Endosomal trafficking of two pore K+ efflux channel TWIK2 to plasmalemma mediates NLRP3 inflammasome activation and inflammatory injury

  1. Long Shuang Huang
  2. Mohammad Anas
  3. Jingsong Xu
  4. Bisheng Zhou
  5. Peter T Toth
  6. Yamuna Krishnan
  7. Anke Di
  8. Asrar B Malik  Is a corresponding author
  1. Shanghai Jiao Tong University, China
  2. University of Illinois at Chicago, United States
  3. University of Chicago, United States
  4. The University of Illinois at Chicago, United States

Abstract

Potassium efflux via the two-pore K+ channel TWIK2 is a requisite step for the activation of NLRP3 inflammasome, however, it remains unclear how K+ efflux is activated in response to select cues. Here we report that during homeostasis, TWIK2 resides in endosomal compartments. TWIK2 is transported by endosomal fusion to the plasmalemma in response to increased extracellular ATP resulting in the extrusion of K+. We showed that ATP-induced endosomal TWIK2 plasmalemma translocation is regulated by Rab11a. Deleting Rab11a or ATP-ligated purinergic receptor P2X7 each prevented endosomal fusion with the plasmalemma and K+ efflux as well as NLRP3 inflammasome activation in macrophages. Adoptive transfer of Rab11a-depleted macrophages into mouse lungs prevented NLRP3 inflammasome activation and inflammatory lung injury. We conclude that Rab11a-mediated endosomal trafficking in macrophages thus regulates TWIK2 localization and activity at the cell surface and the downstream activation of the NLRP3 inflammasome. Results show that endosomal trafficking of TWIK2 to the plasmalemma is a potential therapeutic target in acute or chronic inflammatory states.

Data availability

All data generated or analyzed during this study are included in the manuscript. Source Data have been provided for Figure 3, Figure 4, Figure 5, and Figure 6 for the Western blots.

Article and author information

Author details

  1. Long Shuang Huang

    Shanghai Frontiers Science Center of Drug Target Identification and Delivery, Shanghai Jiao Tong University, Shanghai, China
    Competing interests
    The authors declare that no competing interests exist.
  2. Mohammad Anas

    Department of Pharmacology and Regenerative Medicine, University of Illinois at Chicago, Chicago, United States
    Competing interests
    The authors declare that no competing interests exist.
  3. Jingsong Xu

    Department of Pharmacology and Regenerative Medicine, University of Illinois at Chicago, Chicago, United States
    Competing interests
    The authors declare that no competing interests exist.
  4. Bisheng Zhou

    Department of Pharmacology and Regenerative Medicine, University of Illinois at Chicago, Chicago, United States
    Competing interests
    The authors declare that no competing interests exist.
  5. Peter T Toth

    Fluorescence Imaging Core, University of Illinois at Chicago, Chicago, United States
    Competing interests
    The authors declare that no competing interests exist.
  6. Yamuna Krishnan

    Department of Chemistry, University of Chicago, Chicago, United States
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-5282-8852
  7. Anke Di

    Department of Pharmacology and Regenerative Medicine, University of Illinois at Chicago, Chicago, United States
    Competing interests
    The authors declare that no competing interests exist.
  8. Asrar B Malik

    Department of Pharmacology and Regenerative Medicine, The University of Illinois at Chicago, Chicago, United States
    For correspondence
    abmalik@uic.edu
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-8205-7128

Funding

National Institutes of Health (P01-HL60678)

  • Asrar B Malik

National Institutes of Health (P01-HL077806)

  • Asrar B Malik

National Institutes of Health (R01-HL118068)

  • Asrar B Malik

National Institutes of Health (R01-HL90152)

  • Asrar B Malik

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Reviewing Editor

  1. Sivapriya Vanaja

Ethics

Animal experimentation: All mice were housed in the University of Illinois Animal Care Facility in accordance with institutional and NIH guidelines. Veterinary care and animal experiments were approved by the University of Illinois Animal Care & Use Committee (ACC protocol number: 21-032).

Version history

  1. Received: September 30, 2022
  2. Preprint posted: October 12, 2022 (view preprint)
  3. Accepted: May 7, 2023
  4. Accepted Manuscript published: May 9, 2023 (version 1)
  5. Version of Record published: May 22, 2023 (version 2)

Copyright

© 2023, Huang et al.

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

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  1. Long Shuang Huang
  2. Mohammad Anas
  3. Jingsong Xu
  4. Bisheng Zhou
  5. Peter T Toth
  6. Yamuna Krishnan
  7. Anke Di
  8. Asrar B Malik
(2023)
Endosomal trafficking of two pore K+ efflux channel TWIK2 to plasmalemma mediates NLRP3 inflammasome activation and inflammatory injury
eLife 12:e83842.
https://doi.org/10.7554/eLife.83842

Share this article

https://doi.org/10.7554/eLife.83842

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