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14 August 2013

Efficient liver gene transfer with foamy virus vectors

Dimitris ZacharoulisAEG, Chris RountasBDE, Michalis KatsimpoulasBEF, John MorianosCDF, Ilenia ChatziandreouCD, George VassilopoulosACDEFG

DOI: 10.12659/MSMBR.883996

Med Sci Monit Basic Res 2013; 19:214-220

Abstract

BACKGROUND: Liver gene transfer offers hope for the correction of genetic and acquired disorders. Efficient gene transfer in large animals can be obtained with hydrodynamic gene transfer (HGT), a method that can achieve sufficient levels of gene delivery.

MATERIAL AND METHODS: To test the relative efficiency between plasmid versus foamy virus (FV) vector-based liver gene transfer efficiency, we applied HGT in 4 juvenile pigs, using the same plasmid backbone, either naked or coated as a FV vector particle. Gene transfer efficiency and persistence of expression was assayed by PCR and real-time PCR, respectively, at 1 week and at 1 month after the infusions.

RESULTS: HGT was tolerated well and no adverse reactions were observed. Plasmid injections resulted in no detectable DNA sequences at 1 week. At the 1 month time point, 2/15 liver sections analyzed were positive for the presence of plasmid DNA. When FV vectors were infused under identical conditions, 18/28 (64.3%) of the liver samples were positive for the presence of vector sequences, and the expression levels reached 29.7 and 15.6% of the endogenous GAPDH levels in the injected and the adjacent liver lobes.

CONCLUSIONS: Our results indicate that medium-term therapeutic levels of gene expression can be obtained with FV vectors, an effect that can be attributed to the potential of the HGT procedure and to the natural affinity of FV vectors for hepatocytes.

Keywords: Polymerase Chain Reaction, Plasmids - genetics, Models, Animal, Liver - metabolism, Genetic Vectors, Genetic Therapy - methods, Gene Transfer Techniques, Real-Time Polymerase Chain Reaction, Spumavirus - genetics, Swine

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Medical Science Monitor Basic Research eISSN: 2325-4416
Medical Science Monitor Basic Research eISSN: 2325-4416