Blockade of PDGFR-β activation eliminates morphine analgesic tolerance

Journal name:
Nature Medicine
Volume:
18,
Pages:
385–387
Year published:
DOI:
doi:10.1038/nm.2633
Received
Accepted
Published online

For centuries, opioid drugs have been the mainstay of chronic pain treatment. However, over time analgesic tolerance develops, leaving few treatment options. Here we show that platelet-derived growth factor receptor-β (PDGFR-β)-mediated signaling plays a key role in morphine tolerance. PDGFR-β inhibition selectively eliminates morphine tolerance in rats. PDGFR-β inhibitors are widely used and well tolerated, suggesting that clinical translation of our findings could reduce the suffering endured by individuals with intractable pain.

At a glance

Figures

  1. Morphine activates the PDGFR-[beta], and PDGFR-[beta] inhibition blocks tolerance.
    Figure 1: Morphine activates the PDGFR-β, and PDGFR-β inhibition blocks tolerance.

    (a) Rats were treated for 40 min with 0.6 nmol morphine, 10 μg imatinib, morphine plus imatinib, or vehicle (a 1:1 mixture of aCSF with 20% Captsol). Lumbar spinal cords were then collected, and the substantia gelatinosa was microdissected. Individual lysates were prepared for each rat, and immunoprecipitation was performed with antibody to PDGFR-β. Samples were then run on SDS-PAGE gels and immunoblotted with antibody to phosphotyrosine (pY20). Blots were stripped and reprobed with antibody to PDGFR-β to control for IP efficiency. A representative IP-IB experiment is shown, with irrelevant lanes removed. (b) Quantification of PDGFR-β phosphorylation from IP-IB experiments as in a. Data are shown as mean ± s.d. Overall one-way analysis of variance (ANOVA) F(3,19) = 13.8; P < 0.0001; *P < 0.05 versus all other treatment groups by Bonferroni multiple comparison post-tests. n = 5 or 6 independent rats per treatment group. (c) Rats were treated daily with i.t. injection of either 0.6 nmol morphine alone or morphine plus 10 μg imatinib, with imatinib begun on day 1, 3 or 5 of morphine treatment (morphine + imatinib-1, morphine + imatinib-3 or morphine + imatinib-5, respectively). On day 7, all rats received morphine alone (indicated by discontinuous lines between days 6 and 7). Analgesic responses were monitored using tail flick latency. Treatment F(3,32) = 18.5, day F(6,224) = 160, interaction F(21,224) = 22.0; all P < 0.0001 (two-way ANOVA). n = 9 rats per treatment group. (d) Rats were treated for 4 d with s.c. injection of either 3.5 mg per kg body weight morphine, 5 mg per kg imatinib body weight, both morphine and imatinib, or vehicle (a 1:1 mixture of 0.9% normal saline with 20% Captsol). On day 5, all rats received morphine alone. Treatment F(3,32) = 90.2, day F(5,160) = 44.5, interaction F(15,160) = 41.2. Data in c,d are shown as mean ± s.e.m.; all P < 0.0001 (two-way ANOVA); n = 9 rats per treatment group.

  2. Imatinib reverses morphine tolerance, and its effects are mediated by PDGFR-[beta].
    Figure 2: Imatinib reverses morphine tolerance, and its effects are mediated by PDGFR-β.

    (ac) Three groups of eight opioid-naive rats received s.c. injection of 2 mg per kg body weight morphine. Analgesia was assessed using tail flick latency (TFL) 30 min later. Fifteen minutes after TFL testing, the s.c. morphine dose was doubled and testing repeated until TFL values exceeded the cutoff value of 10 s. (a) Baseline dose-response curves. After baseline testing, two groups of eight rats had continuous-release morphine pellets (2–75 mg) implanted under isoflurane anesthesia, whereas the third group received two placebo pellets. (b) On day 5 after pellet implantation, the rats underwent dose-response testing. Thirty minutes before the initial morphine injection, one group of morphine-pelleted rats received 5 mg per kg imatinib s.c., whereas the other morphine-pelleted group and the placebo-pelleted rats were injected with an equivalent volume of vehicle (a 1:1 mixture of 0.9% normal saline with 20% Captsol). Imatinib substantially reduced an approximately sevenfold ED50 shift in morphine-tolerant rats to approximately 1.5-fold. (c) Day 6 dose-response results. The procedure described above was repeated the following day. Imatinib completely reversed an approximately eightfold ED50 shift in morphine-tolerant rats. (d) Rats were treated daily for 4 d with i.t. injection of either 0.6 nmol morphine, 10 ng PDGFR-β–Fc fragment, morphine plus PDGFR-β–Fc, or vehicle (a 1:1 mixture of aCSF with 20% Captsol). On day 5, the groups treated with PDGFR-β–Fc, vehicle, and morphine plus PDGFR-β–Fc received morphine alone, whereas the morphine group received morphine and PDGFR-β–Fc (indicated by discontinuous lines between days 4 and 5). Treatment F(3,192) = 84.8, day F(5,192) = 64.4, interaction F(15,160) = 37.8; all P < 0.0001 (two-way ANOVA). n = 6 rats for PDGFR-β–Fc and vehicle groups; n = 12 rats for morphine and morphine plus PDGFR-β–Fc groups. (e) Rats received daily i.t. injections for 4 d of either 0.6 nmol morphine; 10 pmol PDGF-BB; morphine and 10 μg imatinib; morphine and 10 pmol PDGF-BB; morphine, imatinib, and PDGF-BB; or vehicle (a 1:1 mixture of aCSF with 20% Captsol). On day 5, the PDGF-BB and vehicle groups received morphine alone, and all other groups continued their previous treatments (indicated by discontinuous lines between days 4 and 5). Treatment F(5,31) = 236, day F(5,155) = 73.8, interaction F(25,155) = 20.4; all P < 0.0001 (two-way ANOVA). n = 5–8 rats per group. Data in ae are shown as mean ± s.e.m.

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Author information

  1. These authors contributed equally to this work.

    • Yan Wang &
    • Katherine Barker

Affiliations

  1. Department of Anesthesiology, The University of Texas–MD Anderson Cancer Center, Houston, Texas, USA.

    • Yan Wang,
    • Katherine Barker,
    • Shanping Shi,
    • Miguel Diaz,
    • Bing Mo &
    • Howard B Gutstein
  2. Department of Biochemistry and Molecular Biology, The University of Texas–MD Anderson Cancer Center, Houston, Texas, USA.

    • Yan Wang,
    • Katherine Barker,
    • Shanping Shi,
    • Miguel Diaz,
    • Bing Mo &
    • Howard B Gutstein
  3. Genes and Development Graduate Program, The University of Texas–MD Anderson Cancer Center, Houston, Texas, USA.

    • Howard B Gutstein
  4. Deceased.

    • Bing Mo

Contributions

Y.W., K.B., S.S., M.D. and B.M. performed experiments and analyzed data. H.B.G. conceived and designed all experiments, interpreted results and wrote the manuscript.

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The authors declare no competing financial interests.

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