Lentiviral gene transfer into the dorsal root ganglion of adult rats. Mol Pain 2011 Aug 23;7:63
Date
08/25/2011Pubmed ID
21861915Pubmed Central ID
PMC3179738DOI
10.1186/1744-8069-7-63Scopus ID
2-s2.0-80052035933 (requires institutional sign-in at Scopus site) 41 CitationsAbstract
BACKGROUND: Lentivector-mediated gene delivery into the dorsal root ganglion (DRG) is a promising method for exploring pain pathophysiology and for genetic treatment of chronic neuropathic pain. In this study, a series of modified lentivector particles with different cellular promoters, envelope glycoproteins, and viral accessory proteins were generated to evaluate the requirements for efficient transduction into neuronal cells in vitro and adult rat DRG in vivo.
RESULTS: In vitro, lentivectors expressing enhanced green fluorescent protein (EGFP) under control of the human elongation factor 1α (EF1α) promoter and pseudotyped with the conventional vesicular stomatitis virus G protein (VSV-G) envelope exhibited the best performance in the transfer of EGFP into an immortalized DRG sensory neuron cell line at low multiplicities of infection (MOIs), and into primary cultured DRG neurons at higher MOIs. In vivo, injection of either first or second-generation EF1α-EGFP lentivectors directly into adult rat DRGs led to transduction rates of 19 ± 9% and 20 ± 8% EGFP-positive DRG neurons, respectively, detected at 4 weeks post injection. Transduced cells included a full range of neuronal phenotypes, including myelinated neurons as well as both non-peptidergic and peptidergic nociceptive unmyelinated neurons.
CONCLUSION: VSV-G pseudotyped lentivectors containing the human elongation factor 1α (EF1α)-EGFP expression cassette demonstrated relatively efficient transduction to sensory neurons following direct injection into the DRG. These results clearly show the potential of lentivectors as a viable system for delivering target genes into DRGs to explore basic mechanisms of neuropathic pain, with the potential for future clinical use in treating chronic pain.
Author List
Yu H, Fischer G, Jia G, Reiser J, Park F, Hogan QHAuthors
Quinn H. Hogan MD Professor in the Anesthesiology department at Medical College of WisconsinHongwei Yu MD Professor in the Anesthesiology department at Medical College of Wisconsin
MESH terms used to index this publication - Major topics in bold
AgingAnimals
Behavior, Animal
Cell Line, Transformed
Ganglia, Spinal
Gene Transfer Techniques
Genetic Vectors
Green Fluorescent Proteins
Humans
Injections
Lentivirus
Membrane Glycoproteins
Muscles
Pain
Peptide Elongation Factor 1
Promoter Regions, Genetic
Rabies virus
Rats
Rats, Sprague-Dawley
Sensory Receptor Cells
Transduction, Genetic
Viral Envelope Proteins
Viral Regulatory and Accessory Proteins