graft disease, and local application of shRNA against CCR2 to the vessel wall prevents vein graft thickening in hypercholesterolemic mice, suggesting that local overexpressing of shRNA using organ-targeted lentiviral gene delivery may be a promising therapeutic tool to improve vein graft disease in bypassed patients. © 2009 Society for Vascular Surgery.