Abstract
Direct injection of double-stranded adeno-associated virus type 2 (dsAAV2) with a μ-opioid receptor (MOR) mutant [S4.45(196)A], and a reporter protein (enhanced green fluorescent protein) into the spinal cord (S2/S3) dorsal horn region of ICR mice resulted in antinociceptive responses to systemic injection of opioid antagonist naloxone without altering the acute agonist morphine responses and no measurable tolerance or dependence development during subchronic naloxone treatment. To develop further such mutant MORs into a therapeutic agent in pain management, a less invasive method for virus delivery is needed. Thus, in current studies, the dsAAV2 was locally injected into the subarachnoid space of the spinal cord by intrathecal administration. Instead of using the MORS196A mutant, we constructed the dsAAV2 vector with the MORS196ACSTA mutant, a receptor mutant in which naloxone has been shown to exhibit full agonistic properties in vitro. After 2 weeks of virus injection, naloxone (10 mg/kg s.c.) elicited antinociceptive effect (determined by tail-flick test) without tolerance (10 mg/kg s.c., b.i.d. for 6 days) and significant withdrawal symptoms. On the other hand, subchronic treatment with morphine (10 mg/kg s.c., b.i.d.) for 6 days induced significant tolerance (4.8-fold) and withdrawal symptoms. Furthermore, we found that morphine, but not naloxone, induced the rewarding effects (determined by conditioned place preference test). These data suggest that local expression of MORS196ACSTA in spinal cord and systemic administration of naloxone has the potential to be developed into a new strategy in the management of pain without addiction liability.
Footnotes
This work was supported by the National Science Council, Taiwan, Republic of China [Grant NSC97-2320-B-016-003]; and the National Institutes of Health National Institute on Drug Abuse [Grants R01-DA023905, R01-DA011806, K05-DA000513 (to P.Y.L.)].
Article, publication date, and citation information can be found at http://jpet.aspetjournals.org.
doi:10.1124/jpet.109.165399.
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ABBREVIATIONS:
- dsAAV2
- double-stranded adeno-associated virus type 2
- MOR
- μ-opioid receptor
- KOR
- κ-opioid receptor
- CPP
- conditioned place preference
- EGFP
- enhanced green fluorescent protein
- ANOVA
- analysis of variance
- %MPE
- percentage of maximum possible effect
- AUC
- area under the curve
- DRG
- dorsal root ganglion
- CGRP
- calcitonin gene-related peptide.
- Received December 31, 2009.
- Accepted June 15, 2010.
- Copyright © 2010 by The American Society for Pharmacology and Experimental Therapeutics
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