Por favor, use este identificador para citar o enlazar este Item:http://hdl.handle.net/20.500.12105/17709
Título
Induction of sustained hypercholesterolemia by single adeno-associated virus-mediated gene transfer of mutant hPCSK9.
Autor(es)
Roche-Molina, Marta CNIC | Sanz-Rosa, David CNIC | Cruz, Francisco M | García-Prieto, Jaime | López, Sergio | Abia, Rocío | Muriana, Francisco J G | Fuster, Valentín | Ibáñez, Borja CNIC | Bernal, Juan A
Fecha de publicación
2015-01
Cita
Arterioscler Thromb Vasc Biol. 2015 Jan;35(1):50-9.
Idioma
Inglés
Tipo de documento
journal article
Resumen
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.
MESH
Gene Transfer Techniques | Genetic Vectors | Mutation | Animals | Aorta | Aortic Diseases | Apolipoproteins E | Atherosclerosis | Biomarkers | Cholesterol | Dependovirus | Diet, High-Fat | Disease Models, Animal | Humans | Hypercholesterolemia | Male | Mice, 129 Strain | Mice, Inbred C57BL | Mice, Knockout | Plaque, Atherosclerotic | Proprotein Convertase 9 | Proprotein Convertases | Serine Endopeptidases | Time Factors
Versión en línea
DOI
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- Induction of sustained hyperch ...
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