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Suppression of WNK1-SPAK/OSR1 Attenuates Bone Cancer Pain by Regulating NKCC1 and KCC2

  • Jian-ling Gao
    Affiliations
    Department of Anesthesiology, Intensive Care Medicine, and Pain Medicine, First Affiliated Hospital of Soochow University, Suzhou, Jiangsu, China
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  • Ke Peng
    Affiliations
    Department of Anesthesiology, Intensive Care Medicine, and Pain Medicine, First Affiliated Hospital of Soochow University, Suzhou, Jiangsu, China
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  • Meng-wei Shen
    Affiliations
    Department of Anesthesiology, Intensive Care Medicine, and Pain Medicine, First Affiliated Hospital of Soochow University, Suzhou, Jiangsu, China

    Department of Anesthesiology, Suzhou Municipal Hospital, Nanjing Medical University Affiliated Suzhou Hospital, Suzhou, Jiangsu, China
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  • Yong-heng Hou
    Affiliations
    Department of Anesthesiology, Intensive Care Medicine, and Pain Medicine, First Affiliated Hospital of Soochow University, Suzhou, Jiangsu, China
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  • Xiao-bo Qian
    Affiliations
    Department of Anesthesiology, Intensive Care Medicine, and Pain Medicine, First Affiliated Hospital of Soochow University, Suzhou, Jiangsu, China
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  • Xiao-wen Meng
    Affiliations
    Department of Anesthesiology, Intensive Care Medicine, and Pain Medicine, First Affiliated Hospital of Soochow University, Suzhou, Jiangsu, China
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  • Fu-hai Ji
    Affiliations
    Department of Anesthesiology, Intensive Care Medicine, and Pain Medicine, First Affiliated Hospital of Soochow University, Suzhou, Jiangsu, China
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  • Li-na Wang
    Affiliations
    Department of Anesthesiology, Intensive Care Medicine, and Pain Medicine, First Affiliated Hospital of Soochow University, Suzhou, Jiangsu, China
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  • Jian-ping Yang
    Correspondence
    Address reprint requests to Jian-ping Yang, Hospital of Soochow University, Department of Anesthesiology, Intensive Care Medicine, and Pain Medicine, No. 188 Shizi Street, Suzhou, Jiangsu 215006, China.
    Affiliations
    Department of Anesthesiology, Intensive Care Medicine, and Pain Medicine, First Affiliated Hospital of Soochow University, Suzhou, Jiangsu, China
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      Highlights

      • WNK1 was upregulated in the bone cancer pain (BCP) rat model.
      • WNK1-SPAK/OSR1-NKCC1/KCC2 signaling contributed to BCP in rats.
      • Intrathecal WNK1 siRNA or closantel attenuated pain behavior of BCP rats.
      • The WNK1 pathway may be a potential therapeutic target for BCP.

      Abstract

      Our preliminary experiment indicated the activation of with-nolysine kinases 1 (WNK1) in bone cancer pain (BCP) rats. This study aimed to investigate the underlying mechanisms via which WNK1 contributed to BCP. A rat model of BCP was induced by Walker-256 tumor cell implantation. WNK1 expression and distribution in the lumbar spinal cord dorsal horn and dorsal root ganglion were examined. SPS1-related proline/alanine-rich kinase (SPAK), oxidative stress-responsive kinase 1 (OSR1), sodium-potassium-chloride cotransporter 1 (NKCC1), and potassium-chloride cotransporter 2 (KCC2) expression were assessed. Pain behaviors including mechanical allodynia and movement-evoked pain were measured. BCP rats exhibited significant mechanical allodynia, with increased WNK1 expression in the dorsal horn and dorsal root ganglion neurons, elevated SPAK/OSR1 and NKCC1 expression in the dorsal root ganglion, and decreased KCC2 expression in the dorsal horn. WNK1 knock-down by small interfering alleviated mechanical allodynia and movement-evoked pain, inhibited WNK1-SPAK/OSR1-NKCC1 activities, and restored KCC2 expression. In addition, closantel (a WNK1-SPAK/OSR1 inhibitor) improved pain behaviors, downregulated SPAK/OSR1 and NKCC1 expression, and upregulated KCC2 expression in BCP rats. Activation of WNK1-SPAK/OSR1 signaling contributed to BCP in rats by modulating NKCC1 and KCC2 expression. Therefore, suppression of WNK1-SPAK/OSR1 may serve as a potential target for BCP therapy.

      Perspective

      Our findings demonstrated that the WNK1-SPAK/OSR1 signaling contributed to BCP in rats via regulating NKCC1 and KCC2. Suppressing this pathway reduced pain behaviors. Based on these findings, the WNK1-SPAK/OSR1 signaling may be a potential target for BCP therapy.

      Key words

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