Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Article
  • Published:

Dextran sodium sulfate potentiates NLRP3 inflammasome activation by modulating the KCa3.1 potassium channel in a mouse model of colitis

Cellular & Molecular Immunology volume 19, pages 925–943 (2022)Cite this article

Subjects

Abstract

Inflammatory bowel disease (IBD), including Crohn’s disease and ulcerative colitis, has increased in incidence and prevalence in recent decades. Both clinical and animal studies are critical for understanding the pathogenesis of this disease. Dextran sodium sulfate (DSS)-induced colitis is a frequently used animal model of IBD, but the underlying mechanism of the model remains incompletely understood. In this study, we found that NOD-like receptor family pyrin containing 3 (NLRP3) depletion markedly mitigated DSS-induced colitis and was accompanied by decreased activation of the inflammasome in the colons of mice. However, in vitro assays showed that DSS did not directly trigger but instead potentiated NLRP3 inflammasome assembly in macrophages in response to suboptimal ATP or nigericin stimulation. Mechanistically, DSS potentiated NLRP3 inflammasome activation in macrophages by augmenting KCa3.1-mediated potassium ion (K+) efflux. Furthermore, we found that pharmacologic blockade of the K+ channel KCa3.1 with TRAM-34 or genetic depletion of the Kcnn4 gene (encoding KCa3.1) not only ameliorated the severity of DSS-induced colitis but also attenuated in vivo inflammasome assembly in the colonic tissues of mice, suggesting a causal link between KCa3.1-mediated augmentation of the NLRP3 inflammasome and DSS-induced inflammatory injuries. Collectively, these results indicate that KCa3.1 plays a critical role in mediating DSS-induced colitis in mice by potentiating NLRP3 inflammasome activation. Our data provide a previously unknown mechanism by which DSS induces colitis in mice and suggests that KCa3.1 is an alternative therapeutic target for treating IBD.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8
Fig. 9

Similar content being viewed by others

References

  1. Graham DB, Xavier RJ. Pathway paradigms revealed from the genetics of inflammatory bowel disease. Nature. 2020;578:527–39.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  2. Ng SC, Shi HY, Hamidi N, Underwood F, Tang W, Benchimol E, et al. Worldwide incidence and prevalence of inflammatory bowel disease in the 21st century: a systematic review of population-based studies. Lancet. 2017;390:2769–78.

    Article  PubMed  Google Scholar 

  3. Kobayashi T, Siegmund B, Le Berre C, Wei S, Ferrante M, Shen B, et al. Ulcerative colitis. Nat Rev Dis primers. 2020;6:74.

    Article  PubMed  Google Scholar 

  4. Atreya R, Neurath MF. Mechanisms of molecular resistance and predictors of response to biological therapy in inflammatory bowel disease. Lancet Gastroenterol Hepatol. 2018;3:790–802.

    Article  PubMed  Google Scholar 

  5. de Souza HSP, Fiocchi C, Iliopoulos D. The IBD interactome: an integrated view of aetiology, pathogenesis and therapy. Nat Rev Gastroenterol Hepatol. 2017;14:739–49.

    Article  PubMed  Google Scholar 

  6. Ananthakrishnan AN, Bernstein C, Iliopoulos D, Macpherson A, Neurath M, Ali R, et al. Environmental triggers in IBD: a review of progress and evidence. Nat Rev Gastroenterol Hepatol. 2018;15:39–49.

    Article  PubMed  Google Scholar 

  7. Chang JT. Pathophysiology of inflammatory bowel diseases. N Engl J Med. 2020;383:2652–64.

    Article  CAS  PubMed  Google Scholar 

  8. Kiesler P, Fuss IJ, Strober W. Experimental models of inflammatory bowel diseases. Cell Mol Gastroenterol Hepatol. 2015;1:154–70.

    Article  PubMed Central  PubMed  Google Scholar 

  9. Okayasu I, Hatakeyama S, Yamada M, Ohkusa T, Inagaki Y, Nakaya R. A novel method in the induction of reliable experimental acute and chronic ulcerative colitis in mice. Gastroenterology. 1990;98:694–702.

    Article  CAS  PubMed  Google Scholar 

  10. Cooper HS, Murthy S, Shah R, Sedergran D. Clinicopathologic study of dextran sulfate sodium experimental murine colitis. Lab Invest. 1993;69:238–49.

