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High‐efficiency growth hormone releasing hormone plasmid vector administration into skeletal muscle mediated by electroporation in pigs
Author(s) -
DraghiaAkli Ruxandra,
Ellis Kenneth M.,
Hill LeighAnne,
Malone P. Brandon,
Fiorotto Marta L.
Publication year - 2003
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fj.02-0671fje
Subject(s) - electroporation , plasmid , endocrinology , chemistry , medicine , skeletal muscle , hormone , bone mineral , weight gain , andrology , biology , microbiology and biotechnology , body weight , dna , gene , biochemistry , osteoporosis
We report here a very efficient method for the in vivo transfer of therapeutic plasmid DNA into porcine muscle fibers by using electric pulses of low field intensity. We evaluated delivery of 0.1–3 mg of plasmid vectors that encode reporter secreted‐embryonic alkaline phosphatase (SEAP) or therapeutic growth hormone releasing hormone (GHRH). Reporter gene studies showed that internal needle electrodes give a 25‐fold increase in expression levels compared with caliper electrodes in skeletal muscle in swine. Dose and time courses were performed. Pigs injected with 0.1 mg plasmid had significantly greater weight gain than controls over 53 days (22.4 ± 0.8 kg vs. 19.7 ± 0.03 kg, respectively; P <0.01). The group treated with GHRH‐expressing plasmid at 14 days of age demonstrated greater weight gain than controls at every time point (25.8 ± 1.5 kg vs. 19.7 ± 0.03 kg; P <0.01). Body composition studies by dual X‐ray absorbitometry showed a 22% decrease in fat deposition ( P <0.05) and a 10% increase in bone mineral density ( P <0.004). Our studies demonstrate that by optimizing the electroporation method, favorable physiological changes, such as enhanced weight gain and improved body composition, can be obtained at extremely low plasmid doses in a large mammal.