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Loss of μ opioid receptor signaling in nociceptors, but not microglia, abrogates morphine tolerance without disrupting analgesia

Nature Medicine volume 23, pages 164173 (2017) | Download Citation

Abstract

Opioid pain medications have detrimental side effects including analgesic tolerance and opioid-induced hyperalgesia (OIH). Tolerance and OIH counteract opioid analgesia and drive dose escalation. The cell types and receptors on which opioids act to initiate these maladaptive processes remain disputed, which has prevented the development of therapies to maximize and sustain opioid analgesic efficacy. We found that μ opioid receptors (MORs) expressed by primary afferent nociceptors initiate tolerance and OIH development. RNA sequencing and histological analysis revealed that MORs are expressed by nociceptors, but not by spinal microglia. Deletion of MORs specifically in nociceptors eliminated morphine tolerance, OIH and pronociceptive synaptic long-term potentiation without altering antinociception. Furthermore, we found that co-administration of methylnaltrexone bromide, a peripherally restricted MOR antagonist, was sufficient to abrogate morphine tolerance and OIH without diminishing antinociception in perioperative and chronic pain models. Collectively, our data support the idea that opioid agonists can be combined with peripheral MOR antagonists to limit analgesic tolerance and OIH.

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References

  1. 1.

    , & Management of perioperative pain in patients chronically consuming opioids. Reg. Anesth. Pain Med. 29, 576–591 (2004).

  2. 2.

    , , & Opioids in chronic non-cancer pain: systematic review of efficacy and safety. Pain 112, 372–380 (2004).

  3. 3.

    Relieving pain in America: a blueprint for transforming prevention, care, education, and research. J. Pain Palliative Care Pharmacother. 26, 197–198 (2012).

  4. 4.

    & Opioid abuse in chronic pain–misconceptions and mitigation strategies. N. Engl. J. Med. 374, 1253–1263 (2016).

  5. 5.

    , & Opioid-induced hyperalgesia in humans: molecular mechanisms and clinical considerations. Clin. J. Pain 24, 479–496 (2008).

  6. 6.

    & Opioid-induced hyperalgesia: a qualitative systematic review. Anesthesiology 104, 570–587 (2006).

  7. 7.

    Opioid tolerance: the clinical perspective. Br. J. Anaesth. 81, 58–68 (1998).

  8. 8.

    et al. Loss of morphine-induced analgesia, reward effect and withdrawal symptoms in mice lacking the μ-opioid-receptor gene. Nature 383, 819–823 (1996).

  9. 9.

    , , & Opioid-receptor mRNA expression in the rat CNS: anatomical and functional implications. Trends Neurosci. 18, 22–29 (1995).

  10. 10.

    et al. Morphine hyperalgesia gated through microglia-mediated disruption of neuronal Cl homeostasis. Nat. Neurosci. 16, 183–192 (2013).

  11. 11.

    et al. Blockade of PDGFR-β activation eliminates morphine analgesic tolerance. Nat. Med. 18, 385–387 (2012).

  12. 12.

    Cellular neuroadaptations to chronic opioids: tolerance, withdrawal and addiction. Br. J. Pharmacol. 154, 384–396 (2008).

  13. 13.

    & The dark side of opioids in pain management: basic science explains clinical observation. Pain Rep. 1, e570 (2016).

  14. 14.

    , , & Opioid-induced hyperalgesia: cellular and molecular mechanisms. Neuroscience 338, 160–182 (2016).

  15. 15.

    et al. Pain and poppies: the good, the bad, and the ugly of opioid analgesics. J. Neurosci. 35, 13879–13888 (2015).

  16. 16.

    , & Attenuation of morphine tolerance, withdrawal-induced hyperalgesia, and associated spinal inflammatory immune responses by propentofylline in rats. Neuropsychopharmacology 29, 327–334 (2004).

  17. 17.

    , , & The “toll” of opioid-induced glial activation: improving the clinical efficacy of opioids by targeting glia. Trends Pharmacol. Sci. 30, 581–591 (2009).

  18. 18.

    Absence of μ opioid receptor mRNA expression in astrocytes and microglia of rat spinal cord. Neuroreport 23, 378–384 (2012).

  19. 19.

    et al. Evidence that opioids may have toll-like receptor 4 and MD-2 effects. Brain Behav. Immun. 24, 83–95 (2010).

  20. 20.

    et al. Toll-like receptor 4 mutant and null mice retain morphine-induced tolerance, hyperalgesia, and physical dependence. PLoS One 9, e97361 (2014).

  21. 21.

