Nonviral gene delivery to the lateral ventricles in rat brain: initial evidence for widespread distribution and expression in the central nervous system.
The use of DNA for nonviral gene expression depends on several factors. These include (i) delivery and accessibility to the targeted tissue, (ii) protection from extracellular degradation, (iii) sufficient uptake by cells of interest, and (iv) protection from intracellular degradation to allow translation of adequate levels of intracellular or secreted proteins. As an initial step in demonstrating the feasibility of nonviral, cationic lipid-mediated gene therapy, we present evidence for the successful delivery and expression of heat shock protein Hsp70 and reporter gene enzymes in the central nervous system (CNS) of the rat after injection into the lateral ventricle. Gene delivery is accomplished using optimized formulations of plasmid DNA, which have been complexed with the cationic lipid MLRI. Results from DNA vectors encoding for green fluorescent protein (GFP), luciferase, and Hsp70 are reported. Standard immunofluorescent methods were used to demonstrate widespread expression of the reporter proteins and of Hsp70. Stereology analysis has been completed on three coronal sections, which illustrates the distribution of expression along the longitudinal axis. These initial findings support the further development of nonviral, lipid-mediated gene delivery technology for transient expression of protective, intracellular proteins and represent an important step leading to in vivo studies to identify potential clinical benefits.
Hecker
J G
JG
Department of Anesthesiology, University of California-Davis, Davis, California 95616, USA. jghecker@ucdavis.edu
Hall
L L
LL
Irion
V R
VR
D000818:Animals; D002412:Cations; D002490:Central Nervous System / Q000737:chemistry; D005240:Feasibility Studies; D005455:Fluorescent Antibody Technique; D005786:Gene Expression Regulation / Q000187:drug effects; D018014:Gene Transfer Techniques; D015316:Genetic Therapy; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins; D018840:HSP70 Heat-Shock Proteins / Q000096:biosynthesis / Q000235:genetics*; D007276:Injections, Intraventricular; D008055:Lipids; D008156:Luciferases / Q000096:biosynthesis / Q000235:genetics; D008164:Luminescent Proteins / Q000096:biosynthesis / Q000235:genetics; D009474:Neurons; D019076:Transgenes
D002412:Cations; D018840:HSP70 Heat-Shock Proteins; D008055:Lipids; D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins; D008156:Luciferases