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The maintenance of cisplatin- and paclitaxel-induced mechanical and cold allodynia is suppressed by cannabinoid CB2 receptor activation and independent of CXCR4 signaling in models of chemotherapy-induced peripheral neuropathy

Liting Deng123, Josée Guindon4, V Kiran Vemuri5, Ganesh A Thakur5, Fletcher A White6, Alexandros Makriyannis5 and Andrea G Hohmann134*

Author Affiliations

1 Program in Neuroscience, Indiana University, Bloomington, IN, USA

2 Department of Molecular and Cellular Biochemistry, Indiana University, Bloomington, IN, USA

3 Interdisciplinary Biochemistry Graduate Program, Indiana University, Bloomington, IN, USA

4 Department of Psychological and Brain Sciences, Indiana University, Bloomington, IN, USA

5 Center for Drug Discovery, Bouvé College of Health Sciences, Northeastern University, Boston, MA, USA

6 Department of Anesthesia, Indiana University School of Medicine, Indianapolis, IN, USA

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Molecular Pain 2012, 8:71  doi:10.1186/1744-8069-8-71

The electronic version of this article is the complete one and can be found online at: http://www.molecularpain.com/content/8/1/71


Received:30 May 2012
Accepted:19 September 2012
Published:22 September 2012

© 2012 Deng et al.; licensee BioMed Central Ltd.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License ( http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

Background

Chemotherapeutic agents produce dose-limiting peripheral neuropathy through mechanisms that remain poorly understood. We previously showed that AM1710, a cannabilactone CB2 agonist, produces antinociception without producing central nervous system (CNS)-associated side effects. The present study was conducted to examine the antinociceptive effect of AM1710 in rodent models of neuropathic pain evoked by diverse chemotherapeutic agents (cisplatin and paclitaxel). A secondary objective was to investigate the potential contribution of alpha-chemokine receptor (CXCR4) signaling to both chemotherapy-induced neuropathy and CB2 agonist efficacy.

Results

AM1710 (0.1, 1 or 5 mg/kg i.p.) suppressed the maintenance of mechanical and cold allodynia in the cisplatin and paclitaxel models. Anti-allodynic effects of AM1710 were blocked by the CB2 antagonist AM630 (3 mg/kg i.p.), but not the CB1 antagonist AM251 (3 mg/kg i.p.), consistent with a CB2-mediated effect. By contrast, blockade of CXCR4 signaling with its receptor antagonist AMD3100 (10 mg/kg i.p.) failed to attenuate mechanical or cold hypersensitivity induced by either cisplatin or paclitaxel. Moreover, blockade of CXCR4 signaling failed to alter the anti-allodynic effects of AM1710 in the paclitaxel model, further suggesting distinct mechanisms of action.

Conclusions

Our results indicate that activation of cannabinoid CB2 receptors by AM1710 suppresses both mechanical and cold allodynia in two distinct models of chemotherapy-induced neuropathic pain. By contrast, CXCR4 signaling does not contribute to the maintenance of chemotherapy-induced established neuropathy or efficacy of AM1710. Our studies suggest that CB2 receptors represent a promising therapeutic target for the treatment of toxic neuropathies produced by cisplatin and paclitaxel chemotherapeutic agents.

Keywords:
Endocannabinoid; Cannabilactone; AM1710; Chemotherapy; Neuropathic pain; Chemokine; CXCR4; Mechanical allodynia; Cold allodynia; Hyperalgesia

Background

More than half of cancer patients are treated with chemotherapeutic agents (e.g. platinum-based compounds (cisplatin), taxanes (paclitaxel) and vinca alkaloids (vincristine)), of which about 30-40% patients develop neuropathic pain [1-4]. Chemotherapy-induced neuropathy is dose-limiting and is the major toxicity responsible for discontinuation of chemotherapy [3,5-7]. Severe peripheral neuropathy can occur at the early stage of chemotherapy and persist for years after cessation of treatment [8]. Sensory abnormalities (such as tingling, numbness) as well as shooting and burning pain due to chemotherapy can impair the quality of life in patients [2]. To date, no medication has been recognized to effectively and safely cure chemotherapy-induced neuropathy [6,9,10].

Cannabinoids suppress pain through activation of cannabinoid CB1 and/or CB2 receptors [11]. Cannabis-based medicines, such as Cesamet® (nabilone) or Sativex® (mixture of Δ9-tetrahydrocannabinol and non-psychoactive cannabidiol), are already used clinically to manage neuropathic pain [12,13]. However, cannabinoids may produce unwanted central nervous system (CNS) side effects associated with CB1 receptors. Efficacy of cannabis medicines in treating chemotherapy-induced neuropathy has yet to be fully evaluated [13]. A small number of preclinical studies have reported a role of CB2-selective agonists in suppressing chemotherapy-evoked neuropathic pain [14-17]. In these studies, CB2-selective agonists suppressed paclitaxel- or vincristine-induced mechanical allodynia [14-18]. Whether CB2 selective agonists suppress cold allodynia after development of chemotherapy-induced neuropathic pain remains poorly understood and effects of CB2-selective agonists on cisplatin-induced neuropathy are unknown.

AM1710, a cannabilactone CB2 agonist with limited blood brain barrier penetration [19], exhibits 54-fold selectivity for CB2 over CB1 receptors [20]. We previously showed that AM1710 produces antinociception in the plantar test in naïve animals without producing CNS side effects, such as hypothermia, hypoactivity, tail-flick antinociception or motor ataxia at doses 100 times higher than the lowest maximally effective antinociceptive dose [19]. In the present study, we evaluated effects of AM1710 (Figure  1) in two distinct animal models of chemotherapy-induced neuropathy (cisplatin and paclitaxel models) and characterized its mechanism of action. Pharmacological specificity was established using cannabinoid CB2 (AM630) and CB1 (AM251) antagonists (Figure  1).

thumbnailFigure 1. Chemical stuctures of compounds employed. Chemical structure of the cannabilactone CB2 agonist AM1710, the CXCR4 antagonist AMD3100, the CB2 receptor antagonist AM630 and the CB1 receptor antagonist AM251.

Mechanisms underlying chemotherapy-induced neuropathy remain poorly understood [21]. An emerging body of literature implicates a role for chemokine stromal derived factor-1 (SDF-1/CXCL12) and its receptor CXCR4 in mechanisms of other distinct neuropathic pain states [22]. For instance, blockade of CXCR4 signaling by its antagonist AMD3100 reversed the maintenance of neuropathic pain induced by either chronic constriction injury (CCI) of the sciatic nerve [23] or HIV-associated neuropathy [24,25]. However, whether CXCR4 signaling is also involved in chemotherapy-induced neuropathy has not been studied. In the present study, we investigated the role of CXCR4 signaling in established chemotherapy-induced neuropathic pain and examined its potential interaction with CB2 signaling.

Results

Established chemotherapy-induced neuropathy

Prior to cisplatin or paclitaxel treatment, there were no differences between groups in either paw withdrawal thresholds to mechanical stimulation or paw withdrawal frequencies to cold stimulation (P > 0.15 for all studies).

