Gene therapy by electroporation for the treatment of chronic renal failure in companion animals

<p>Abstract</p> <p>Background</p> <p>Growth hormone-releasing hormone (GHRH) plasmid-based therapy for the treatment of chronic renal failure and its complications was examined. Companion dogs (13.1 ± 0.8 years, 29.4 ± 5.01 kg) and cats (13.2 ± 0.9 years, 8.5 ± 0.37 kg) received a single 0.4 mg or 0.1 mg species-specific plasmid injection, respectively, intramuscularly followed by electroporation, and analyzed up to 75 days post-treatment; controls underwent electroporation without plasmid administration.</p> <p>Results</p> <p>Plasmid-treated animals showed an increase in body weight (dogs 22.5% and cats 3.2%) compared to control animals, and displayed improved quality of life parameters including significant increases in appetite, activity, mentation and exercise tolerance levels. Insulin-like growth factor I (IGF-I, the downstream effector of GHRH) levels were increased in the plasmid treated animals. Hematological parameters were also significantly improved. Protein metabolism changes were observed suggesting a shift from a catabolic to an anabolic state in the treated animals. Blood urea nitrogen and creatinine did not show any significant changes suggesting maintenance of kidney function whereas the control animal's renal function deteriorated. Treated animals survived longer than control animals with 70% of dogs and 80% of cats surviving until study day 75. Only 17% and 40% of the control dogs and cats, respectively, survived to day 75.</p> <p>Conclusion</p> <p>Improved quality of life, survival and general well-being indicate that further investigation is warranted, and show the potential of a plasmid-based therapy by electroporation in preventing and managing complications of renal insufficiency.</p

Immune-enhancing effects of growth hormone-releasing hormone delivered by plasmid injection and electroporation

Growth hormone-releasing hormone (GHRH) is a hypothalamic hormone with both direct and indirect functions in the maintenance of immune status under physiological and pathological conditions. In this study, 52 Holstein heifers were evaluated for the effects of a plasmid-mediated GHRH treatment on their immune function and on the morbidity and mortality of treated animals. In the third trimester of pregnancy, 32 heifers received 2.5 mg of a myogenic GHRH-expressing plasmid by intramuscular injection followed by electroporation, while 20 heifers were used as controls. No adverse effects were associated with either the plasmid delivery or GHRH expression. At 18 days after plasmid administration, GHRH-treated animals had increased numbers of CD2(+) alphabeta T-cells (P < 0.004), CD25(+)CD4(+) cells (P < 0.0007), and CD4(+)CD45R(+) cells (P < 0.016) compared to controls. These increases were maintained long term after treatment and correlated with plasmid expression. At 300 days post-GHRH therapy, CD45R(+)/CD45R0(-) naïve lymphocytes were significantly increased in frequency (P < 0.05). Natural killer lymphocytes (CD3(-)CD2(+)) were also increased. As a consequence of improved health status, body condition scores of treated animals improved (3.55 vs. 3.35, P < 0.0001). Hoof pathology was also reduced with treatment. The mortality of heifers was decreased (3% vs. 20% in controls, P < 0.003). Collectively, these results indicate that the myogenic GHRH plasmid can be successfully electrotransferred into a 500-kg mammal and expressed for prolonged periods of time, ensuring physiological levels of GHRH. The plasmid injection followed by electroporation could prove an efficient method for the systemic production of therapeutic proteins and may provide a useful means for basic research in relevant animal models