Expression of Reverse Cholesterol Transport Pathway Associated Protein Genes in Skeletal Muscle

Viral and nonviral vectors containing apoAI, apoE or LCAT genes were constructed and transfected into myogenic cells in vitro or injected directly into mouse skeletal muscle. The expression efficiencies of these vectors were assaied to investigate the possibility of ectopic expression of these genes in skeletal muscle and to develop a safe and convenient gene therapy method for atherosclerosis. The results showed that the primary cultured mouse myoblasts, C2C12 cells transfected with pCMVapoE3 expressed human apoE3 successfully and the expressed product was secreted into the medium. Mouse skeletal muscle efficiently expressed apoE3 in vivo after direct plasmid injection. The expression level of Ad-RSV-apoAI in primary cultured mouse myoblasts was correlated with virus titer. Human apoAI was synthesized in mouse skeletal muscle by direct injection of recombinant virus and was secreted into blood continuously up to 30 days functional LCAT was expressed by C2C12 and 293 cells transfected with conventional vector or recombinant AAV plasmid DNA. The expression efficiency of recombinant AAV plasmid DNA was 2-5 times higher than that of conventional plasmid vector. The above results provided experimental data for further studying and developing a gene therapy method for atherosclerosis by enhancement of reverse cholesterol transport using skeletal muscle as target.