Cisplatin or paclitaxel treatment established and maintained neuropathic states characterized by hypersensitivities to mechanical and cold stimulation. Cisplatin decreased mechanical paw withdrawal thresholds (F1,40 = 1565.23, P < 0.0001; Figure  2A) and increased cold withdrawal frequencies (F1,40 = 632.24, P < 0.0001; Figure  2C). Mechanical (P < 0.0001) and cold (P < 0.0001) allodynia were maintained from day 4 to day 28 in cisplatin-treated group (Figure  2A and C). Similarly, paclitaxel decreased paw withdrawal thresholds to mechanical stimulation (F1,79 = 290.19, P < 0.0001; Figure  2B) and increased frequencies of withdrawal to cold stimulation (F1,79 = 37.11, P < 0.0001; Figure  2D). Mechanical (P < 0.0001) and cold (P < 0.03) allodynia were present from day 4 to day 20 in the paclitaxel-treated group (Figure  2B and D).

thumbnailFigure 2. Time course of development of chemotherapy-induced peripheral neuropathy evoked by cisplatin and paclitaxel treatment. Mechanical (A, B) and cold (C, D) allodynia developed following cisplatin (A, C) or paclitaxel (B, D) treatment. Arrows show timing of injections of chemotherapeutic agents. Data are expressed as mean ± SEM (paclitaxel, n = 76; cremophor, n = 5; cisplatin, n = 36; saline, n = 6). ***P < 0.001, **P < 0.01, *P < 0.05 vs. vehicle. Non-chemotherapy controls received saline in lieu of cisplatin (A, C) or cremophor vehicle in lieu of paclitaxel (B, D), repeated measures ANOVA and One-way ANOVA at each time point.

AM1710 suppressed the maintenance of mechanical and cold allodynia produced by either cisplatin or paclitaxel treatment

The cannabilactone AM1710 (0.1, 1 and 5 mg/kg i.p.) suppressed cisplatin-evoked mechanical (F4,21 = 547.02, P < 0.0001) and cold (F4,21 = 59.10, P < 0.0001) allodynia compared to vehicle treatment (Figure  3A and C). AM1710 (1 or 5 mg/kg i.p.) fully reversed cisplatin-evoked neuropathy and normalized responses to pre-drug levels for both modalities (P = 0.10 mechanical, Figure  3A; P = 0.17 cold, Figure  3C). The lowest dose of AM1710 (0.1 mg/kg i.p.) suppressed mechanical (P < 0.0001 vs. 1 or 5 mg/kg i.p.) and cold (P < 0.002 vs. 1 or 5 mg/kg i.p.) allodynia to a lesser extent than either the middle or the high doses at each time point.

thumbnailFigure 3. Effect of AM1710 on chemotherapy-induced mechanical and cold allodynia. AM1710 suppressed both mechanical (A, B) and cold (C, D) allodynia evoked by cisplatin (A, C) or paclitaxel (B, D) treatment. Data are expressed as mean ± SEM (n = 5-13 per group). ***P < 0.001, **P < 0.01, *P < 0.05 vs. DMSO vehicle, One-way ANOVA followed by Dunnett post hoc test. xxxP < 0.001 vs. AM1710 (1 and 5 mg/kg i.p.), One-way ANOVA followed by Bonferroni post hoc test. +P < 0.05 vs. DMSO vehicle, planned comparison t-test. $$P < 0.01, $P < 0.05 vs. pre-drug baseline (−60 min), paired t-test. ###P < 0.001 vs. pre-cisplatin/paclitaxel baseline (BL), repeated measures ANOVA.

AM1710 produced time-dependent attenuations of cisplatin-evoked mechanical (F16,84 = 62.38, P < 0.0001) and cold (F16,84 = 15.52, P < 0.0001) allodynia (Figure  3A and C). Anti-allodynic effects of AM1710 on cisplatin-evoked mechanical (P < 0.0001) and cold (P < 0.02) responsiveness lasted at least 150 min post injection. AM1710 failed to alter responsiveness to mechanical (P = 0.13) or cold (P = 0.94) stimulation in animals treated with saline vehicle in lieu of cisplatin (Figure  3A and C).

AM1710 (1 and 5 mg/kg i.p.) also suppressed paclitaxel-evoked mechanical (F4,37 = 13.75, P < 0.0001) and cold (F4,37 = 4.41, P < 0.01) allodynia compared to the vehicle group (Figure  3B and D). The low dose of AM1710 (0.1 mg/kg i.p.) suppressed paclitaxel-evoked mechanical (P < 0.05) allodynia but did not reliably attenuate cold allodynia (P = 0.44). AM1710 produced time-dependent suppressions of paclitaxel-induced mechanical (F12,111 = 7.09, P < 0.0001) and cold (F12,111 = 3.15, P < 0.001) hypersensitivities. Suppression of mechanical allodynia was observed relative to vehicle (Figure  3B) throughout the 180 min post injection observation interval (P < 0.05). AM1710 (1–5 mg/kg i.p.) attenuated paclitaxel-evoked cold allodynia relative to vehicle at 30 min post injection (P < 0.04; Figure  3D). AM1710 (5 mg/kg i.p.) also reliably attenuated paclitaxel-evoked cold allodynia relative to pre-drug baseline levels at both 30 (P < 0.03) and 180 (P < 0.04) min post injection (Figure  3D). AM1710 failed to alter responsiveness to mechanical (P = 0.06) or cold (P = 0.72) stimulation in animals treated with cremophor vehicle in lieu of paclitaxel (Figure  3B and D).

To examine the duration of action of AM1710 (5 mg/kg i.p.), a subset of paclitaxel-treated animals was tested at 6 and 24 h post injection. AM1710 reliably suppressed mechanical (P < 0.0001) and cold (P < 0.03) allodynia over 180 min post injection (Figure  4B and D). Mechanical (P = 0.43) and cold (P = 0.76) allodynia was reinstated 6 h post injection (Figure  4B and D). Similarly, no evidence for anti-allodynic efficacy of AM1710 (0.1-5 mg/kg i.p.) was found at 6 h following injection in the subset of cisplatin-treated animals used to further characterize the time course of AM1710 antinociception (data not shown).

thumbnailFigure 4. The cannabilactone AM1710 suppresses chemotherapy-induced mechanical and cold allodynia through a CB2-specific mechanism. AM1710-induced suppressions of cisplatin- (A, C) and paclitaxel- (B, D) evoked mechanical (A, B) and cold (C, D) allodynia were blocked by the CB2 antagonist AM630 (3 mg/kg i.p.) but not the CB1 antagonist AM251 (3 mg/kg i.p.). Data are expressed as mean ± SEM (n = 5-6 per group). ***P < 0.001, **P < 0.01, *P < 0.05 vs. DMSO vehicle, One-way ANOVA followed by Dunnett post hoc test. xxxP < 0.01, xxP < 0.01, xP < 0.05 vs. AM1710 (5 mg/kg i.p.), Bonferroni post hoc test. $P < 0.05 vs. AM1710 (5 mg/kg i.p.), planned t-test. ###P < 0.001 vs. pre-cisplatin/paclitaxel baseline (BL), repeated measures ANOVA.

Anti-allodynic effects of AM1710 were mediated by cannabinoid CB2 receptors

To evaluate pharmacological specificity, the highest behaviorally active dose of AM1710 (5 mg/kg i.p.) was co-administered with either the CB2 antagonist AM630 (3 mg/kg i.p.) or the CB1 antagonist AM251 (3 mg/kg i.p.) in cisplatin- or paclitaxel-treated animals.

Anti-allodynic effects of AM1710 on cisplatin-evoked mechanical (F3,22 = 311.81, P < 0.0001) and cold (F3,22 = 39.03, P < 0.0001) hypersensitivities were blocked by the CB2 antagonist AM630 throughout the 150 min post injection observation interval (P < 0.0001 mechanical and P < 0.0001 cold; Figure  4A and C). By contrast, the CB1 antagonist AM251 failed to block the mechanical (P = 1.00) and cold (P = 1.00) anti-allodynic effects of AM1710 (Figure  4A and C).