    CAS  PubMed  Google Scholar 

  11. Chassaing B, Aitken JD, Malleshappa M, Vijay-Kumar M. Dextran sulfate sodium (DSS)-induced colitis in mice. Curr Protoc Immunol. 2014;104:15.25.11–15.25.14.

    Article  Google Scholar 

  12. Huang B, Chen Z, Geng L, Wang J, Liang H, Cao Y, et al. Mucosal profiling of pediatric-onset colitis and IBD reveals common pathogenics and therapeutic pathways. Cell. 2019;179:1160–76.

    Article  CAS  PubMed  Google Scholar 

  13. Pandey A, Shen C, Feng S, Man SM. Cell biology of inflammasome activation. Trends Cell Biol. 2021;31:924–39.

    Article  CAS  PubMed  Google Scholar 

  14. Broz P, Dixit VM. Inflammasomes: mechanism of assembly, regulation and signalling. Nat Rev Immunol. 2016;16:407–20.

    Article  CAS  PubMed  Google Scholar 

  15. Mangan MSJ, Olhava EJ, Roush WR, Seidel HM, Glick GD, Latz E. Targeting the NLRP3 inflammasome in inflammatory diseases. Nat Rev Drug Discov. 2018;17:688.

    Article  CAS  PubMed  Google Scholar 

  16. Li Y, Huang H, Liu B, Zhang Y, Pan X, Yu XY, et al. Inflammasomes as therapeutic targets in human diseases. Signal Transduct Target Ther. 2021;6:247.

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  17. Zhen Y & Zhang H, NLRP3 Inflammasome and inflammatory bowel disease. Front Immunol. 2019;10:276.

  18. Villani AC, Lemire M, Fortin G, Louis E, Silverberg M, Collette C, et al. Common variants in the NLRP3 region contribute to Crohn’s disease susceptibility. Nat Genet. 2009;41:71–76.

    Article  CAS  PubMed  Google Scholar 

  19. Bauer C, Duewell P, Mayer C, Lehr HA, Fitzgerald KA, Dauer M, et al. Colitis induced in mice with dextran sulfate sodium (DSS) is mediated by the NLRP3 inflammasome. Gut. 2010;59:1192–9.

    Article  CAS  PubMed  Google Scholar 

  20. Neudecker V, Haneklaus M, Jensen O, Khailova L, Masterson JC, Tye H, et al. Myeloid-derived miR-223 regulates intestinal inflammation via repression of the NLRP3 inflammasome. J Exp Med. 2017;214:1737–52.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  21. Perera AP, Fernando R, Shinde T, Gundamaraju R, Southam B, Sohal SS, et al. MCC950, a specific small molecule inhibitor of NLRP3 inflammasome attenuates colonic inflammation in spontaneous colitis mice. Sci Rep. 2018;8:8618.

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  22. Cosin-Roger J, Simmen S, Melhem H, Atrott K, Frey-Wagner I, Hausmann M, et al. Hypoxia ameliorates intestinal inflammation through NLRP3/mTOR downregulation and autophagy activation. Nat Commun. 2017;8:98.

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  23. Mao L, Kitani A, Hiejima E, Montgomery-Recht K, Zhou W, Fuss I, et al. Bruton tyrosine kinase deficiency augments NLRP3 inflammasome activation and causes IL-1β-mediated colitis. J Clin Invest. 2020;130:1793–807.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  24. Allen IC, TeKippe EM, Woodford RM, Uronis JM, Holl EK, Rogers AB, et al. The NLRP3 inflammasome functions as a negative regulator of tumorigenesis during colitis-associated cancer. J Exp Med. 2010;207:1045–56.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  25. Zaki MH, Boyd KL, Vogel P, Kastan MB, Lamkanfi M, Kanneganti TD. The NLRP3 inflammasome protects against loss of epithelial integrity and mortality during experimental colitis. Immunity. 2010;32:379–91.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  26. Rühl S, Broz P. Caspase-11 activates a canonical NLRP3 inflammasome by promoting K(+) efflux. Eur J Immunol. 2015;45:2927–36.

