Virus-mediated shRNA knockdown of Na(v)1.3 in rat dorsal root ganglion attenuates nerve injury-induced neuropathic pain.
Neuropathic pain is a chronic condition that is often refractory to treatment with available therapies and thus an unmet medical need. We have previously shown that the voltage-gated sodium channel Na(v)1.3 is upregulated in peripheral and central nervous system (CNS) of rats following nerve injury, and that it contributes to nociceptive neuron hyperexcitability in neuropathic conditions. To evaluate the therapeutic potential of peripheral Na(v)1.3 knockdown at a specific segmental level, we constructed adeno-associated viral (AAV) vector expressing small hairpin RNA against rat Na(v)1.3 and injected it into lumbar dorsal root ganglion (DRG) of rats with spared nerve injury (SNI). Our data show that direct DRG injection provides a model that can be used for proof-of-principle studies in chronic pain with respect to peripheral delivery route of gene transfer constructs, high transduction efficiency, flexibility in terms of segmental localization, and limited behavioral effects of the surgical procedure. We show that knockdown of Na(v)1.3 in lumbar 4 (L4) DRG results in an attenuation of nerve injury-induced mechanical allodynia in the SNI model. Taken together, our studies support the contribution of peripheral Na(v)1.3 to pain in adult rats with neuropathic pain, validate Na(v)1.3 as a target, and provide validation for this approach of AAV-mediated peripheral gene therapy.
Department of Neurology, Yale University School of Medicine, New Haven, Connecticut, USA.
Omar A
OA
Samad
Andrew M
AM
Tan
Xiaoyang
X
Cheng
Edmund
E
Foster
Sulayman D
SD
Dib-Hajj
Stephen G
SG
Waxman
D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D017931:DNA Primers; D000229:Dependovirus / Q000235:genetics*; D015536:Down-Regulation; D005727:Ganglia, Spinal / Q000378:metabolism*; D055785:Gene Knockdown Techniques; D005822:Genetic Vectors; D008297:Male; D062552:NAV1.3 Voltage-Gated Sodium Channel / Q000235:genetics / Q000502:physiology*; D018408:Patch-Clamp Techniques; D010523:Peripheral Nervous System Diseases / Q000517:prevention & control*; D034741:RNA, Small Interfering / Q000235:genetics*; D051381:Rats; D017207:Rats, Sprague-Dawley; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction
D017931:DNA Primers; D062552:NAV1.3 Voltage-Gated Sodium Channel; D034741:RNA, Small Interfering; C478638:Scn3a protein, rat