Similarly, in the paclitaxel model, the CB2 antagonist AM630 blocked the AM1710-induced suppressions of mechanical (F3,17 =12.73, P < 0.0001) and cold (F3,17 = 3.20, P < 0.05) allodynia from 30 to 180 min post injection (P < 0.001 mechanical, Figure  4B; and P < 0.03 cold, Figure  4D). Hypersensitivities to mechanical (P = 0.87) and cold (P = 0.41) stimulation were reinstated by 6 h post injection (Figure  4B and D). By contrast, the CB1 antagonist AM251 failed to block the AM1710-induced suppressions of mechanical (P = 1.00) and cold (P = 1.00) allodynia (Figure  4B and D).

Antagonist treatment alone failed to alter nociceptive thresholds in cisplatin- or paclitaxel-treated animals. Neither the CB2 antagonist AM630 (3 mg/kg i.p.) nor the CB1 antagonist AM251 (3 mg/kg i.p.), administered alone, altered mechanical paw withdrawal thresholds (F2,13 = 0.38, P = 0.69) or cold withdrawal frequencies (F2,13 = 3.32, P = 0.07) relative to vehicle in cisplatin-treated animals at any time point (F8,52 =1.35, P = 0.24 mechanical, Figure  5A; F8,52 =1.10, P = 0.38 cold, Figure  5C). Similarly, the same doses of AM630 and AM251, administered alone, failed to alter paclitaxel-evoked mechanical (F2,19 = 0.89, P = 0.43) and cold (F2,19 = 0.88, P = 0.43) allodynia relative to vehicle at any time point (F6,57 =1.19, P = 0.33 mechanical, Figure  5B; F6,57 =1.44, P = 0.22 cold, Figure  5D).

thumbnailFigure 5. Pharmacological blockade of CB2 or CB1 receptors does not alter cisplatin- or paclitaxel-induced neuropathic nociception. Neither the CB2 antagonist AM630 (3 mg/kg i.p.) nor the CB1 antagonist AM251 (3 mg/kg i.p.) altered cisplatin- (A, C) or paclitaxel- (B, D) evoked mechanical (A, B) and cold (C, D) allodynia. ###P < 0.001 vs. baseline (BL) prior to chemotherapy treatment, repeated measures ANOVA.

Blockade of CXCR4 signaling with AMD3100 failed to reverse established chemotherapy-evoked neuropathy

We asked whether blockade of CXCR4 signaling with its antagonist AMD3100 would reverse established neuropathy induced by cisplatin and paclitaxel treatment. Whereas AM1710 (5 mg/kg i.p.) attenuated mechanical and cold allodynia in both neuropathy models, AMD3100 (10 mg/kg i.p.) failed to do so. In the cisplatin model, AMD3100 (10 mg/kg i.p.) failed to alter mechanical (P = 0.97, P = 0.99 and P = 0.59 at 30, 90 and 150 min, respectively) or cold (P = 1.00, P = 1.00 and P = 0.84 at 30, 90 and 150 min, respectively) allodynia relative to vehicle (Figure  6A and C). Similarly, in paclitaxel-treated animals, the same dose of AMD3100 failed to alter mechanical withdrawal thresholds (P = 0.93 and P = 0.99 at 30 and 180 min, respectively) or cold withdrawal frequencies (P = 0.13 and P = 1.00 at 30 and 180 min, respectively) compared to vehicle (Figure  6B and D).

thumbnailFigure 6. CXCR4 signaling did not contribute to either chemotherapy-induced neuropathy or CB2 agonist efficacy. The CXCR4 antagonist AMD3100 (10 mg/kg i.p.) failed to suppress the maintenance of cisplatin- (A, C) or paclitaxel- (B, D) evoked mechanical (A, B) and cold (C, D) allodynia. The CXCR4 antagonist AMD3100 (10 mg/kg i.p.) did not alter antinociceptive efficacy of the CB2 agonist AM1710 (5 mg/kg i.p.) in suppressing paclitaxel-induced mechanical (B) and cold (D) allodynia. ***P < 0.001, **P < 0.01, *P < 0.05 vs. DMSO vehicle, One-way ANOVA followed by Dunnett post hoc test. +P < 0.05 vs. DMSO vehicle, planned t-test. xxxP < 0.001, xxP < 0.01, xP < 0.05 vs. AM1710 (5 mg/kg), Bonferroni post hoc test. $$P < 0.01, $P < 0.05 vs. pre-drug baseline (−60 min), paired t-test. ###P < 0.001 vs. baseline (BL) prior to cisplatin/paclitaxel treatment, repeated measures ANOVA.

Blockade of CXCR4 signaling with AMD3100 failed to enhance CB2 agonist efficacy

We asked whether blockade of CXCR4 signaling in the paclitaxel model would enhance the efficacy of a CB2 agonist by assessing the effects of co-administration of AMD3100 (10 mg/kg i.p.) with a sub-maximal dose of AM1710 (5 mg/kg i.p.). Responsiveness to mechanical (P = 0.61 and P = 0.23 at 30 and 180 min, respectively) and cold (P = 1.00 and P = 0.86 at 30 and 180 min, respectively) stimulations did not differ in animals receiving AM1710 in either the presence or absence of AMD3100 (Figure  6B and D).

Discussion

To our knowledge, this is the first study to demonstrate antinociceptive efficacy of a CB2-selective agonist in the cisplatin model and the first to evaluate a potential role for CXCR4 signaling in contributing to mechanisms of chemotherapy-induced peripheral neuropathy. Our studies demonstrate that activation of cannabinoid CB2 receptors suppresses both mechanical and cold allodynia induced by either cisplatin or paclitaxel treatment. Effects of the cannabilactone CB2-selective agonist AM1710 on established neuropathy induced by chemotherapeutic treatment have not previously been evaluated. We recently showed that systemic administration of AM1710 in naïve animals produces antinociception to heat, but not to mechanical stimulation, in the plantar test in otherwise naïve animals [19]. Suppression of thermal nociception was also observed following local (i.paw) administration of AM1710 [20]. In the present study, we extended our investigation of the antinociception profile of AM1710 by showing that AM1710 suppressed both mechanical and cold allodynia in two distinct models of chemotherapy-induced neuropathy. Strikingly, anti-allodynic effects of AM1710 were observed at low doses (0.1 mg/kg i.p.) and lasted at least 2.5-3 h following systemic administration. In our previous work, higher doses of AM1710 (10 mg/kg i.p.; a dose 100 times higher than the lowest effective antinociceptive dose identified here) failed to produce CNS side-effects in otherwise naïve animals [19]. These observations suggest that AM1710 exhibits a very favorable efficacy to toxicity ratio.

In the present study, AM1710-induced suppressions of chemotherapy-induced mechanical and cold allodynia were completely blocked by the CB2 antagonist AM630 (3 mg/kg i.p.), but not the CB1 antagonist AM251 (3 mg/kg i.p.), following either cisplatin or paclitaxel treatment. Thus, selective activation of cannabinoid CB2 receptors attenuates the maintenance of neuropathic pain induced by diverse chemotherapeutic agents. These findings are consistent with other studies showing that CB2 selective agonists are antinociceptive in animal models of toxic neuropathies [14-17,26]. In particular, AM1714, which also belongs to the cannabilactone class of CB2 selective agonists [20], suppresses paclitaxel-induced mechanical allodynia via a CB2-specific mechanism [14]. In this latter study, responses to cold stimulation were not previously characterized [14]. CB2 receptors are found primarily, although not exclusively, in cells of the immune system and reside centrally at low levels relative to CB1 receptors [27,28]; however, CB2 receptor expression is highly inducible in response to injury (for review see [29]). CB2 receptors have also been localized to primary afferent terminals in humans [30]. Thus, it may be possible to target the cannabinoid CB2 signaling system to produce antinociception without adverse CNS side effects associated with CB1 receptors (e.g. HU308 [31], AM1241 [32], and AM1710 [19,33]). Our studies suggest that cannabinoid CB2 receptors represent a promising target for the treatment of toxic neuropathies induced by chemotherapeutic agents.

