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Microtubule-driven spatial arrangement of mitochondria promotes activation of the NLRP3 inflammasome

Abstract

NLRP3 forms an inflammasome with its adaptor ASC, and its excessive activation can cause inflammatory diseases. However, little is known about the mechanisms that control assembly of the inflammasome complex. Here we show that microtubules mediated assembly of the NLRP3 inflammasome. Inducers of the NLRP3 inflammasome caused aberrant mitochondrial homeostasis to diminish the concentration of the coenzyme NAD+, which in turn inactivated the NAD+-dependent α-tubulin deacetylase sirtuin 2; this resulted in the accumulation of acetylated α-tubulin. Acetylated α-tubulin mediated the dynein-dependent transport of mitochondria and subsequent apposition of ASC on mitochondria to NLRP3 on the endoplasmic reticulum. Therefore, in addition to direct activation of NLRP3, the creation of optimal sites for signal transduction by microtubules is required for activation of the entire NLRP3 inflammasome.

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Figure 1: Inhibitors of tubulin polymerization specifically suppress activation of the NLRP3 inflammasome.
Figure 2: Microtubules mediate the approximation of ASC on mitochondria to NLRP3 on the endoplasmic reticulum in response to inducers of the NLRP3 inflammasome.
Figure 3: Dynein mediates the approximation of ASC on mitochondria to NLRP3 on the endoplasmic reticulum in response to inducers of the NLRP3 inflammasome.
Figure 4: Inducers of the NLRP3 inflammasome cause accumulation of acetylated α-tubulin in the perinuclear region.
Figure 5: Involvement of MEC-17 in activation of the NLRP3 inflammasome.
Figure 6: Acetylated α-tubulin and mitochondria accumulate in the perinuclear region after inactivation of SIRT2.
Figure 7: Loss of intracellular NAD+ promotes activation of the NLRP3 inflammasome.
Figure 8: Mitochondrial damage results in a lower abundance of intracellular NAD+.

References

  1. Núñez, G. Intracellular sensors of microbes and danger. Immunol. Rev. 243, 5–8 (2011).

    Article  PubMed  Google Scholar 

  2. Strowig, T., Henao-Mejia, J., Elinav, E. & Flavell, R. Inflammasomes in health and disease. Nature 481, 278–286 (2012).

    Article  CAS  PubMed  Google Scholar 

  3. Martinon, F. et al. Gout-associated uric acid crystals activate the NALP3 inflammasome. Nature 440, 237–241 (2006).

    Article  CAS  PubMed  Google Scholar 

  4. Duewell, P. et al. NLRP3 inflammasomes are required for atherogenesis and activated by cholesterol crystals. Nature 464, 1357–1361 (2010).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Masters, S.L. et al. Activation of the NLRP3 inflammasome by islet amyloid polypeptide provides a mechanism for enhanced IL-1β in type 2 diabetes. Nat. Immunol. 11, 897–904 (2010).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Vandanmagsar, B. et al. The NLRP3 inflammasome instigates obesity-induced inflammation and insulin resistance. Nat. Med. 17, 179–188 (2011).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Wen, H. et al. Fatty acid-induced NLRP3-ASC inflammasome activation interferes with insulin signaling. Nat. Immunol. 12, 408–415 (2011).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Cronstein, B.N. & Terkeltaub, R. The inflammatory process of gout and its treatment. Arthritis Res. Ther. 8, S3 (2006).

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  9. Allen, J.N. et al. Colchicine has opposite effects on interleukin-1β and tumor necrosis factor-alpha production. Am. J. Physiol. 26, 315–21 (1991).

    Google Scholar 

  10. Carta, S. et al. Histone deacetylase inhibitors prevent exocytosis of interleukin-1β-containing secretory lysosomes: role of microtubules. Blood 108, 1618–1626 (2006).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Zhou, R., Yazdi, A.S., Menu, P. & Tschopp, J. A role for mitochondria in NLRP3 inflammasome activation. Nature 469, 221–225 (2011).

    Article  CAS  PubMed  Google Scholar 

  12. Howard, J. & Hyman, A.A. Dynamics and mechanics of the microtubule plus end. Nature 422, 753–758 (2003).

    Article  CAS  PubMed  Google Scholar 

  13. Kardon, J.R. & Vale, R.D. Regulators of the cytoplasmic dynein motor. Nat. Rev. Mol. Cell Biol. 10, 854–865 (2009).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Firestone, A.J. et al. Small-molecule inhibitors of the AAA+ ATPase motor cytoplasmic dynein. Nature 484, 125–129 (2012).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Naik, E. & Dixit, V.M. Mitochondrial reactive oxygen species drive proinflammatory cytokine production. J. Exp. Med. 208, 417–420 (2011).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Wloga, D. & Gaertig, J. Post-translational modifications of microtubules. J. Cell Sci. 123, 3447–3455 (2010).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Reed, N.A. et al. Microtubule acetylation promotes kinesin-1 binding and transport. Curr. Biol. 16, 2166–2172 (2006).

