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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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References

  1. Walker TL, Yasuda T, Adams DJ, Bartlett PF. The doublecortin-expressing population in the developing and adult brain contains multipotential precursors in addition to neuronal-lineage cells. J Neurosci. 2007;27:3734–42.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  2. Vreugdenhil E, Kolk SM, Boekhoorn K, Fitzsimons CP, Schaaf M, Schouten T, et al. Doublecortin-like, a microtubule-associated protein expressed in radial glia, is crucial for neuronal precursor division and radial process stability. Eur J Neurosci. 2007;25:635–48.

    Article  PubMed  Google Scholar 

  3. Sossey-Alaoui K, Srivastava AK. DCAMKL1, a brain-specific transmembrane protein on 13q12.3 that is similar to doublecortin (DCX). Genomics. 1999;56:121–6.

    Article  CAS  PubMed  Google Scholar 

  4. Reiner O, Coquelle FM, Peter B, Levy T, Kaplan A, Sapir T, et al. The evolving doublecortin (DCX) superfamily. BMC Genomics. 2006;7:188.

    Article  PubMed  PubMed Central  Google Scholar 

  5. Cao Z, Weygant N, Chandrakesan P, Houchen CW, Peng J, Qu D. Tuft and Cancer Stem Cell Marker DCLK1: A New Target to Enhance Anti-Tumor Immunity in the Tumor Microenvironment. Cancers. 2020;12:3801.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Lu Q, Feng H, Chen H, Weygant N, Du J, Yan Z, et al. Role of DCLK1 in oncogenic signaling (Review). Int J Oncol. 2022;61:137.

    Article  CAS  PubMed  Google Scholar 

  7. Yi J, Bergstrom K, Fu J, Shan X, McDaniel JM, McGee S, et al. Dclk1 in tuft cells promotes inflammation-driven epithelial restitution and mitigates chronic colitis. Cell Death Differ. 2019;26:1656–69.

    Article  CAS  PubMed  Google Scholar 

  8. Qu D, Weygant N, May R, Chandrakesan P, Madhoun M, Ali N, et al. Ablation of Doublecortin-Like Kinase 1 in the Colonic Epithelium Exacerbates Dextran Sulfate Sodium-Induced Colitis. PLoS One. 2015;10:e0134212.

    Article  PubMed  PubMed Central  Google Scholar 

  9. Wu X, Qu D, Weygant N, Peng J, Houchen CW. Cancer Stem Cell Marker DCLK1 Correlates with Tumorigenic Immune Infiltrates in the Colon and Gastric Adenocarcinoma Microenvironments. Cancers. 2020;12:274.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Remick DG, Newcomb DE, Bolgos GL, Call DR. Comparison of the mortality and inflammatory response of two models of sepsis: lipopolysaccharide vs. cecal ligation and puncture. Shock. 2000;13:110–6.

    Article  CAS  PubMed  Google Scholar 

  11. Page MJ, Kell DB, Pretorius E. The role of lipopolysaccharide-induced cell signalling in chronic inflammation. Chronic Stress. 2022;6:24705470221076390.

    Article  PubMed  PubMed Central  Google Scholar 

  12. Gay NJ, Symmons MF, Gangloff M, Bryant CE. Assembly and localization of Toll-like receptor signalling complexes. Nat Rev Immunol. 2014;14:546–58.

    Article  CAS  PubMed  Google Scholar 

  13. Oeckinghaus A, Hayden MS, Ghosh S. Crosstalk in NF-kappaB signaling pathways. Nat Immunol. 2011;12:695–708.

    Article  CAS  PubMed  Google Scholar 

  14. Pan Y, Wang Y, Cai L, Cai Y, Hu J, Yu C, et al. Inhibition of high glucose-induced inflammatory response and macrophage infiltration by a novel curcumin derivative prevents renal injury in diabetic rats. Br J Pharm. 2012;166:1169–82.

    Article  CAS  Google Scholar 

  15. Chen L, Chen H, Chen P, Zhang W, Wu C, Sun C, et al. Development of 2-amino-4-phenylthiazole analogues to disrupt myeloid differentiation factor 88 and prevent inflammatory responses in acute lung injury. Eur J Med Chem. 2019;161:22–38.

    Article  CAS  PubMed  Google Scholar 

  16. Halder L, Jo E, Hasan M, Ferreira-Gomes M, Krüger T, Westermann M, et al. Immune modulation by complement receptor 3-dependent human monocyte TGF-β1-transporting vesicles. Nat Commun. 2020;11:2331.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Matute-Bello G, Downey G, Moore BB, Groshong SD, Matthay MA, Slutsky AS, et al. An official American Thoracic Society workshop report: features and measurements of experimental acute lung injury in animals. Am J Respir Cell Mol Biol. 2011;44:725–38.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. Engels BM, Schouten TG, van Dullemen J, Gosens I, Vreugdenhil E. Functional differences between two DCLK splice variants. Brain Res Mol Brain Res. 2004;120:103–14.