Cisplatin, a platinum-based compound, produces sensory axonal nerve damage and paresthesias in the distal extremities in humans [34,35]. Paclitaxel belongs to the taxane class of chemotherapeutic agents and also produces peripheral nerve damage and sensory neuropathies such as numbness, tingling and burning pain in patients [36]. Different mechanisms may underly the maintenance of neuropathy induced by different classes of chemotherapeutic agents (for review see [21]); however, similarities are also shared between the models. For example, glutamatergic neurotransmission and N-methyl-D-aspartate (NMDA) receptors are involved in both cisplatin- and paclitaxel-induced neuropathic pain [21]. Peripheral nerve damage results in glutamate/NMDA receptor-mediated sensitization and spontaneous activity of primary afferents, and causes hyper-excitability of dorsal horn neurons [37]. Decreased glutamate levels facilitate nerve conduction in the cisplatin and paclitaxel models [38]. The NMDA receptor antagonist ketamine also produces antinociceptive effects in paclitaxel-treated rats [39]. Paclitaxel treatment also down-regulates glial glutamate transporters (i.e. GLAST and GLT-1) in the spinal dorsal horn [40]. The transient receptor potential channel family is also implicated in mechanisms of nociception in both models. Cisplatin increases transient receptor potential vanilloid 1 (TRPV1) and transient receptor potential ankyrin 1 (TRPA1) expression levels and nociceptor responsiveness in dorsal root ganglion (DRG) neurons [41]. Mechanical hypersensitivity is preserved in cisplatin-treated animals lacking TRPV1 [41] and reduced in paclitaxel-treated animals lacking TRPV4 [42]. In addition, according to the mitotoxicity hypothesis, both cisplatin and paclitaxel induce morphological changes (swollen and vacuolated mitochondria) and dysfunction (reduced respiration and energy production) of mitochondria in axons, which then alters intracellular calcium levels and initiates apoptosis pathways [43-46]. CB2 agonists may interfere with pro-nociceptive signaling cascades (i.e. NMDA, TRPV1, TRPA1) activated by the chemotherapy. More work is necessary to determine whether CB2 agonists such as AM1710 directly reduce nociceptor excitability (i.e. at the level of the primary afferent terminal or DRG) and/or attenuate the mitotoxicity and structural damage to DRG or axons that are produced by chemotherapy and result in aberrant neuronal excitability [47-50].

In the present study, AM1710 suppressed the hypersensitivities induced by both cisplatin and paclitaxel through a CB2-specific mechanism, suggesting a shared role for CB2 in modulating hypersensitivity in both models. CB2 modulation of chemotherapy-induced neuropathy may result from suppression of central sensitization. In animal models of inflammatory pain and nerve injury, CB2 agonists (such as AM1241 and JWH133) decrease windup and central sensitization [48] as well as mechanically-evoked responses [49,50] in spinal dorsal horn neurons (for review see [29]). Peripheral nerve injury also leads to secretion of chemokines, increased release of proinflammatory cytokines, and increased activation of microglia and astrocytes, which facilitate responses to noxious stimulations and contribute to central sensitization (for review see [51]). Hence, it is possible that activation of CB2 receptors attenuates chemotherapy-induced neuropathy by interfering with astrocyte and/or microglial activation and pro-inflammatory cytokine signaling [16,40,52,53].

To better explore the maintenance of chemotherapy-induced neuropathic pain and its modulation by CB2 agonists, we investigated the possible contribution of the alpha-chemokine receptor CXCR4 to cisplatin and paclitaxel-induced neuropathies. CXCR4 signaling has been implicated in the mechanisms underlying several neuropathic pain states. Notably, intradermal injection of SDF-1α in rats produces onset tactile allodynia, suggesting a direct role in pain [54]. Blockade of CXCR4 signaling by its antagonist AMD3100 also suppresses established mechanical allodynia in HIV-associated peripheral neuropathy [24,25] and reverses heat hyperalgesia, but not mechanical allodynia, induced by chronic constriction injury of sciatic nerve (CCI) [23]. Contrary to HIV-associated neuropathy, blockade of CXCR4 signaling by AMD3100 in our study did not inhibit the maintenance of mechanical or cold allodynia evoked by either cisplatin or paclitaxel. Thus, CXCR4 signaling is unlikely to contribute to the maintenance of neuropathic pain induced by chemotherapeutic agents. Moreover, chemotherapy-induced neuropathy is thus likely to recruit pain mechanisms distinct from traumatic nerve injury (e.g. CCI) and HIV-associated neuropathy. Physiological studies indicate that neuropathic pains induced by traumatic nerve injury produce axonal degeneration with an increased discharge in A-fiber and C-fiber nociceptors [55,56]. By contrast, this degeneration is not observed in animals treated with paclitaxel or vincristine [43,57,58], although hypersensitivities of C-fiber nociceptors are nonetheless observed [59,60]. Indeed, paclitaxel increases spontaneous discharge in both A-fibers and C-fibers [61]. Diverse second messengers, including protein kinase Cɛ and protein kinase A, also contribute to the maintenance of paclitaxel-induced hyperalgesia. For example, intradermal injection of antagonists for protein kinase A attenuates hyperalgesia evoked by both acute and chronic paclitaxel treatments [62]. More work is necessary to elucidate mechanisms of chemotherapy-induced neuropathic pain at the molecular and neurophysiological levels and characterize effects of CB2 agonists such as AM1710 on nociceptor excitability in these models.

Our study is the first to test the hypothesis that CB2 modulation of chemotherapy-induced neuropathy may interact with CXCR4 signaling. Several publications have suggested that CXCR4 signaling crosstalks with the cannabinoid system. Behavioral and physiological studies suggest that both antinociceptive and hypothermic effects of the mixed CB1/CB2 agonist WIN55,212-2 is inhibited by CXCR4 activation with SDF-1α [63,64]. The interaction between the CXCR4 and the cannabinoid CB2 receptor signaling also modulates chemotaxis of CD4+ T lymphocytes [65] as well as growth and metastasis of breast cancer [66]. Although there is support for spinal cord-infiltrating CD4+ T lymphocytes in contributing to neuropathic pain due to spinal nerve transection injury [67], chemotherapy-induced neuropathy does not appear to be influenced by either this cell type or the CXCR4 antagonist AMD3100. Furthermore, blockade of CXCR4 signaling did not reliably enhance (or inhibit) the anti-allodynic efficacy of AM1710. These results imply that CXCR4 signaling does not contribute to CB2-modulation of established chemotherapy-induced neuropathy. More work is necessary to determine whether CXCR4 signaling contributes to the development of chemotherapy-induced neuropathy.

Conclusions

In conclusion, the present study demonstrates that selective activation of cannabinoid CB2 receptors suppresses neuropathic nociception to multiple stimulus modalities that is evoked by different classes of chemotherapeutic agents. The cannabilactone CB2 selective agonist AM1710 produces CB2-mediated suppressions of mechanical and cold allodynia induced by chemotherapeutic treatment with cisplatin or paclitaxel. In addition, our data indicate that neither the maintenance of chemotherapy-induced neuropathy nor the anti-allodynic efficacy of CB2 agonist is dependent upon CXCR4 signaling.

Methods

Subjects

One hundred and thirty-seven adult male Sprague–Dawley rats (Harlan, Indianapolis, IN, USA), weighing 305 to 400 g, were used in these experiments. All procedures were approved by Bloomington Institutional Animal Care and Use Committee (BIACUC) of Indiana University Bloomington and followed the guidelines for the treatment of animals of the International Association for the Study of Pain [68]. All animals were single housed in a temperature-controlled facility, with food and water ad libitum. Animals were maintained on a regular 12 h light/ 12 h dark cycle (lights on at 7 am).

