Bjørklund, Martin Mæng2; Hollensen, Anne Kruse3; Hagensen, Mette Kallestrup4; Dagnæs-Hansen, Frederik4; Christoffersen, Christina5; Mikkelsen, Jacob Giehm4; Bentzon, Jacob Fog4
1 Section of Heart and Circulatory Research, Department of Biomedical Sciences, Faculty of Health and Medical Sciences, Københavns Universitet2 Hjertemedicinsk Afdeling B, SKS3 Aarhus Universitet, Institut for Biomedicin4 From the Departments of Clinical Medicine (M.M.B., M.K.H., J.F.B.) and Biomedicine (A.K.H., F.D.-H., J.G.M.), Aarhus University, Aarhus, Denmark; Department of Cardiology, Aarhus University Hospital, Aarhus, Denmark (M.M.B., M.K.H., J.F.B.); and Department of Clinical Biochemistry, Rigshospitalet, Copenhagen, Denmark (C.C.).5 Section of Heart and Circulatory Research, Department of Biomedical Sciences, Faculty of Health and Medical Sciences, Københavns Universitet
RATIONALE: Atherosclerosis can be achieved in animals by germline genetic engineering, leading to hypercholesterolemia, but such models are constrained to few species and strains, and they are difficult to combine with other powerful techniques involving genetic manipulation or variation. OBJECTIVE: To develop a method for induction of atherosclerosis without germline genetic engineering. METHODS AND RESULTS: Recombinant adeno-associated viral vectors were engineered to encode gain-of-function proprotein convertase subtilisin/kexin type 9 mutants, and mice were given a single intravenous vector injection followed by high-fat diet feeding. Plasma proprotein convertase subtilisin/kexin type 9 and total cholesterol increased rapidly and were maintained at high levels, and after 12 weeks, mice had atherosclerotic lesions in the aorta. Histology of the aortic root showed progression of lesions to the fibroatheromatous stage. To demonstrate the applicability of this method for rapid analysis of the atherosclerosis susceptibility of a mouse strain and for providing temporal control over disease induction, we demonstrated the accelerated atherosclerosis of mature diabetic Akita mice. Furthermore, the versatility of this approach for creating atherosclerosis models also in nonmurine species was demonstrated by inducing hypercholesterolemia and early atherosclerosis in Golden Syrian hamsters. CONCLUSIONS: Single injections of proprotein convertase subtilisin/kexin type 9-encoding recombinant adeno-associated viral vectors are a rapid and versatile method to induce atherosclerosis in animals. This method should prove useful for experiments that are high-throughput or involve genetic techniques, strains, or species that do not combine well with current genetically engineered models.
Circulation Research, 2014, Vol 114, Issue 11, p. 1684-9