Wnt5b/Ryk-mediated membrane trafficking of P2X3 receptors contributes to bone cancer pain
Wnt5b/Ryk-mediated membrane trafficking of P2X3 receptors contributes to bone cancer pain
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Wnt5b/Ryk 介导的 P2X3 受体膜运输导致骨癌疼痛
DOI：
10.1016/j.expneurol.2020.113482
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发表时间：
2020-12-01
影响因子：
5.3
通讯作者：
Fang, Dong
中科院分区：
文献类型：
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作者：
He, Jin-Jin;Wang, Xiao;Fang, Dong
Wnt 5 b是Wnt家族的成员之一，在肿瘤的发生、发展和转移过程中发挥着重要作用。然而，Wnt 5 b是否参与背根神经节（DRG）神经元的敏化和骨癌痛的发病机制仍不清楚。在这里，我们发现Wnt 5 b及其非典型酪氨酸蛋白激酶受体Ryk的蛋白表达上调，在同侧背根神经节在荷瘤小鼠。Wnt 5 b的应用引起了分离的背根节神经元放电频率的增加和幼稚小鼠的疼痛超敏反应，而抗Ryk抗体几乎完全阻止了这一现象。此外，鞘内注射抗Ryk抗体的荷瘤小鼠显着抑制骨癌引起的机械异常性疼痛和热痛觉过敏。随后，我们还证明了将Wnt 5 b应用于培养的DRG神经元可以增强膜P2 X3受体和α，β-meATP诱导的电流。鞘内注射钙调素依赖性蛋白激酶II（CaMK II）抑制剂KN 93或P2 X3受体拮抗剂A317491几乎完全消除了Wnt 5诱导的小鼠机械异常性疼痛和热痛觉过敏。同时，抗Ryk抗体或CaMK Ⅱ抑制剂KN 93预处理可减轻骨癌诱导的P2 X3膜蛋白上调以及疼痛敏感性。这些发现表明，Wnt 5 b/Ryk通过激活初级感觉神经元中的CaMKII促进P2 X3受体向膜的运输，导致外周致敏和骨癌诱导的疼痛。我们的研究结果可能为骨癌疼痛提供一种潜在的治疗策略。
Wnt5b, a member of Wnt family, plays multiple roles in tumor progression and metastasis. However, whether Wnt5b contributes to the sensitization of dorsal root ganglia (DRG) neurons and pathogenesis of bone cancer pain still remains unclear. Here, we found that the protein expression of Wnt5b and its atypical tyrosine protein kinase receptor Ryk was upregulated in ipsilateral DRGs in tumor-bearing mice. Application of Wnt5b evoked an increased discharge frequency in isolated DRG neurons and pain hypersensitivity in naive mice which were almost completely prevented by anti-Ryk antibody. Moreover, intrathecal injection of anti-Ryk antibody to tumor-bearing mice significantly inhibited bone cancer-induced mechanic allodynia and thermal hyperalgesia. Subsequently, we also demonstrated that application of Wnt5b to cultured DRG neurons could enhance membrane P2X3 receptors and alpha,beta-meATP-induced currents. Intrathecal injection of calmodulin-dependent protein kinase II (CaMKII) inhibitor KN93 or P2X3 receptors antagonist A317491 almost completely abolished Wnt5binduced mechanical allodynia and thermal hyperalgesia in mice. Meanwhile, pretreatment with anti-Ryk antibody or CaMKII inhibitor KN93 can attenuate bone-cancer induced the upregulation of P2X3 membrane protein as well as pain hypersensitivity. These findings suggested that Wnt5b/Ryk promoted the trafficking of P2X3 receptors to the membrane via the activation of CaMKII in primary sensory neurons, resulting in peripheral sensitization and bone cancer-induced pain. Our results may offer a potential therapeutic strategy for bone cancer pain.