Induction of Sustained Hypercholesterolemia by Single Adeno-Associated Virus–Mediated Gene Transfer of Mutant hPCSK9. Issue 1 (January 2015)

• Record Type: Journal Article
• Authors: Roche-Molina, Marta; Sanz-Rosa, David; Cruz, Francisco M.; García-Prieto, Jaime; López, Sergio; Abia, Rocío; Muriana, Francisco J.G.; Fuster, Valentín; Ibáñez, Borja; Bernal, Juan A.
• Is Part Of: Arteriosclerosis, thrombosis, and vascular biology. Volume 35:Issue 1(2015)
• Publication Date: 2015-01
• Subjects: atherosclerosis -- hypercholesterolemia -- PCSK9; Arteriosclerosis -- Periodicals; Thrombosis -- Periodicals; Blood-vessels -- Pathophysiology -- Periodicals; Electronic journals; 616.13
• DOI: 10.1161/ATVBAHA.114.303617
• Languages: English
• ISSNs: 1079-5642

Abstract

Abstract : Objectives—: Patients with mutations in the proprotein convertase subtilisin/kexin type 9 ( PCSK9 ) gene have hypercholesterolemia and are at high risk of adverse cardiovascular events. We aimed to stably express the pathological human D374Y gain-of-function mutant form of PCSK9 ( PCSK9 DY ) in adult wild-type mice to generate a hyperlipidemic and proatherogenic animal model, achieved with a single systemic injection with adeno-associated virus (AAV). Approach and Results—: We constructed an AAV-based vector to support targeted transfer of the PCSK9 DY gene to liver. After injection with 3.5×10 10 viral particles, mice in the C57BL/6J, 129/SvPasCrlf, or FVB/NCrl backgrounds developed long-term hyperlipidemia with a strong increase in serum low-density lipoprotein. Macroscopic and histological analysis showed atherosclerotic lesions in the aortas of AAV- PCSK9 DY mice fed a high-fat-diet. Advanced lesions in these high-fat-diet–fed mice also showed evidence of macrophage infiltration and fibrous cap formation. Hepatic AAV- PCSK9 DY infection did not result in liver damage or signs of immunologic response. We further tested the use of AAV- PCSK9 DY to study potential genetic interaction with the ApoE gene. Histological analysis of ApoE −/− AAV- PCSK9 DY mice showed a synergistic response to ApoE deficiency, with aortic lesions twice as extensive in ApoE −/− AAV- PCSK9 DY -transexpressing mice as in ApoE −/− AAV- Luc controls without altering serum cholesterol levels. Conclusions—: Single intravenous AAV- PCSK9 DY injection is a fast, easy, and cost-effective approach, resulting in rapid and long-term sustained hyperlipidemia and atherosclerosis. We demonstrate as a proof of concept the synergy between PCSK9 DY gain-of-function and ApoE deficiency. This methodology could allow testing of the genetic interaction of several mutations without the need for complex and time-consuming backcrosses. Abstract : Supplemental Digital Content is available in the text.