title: Dose response in rodents and nonhuman primates after hydrodynamic limb vein delivery of naked plasmid DNA.
abstract: The efficacy of gene therapy mediated by plasmid DNA (pDNA) depends on the selection of suitable vectors and doses. Using hydrodynamic limb vein (HLV) injection to deliver naked pDNA to skeletal muscles of the limbs, we evaluated key parameters that affect expression in muscle from genes encoded in pDNA. Short-term and long-term promoter comparisons demonstrated that kinetics of expression differed between cytomegalovirus (CMV), muscle creatine kinase, and desmin promoters, but all gave stable expression from 2 to 49 weeks after delivery to mouse muscle. Expression from the CMV promoter was highest. For mice, rats, and rhesus monkeys, the linear range for pDNA dose response could be defined by the mass of pDNA relative to the mass of target muscle. Correlation between pDNA dose and expression was linear between a threshold dose of 75 μg/g and maximal expression at approximately 400 μg/g. One HLV injection into rats of a dose of CMV-LacZ yielding maximal expression resulted in an average transfection of 28% of all hind leg muscle and 40% of the gastrocnemius and soleus. Despite an immune reaction to the reporter gene in monkeys, a single injection transfected an average of 10% of all myofibers in the targeted muscle of the arms and legs and an average of 15% of myofibers in the gastrocnemius and soleus.
authors.affiliation: Roche Madison, Madison, WI 53711, USA. chris.wooddell@roche.com
authors.forename: Christine I
authors.initials: CI
authors.lastname: Wooddell
authors.forename: Julia O
authors.initials: JO
authors.lastname: Hegge
authors.forename: Guofeng
authors.initials: G
authors.lastname: Zhang
authors.forename: Magdolna G
authors.initials: MG
authors.lastname: Sebestyén
authors.forename: Mark
authors.initials: M
authors.lastname: Noble
authors.forename: Jacob B
authors.initials: JB
authors.lastname: Griffin
authors.forename: Loretta V
authors.initials: LV
authors.lastname: Pfannes
authors.forename: Hans
authors.initials: H
authors.lastname: Herweijer
authors.forename: James E
authors.initials: JE
authors.lastname: Hagstrom
authors.forename: Serge
authors.initials: S
authors.lastname: Braun
authors.forename: Thierry
authors.initials: T
authors.lastname: Huss
authors.forename: Jon A
authors.initials: JA
authors.lastname: Wolff
mesh_terms: D000818:Animals; D052283:Creatine Kinase, MM Form / Q000235:genetics / Q000378:metabolism; D003587:Cytomegalovirus / Q000235:genetics; D004247:DNA / Q000008:administration & dosage* / Q000235:genetics; D005121:Extremities; D005260:Female; D015870:Gene Expression; D018014:Gene Transfer Techniques; D017930:Genes, Reporter; D015316:Genetic Therapy; D005822:Genetic Vectors; D057446:Hydrodynamics; D007700:Kinetics; D007763:Lac Operon; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D008253:Macaca mulatta; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008813:Mice, Inbred ICR; D018482:Muscle, Skeletal / Q000378:metabolism; D010957:Plasmids / Q000008:administration & dosage* / Q000235:genetics; D011401:Promoter Regions, Genetic; D051381:Rats; D017207:Rats, Sprague-Dawley; D014162:Transfection; D014680:Veins / Q000378:metabolism*; D001616:beta-Galactosidase / Q000032:analysis
chemical_list: D004247:DNA; D008156:Luciferases; D052283:Creatine Kinase, MM Form; D001616:beta-Galactosidase