Drugs and chemicals

Cisplatin was purchased from Tocris Bioscience (Ellisville, MO, USA) and was dissolved in saline (0.9% sodium chloride). Paclitaxel was obtained from Tecoland Corporation (Edison, NJ, USA) and was dissolved in cremophor vehicle (1:1:4 ratio of cremophor® EL/ ethanol/ saline). AM1710, AM630 and AM251 were synthesized by the Makriyannis laboratory. AMD3100 octahydrochloride hydrate was purchased from Sigma-Aldrich (St. Louis, MO, USA). AM1710, AM630, AM251 and AMD3100 were dissolved in 100% dimethyl sulfoxide (DMSO). DMSO, cremophor® EL and acetone were purchased from Sigma-Aldrich (St. Louis, MO, USA). Saline was purchased from Aqualite System (Lake Forest, IL, USA).

General experimental protocol

All experiments were conducted double-blinded with animals randomly assigned into groups. Cisplatin and paclitaxel were used to produce chemotherapy-induced neuropathy. Cisplatin (3 mg/kg i.p.) or saline vehicle was injected four times once weekly [69] in a volume of 10 ml/kg. Cisplatin/saline-treated animals were assessed for mechanical paw withdrawal thresholds and cold withdrawal frequencies every four days. Paclitaxel (2 mg/kg i.p.) or cremophor EL: ethanol: saline (1: 1: 4) vehicle was administered to rats four times every two days [70] in a volume of 1 ml/kg. Animals with paclitaxel/cremophor treatment were assessed for paw withdrawal thresholds to mechanical stimulation every two days and paw withdrawal frequencies to cold stimulation every four days. On the days animals received cisplatin/saline or paclitaxel/cremophor treatments, behavioral testing was performed prior to pharmacological manipulations.

Effects of pharmacological manipulations on mechanical and cold allodynia were assessed on day 28 in animals receiving cisplatin/saline treatments or day 20 in paclitaxel/cremophor-treated animals. On the test days, animals received either vehicle (DMSO), AM1710 either alone or in combination with the CB2 antagonist AM630 or the CB1 antagonist AM251, or the CXCR4 antagonist AMD3100. Withdrawal thresholds to mechanical stimulation and withdrawal frequencies to cold stimulation were measured before drug administration (−60 min) and at 30, 90, 150 min post drug administration in cisplatin/saline-treated animals, or at 30 min and 3 h post drug in paclitaxel/cremophor-treated animals. A subset of cisplatin- and paclitaxel-treated animals was additionally tested at 6 h and 24 h post drug administration.

In Experiments 1 and 2, antinociceptive effects of AM1710 in chemotherapy-induced neuropathy evoked by cisplatin or paclitaxel treatments were studied. Effects of AM1710 (0.1, 1 or 5 mg/kg i.p.) [19] or vehicle were assessed in animals receiving cisplatin or paclitaxel treatment. The high dose of AM1710 was also administered to animals that received saline or cremophor-vehicle in lieu of cisplatin or paclitaxel, respectively. To further evaluate the duration of action of the compound, a subset of paclitaxel-treated animals receiving AM1710 (5 mg/kg i.p.) or DMSO vehicle were tested from 30 min to 24 h post injection. Pharmacological specificity of anti-allodynic effects of AM1710 was assessed in both models by co-administering AM1710 (5 mg/kg i.p.) with the CB2 antagonist AM630 (3 mg/kg i.p.) [71] or CB1 antagonist AM251 (3 mg/kg i.p.) [72]. Separate groups received AM630 (3 mg/kg i.p.) or AM251 (3 mg/kg i.p.) alone. In Experiments 3 and 4, the CXCR4 antagonist AMD3100 (10 mg/kg i.p.) [73] was administered to animals to examine the impact of blockade of CXCR4 signaling on established neuropathy produced by cisplatin or paclitaxel treatment. AMD3100 (10 mg/kg i.p.) was administered to paclitaxel-treated animals either in absence or presence of AM1710 (5 mg/kg i.p.) to evaluate whether blockade of CXCR4 signaling would enhance CB2 agonist efficacy.

Assessment of paw withdrawal thresholds to mechanical stimulation

Paw withdrawal thresholds to mechanical stimulation were measured using an electronic von Frey anesthesiometer (IITC model Alemo 2390–5, Woodland Hills, CA) as described previously [14]. Animals were placed on an elevated metal mesh table and habituated under inverted transparent plastic cages individually for at least 15 min prior to testing. After habituation to the testing environment (i.e. when animals ceased exploratory behaviors), a force was applied to the midplantar region of the hind paw by a rigid tip connected to the anethesiometer. Mechanical stimulation was terminated when the animal withdrew its paw and the value of the force was recorded in units of grams. Mechanical paw withdrawal thresholds were measured in duplicate for each paw and were reported as the mean of duplicate determinations averaged across paws.

Assessment of paw withdrawal frequencies to cold stimulation

Paw withdrawal frequencies to cold stimulation were measured in the same animals assessed for mechanical hypersensitivity using the acetone method [74]. Rats were placed underneath inverted plastic cages on an elevated metal mesh table, the same environment used in the mechanical tests. After habituation, an acetone bubble that formed at the end of a blunt one C.C. syringe was gently presented to the plantar surface of the hind paw with care that application of acetone did not inadvertently result in mechanical stimulation of the paw with the syringe hub. Animals were observed for 20 seconds after acetone application. Acetone was applied to each paw of the animal 5 times alternately with a 3 min interval between applications. Paw withdrawal on a given trial was deemed present if animals showed one or more forms of unilateral nocifensive behavior. Trials on which an animal did not show unilateral behavior during the observation time were counted as zero. Unilateral behaviors that qualified as nocifensive behavior included withdrawing, raising, licking, shaking or repetitive stepping on the stimulated paw. Paw withdrawal frequencies were recorded as the percentage of the total number of occurrences of paw withdrawal out of 10 trials per animal.

Statistical analyses

Paw withdrawal thresholds (mechanical) and frequencies (cold) were calculated for each paw and averaged. Analysis of variance (ANOVA) for repeated measures was used to determine the time course of paclitaxel and cisplatin-induced neuropathy as well as drug effects. One-way ANOVA was used to identify the source of significant interactions at each time point, followed by Dunnett post hoc tests (for comparisons to vehicle), Bonferroni post hoc tests (for comparisons between groups). A priori comparisons were also made using planned comparison and paired t-tests, as appropriate. All statistical analyses were performed using IBM-SPSS Statistics version 19.0 (SPSS inc., an IBM company, Chicago, IL, USA). P < 0.05 was considered statistically significant.

Abbreviations

ANOVA: Analysis of variance; CB1: Cannabinoid receptor 1; CB2: Cannabinoid receptor 2; CCI: Chronic constriction injury; CNS: Central nervous system; CXCR4: Alpha-chemokine receptor; DMSO: Dimethylsulfoxide; DRG: Dorsal root ganglion; GLAST: Glutamate aspartate transporter; GLT-1: Glutamate transporter 1; HIV: Human immunodeficiency virus; i.p: Intraperitoneal; NMDA: N-methyl-D-aspartate; SDF-1/CXCL12: Chemokine stromal derived factor-1; s.c: Subcutaneous; TRPA: Transient receptor potential ankyrin; TRPV: Transient receptor potential vanilloid.

Competing interests

The authors declared that they have no competing interest.

Authors’ contributions

LD and JG contributed equally to this work. LD and JG conducted all behavioral tests, data analyses, and wrote the manuscript. KV synthesized AM630 and AM251 and GAT synthesized AM1710. AM630, AM251 and AM1710 were developed by AM. FAW contributed to the experimental design, provided AMD3100 used in an early study and revised the manuscript. AGH designed the studies, oversaw the project and wrote the manuscript. All authors read and approved the final manuscript.