    Article  CAS  PubMed  Google Scholar 

  27. He Y, Zeng MY, Yang D, Motro B, Núñez G. NEK7 is an essential mediator of NLRP3 activation downstream of potassium efflux. Nature. 2016;530:354–7.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  28. Di A, Xiong S, Ye Z, Malireddi RKS, Kometani S, Zhong M, et al. The TWIK2 potassium efflux channel in macrophages mediates NLRP3 inflammasome-induced inflammation. Immunity. 2018;49:56–65.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  29. Ye J, Zeng B, Zhong M, Li H, Xu L, Shu J, et al. Scutellarin inhibits caspase-11 activation and pyroptosis in macrophages via regulating PKA signaling. Acta Pharm Sin B. 2021;11:112–26.

    Article  CAS  PubMed  Google Scholar 

  30. Zhong CS, Zeng B, Qiu JH, Xu LH, Zhong MY, Huang YT, et al. Gout-associated monosodium urate crystal-induced necrosis is independent of NLRP3 activity but can be suppressed by combined inhibitors for multiple signaling pathways. Acta Pharmacol Sin. 2021;43:1324–36.

    Article  CAS  PubMed  Google Scholar 

  31. Shu JX, Zhong C, Shi Z, Zeng B, Xu L, Ye J, et al. Berberine augments hypertrophy of colonic patches in mice with intraperitoneal bacterial infection. Int Immunopharmacol. 2021;90:107242.

    Article  CAS  PubMed  Google Scholar 

  32. Liu L, Li X. NLRP3 inflammasome in inflammatory bowel disease: friend or foe? Dig Dis Sci. 2017;62:2211–4.

    Article  PubMed  Google Scholar 

  33. Bauer C, Duewell P, Lehr H, Endres S, Schnurr M. Protective and aggravating effects of Nlrp3 inflammasome activation in IBD models: influence of genetic and environmental factors. Dig Dis. 2012;30:82–90.

    Article  PubMed  Google Scholar 

  34. Hara H, Seregin S, Yang D, Fukase K, Chamaillard M, Alnemri E, et al. The NLRP6 inflammasome recognizes lipoteichoic acid and regulates gram-positive pathogen infection. Cell. 2018;175:1651–64. e1614.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  35. Zhao Y, Yang J, Shi J, Gong Y, Lu Q, Xu H, et al. The NLRC4 inflammasome receptors for bacterial flagellin and type III secretion apparatus. Nature. 2011;477:596–600.

    Article  CAS  PubMed  Google Scholar 

  36. Hornung V, Ablasser A, Charrel-Dennis M, Bauernfeind F, Horvath G, Caffrey D, et al. AIM2 recognizes cytosolic dsDNA and forms a caspase-1-activating inflammasome with ASC. Nature. 2009;458:514–8.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  37. Coll RC, Robertson A, Chae J, Higgins S, Muñoz-Planillo R, Inserra M, et al. A small-molecule inhibitor of the NLRP3 inflammasome for the treatment of inflammatory diseases. Nat Med. 2015;21:248–55.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  38. Groß CJ, Mishra R, Schneider KS, Médard G, Wettmarshausen J, Dittlein DC, et al. K(+) efflux-independent NLRP3 inflammasome activation by small molecules targeting mitochondria. Immunity. 2016;45:761–73.

    Article  CAS  PubMed  Google Scholar 

  39. Xu Z, Chen ZM, Wu X, Zhang L, Cao Y & Zhou P. Distinct molecular mechanisms underlying potassium efflux for NLRP3 inflammasome activation. Front Immunol. 2020;11:609441.

  40. Dudem S, Sergeant GP, Thornbury KD & Hollywood MA. Calcium-activated K(+) Channels (K(Ca)) and therapeutic implications. Handb Exp Pharmacol. 2021;267:379–416.

  41. Nielsen OH, New strategies for treatment of inflammatory bowel disease. Front Med (Lausanne). 2014;1:3.

  42. Kaplan GG. The global burden of IBD: from 2015 to 2025. Nat Rev Gastroenterol Hepatol. 2015;12:720–7.

    Article  PubMed  Google Scholar 

  43. Hu JJ, Liu X, Xia S, Zhang Z, Zhang Y, Zhao J, et al. FDA-approved disulfiram inhibits pyroptosis by blocking gasdermin D pore formation. Nat Immunol. 2020;21:736–45.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  44. Kayagaki N, Stowe IB, Lee BL, O’Rourke K, Anderson K, Warming S, et al. Caspase-11 cleaves gasdermin D for non-canonical inflammasome signalling. Nature. 2015;526:666–71.