    Article  CAS  PubMed  Google Scholar 

  19. Burgess HA, Reiner O. Alternative splice variants of doublecortin-like kinase are differentially expressed and have different kinase activities. J Biol Chem. 2002;277:17696–705.

    Article  CAS  PubMed  Google Scholar 

  20. Weygant N, Qu D, Berry WL, May R, Chandrakesan P, Owen DB, et al. Small molecule kinase inhibitor LRRK2-IN-1 demonstrates potent activity against colorectal and pancreatic cancer through inhibition of doublecortin-like kinase 1. Mol Cancer. 2014;13:103.

    Article  PubMed  PubMed Central  Google Scholar 

  21. Koizumi H, Fujioka H, Togashi K, Thompson J, Yates JR 3rd, Gleeson JG, et al. DCLK1 phosphorylates the microtubule-associated protein MAP7D1 to promote axon elongation in cortical neurons. Dev Neurobiol. 2017;77:493–510.

    Article  CAS  PubMed  Google Scholar 

  22. Jean DC, Baas PW, Black MM. A novel role for doublecortin and doublecortin-like kinase in regulating growth cone microtubules. Hum Mol Genet. 2012;21:5511–27.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  23. Delhase M, Hayakawa M, Chen Y, Karin M. Positive and negative regulation of IkappaB kinase activity through IKKbeta subunit phosphorylation. Science. 1999;284:309–13.

    Article  CAS  PubMed  Google Scholar 

  24. Sureban SM, May R, Weygant N, Qu D, Chandrakesan P, Bannerman-Menson E, et al. XMD8-92 inhibits pancreatic tumor xenograft growth via a DCLK1-dependent mechanism. Cancer Lett. 2014;351:151–61.

    Article  CAS  PubMed  Google Scholar 

  25. Shu T, Tseng HC, Sapir T, Stern P, Zhou Y, Sanada K, et al. Doublecortin-like kinase controls neurogenesis by regulating mitotic spindles and M phase progression. Neuron. 2006;49:25–39.

    Article  CAS  PubMed  Google Scholar 

  26. Koizumi H, Tanaka T, Gleeson JG. Doublecortin-like kinase functions with doublecortin to mediate fiber tract decussation and neuronal migration. Neuron. 2006;49:55–66.

    Article  CAS  PubMed  Google Scholar 

  27. Deuel TA, Liu JS, Corbo JC, Yoo SY, Rorke-Adams LB, Walsh CA. Genetic interactions between doublecortin and doublecortin-like kinase in neuronal migration and axon outgrowth. Neuron. 2006;49:41–53.

    Article  CAS  PubMed  Google Scholar 

  28. Chhetri D, Vengadassalapathy S, Venkadassalapathy S, Balachandran V, Umapathy VR, Veeraraghavan VP, et al. Pleiotropic effects of DCLK1 in cancer and cancer stem cells. Front Mol Biosci. 2022;9:965730.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  29. Ali N, Chandrakesan P, Nguyen CB, Husain S, Gillaspy AF, Huycke M, et al. Inflammatory and oncogenic roles of a tumor stem cell marker doublecortin-like kinase (DCLK1) in virus-induced chronic liver diseases. Oncotarget. 2015;6:20327–44.

    Article  PubMed  PubMed Central  Google Scholar 

  30. Patel O, Dai W, Mentzel M, Griffin M, Serindoux J, Gay Y, et al. Biochemical and Structural Insights into Doublecortin-like Kinase Domain 1. Structure. 2016;24:1550–61.

    Article  CAS  PubMed  Google Scholar 

  31. Kuribara M, Jenks BG, Dijkmans TF, de Gouw D, Ouwens DT, Roubos EW, et al. ERK-regulated double cortin-like kinase (DCLK)-short phosphorylation and nuclear translocation stimulate POMC gene expression in endocrine melanotrope cells. Endocrinology. 2011;152:2321–9.

    Article  CAS  PubMed  Google Scholar 

  32. Burgoyne RD, Helassa N, McCue HV, Haynes LP. Calcium sensors in neuronal function and dysfunction. Cold Spring Harb Perspect Biol. 2019;11:a035154.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  33. Lin PT, Gleeson JG, Corbo JC, Flanagan L, Walsh CA. DCAMKL1 encodes a protein kinase with homology to doublecortin that regulates microtubule polymerization. J Neurosci. 2000;20:9152–61.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  34. Kim MH, Cierpicki T, Derewenda U, Krowarsch D, Feng Y, Devedjiev Y, et al. The DCX-domain tandems of doublecortin and doublecortin-like kinase. Nat Struct Biol. 2003;10:324–33.

    Article  CAS  PubMed  Google Scholar 

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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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