Acknowledgments

This study was supported by DA028200 and DA021644 (to AGH); DA026040 (to FAW); and DA9158, DA3801 (to AM).

References

  1. Cavaletti G, Alberti P, Frigeni B, Piatti M, Susani E: Chemotherapy-induced neuropathy.

    Curr Treat Options Neurol 2011, 13:180-190. PubMed Abstract | Publisher Full Text OpenURL

  2. Wolf S, Barton D, Kottschade L, Grothey A, Loprinzi C: Chemotherapy-induced peripheral neuropathy: prevention and treatment strategies.

    Eur J Cancer 2008, 44:1507-1515. PubMed Abstract | Publisher Full Text OpenURL

  3. Rowinsky EK, Eisenhauer EA, Chaudhry V, Arbuck SG, Donehower RC: Clinical toxicities encountered with paclitaxel (Taxol).

    Semin Oncol 1993, 20:1-15. PubMed Abstract OpenURL

  4. Windebank AJ, Grisold W: Chemotherapy-induced neuropathy.

    J Peripher Nerv Syst 2008, 13:27-46. PubMed Abstract | Publisher Full Text OpenURL

  5. Cavaletti G, Marzorati L, Bogliun G, Colombo N, Marzola M, Pittelli MR, Tredici G: Cisplatin-induced peripheral neurotoxicity is dependent on total-dose intensity and single-dose intensity.

    Cancer 1992, 69:203-207. PubMed Abstract | Publisher Full Text OpenURL

  6. Uhm JH, Yung WK: Neurologic Complications of Cancer Therapy.

    Curr Treat Options Neurol 1999, 1:428-437. PubMed Abstract | Publisher Full Text OpenURL

  7. Polomano RC, Bennett GJ: Chemotherapy-evoked painful peripheral neuropathy.

    Pain Med 2001, 2:8-14. PubMed Abstract | Publisher Full Text OpenURL

  8. Authier N, Balayssac D, Marchand F, Ling B, Zangarelli A, Descoeur J, Coudore F, Bourinet E, Eschalier A: Animal models of chemotherapy-evoked painful peripheral neuropathies.

    Neurotherapeutics 2009, 6:620-629. PubMed Abstract | Publisher Full Text OpenURL

  9. Pachman DR, Barton DL, Watson JC, Loprinzi CL: Chemotherapy-induced peripheral neuropathy: prevention and treatment.

    Clin Pharmacol Ther 2011, 90:377-387. PubMed Abstract | Publisher Full Text OpenURL

  10. Hohmann AG: A cannabinoid pharmacotherapy for chemotherapy-evoked painful peripheral neuropathy.

    Pain 2005, 118:3-5. PubMed Abstract | Publisher Full Text OpenURL

  11. Guindon J, Hohmann AG: The endocannabinoid system and pain.

    CNS Neurol Disord Drug Targets 2009, 8:403-421. PubMed Abstract | Publisher Full Text | PubMed Central Full Text OpenURL

  12. Leung L: Cannabis and its derivatives: review of medical use.

    J Am Board Fam Med 2011, 24:452-462. PubMed Abstract | Publisher Full Text OpenURL

  13. Pertwee RG: Emerging strategies for exploiting cannabinoid receptor agonists as medicines.

    Br J Pharmacol 2009, 156:397-411. PubMed Abstract | Publisher Full Text | PubMed Central Full Text OpenURL

  14. Rahn EJ, Zvonok AM, Thakur GA, Khanolkar AD, Makriyannis A, Hohmann AG: Selective activation of cannabinoid CB2 receptors suppresses neuropathic nociception induced by treatment with the chemotherapeutic agent paclitaxel in rats.

    J Pharmacol Exp Ther 2008, 327:584-591. PubMed Abstract | Publisher Full Text | PubMed Central Full Text OpenURL

  15. Naguib M, Diaz P, Xu JJ, Astruc-Diaz F, Craig S, Vivas-Mejia P, Brown DL: MDA7: a novel selective agonist for CB2 receptors that prevents allodynia in rat neuropathic pain models.

    Br J Pharmacol 2008, 155:1104-1116. PubMed Abstract | Publisher Full Text | PubMed Central Full Text OpenURL

  16. Naguib M, Xu JJ, Diaz P, Brown DL, Cogdell D, Bie B, Hu J, Craig S, Hittelman WN: Prevention of Paclitaxel-induced neuropathy through activation of the central cannabinoid type 2 receptor system.

    Anesth Analg 2012, 114:1104-1120. PubMed Abstract | Publisher Full Text OpenURL

  17. Xu JJ, Diaz P, Astruc-Diaz F, Craig S, Munoz E, Naguib M: Pharmacological characterization of a novel cannabinoid ligand, MDA19, for treatment of neuropathic pain.

    Anesth Analg 2010, 111:99-109. PubMed Abstract | Publisher Full Text | PubMed Central Full Text OpenURL

  18. Rahn EJ, Makriyannis A, Hohmann AG: Activation of cannabinoid CB1 and CB2 receptors suppresses neuropathic nociception evoked by the chemotherapeutic agent vincristine in rats.

    Br J Pharmacol 2007, 152:765-777. PubMed Abstract | Publisher Full Text | PubMed Central Full Text OpenURL

  19. Rahn EJ, Thakur GA, Wood JA, Zvonok AM, Makriyannis A, Hohmann AG: Pharmacological characterization of AM1710, a putative cannabinoid CB2 agonist from the cannabilactone class: antinociception without central nervous system side-effects.

    Pharmacol Biochem Behav 2011, 98:493-502. PubMed Abstract | Publisher Full Text | PubMed Central Full Text OpenURL

  20. Khanolkar AD, Lu D, Ibrahim M, Duclos RI Jr, Thakur GA, Malan TP Jr, Porreca F, Veerappan V, Tian X, George C, et al.: Cannabilactones: a novel class of CB2 selective agonists with peripheral analgesic activity.

    J Med Chem 2007, 50:6493-6500. PubMed Abstract | Publisher Full Text OpenURL

  21. Jaggi AS, Singh N: Mechanisms in cancer-chemotherapeutic drugs-induced peripheral neuropathy.

    Toxicology 2012, 291:1-9. PubMed Abstract | Publisher Full Text OpenURL

  22. Gao YJ, Ji RR: Chemokines, neuronal-glial interactions, and central processing of neuropathic pain.

    Pharmacol Ther 2010, 126:56-68. PubMed Abstract | Publisher Full Text | PubMed Central Full Text OpenURL

  23. Dubovy P, Klusakova I, Svizenska I, Brazda V: Spatio-temporal changes of SDF1 and its CXCR4 receptor in the dorsal root ganglia following unilateral sciatic nerve injury as a model of neuropathic pain.

    Histochem Cell Biol 2010, 133:323-337. PubMed Abstract | Publisher Full Text OpenURL

  24. Bhangoo SK, Ren D, Miller RJ, Chan DM, Ripsch MS, Weiss C, McGinnis C, White FA: CXCR4 chemokine receptor signaling mediates pain hypersensitivity in association with antiretroviral toxic neuropathy.

    Brain Behav Immun 2007, 21:581-591. PubMed Abstract | Publisher Full Text | PubMed Central Full Text OpenURL

  25. Bhangoo SK, Ripsch MS, Buchanan DJ, Miller RJ, White FA: Increased chemokine signaling in a model of HIV1-associated peripheral neuropathy.

    Mol Pain 2009, 5:48. PubMed Abstract | BioMed Central Full Text | PubMed Central Full Text OpenURL

  26. Rahn EJ, Thakur GA, Zvonok AM, Vemuri VK, Makriyannis A, Hohmann AG: Prophylactic treatment with cannabinoids suppresses the development of neuropathic nociception resulting from treatment with the chemotherapeutic agent paclitaxel in rats.

