title: Image-guided, intravascular hydrodynamic gene delivery to skeletal muscle in pigs.
abstract: Development of an effective, safe, and convenient method for gene delivery to muscle is a critical step toward gene therapy for muscle-associated diseases. Toward this end, we have explored the possibility of combining the image-guided catheter insertion technique with the principle of hydrodynamic delivery to achieve muscle-specific gene transfer in pigs. We demonstrate that gene transfer efficiency of the procedure is directly related to flow rate, injection pressure, and injection volume. The optimal gene delivery was achieved at a flow rate of 15 ml/second with injection pressure of 300 psi and injection volume equal to 1.5% of body weight. Under such a condition, hydrodynamic injection of saline containing pCMV-Luc (100 microg/ml) resulted in luciferase activity of 10(6) to 10(7) relative light units (RLU)/mg of proteins extracted from the targeted muscle 5 days after hydrodynamic gene delivery. Result from immunohistochemical analysis revealed 70-90% transfection efficiency of muscle groups in the hindlimb and persistent reporter gene expression for 2 months in transfected cells. With an exception of transient edema and elevation of creatine phosphokinase, no permanent tissue damage was observed. These results suggest that the image-guided, intravenous hydrodynamic delivery is an effective and safe method for gene delivery to skeletal muscle.
authors.affiliation: Department of Pharmaceutical Sciences, School of Pharmacy, University of Pittsburgh, Pittsburgh, Pennsylvania 15261, USA.
authors.forename: Kenya
authors.initials: K
authors.lastname: Kamimura
authors.forename: Guisheng
authors.initials: G
authors.lastname: Zhang
authors.forename: Dexi
authors.initials: D
authors.lastname: Liu
mesh_terms: D000818:Animals; D015316:Genetic Therapy / Q000379:methods; D006614:Hindlimb / Q000378:metabolism; D007150:Immunohistochemistry; D007275:Injections, Intravenous / Q000379:methods*; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D018482:Muscle, Skeletal / Q000378:metabolism*; D010690:Phlebography / Q000379:methods*; D010957:Plasmids / Q000008:administration & dosage / Q000235:genetics; D013552:Swine
chemical_list: D008156:Luciferases