Accelerated atherosclerosis development in C57Bl6 mice by overexpressing AAV-mediated PCSK9 and partial carotid ligation

• Type of Material: Article
• Authors: Sandeep Kumar, Emory University; Dong Won Kang, Georgia Institute of Technology; Amir Rezvan, Emory University; Hanjoong Jo, Emory University
• Language: English
• Date: 2017-08-01
• Publisher: Nature Publishing Group: Open Access Hybrid Model Option B
• Publication Version: Author Accepted Manuscript (After Peer Review)
• Copyright Statement: © 2017 USCAP, Inc All rights reserved.
• Final Published Version (URL): http://dx.doi.org/10.1038/labinvest.2017.47
• Title of Journal or Parent Work: Laboratory Investigation
• ISSN: 0023-6837
• Volume: 97
• Issue: 8
• Start Page: 935
• End Page: 945
• Grant/Funding Information: This work was supported by funding from National Institutes of Health Grants HL119798, HL113451, HL095070, and HL124879 to HJ. HJ is John and Jan Portman Professor.; This project was supported in part by the Viral Vector Core of the Emory Neuroscience NINDS Core Facilities Grant, P30NS055077.
• Keywords: Research & Experimental Medicine; GENE-EXPRESSION; E-NULL MICE; MOUSE MODELS; FAMILIAL HYPERCHOLESTEROLEMIA; Pathology; APOLIPOPROTEIN-E; DISTURBED FLOW; RECEPTOR KNOCKOUT MICE; APOE(-/-) MICE; KEY ROLE; Medicine, Research & Experimental; ENDOTHELIAL DYSFUNCTION; Life Sciences & Biomedicine; Science & Technology
• Research Categories: Engineering, Biomedical

Abstract

Studying the role of a particular gene in atherosclerosis typically requires a time-consuming and often difficult process of generating double knockouts or transgenics on ApoE -/- or LDL receptor (LDLR) -/- background. Recently, it was reported that adeno-associated-virus-8 (AAV8)-mediated overexpression of PCSK9 (AAV8-PCSK9) rapidly induced hyperlipidemia. However, using this method in C57BL6 wild-type (C57) mice, it took ∼3 months to develop atherosclerosis. Our partial carotid ligation model is used to rapidly develop atherosclerosis by inducing disturbed flow in the left common carotid artery within 2 weeks in ApoE -/- or LDLR -/- mice. Here, we combined these two approaches to develop an accelerated model of atherosclerosis in C57 mice. C57 mice were injected with AAV9-PCSK9 or AAV9-luciferase (control) and high-fat diet was initiated. A week later, partial ligation was performed. Compared to the control , AAV-PCSK9 led to elevated serum PCSK9, hypercholesterolemia, and rapid atherosclerosis development within 3 weeks as determined by gross plaque imaging, and staining with Oil-Red-O, Movat's pentachrome, and CD45 antibody. These plaque lesions were comparable to the atherosclerotic lesions that have been previously observed in ApoE -/- or LDLR -/- mice that were subjected to partial carotid ligation and high-fat diet. Next, we tested whether our method can be utilized to rapidly determine the role of a particular gene in atherosclerosis. Using eNOS -/- and NOX1 â y mice on C57 background, we found that the eNOS -/- mice developed more advanced lesions, while the NOX1 â y mice developed less atherosclerotic lesions as compared to the C57 controls. These results are consistent with the previous findings using double knockouts (eNOS -/- -ApoE -/- and NOX1 â y -ApoE -/- ). AAV9-PCSK9 injection followed by partial carotid ligation is an effective and time-saving approach to rapidly induce atherosclerosis. This accelerated model is well-suited to quickly determine the role of gene(s) interest without generating double or triple knockouts.