    Society for Neuroscience Abstract Viewer/Itinerary Planner CD-ROM 2010., 681.3 OpenURL

  27. Jhaveri MD, Sagar DR, Elmes SJ, Kendall DA, Chapman V: Cannabinoid CB2 receptor-mediated anti-nociception in models of acute and chronic pain.

    Mol Neurobiol 2007, 36:26-35. PubMed Abstract | Publisher Full Text OpenURL

  28. Van Sickle MD, Duncan M, Kingsley PJ, Mouihate A, Urbani P, Mackie K, Stella N, Makriyannis A, Piomelli D, Davison JS, et al.: Identification and functional characterization of brainstem cannabinoid CB2 receptors.

    Science 2005, 310:329-332. PubMed Abstract | Publisher Full Text OpenURL

  29. Guindon J, Hohmann AG: Cannabinoid CB2 receptors: a therapeutic target for the treatment of inflammatory and neuropathic pain.

    Br J Pharmacol 2008, 153:319-334. PubMed Abstract | Publisher Full Text | PubMed Central Full Text OpenURL

  30. Anand U, Otto WR, Sanchez-Herrera D, Facer P, Yiangou Y, Korchev Y, Birch R, Benham C, Bountra C, Chessell IP, Anand P: Cannabinoid receptor CB2 localisation and agonist-mediated inhibition of capsaicin responses in human sensory neurons.

    Pain 2008, 138:667-680. PubMed Abstract | Publisher Full Text OpenURL

  31. Hanus L, Breuer A, Tchilibon S, Shiloah S, Goldenberg D, Horowitz M, Pertwee RG, Ross RA, Mechoulam R, Fride E: HU-308: a specific agonist for CB(2), a peripheral cannabinoid receptor.

    Proc Natl Acad Sci USA 1999, 96:14228-14233. PubMed Abstract | Publisher Full Text | PubMed Central Full Text OpenURL

  32. Ibrahim MM, Deng H, Zvonok A, Cockayne DA, Kwan J, Mata HP, Vanderah TW, Lai J, Porreca F, Makriyannis A, Malan TP Jr: Activation of CB2 cannabinoid receptors by AM1241 inhibits experimental neuropathic pain: pain inhibition by receptors not present in the CNS.

    Proc Natl Acad Sci USA 2003, 100:10529-10533. PubMed Abstract | Publisher Full Text | PubMed Central Full Text OpenURL

  33. Wilkerson JL, Gentry KR, Dengler EC, Wallace JA, Kerwin AA, Armijo LM, Kuhn MN, Thakur GA, Makriyannis A, Milligan ED: Intrathecal cannabilactone CB(2)R agonist, AM1710, controls pathological pain and restores basal cytokine levels.

    Pain 2012, 153:1091-1106. PubMed Abstract | Publisher Full Text OpenURL

  34. Thompson SW, Davis LE, Kornfeld M, Hilgers RD, Standefer JC: Cisplatin neuropathy. Clinical, electrophysiologic, morphologic, and toxicologic studies.

    Cancer 1984, 54:1269-1275. PubMed Abstract | Publisher Full Text OpenURL

  35. Roelofs RI, Hrushesky W, Rogin J, Rosenberg L: Peripheral sensory neuropathy and cisplatin chemotherapy.

    Neurology 1984, 34:934-938. PubMed Abstract | Publisher Full Text OpenURL

  36. Lipton RB, Apfel SC, Dutcher JP, Rosenberg R, Kaplan J, Berger A, Einzig AI, Wiernik P, Schaumburg HH: Taxol produces a predominantly sensory neuropathy.

    Neurology 1989, 39:368-373. PubMed Abstract | Publisher Full Text OpenURL

  37. Petrenko AB, Yamakura T, Baba H, Shimoji K: The role of N-methyl-D-aspartate (NMDA) receptors in pain: a review.

    Anesth Analg 2003, 97:1108-1116. PubMed Abstract | Publisher Full Text OpenURL

  38. Carozzi VA, Chiorazzi A, Canta A, Lapidus RG, Slusher BS, Wozniak KM, Cavaletti G: Glutamate carboxypeptidase inhibition reduces the severity of chemotherapy-induced peripheral neurotoxicity in rat.

    Neurotox Res 2010, 17:380-391. PubMed Abstract | Publisher Full Text OpenURL

  39. Pascual D, Goicoechea C, Burgos E, Martin MI: Antinociceptive effect of three common analgesic drugs on peripheral neuropathy induced by paclitaxel in rats.

    Pharmacol Biochem Behav 2010, 95:331-337. PubMed Abstract | Publisher Full Text OpenURL

  40. Zhang H, Yoon SY, Dougherty PM: Evidence that spinal astrocytes but not microglia contribute to the pathogenesis of Paclitaxel-induced painful neuropathy.

    J Pain 2012, 13:293-303. PubMed Abstract | Publisher Full Text OpenURL

  41. Ta LE, Bieber AJ, Carlton SM, Loprinzi CL, Low PA, Windebank AJ: Transient Receptor Potential Vanilloid 1 is essential for cisplatin-induced heat hyperalgesia in mice.

    Mol Pain 2010, 6:15. PubMed Abstract | BioMed Central Full Text | PubMed Central Full Text OpenURL

  42. Alessandri-Haber N, Dina OA, Joseph EK, Reichling DB, Levine JD: Interaction of transient receptor potential vanilloid 4, integrin, and SRC tyrosine kinase in mechanical hyperalgesia.

    J Neurosci 2008, 28:1046-1057. PubMed Abstract | Publisher Full Text OpenURL

  43. Flatters SJ, Bennett GJ: Studies of peripheral sensory nerves in paclitaxel-induced painful peripheral neuropathy: evidence for mitochondrial dysfunction.

    Pain 2006, 122:245-257. PubMed Abstract | Publisher Full Text | PubMed Central Full Text OpenURL

  44. Melli G, Taiana M, Camozzi F, Triolo D, Podini P, Quattrini A, Taroni F, Lauria G: Alpha-lipoic acid prevents mitochondrial damage and neurotoxicity in experimental chemotherapy neuropathy.

    Exp Neurol 2008, 214:276-284. PubMed Abstract | Publisher Full Text OpenURL

  45. Xiao WH, Zheng H, Zheng FY, Nuydens R, Meert TF, Bennett GJ: Mitochondrial abnormality in sensory, but not motor, axons in paclitaxel-evoked painful peripheral neuropathy in the rat.

    Neuroscience 2011, 199:461-469. PubMed Abstract | Publisher Full Text OpenURL

  46. Zheng H, Xiao WH, Bennett GJ: Functional deficits in peripheral nerve mitochondria in rats with paclitaxel- and oxaliplatin-evoked painful peripheral neuropathy.

    Exp Neurol 2011, 232:154-161. PubMed Abstract | Publisher Full Text OpenURL

  47. Khasabova IA, Khasabov S, Paz J, Harding-Rose C, Simone DA, Seybold VS: Cannabinoid type-1 receptor reduces pain and neurotoxicity produced by chemotherapy.

    J Neurosci 2012, 32:7091-7101. PubMed Abstract | Publisher Full Text | PubMed Central Full Text OpenURL

  48. Nackley AG, Zvonok AM, Makriyannis A, Hohmann AG: Activation of cannabinoid CB2 receptors suppresses C-fiber responses and windup in spinal wide dynamic range neurons in the absence and presence of inflammation.

    J Neurophysiol 2004, 92:3562-3574. PubMed Abstract | Publisher Full Text OpenURL

  49. Sagar DR, Kelly S, Millns PJ, O'Shaughnessey CT, Kendall DA, Chapman V: Inhibitory effects of CB1 and CB2 receptor agonists on responses of DRG neurons and dorsal horn neurons in neuropathic rats.

