Could plasmid-mediated gene transfer into the myocardium be augmented by left ventricular guided laser myocardial injury?
Early studies have indicated no correlation between the amount of mechanical injury and the level of myocardial gene expression following direct plasmid vector injection. Recently, however, evidence suggests that combined laser myocardial injury and plasmid-based gene delivery exert synergistic effects on gene expression and activity. The purpose of the study was to determine whether laser-induced myocardial injury followed by transendocardial gene transfer increases gene expression compared to gene transfer alone. We assessed the ability of a plasmid vector to express its transgene after injection into porcine ischemic myocardium with and without preceding laser myocardial injury. Thirteen animals had transendocardial injections of the luciferase reporter gene in a plamid vector using a catheter-based injection system. Injections (0.5 mg per animal, 50 microg per injection site) were divided into 10 sites in the ischemic territory. Eight animals underwent transendocardial laser injury of the ischemic region (2 Joule per pulse x 10 sites) prior to gene delivery. In five animals, gene injection sites were dispersed between laser channels, and in three animals laser and gene delivery were applied in close proximity (< 5 mm) or at the same location. Luciferase activity was measured at 3 and 7 days. Luciferase expression in ischemic zones was markedly elevated at day 3 and 7, and similar whether animals were pretreated using laser injury followed by gene transfer compared to gene transfer alone. Neither same-spot injection nor dispersed gene delivery were associated with augmented gene expression compared to gene transfer alone. Using the above-described catheter-based approach to combine localized laser injury and injection of naked DNA into ischemic myocardium, laser injury did not augment gene expression above levels present with gene transfer alone.
Fuchs
S
S
Cardiolovascular Research Institute, Washington, DC 20010, USA. sxf6@mhg.edu
Baffour
R
R
Shou
M
M
Stabile
E
E
Singh
S
S
Schwartz
B
B
Branellec
D
D
Epstein
S E
SE
Kornowski
R
R
D000818:Animals; D022062:Electrophysiologic Techniques, Cardiac; D015870:Gene Expression / Q000235:genetics; D018014:Gene Transfer Techniques; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics / Q000627:therapeutic use; D006335:Heart Injuries / Q000209:etiology*; D006352:Heart Ventricles / Q000293:injuries; D053685:Laser Therapy; D008156:Luciferases / Q000096:biosynthesis / Q000235:genetics; D008955:Models, Cardiovascular; D017202:Myocardial Ischemia / Q000235:genetics* / Q000601:surgery*; D009206:Myocardium / Q000378:metabolism*; D010957:Plasmids / Q000235:genetics* / Q000627:therapeutic use*; D013552:Swine; D016896:Treatment Outcome
D008156:Luciferases