    Article  CAS  PubMed  Google Scholar 

  45. Shi J, Zhao Y, Wang K, Shi X, Wang Y, Huang H, et al. Cleavage of GSDMD by inflammatory caspases determines pyroptotic cell death. Nature. 2015;526:660–5.

    Article  CAS  PubMed  Google Scholar 

  46. Wegiel B, Larsen R, Gallo D, Chin B, Harris C, Mannam P, et al. Macrophages sense and kill bacteria through carbon monoxide-dependent inflammasome activation. J Clin Invest. 2014;124:4926–40.

    Article  PubMed Central  PubMed  Google Scholar 

  47. Piccini A, Carta S, Tassi S, Lasiglié D, Fossati G, Rubartelli A. ATP is released by monocytes stimulated with pathogen-sensing receptor ligands and induces IL-1beta and IL-18 secretion in an autocrine way. Proc Natl Acad Sci USA. 2008;105:8067–72.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  48. Hansson GC. Mucins and the microbiome. Annu Rev Biochem. 2020;89:769–93.

  49. Yin J, Sheng B, Yang K, Sun L, Xiao W, Yang H. The protective roles of NLRP6 in intestinal epithelial cells. Cell Prolif. 2019;52:e12555.

    Article  CAS  PubMed  Google Scholar 

  50. Zhu S, Ding S, Wang P, Wei Z, Pan W, Palm N, et al. Nlrp9b inflammasome restricts rotavirus infection in intestinal epithelial cells. Nature. 2017;546:667–70.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  51. Yao X, Zhang C, Xing Y, Xue G, Zhang Q, Pan F, et al. Remodelling of the gut microbiota by hyperactive NLRP3 induces regulatory T cells to maintain homeostasis. Nat Commun. 2017;8:1896.

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  52. Song-Zhao G, Srinivasan N, Pott J, Baban D, Frankel G, Maloy K. Nlrp3 activation in the intestinal epithelium protects against a mucosal pathogen. Mucosal Immunol. 2014;7:763–74.

    Article  CAS  PubMed  Google Scholar 

  53. Valera S, Hussy N, Evans RJ, Adami N, North RA, Surprenant A, et al. A new class of ligand-gated ion channel defined by P2x receptor for extracellular ATP. Nature. 1994;371:516–9.

    Article  CAS  PubMed  Google Scholar 

  54. Feske S, Wulff H & Skolnik EY. Ion channels in innate and adaptive immunity. Annu Rev Immunol. 2015;33:291–353.

  55. Toyama K, Wulff H, Chandy KG, Azam P, Raman G, Saito T, et al. The intermediate-conductance calcium-activated potassium channel KCa3.1 contributes to atherogenesis in mice and humans. J Clin Invest. 2008;118:3025–37.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  56. Thompson-Vest N, Shimizu Y, Hunne B, Furness JB. The distribution of intermediate-conductance, calcium-activated, potassium (IK) channels in epithelial cells. J Anat. 2006;208:219–29.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  57. Nguyen HM, Singh V, Pressly B, Jenkins DP, Wulff H, Yarov-Yarovoy V. Structural insights into the atomistic mechanisms of action of small molecule inhibitors targeting the KCa3.1 channel pore. Mol Pharmacol. 2017;91:392–402.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  58. Ohya S, Fukuyo Y, Kito H, Shibaoka R, Matsui M, Niguma H, et al. Upregulation of KCa3.1 K(+) channel in mesenteric lymph node CD4(+) T lymphocytes from a mouse model of dextran sodium sulfate-induced inflammatory bowel disease. Am J Physiol Gastrointest Liver Physiol. 2014;306:G873–G885.

    Article  CAS  PubMed  Google Scholar 

  59. Di L, Srivastava S, Zhdanova O, Ding Y, Li Z, Wulff H, et al. Inhibition of the K+ channel KCa3.1 ameliorates T cell-mediated colitis. Proc Natl Acad Sci USA. 2010;107:1541–6.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  60. Dieleman LA, Ridwan BU, Tennyson GS, Beagley KW, Bucy RP, Elson CO. Dextran sulfate sodium-induced colitis occurs in severe combined immunodeficient mice. Gastroenterology. 1994;107:1643–52.

