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  • Inherited Disease
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Treatment of severe hypercholesterolemia in apolipoprotein E-deficient mice by intramuscular injection of plasmid DNA

Gene Therapy volume 7, pages 1795–1801 (2000)Cite this article

Abstract

We report on systemic delivery and long-term biological effects of apolipoprotein E (apoE) obtained by intramuscular (i.m.) plasmid DNA injection. ApoE plays an important role in lipoprotein catabolism and apoE knock-out mice develop severe hypercholesterolemia and diffuse atherosclerosis. We have injected apoE-deficient mice with 80 μg of a plasmid vector (pCMV-E3) encoding the human apoE3 cDNA under the control of the CMV promoter-enhancer in both posterior legs. Local expression of the transgene was demonstrated throughout 16 weeks. Human apoE3 recombinant protein reached 0.6 ng/ml serum level. After i.m. injection of pCMV-E3 expression vector the mean serum cholesterol concentrations decreased from 439 ± 57 mg/dl to 253 ± 99 mg/dl (P < 0.05) 2 weeks after injection and persisted at a significantly reduced level throughout the 16 weeks observation period (P < 0.005). Serum cholesterol was unaffected and reached an absolute level of 636 ± 67 mg/dl in control groups. Finally, injection of pCMV-E3 into apoE-deficient mice resulted in a redistribution of cholesterol content between lipoprotein fractions, with a marked decrease in VLDL, IDL and LDL cholesterol content and an increase in HDL cholesterol. These results demonstrate that severe hypercholesterolemia in apoE-deficient mice can be effectively reversed by i.m. DNA injection, and indicate that this approach could represent a useful tool to correct several hyperlipidemic conditions resulting in atherosclerosis.

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Acknowledgements

This research was partially supported by Italian Ministero della Sanità, RC-IRCCS and MURST 60% to VMF. We thank the staff of ‘Servizio Stabulario’, Catholic University of Rome, for expert and excellent technical assistance with animals.

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Authors and Affiliations

  1. Laboratory for Molecular Medicine and Biotechnology, University Campus Bio-Medico, School of Medicine, Rome, Italy

    M Rinaldi, P Parrella & V M Fazio

  2. Institute of Experimental Medicine, CNR, Rome, Italy

    M Rinaldi, P Parrella, E Signori & V M Fazio

  3. Institute of Pharmacological Sciences, University of Milan, Italy

    A L Catapano & P Uboldi

  4. Department of Experimental Medicine and Biochemical Sciences, University of Rome ‘Tor Vergata’, Italy

    S A Ciafrè & M G Farace

  5. Laboratory of Molecular Pathology and Gene Therapy, IRCCS H ‘Casa Sollievo della Sofferenza’, San Giovanni Rotondo (FG), Italy

    E Signori, D Seripa & V M Fazio

  6. Istituto Terapia Medica Sistematica, University of Rome ‘La Sapienza’, Italy

    R Antonini & G Ricci

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  1. M Rinaldi
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Rinaldi, M., Catapano, A., Parrella, P. et al. Treatment of severe hypercholesterolemia in apolipoprotein E-deficient mice by intramuscular injection of plasmid DNA. Gene Ther 7, 1795–1801 (2000). https://doi.org/10.1038/sj.gt.3301310

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