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Doublecortin-like kinase 1 activates NF-κB to induce inflammatory responses by binding directly to IKKβ

Cell Death & Differentiation volume 30, pages 1184–1197 (2023)Cite this article

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Abstract

Doublecortin-like kinase 1 (DCLK1), a microtubule-associated protein kinase, is involved in neurogenesis, and its levels are elevated in various human cancers. Recent studies suggest that DCLK1 may relate to inflammatory responses in the mouse model of colitis. However, cellular pathways engaged by DCLK1, and potential substrates of the kinase remain undefined. To understand how DCLK1 regulates inflammatory responses, we utilized the well-established lipopolysaccharide (LPS)-stimulated macrophages and mouse model. Through a range of macrophage-based and cell-free platforms, we discovered that DCLK1 binds directly with the inhibitor of κB kinase β (IKKβ) and induces IKKβ phosphorylation on Ser177/181 to initiate nuclear factor-κB (NF-κB) pathway. Deficiency in DCLK1, achieved by silencing or through pharmacological inhibition, prevented LPS-induced NF-κB activation and cytokine production in macrophages. We further show that mice with myeloid-specific DCLK1 knockout or DCLK1 inhibitor treatment are protected against LPS-induced acute lung injury and septic death. Our studies report a novel functional role of macrophage DCLK1 as a direct IKKβ regulator in inflammatory signaling and suggest targeted therapy against DCLK1 for inflammatory diseases.

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Fig. 1: DCLK1 is elevated in LPS-challenged macrophages and mediates inflammatory responses.
Fig. 2: DCLK1 mediates inflammatory responses through NF-κB.
Fig. 3: DCLK1 regulates NF-κB activity, independent of MAP7D1 and DCX.
Fig. 4: DCLK1 directly binds to IKKβ.
Fig. 5: DCLK1 promotes IKKβ phosphorylation at S177/181 to mediate inflammatory responses.
Fig. 6: Myeloid DCLK1 knockout reduces LPS-induced inflammatory responses and lung injury.
Fig. 7: Pharmacological inhibition of DCLK1 prevents LPS-induced lung injury in mice.
Fig. 8: DCLK1 deficiency increases survival in septic mice.

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Acknowledgements

This study was supported by the National Natural Science Foundation of China (81930108 to GL, 82000793 to WL, and 82170373 to YW), Natural Science Foundation of Zhejiang Province (LY22H070004 to WL), and Zhejiang Provincial Key Scientific Project (2021C03041 to GL).

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Authors and Affiliations

  1. Chemical Biology Research Center, School of Pharmaceutical Sciences, Wenzhou Medical University, Wenzhou, Zhejiang, 325035, China

    Wu Luo, Yiyi Jin, Yuchen Jiang, Libin Yang, Haowen Xu, Di Wu, Guang Liang & Yi Wang

  2. Department of Cardiology and Medical Research Center, the First Affiliated Hospital, Wenzhou Medical University, Wenzhou, Zhejiang, 325035, China

    Wu Luo & Guang Liang

  3. School of Pharmaceutical Sciences, Hangzhou Medical College, Hangzhou, Zhejiang, 311399, China

    Wu Luo, Yanmei Zhang, Lina Yin & Guang Liang

  4. Department of Pathology and Laboratory Medicine, University of Western Ontario, London, ON, N6A 5C1, Canada

    Zia Ali Khan

  5. School of Pharmaceutical Sciences, Hangzhou Normal University, Hangzhou, Zhejiang, 311121, China

    Yi Wang

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Contributions

GL, YW, and WL contributed to the literature search and study design. GL and WL participated in the drafting of the article. WL, YJin, YJiang, LYang, HX, DW, YZ, and LYin carried out the experiments. GL, ZAK, and YW revised the manuscript. LYin, YW, and WL contributed to data collection and analysis.

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Correspondence to Guang Liang or Yi Wang.

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Luo, W., Jin, Y., Jiang, Y. et al. Doublecortin-like kinase 1 activates NF-κB to induce inflammatory responses by binding directly to IKKβ. Cell Death Differ 30, 1184–1197 (2023). https://doi.org/10.1038/s41418-023-01147-8

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