title: Lentiviral gene transfer into the dorsal root ganglion of adult rats.
abstract: 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.
  
  
  CONCLUSIONS
  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.
authors.affiliation: Department of Anesthesiology, Medical College of Wisconsin, 8100 Watertown Plank Rd, Milwaukee, Wisconsin 53226, USA.
authors.forename: Hongwei
authors.initials: H
authors.lastname: Yu
authors.forename: Greg
authors.initials: G
authors.lastname: Fischer
authors.forename: Guangfu
authors.initials: G
authors.lastname: Jia
authors.forename: Jakob
authors.initials: J
authors.lastname: Reiser
authors.forename: Frank
authors.initials: F
authors.lastname: Park
authors.forename: Quinn H
authors.initials: QH
authors.lastname: Hogan
mesh_terms: D000375:Aging / Q000378:metabolism*; D000818:Animals; D001522:Behavior, Animal; D002461:Cell Line, Transformed; D005727:Ganglia, Spinal / Q000378:metabolism*; D018014:Gene Transfer Techniques; D005822:Genetic Vectors / Q000008:administration & dosage / Q000235:genetics; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006801:Humans; D007267:Injections; D016086:Lentivirus / Q000235:genetics*; D008562:Membrane Glycoproteins / Q000378:metabolism; D009132:Muscles / Q000378:metabolism; D010146:Pain / Q000378:metabolism / Q000473:pathology; D020648:Peptide Elongation Factor 1 / Q000235:genetics / Q000378:metabolism; D011401:Promoter Regions, Genetic / Q000235:genetics; D011820:Rabies virus / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D011984:Sensory Receptor Cells / Q000378:metabolism; D014161:Transduction, Genetic; D014759:Viral Envelope Proteins / Q000378:metabolism; D054334:Viral Regulatory and Accessory Proteins / Q000378:metabolism
chemical_list: C028391:G protein, vesicular stomatitis virus; D008562:Membrane Glycoproteins; D020648:Peptide Elongation Factor 1; D014759:Viral Envelope Proteins; D054334:Viral Regulatory and Accessory Proteins; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins