Lentiviral gene transfer into the dorsal root ganglion of adult rats

Hongwei Yu, Greg Fischer, Guangfu Jia, Jakob Reiser, Frank Park, Quinn H. Hogan

Research output: Contribution to journalArticle

29 Citations (Scopus)

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.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.

Original languageEnglish (US)
Article number63
JournalMolecular Pain
Volume7
DOIs
StatePublished - Aug 23 2011

Fingerprint

Spinal Ganglia
Peptide Elongation Factor 1
Genes
Diagnosis-Related Groups
Neuralgia
Sensory Receptor Cells
Neurons
Chronic Pain
Injections
Viral Regulatory and Accessory Proteins
Nociceptors
Infection
Glycoproteins
enhanced green fluorescent protein
Phenotype
Pain
Cell Line

All Science Journal Classification (ASJC) codes

  • Molecular Medicine
  • Cellular and Molecular Neuroscience
  • Anesthesiology and Pain Medicine

Cite this

Lentiviral gene transfer into the dorsal root ganglion of adult rats. / Yu, Hongwei; Fischer, Greg; Jia, Guangfu; Reiser, Jakob; Park, Frank; Hogan, Quinn H.

In: Molecular Pain, Vol. 7, 63, 23.08.2011.

Research output: Contribution to journalArticle

Yu, Hongwei ; Fischer, Greg ; Jia, Guangfu ; Reiser, Jakob ; Park, Frank ; Hogan, Quinn H. / Lentiviral gene transfer into the dorsal root ganglion of adult rats. In: Molecular Pain. 2011 ; Vol. 7.
@article{6c32e8af76e1482daf56a4368e92c0a1,
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.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 = "Hongwei Yu and Greg Fischer and Guangfu Jia and Jakob Reiser and Frank Park and Hogan, {Quinn H.}",
year = "2011",
month = "8",
day = "23",
doi = "10.1186/1744-8069-7-63",
language = "English (US)",
volume = "7",
journal = "Molecular Pain",
issn = "1744-8069",
publisher = "BioMed Central",

}

TY - JOUR

T1 - Lentiviral gene transfer into the dorsal root ganglion of adult rats

AU - Yu, Hongwei

AU - Fischer, Greg

AU - Jia, Guangfu

AU - Reiser, Jakob

AU - Park, Frank

AU - Hogan, Quinn H.

PY - 2011/8/23

Y1 - 2011/8/23

N2 - 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.

AB - 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.

UR - http://www.scopus.com/inward/record.url?scp=80052035933&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=80052035933&partnerID=8YFLogxK

U2 - 10.1186/1744-8069-7-63

DO - 10.1186/1744-8069-7-63

M3 - Article

VL - 7

JO - Molecular Pain

JF - Molecular Pain

SN - 1744-8069

M1 - 63

ER -