    Eur J Neurosci 2005, 22:371-379. PubMed Abstract | Publisher Full Text OpenURL

  50. Elmes SJ, Jhaveri MD, Smart D, Kendall DA, Chapman V: Cannabinoid CB2 receptor activation inhibits mechanically evoked responses of wide dynamic range dorsal horn neurons in naive rats and in rat models of inflammatory and neuropathic pain.

    Eur J Neurosci 2004, 20:2311-2320. PubMed Abstract | Publisher Full Text OpenURL

  51. Basbaum AI, Bautista DM, Scherrer G, Julius D: Cellular and molecular mechanisms of pain.

    Cell 2009, 139:267-284. PubMed Abstract | Publisher Full Text | PubMed Central Full Text OpenURL

  52. Anand P, Whiteside G, Fowler CJ, Hohmann AG: Targeting CB2 receptors and the endocannabinoid system for the treatment of pain.

    Brain Res Rev 2009, 60:255-266. PubMed Abstract | Publisher Full Text OpenURL

  53. Burgos E, Gomez-Nicola D, Pascual D, Martin MI, Nieto-Sampedro M, Goicoechea C: Cannabinoid agonist WIN 55,212-2 prevents the development of paclitaxel-induced peripheral neuropathy in rats. Possible involvement of spinal glial cells.

    Eur J Pharmacol 2012, 682:62-72. PubMed Abstract | Publisher Full Text OpenURL

  54. Oh JW, Drabik K, Kutsch O, Choi C, Tousson A, Benveniste EN: CXC chemokine receptor 4 expression and function in human astroglioma cells.

    J Immunol 2001, 166:2695-2704. PubMed Abstract | Publisher Full Text OpenURL

  55. Kajander KC, Bennett GJ: Onset of a painful peripheral neuropathy in rat: a partial and differential deafferentation and spontaneous discharge in A beta and A delta primary afferent neurons.

    J Neurophysiol 1992, 68:734-744. PubMed Abstract | Publisher Full Text OpenURL

  56. Wu G, Ringkamp M, Murinson BB, Pogatzki EM, Hartke TV, Weerahandi HM, Campbell JN, Griffin JW, Meyer RA: Degeneration of myelinated efferent fibers induces spontaneous activity in uninjured C-fiber afferents.

    J Neurosci 2002, 22:7746-7753. PubMed Abstract | Publisher Full Text OpenURL

  57. Tanner KD, Levine JD, Topp KS: Microtubule disorientation and axonal swelling in unmyelinated sensory axons during vincristine-induced painful neuropathy in rat.

    J Comp Neurol 1998, 395:481-492. PubMed Abstract | Publisher Full Text OpenURL

  58. Topp KS, Tanner KD, Levine JD: Damage to the cytoskeleton of large diameter sensory neurons and myelinated axons in vincristine-induced painful peripheral neuropathy in the rat.

    J Comp Neurol 2000, 424:563-576. PubMed Abstract | Publisher Full Text OpenURL

  59. Tanner KD, Reichling DB, Levine JD: Nociceptor hyper-responsiveness during vincristine-induced painful peripheral neuropathy in the rat.

    J Neurosci 1998, 18:6480-6491. PubMed Abstract | Publisher Full Text OpenURL

  60. Siau C, Xiao W, Bennett GJ: Paclitaxel- and vincristine-evoked painful peripheral neuropathies: loss of epidermal innervation and activation of Langerhans cells.

    Exp Neurol 2006, 201:507-514. PubMed Abstract | Publisher Full Text | PubMed Central Full Text OpenURL

  61. Xiao WH, Bennett GJ: Chemotherapy-evoked neuropathic pain: Abnormal spontaneous discharge in A-fiber and C-fiber primary afferent neurons and its suppression by acetyl-L-carnitine.

    Pain 2008, 135:262-270. PubMed Abstract | Publisher Full Text | PubMed Central Full Text OpenURL

  62. Dina OA, Chen X, Reichling D, Levine JD: Role of protein kinase Cepsilon and protein kinase A in a model of paclitaxel-induced painful peripheral neuropathy in the rat.

    Neuroscience 2001, 108:507-515. PubMed Abstract | Publisher Full Text OpenURL

  63. Benamar K, Geller EB, Adler MW: First in vivo evidence for a functional interaction between chemokine and cannabinoid systems in the brain.

    J Pharmacol Exp Ther 2008, 325:641-645. PubMed Abstract | Publisher Full Text OpenURL

  64. Benamar K, Yondorf M, Geller EB, Eisenstein TK, Adler MW: Physiological evidence for interaction between the HIV-1 co-receptor CXCR4 and the cannabinoid system in the brain.

    Br J Pharmacol 2009, 157:1225-1231. PubMed Abstract | Publisher Full Text | PubMed Central Full Text OpenURL

  65. Ghosh S, Preet A, Groopman JE, Ganju RK: Cannabinoid receptor CB2 modulates the CXCL12/CXCR4-mediated chemotaxis of T lymphocytes.

    Mol Immunol 2006, 43:2169-2179. PubMed Abstract | Publisher Full Text OpenURL

  66. Nasser MW, Qamri Z, Deol YS, Smith D, Shilo K, Zou X, Ganju RK: Crosstalk between chemokine receptor CXCR4 and cannabinoid receptor CB2 in modulating breast cancer growth and invasion.

    PLoS One 2011, 6:e23901. PubMed Abstract | Publisher Full Text | PubMed Central Full Text OpenURL

  67. Cao L, DeLeo JA: CNS-infiltrating CD4+ T lymphocytes contribute to murine spinal nerve transection-induced neuropathic pain.

    Eur J Immunol 2008, 38:448-458. PubMed Abstract | Publisher Full Text | PubMed Central Full Text OpenURL

  68. Zimmermann M: Ethical guidelines for investigations of experimental pain in conscious animals.

    Pain 1983, 16:109-110. PubMed Abstract | Publisher Full Text OpenURL

  69. Authier N, Gillet JP, Fialip J, Eschalier A, Coudore F: An animal model of nociceptive peripheral neuropathy following repeated cisplatin injections.

    Exp Neurol 2003, 182:12-20. PubMed Abstract | Publisher Full Text OpenURL

  70. Polomano RC, Mannes AJ, Clark US, Bennett GJ: A painful peripheral neuropathy in the rat produced by the chemotherapeutic drug, paclitaxel.

    Pain 2001, 94:293-304. PubMed Abstract | Publisher Full Text OpenURL

  71. Rousseaux C, Thuru X, Gelot A, Barnich N, Neut C, Dubuquoy L, Dubuquoy C, Merour E, Geboes K, Chamaillard M, et al.: Lactobacillus acidophilus modulates intestinal pain and induces opioid and cannabinoid receptors.

    Nat Med 2007, 13:35-37. PubMed Abstract | Publisher Full Text OpenURL

  72. Guindon J, Desroches J, Dani M, Beaulieu P: Pre-emptive antinociceptive effects of a synthetic cannabinoid in a model of neuropathic pain.

    Eur J Pharmacol 2007, 568:173-176. PubMed Abstract | Publisher Full Text OpenURL

  73. Wilson NM, Jung H, Ripsch MS, Miller RJ, White FA: CXCR4 signaling mediates morphine-induced tactile hyperalgesia.

    Brain Behav Immun 2011, 25:565-573. PubMed Abstract | Publisher Full Text | PubMed Central Full Text OpenURL

  74. Choi Y, Yoon YW, Na HS, Kim SH, Chung JM: Behavioral signs of ongoing pain and cold allodynia in a rat model of neuropathic pain.

    Pain 1994, 59:369-376. PubMed Abstract | Publisher Full Text OpenURL