Electroporation-mediated pain-killer gene therapy for mononeuropathic rats.
The relatively low expression levels achieved from transferred genes have limited the application of nonviral vectors for gene transfer into the spinal cord in vivo. Thus, the aim of this study was to evaluate the efficacy of electroporation-mediated pro-opiomelanocortin (POMC) gene therapy for neuropathic pain using an animal model of chronic constrictive injury (CCI). Firstly, the optimal pulse characteristics (voltage, pulse duration, number of shocks) were investigated for in vivo electroporation-mediated gene transfer into the spinal cord. The electroporation process makes use of plasmid DNA, which expresses the POMC gene. Expression levels were evaluated in this study by Western blot. We conclude that the optimal conditions for electroporation are a pulse voltage of 200 V, 75-ms duration, 925-ms interval, for five iterations. Secondly, electroporation treatment for neuropathic pain was attempted on CCI rats using plasmid DNA that expresses the POMC gene. Intrathecal administrations of the POMC vector elevated spinal beta-endorphin levels, as manifested in a significantly elevated pain threshold for the CCI limbs. This result suggests that gene therapy for neuropathic pain using this novel technique is very efficacious, and thus shows promise for further clinical trials.
Lin
C-R
CR
Department of Biological Sciences, National Sun Yat-Sen University, Taiwan.
Yang
L-C
LC
Lee
T-H
TH
Lee
C-T
CT
Huang
H-T
HT
Sun
W-Z
WZ
Cheng
J-T
JT
D000704:Analysis of Variance; D000818:Animals; D015153:Blotting, Western / Q000379:methods; D018274:Electroporation / Q000379:methods*; D015316:Genetic Therapy / Q000379:methods*; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods; D008159:Lumbar Vertebrae; D008297:Male; D009270:Naloxone; D009292:Narcotic Antagonists; D010146:Pain / Q000209:etiology; D059408:Pain Management; D010147:Pain Measurement; D011333:Pro-Opiomelanocortin / Q000032:analysis / Q000235:genetics*; D012333:RNA, Messenger / Q000032:analysis; D051381:Rats; D017207:Rats, Sprague-Dawley; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013117:Spinal Cord Compression / Q000150:complications / Q000628:therapy*
D009292:Narcotic Antagonists; D012333:RNA, Messenger; D009270:Naloxone; D011333:Pro-Opiomelanocortin