In vivo NMR imaging evaluation of efficiency and toxicity of gene electrotransfer in rat muscle.
In vivo gene electrotransfer (ET) is a simple method of gene delivery in various tissues relying on the injection of plasmid DNA followed by application of electric pulses. Noninvasive tools are needed to evaluate the ET efficiency and the resulting tissue damages. In this study, we performed ET of rat tibialis muscle after injection of either a plasmid coding for luciferase or a contrast agent (CA) detected by using magnetic resonance imaging (MRI). Plasmid expression and CA intracellular trapped quantity were compared throughout the electric field intensity range 0-300 V/cm. Although the CA trapped quantity reflects only the electropermeabilization step, both measurements were correlated. MRI measurements gave easy access to tridimensional visualization of the labelled zones where the CA has been injected and the applied electric field had a value allowing permeabilization. We also performed MRI measurements of the water transverse relaxation time T2 as an indicator of tissue modification, and tested whether another CA specific for necrosis could be used to detect muscle necrosis at high electric field intensity. In conclusion, MRI measurements may bring multiparametric information upon the efficiency and tissue toxicity of an ET protocol by using a simple and safe CA.
Leroy-Willig
A
A
NMR Unit (AFM-CEA), Institute of Myology, IFR14, Pitié-Salpêtrière University Hospital, Paris, France.
Bureau
M F
MF
Scherman
D
D
Carlier
P G
PG
D000818:Animals; D003287:Contrast Media / Q000032:analysis; D018274:Electroporation / Q000379:methods*; D015870:Gene Expression; D015316:Genetic Therapy / Q000009:adverse effects / Q000379:methods*; D006571:Heterocyclic Compounds / Q000032:analysis; D007273:Injections, Intramuscular; D008156:Luciferases / Q000032:analysis / Q000235:genetics*; D008279:Magnetic Resonance Imaging; D008297:Male; D018482:Muscle, Skeletal / Q000201:enzymology*; D009942:Organometallic Compounds / Q000032:analysis; D010957:Plasmids / Q000008:administration & dosage; D051381:Rats; D017208:Rats, Wistar
D003287:Contrast Media; D006571:Heterocyclic Compounds; D009942:Organometallic Compounds; C050823:gadolinium 1,4,7,10-tetraazacyclododecane-N,N',N'',N'''-tetraacetate; D008156:Luciferases