    , , & Microglial activation involved in morphine tolerance is not mediated by toll-like receptor 4. J. Anesth. 27, 93–97 (2013).

  22. 22.

    Opioid-induced abnormal pain sensitivity: implications in clinical opioid therapy. Pain 100, 213–217 (2002).

  23. 23.

    , & Shared mechanisms for opioid tolerance and a transition to chronic pain. J. Neurosci. 30, 4660–4666 (2010).

  24. 24.

    , , , & Long-term potentiation in spinal nociceptive pathways as a novel target for pain therapy. Mol. Pain 7, 20 (2011).

  25. 25.

    & Critical role of nociceptor plasticity in chronic pain. Trends Neurosci. 32, 611–618 (2009).

  26. 26.

    , & Multiple targets of μ-opioid receptor-mediated presynaptic inhibition at primary afferent Aδ- and C-fibers. J. Neurosci. 31, 1313–1322 (2011).

  27. 27.

    , , & Induction of synaptic long-term potentiation after opioid withdrawal. Science 325, 207–210 (2009).

  28. 28.

    , , & Opioid-induced long-term potentiation in the spinal cord is a presynaptic event. J. Neurosci. 30, 4460–4466 (2010).

  29. 29.

    , , , & Resistance to morphine analgesic tolerance in rats with deleted transient receptor potential vanilloid type 1-expressing sensory neurons. Neuroscience 145, 676–685 (2007).

  30. 30.

    et al. Dissociation of the opioid receptor mechanisms that control mechanical and heat pain. Cell 137, 1148–1159 (2009).

  31. 31.

    et al. Unbiased classification of sensory neuron types by large-scale single-cell RNA sequencing. Nat. Neurosci. 18, 145–153 (2015).

  32. 32.

    Opioids and their receptors: are we there yet? Neuropharmacology 76Part B, 198–203 (2014).

  33. 33.

    et al. Restriction of transient receptor potential vanilloid-1 to the peptidergic subset of primary afferent neurons follows its developmental downregulation in nonpeptidergic neurons. J. Neurosci. 31, 10119–10127 (2011).

  34. 34.

    , , & The μ-opioid receptor (MOR1) is mainly restricted to neurons that do not contain GABA or glycine in the superficial dorsal horn of the rat spinal cord. Neuroscience 75, 1231–1238 (1996).

  35. 35.

    , , & Distinct mechanisms underlying pronociceptive effects of opioids. J. Neurosci. 31, 16748–16756 (2011).

  36. 36.

    , , & Chronic opioid potentiates presynaptic but impairs postsynaptic N-methyl-D-aspartic acid receptor activity in spinal cords: implications for opioid hyperalgesia and tolerance. J. Biol. Chem. 287, 25073–25085 (2012).

  37. 37.

    , , , & Antagonism of gut, but not central effects of morphine with quaternary narcotic antagonists. Eur. J. Pharmacol. 78, 255–261 (1982).

  38. 38.

    , & Regional, developmental and cell-cycle-dependent differences in mu, delta, and kappa-opioid receptor expression among cultured mouse astrocytes. Glia 22, 249–259 (1998).

  39. 39.

    et al. Cell-specific actions of HIV-Tat and morphine on opioid receptor expression in glia. J. Neurosci. Res. 86, 2100–2110 (2008).

  40. 40.

    , & Inhibition of microglial P2X4 receptors attenuates morphine tolerance, Iba1, GFAP and μ opioid receptor protein expression while enhancing perivascular microglial ED2. Pain 150, 401–413 (2010).

  41. 41.

    et al. Microglia change from a reactive to an age-like phenotype with the time in culture. Front. Cell. Neurosci. 8, 152 (2014).

  42. 42.

    et al. Development of a method for the purification and culture of rodent astrocytes. Neuron 71, 799–811 (2011).

  43. 43.

    et al. An RNA-sequencing transcriptome and splicing database of glia, neurons, and vascular cells of the cerebral cortex. J. Neurosci. 34, 11929–11947 (2014).

  44. 44.

    et al. Essential role of toll-like receptor 2 in morphine-induced microglia activation in mice. Neurosci. Lett. 489, 43–47 (2011).

  45. 45.

    et al. Injured sensory neuron-derived CSF1 induces microglial proliferation and DAP12-dependent pain. Nat. Neurosci. 19, 94–101 (2016).

  46. 46.

    et al. Gliogenic LTP spreads widely in nociceptive pathways. Science 354, 1144–1148 (2016).

  47. 47.

    et al. Tolerance develops to the antiallodynic effects of the peripherally acting opioid loperamide hydrochloride in nerve-injured rats. Pain 154, 2477–2486 (2013).

