Adeno-associated virus serotypes 7 and 8 outperform serotype 9 in expressing atheroprotective human apoE3 from mouse skeletal muscle.

Abstract

Intramuscular injection of adeno-associated viral (AAV) vectors is potentially a safe, minimally invasive procedure for the long-term gene expression of circulating antiatherogenic proteins. Here, we compare secretion and atheroprotective effects of human apoE3 after injection of 3 pseudotyped AAV vectors (AAV2/7, AAV2/8, or AAV2/9), driven by the CMV enhancer/chicken β-actin (CAG) promoter, into skeletal muscle of hyperlipidemic apolipoprotein E-deficient (apoE⁻/⁻) mice. Vector viabilities were verified by transducing cultured C2C12 mouse myotubes and assessing secretion of human apoE3 protein. Both hind limb tibialis anterior muscles of female C57BL/6 apoE⁻/⁻ mice, 2 months old and fed a high-fat diet, were each injected with 1 x 10¹⁰ vector genomes of AAV vector. Identical noninjected mice served as controls; and blood was collected at weeks 0, 1, 2, 4, and 13. At termination (13 weeks), the brachiocephalic artery was excised; and after staining sections, plaque morphometry and fractional lipid content were quantified by computerized image analysis. Intramuscular injection of AAV2/7 and AAV2/8 vectors produced up to 2 μg human apoE3 per milliliter plasma, just below the threshold to reverse dyslipoproteinemia. AAV2/9 was notably less effective, mice having a 3-fold lower level of plasma apoE3 at 13 weeks and a 50% greater burden of atherosclerotic plaque lipid in their brachiocephalic arteries. We conclude that although vector refinement is needed to exploit fully apoE3 atheroprotective functions, AAV2/7 and AAV2/8 are promising gene transfer vectors for muscle-based expression of antiatherogenic circulating proteins.

• Authors: Vanessa C Evans, Ian R Graham, Takis Athanasopoulos, Deborah J Galley, Christopher L Jackson, Jonathan Paul Simons, George Dickson, James S Owen
• Journal: Metabolism: clinical and experimental (Metabolism) Vol. 60 Issue 4 Pg. 491-8 (Apr 2011) ISSN: 1532-8600 [Electronic] United States
• PMID: 20580777 (Publication Type: Comparative Study, Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: Copyright © 2011 Elsevier Inc. All rights reserved.

Chemical References

• Apolipoprotein E3
• Culture Media

Topics

• Animals
• Apolipoprotein E3 (genetics, metabolism)
• Atherosclerosis (metabolism, prevention & control)
• Cell Line
• Cerebral Arteries (pathology)
• Culture Media
• Dependovirus (classification, genetics, immunology)
• Enzyme-Linked Immunosorbent Assay
• Genetic Therapy
• Genetic Vectors
• Humans
• Hypercholesterolemia (therapy)
• Lipid Metabolism (genetics)
• Mice
• Mice, Inbred C57BL
• Mice, Knockout
• Muscle, Skeletal (metabolism)
• Serotyping
• Transduction, Genetic