    Article  CAS  PubMed  Google Scholar 

  61. Hansen L. The role of T cell potassium channels, KV1.3 and KCa3.1, in the inflammatory cascade in ulcerative colitis. Dan Med J. 2014;61:B4946.

    PubMed  Google Scholar 

  62. Stocker J, De Franceschi L, McNaughton-Smith G, Corrocher R, Beuzard Y, Brugnara C. ICA-17043, a novel Gardos channel blocker, prevents sickled red blood cell dehydration in vitro and in vivo in SAD mice. Blood. 2003;101:2412–8.

    Article  CAS  PubMed  Google Scholar 

  63. Brown B, Pressley B, Wulff H. KCa3.1 channel modulators as potential therapeutic compounds for glioblastoma. Curr Neuropharmacol. 2018;16:618–26.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  64. Liu L, Dong Y, Ye M, Jin S, Yang J, Joosse M, et al. The pathogenic role of NLRP3 inflammasome activation in inflammatory bowel diseases of both mice and humans. J Crohns Colitis. 2017;11:737–50.

    PubMed Central  PubMed  Google Scholar 

  65. Umiker B, Lee H, Cope J, Ajami N, Laine J, Fregeau C, et al. The NLRP3 inflammasome mediates DSS-induced intestinal inflammation in Nod2 knockout mice. Innate Immun. 2019;25:132–43.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  66. Zhang Y, Huang R, Cheng M, Wang L, Chao J, Li J, et al. Gut microbiota from NLRP3-deficient mice ameliorates depressive-like behaviors by regulating astrocyte dysfunction via circHIPK2. Microbiome. 2019;7:116.

    Article  PubMed Central  PubMed  Google Scholar 

  67. Friebel K, Schönherr R, Kinne RW, Kunisch E. Functional role of the KCa3.1 potassium channel in synovial fibroblasts from rheumatoid arthritis patients. J Cell Physiol. 2015;230:1677–88.

    Article  CAS  PubMed  Google Scholar 

  68. Vega G, Guequén A, Philp A, Gianotti A, Arzola L, Villalón M, et al. Lack of Kcnn4 improves mucociliary clearance in muco-obstructive lung disease. JCI insight. 2020;5:e140076.

    Article  PubMed Central  Google Scholar 

Download references

Funding

This work was supported by the National Natural Science Foundation of China (grant numbers 81773965 to X.H., 81873064 to DO, and 81673664 to QZ).

Author information

Authors and Affiliations

  1. Department of Clinical Laboratory, The Fifth Affiliated Hospital of Jinan University, Heyuan, China

    Bo Zeng, Qingbing Zha & Xianhui He

  2. Department of Immunobiology, College of Life Science and Technology, Jinan University, Guangzhou, China

    Bo Zeng, Yuanting Huang, Siyuan Chen, Rong Xu, Jiahao Qiu, Fuli Shi, Siying Liu, Dongyun Ouyang & Xianhui He

  3. Department of Cell Biology, College of Life Science and Technology, Jinan University, Guangzhou, China

    Lihui Xu

  4. Department of Fetal Medicine, The First Affiliated Hospital of Jinan University, Guangzhou, China

    Qingbing Zha

Authors
  1. Bo Zeng
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Yuanting Huang
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Siyuan Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Rong Xu
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Lihui Xu
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Jiahao Qiu
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Fuli Shi
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Siying Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Qingbing Zha
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Dongyun Ouyang
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Xianhui He
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

X.H, DO, QZ, and BZ designed the experiments. BZ, YH, SC, RX, LX, and JQ performed the experiments. BZ, FS, and SL acquired and analyzed the experimental data. BZ, DO, and XH prepared the manuscript. All authors have read and approved the final manuscript.

Corresponding authors

Correspondence to Qingbing Zha, Dongyun Ouyang or Xianhui He.

Ethics declarations

Conflict of interest

The authors declare no competing interests.

Supplementary information

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Zeng, B., Huang, Y., Chen, S. et al. Dextran sodium sulfate potentiates NLRP3 inflammasome activation by modulating the KCa3.1 potassium channel in a mouse model of colitis. Cell Mol Immunol 19, 925–943 (2022). https://doi.org/10.1038/s41423-022-00891-0

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/s41423-022-00891-0

Keywords

This article is cited by

Search

Advanced search

Quick links