  48. 48.

    , & Drug-poisoning deaths involving opioid analgesics: United States, 1999–2011. NCHS Data Brief 166, 1–8 (2014).

  49. 49.

    et al. Methylnaltrexone for opioid-induced constipation in advanced illness. N. Engl. J. Med. 358, 2332–2343 (2008).

  50. 50.

    et al. Safety and efficacy of methylnaltrexone in shortening the duration of postoperative ileus following segmental colectomy: results of two randomized, placebo-controlled phase 3 trials. Dis. Colon Rectum 54, 570–578 (2011).

  51. 51.

    et al. Analysis of opioid-mediated analgesia in Phase III studies of methylnaltrexone for opioid-induced constipation in patients with chronic noncancer pain. J. Pain Res. 8, 771–780 (2015).

  52. 52.

    et al. Treatment with prolonged-release oxycodone/naloxone improves pain relief and opioid-induced constipation compared with prolonged-release oxycodone in patients with chronic severe pain and laxative-refractory constipation. Clin. Ther. 37, 784–792 (2015).

  53. 53.

    , & Peripheral opioid receptor blockade increases postoperative morphine demands--a randomized, double-blind, placebo-controlled trial. Pain 155, 2056–2062 (2014).

  54. 54.

    , & Methylnaltrexone: its pharmacological effects alone and effects on morphine in healthy volunteers. Psychopharmacology (Berl.) 232, 63–73 (2015).

  55. 55.

    et al. Reversal of opioid-induced bladder dysfunction by intravenous naloxone and methylnaltrexone. Clin. Pharmacol. Ther. 82, 48–53 (2007).

  56. 56.

    et al. Peripheral mechanisms of pain and analgesia. Brain Res. Rev. 60, 90–113 (2009).

  57. 57.

    et al. Constitutive μ-opioid receptor activity leads to long-term endogenous analgesia and dependence. Science 341, 1394–1399 (2013).

  58. 58.

    et al. μ opioid receptors on primary afferent nav1.8 neurons contribute to opiate-induced analgesia: insight from conditional knockout mice. PLoS One 8, e74706 (2013).

  59. 59.

    et al. Olfactory exposure to males, including men, causes stress and related analgesia in rodents. Nat. Methods 11, 629–632 (2014).

  60. 60.

    Animal models of pain: progress and challenges. Nat. Rev. Neurosci. 10, 283–294 (2009).

  61. 61.

    et al. Structure-based discovery of opioid analgesics with reduced side effects. Nature 537, 185–190 (2016).

  62. 62.

    Long term alterations in the excitability of the flexion reflex produced by peripheral tissue injury in the chronic decerebrate rat. Pain 18, 325–343 (1984).

  63. 63.

    Pavlovian conditioned fear in Sidman avoidance learning. J. Comp. Physiol. Psychol. 65, 55–60 (1968).

  64. 64.

    & Passive and active reactions to fear-eliciting stimuli. J. Comp. Physiol. Psychol. 68, 129–135 (1969).

  65. 65.

    Species-specific defense reactions and avoidance learning. Psychol. Rev. 77, 32–48 (1970).

  66. 66.

    & A perceptual-defensive-recuperative model of fear and pain. Behav. Brain Sci. 3, 291–301 (1980).

  67. 67.

    The postshock activity burst. Anim. Learn. Behav. 10, 448–454 (1982).

  68. 68.

    The Expression of The Emotions in Man and Animals (Albemarle, 1872).

  69. 69.

    , , , & Quantitative assessment of tactile allodynia in the rat paw. J. Neurosci. Methods 53, 55–63 (1994).

  70. 70.

    , , , & Tonic inhibition of chronic pain by neuropeptide Y. Proc. Natl. Acad. Sci. USA 108, 7224–7229 (2011).

  71. 71.

    et al. A toolbox of Cre-dependent optogenetic transgenic mice for light-induced activation and silencing. Nat. Neurosci. 15, 793–802 (2012).

  72. 72.

    et al. Delta opioid receptors presynaptically regulate cutaneous mechanosensory neuron input to the spinal cord dorsal horn. Neuron 81, 1312–1327 (2014).

  73. 73.

    et al. Smart-seq2 for sensitive full-length transcriptome profiling in single cells. Nat. Methods 10, 1096–1098 (2013).

  74. 74.

    , & HISAT: a fast spliced aligner with low memory requirements. Nat. Methods 12, 357–360 (2015).

  75. 75.

    & BEDTools: a flexible suite of utilities for comparing genomic features. Bioinformatics 26, 841–842 (2010).

  76. 76.

    et al. Transcript assembly and quantification by RNA-Seq reveals unannotated transcripts and isoform switching during cell differentiation. Nat. Biotechnol. 28, 511–515 (2010).

  77. 77.

    & A peripheral mononeuropathy in rat that produces disorders of pain sensation like those seen in man. Pain 33, 87–107 (1988).

  78. 78.

    et al. Autotomy following peripheral nerve lesions: experimental anaesthesia dolorosa. Pain 7, 103–111 (1979).

  79. 79.

    et al. Stimulation of the α7 nicotinic acetylcholine receptor protects against neuroinflammation after tibia fracture and endotoxemia in mice. Mol. Med. 20, 667–675 (2015).

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Acknowledgements

We thank B. Kieffer (McGill University) for providing the Oprm1 floxed mice and MOR-mCherry mice, C. Evans (University of California, Los Angeles) and F. Simonin (Université de Strasbourg) for providing critical immunohistochemical and pharmacological reagents, J. Kaslow and A. Yu-Chun Wong for assistance with data analysis, and H. Nguyen (Stanford University) for assistance with illustration design. This work was supported by US National Institutes of Health (NIH) grant DA031777 (G.S.), the Rita Allen Foundation and American Pain Society Award in Pain (G.S.), NIH Fellowships F32DA041029 (G.C.), T32GM089626 (D.W.), and T32DA35165 (G.C.), Foundation for Anesthesia Education and Research (FAER) Mentored Research Training grant (V.L.T.), DoD National Defense Science and Engineering Graduate (NDSEG) Fellowship (E.I.S.), NSF Graduate Research Fellowship and Stanford Bio-X Graduate Fellowship (J.R.D.), NIH grant R37DA15043 (B.A.B.), and Damon Runyon Cancer Research Foundation postdoctoral fellowship (C.J.B.).

Author information

Author notes

    • Gregory Corder
    • , Vivianne L Tawfik
    • , Dong Wang
    •  & Elizabeth I Sypek

    These authors contributed equally to this work.

Affiliations

  1. Department of Anesthesiology, Perioperative and Pain Medicine, Stanford University, Stanford, California, USA.

    • Gregory Corder
    • , Vivianne L Tawfik
    • , Dong Wang
    • , Elizabeth I Sypek
    • , Sarah A Low
    • , Jasmine R Dickinson
    • , Chaudy Sotoudeh
    • , J David Clark
    •  & Grégory Scherrer
  2. Department of Molecular and Cellular Physiology, Stanford University, Stanford, California, USA.

    • Gregory Corder
    • , Vivianne L Tawfik
    • , Dong Wang
    • , Sarah A Low
    • , Chaudy Sotoudeh
    •  & Grégory Scherrer
  3. Department of Neurosurgery, Stanford University, Stanford, California, USA.

    • Gregory Corder
    • , Vivianne L Tawfik
    • , Dong Wang
    • , Sarah A Low
    • , Chaudy Sotoudeh
    •  & Grégory Scherrer
  4. Stanford Neurosciences Institute, Stanford, California, USA.

    • Gregory Corder
    • , Vivianne L Tawfik
    • , Dong Wang
    • , Sarah A Low
    • , Chaudy Sotoudeh
    • , Ben A Barres
    • , Christopher J Bohlen
    •  & Grégory Scherrer
  5. Stanford University Neuroscience Graduate Program, Stanford, California, USA.

    • Elizabeth I Sypek
  6. Stanford University Biology Graduate Program, Stanford, California, USA.

    • Jasmine R Dickinson
  7. Anesthesiology Service, Veteran's Affairs Palo Alto Health Care System, Palo Alto, California, USA.

    • J David Clark
  8. Department of Neurobiology, Stanford University, Stanford, California, USA.

    • Ben A Barres
    •  & Christopher J Bohlen

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Contributions

G.C. and J.R.D. performed behavioral pharmacology. G.C., V.L.T., D.W., E.I.S., S.A.L. and C.S. performed histology. D.W. performed spinal cord slice electrophysiology. E.I.S., C.J.B. and B.A.B. designed and performed RNA transcriptome sequencing. J.D.C. provided critical input on study design and interpretation. G.C., V.L.T., D.W., E.I.S., J.R.D. and G.S. designed studies and wrote the manuscript. G.S., G.C., V.L.T. and D.W. conceived the project, and G.S. supervised all experiments. All of the authors contributed to data analysis, interpretation and editing of the manuscript.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Grégory Scherrer.

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https://doi.org/10.1038/nm.4262

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