    Article  CAS  PubMed  Google Scholar 

  18. Dompierre, J.P. et al. Histone deacetylase 6 inhibition compensates for the transport deficit in Huntington's disease by increasing tubulin acetylation. J. Neurosci. 27, 3571–3583 (2007).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. Friedman, J. et al. ER sliding dynamics and ER-mitochondrial contacts occur on acetylated microtubules. J. Cell Biol. 190, 363–375 (2010).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. Akella, J. et al. MEC-17 is an α-tubulin acetyltransferase. Nature 467, 218–222 (2010).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. North, B. et al. The human ortholog, SIRT2, is an NAD+-dependent tubulin deacetylase. Mol. Cell 11, 437–444 (2003).

    Article  CAS  PubMed  Google Scholar 

  22. Hubbert, C. et al. HDAC6 is a microtubule-associated deacetylase. Nature 417, 455–458 (2002).

    Article  CAS  PubMed  Google Scholar 

  23. Nunnari, J. & Suomalainen, A. Mitochondria: in sickness and in health. Cell 148, 1145–1159 (2012).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  24. Hentze, H., Lin, X.Y., Choi, M.S.K. & Porter, A.G. Critical role of cathepsin B in mediating caspase-1-dependent interleukin-18 maturation and caspase-1-independent necrosis triggered by the microbial toxin negericin. Cell Death Differ. 10, 956–968 (2003).

    Article  CAS  PubMed  Google Scholar 

  25. Gracia-Marcos, M. et al. Role of sodium in mitochondrial depolarization induced by P2X7 receptor activation in submandibular glands. FEBS Lett. 579, 5407–5413 (2005).

    Article  CAS  Google Scholar 

  26. Li, Z. et al. The lysosomal-mitochondria axis in free fatty acid-induced hepatic lipotoxicity. Hepatology 47, 1495–1503 (2011).

    Article  CAS  Google Scholar 

  27. McGuire, K. et al. Adenovirus type 5 rupture of lysosome leads to cathepsin B-dependent mitochondrial stress and production of reactive oxygen species. J. Virol. 85, 10806–10813 (2011).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  28. Tanaka, A. Parkin-mediated selective mitochondrial autophagy, mitophagy: Parkin purges damaged organelles from the vital mitochondrial network. FEBS Lett. 584, 1386–1392 (2010).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  29. Hornung, V. et al. Silica crystals and aluminum salts activate the NALP3 inflammasome through phagosomal destabilization. Nat. Immunol. 9, 847–856 (2008).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  30. Dostert, C. et al. Innate immune activation through Nalp3 inflammasome sensing of asbestos and silica. Science 320, 674–677 (2008).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  31. Ichinohe, T. et al. Influenza virus activates inflammasomes via its intracellular M2 ion channel. Nat. Immunol. 11, 404–410 (2010).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  32. Shenoy, A. et al. GBP5 promotes NLRP3 inflammasome assembly and immunity in mammals. Science 336, 481–485 (2012).

    Article  CAS  PubMed  Google Scholar 

  33. Hornung, V. et al. AIM2 recognizes cytosolic dsDNA and forms a caspase-1-activating inflammasome with ASC. Nature 458, 514–518 (2009).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  34. Mariathasan, S. et al. Differential activation of the inflammasome by caspase-1 adaptors ASC and Ipaf. Nature 430, 213–218 (2004).

    Article  CAS  PubMed  Google Scholar 

  35. Broz, P. et al. Redundant roles for inflammasome receptors NLRP3 and NLRC4 in host defense against Salmonella. J. Exp. Med. 207, 1745–1755 (2010).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  36. Li., Z. et al. The lysosomal-mitochondria axis in free fatty acid-induced hepatic lipotoxicity. Hepatology 47, 1495–503 (2008).

    Article  CAS  PubMed  Google Scholar 

  37. Yoshino, J., Mills, K.F., Yoon, M.J. & Imai, S. Nicotinamide mononucleotide, a key NAD+ intermediate, treats pathophysiology of diet- and age-induced diabetes in mice. Cell Metab. 14, 528–536 (2011).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  38. Sorbara, M.T. & Girardin, S.E. Mitochondrial ROS fuel the inflammasome. Cell Res. 21, 558–560 (2011).

    Article  PubMed  PubMed Central  Google Scholar 

  39. Saitoh, T. & Akira, S. Regulation of innate immune responses by autophagy-related proteins. J. Cell Biol. 189, 925–935 (2010).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  40. Itakura, E. et al. Structures containing Atg9A and the ULK1 complex independently target depolarized mitochondria at initial stages of Parkin-mediated mitophagy. J. Cell Sci. 125, 1488–1499 (2012).

    CAS  PubMed  Google Scholar 

  41. Zhao, Y. et al. Cytosolic FoxO1 is essential for the induction of autophagy and tumour suppressor activity. Nat. Cell Biol. 12, 665–675 (2010).

    Article  CAS  PubMed  Google Scholar 

  42. Kume, S. et al. Calorie restriction enhances cell adaption to hypoxia through Sirt1-dependent mitochondrial autophagy in mouse aged kidney. J. Clin. Invest. 120, 1043–1055 (2010).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  43. Chen, D. et al. Tissue specific regulation of SIRT1 by calorie restriction. Genes Dev. 22, 1753–1757 (2008).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  44. Saitoh, T. et al. Loss of the autophagy protein Atg16L1 enhances endotoxin-induced IL-1β production. Nature 456, 264–268 (2008).

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

We thank T. Kitamura (Tokyo University) for Plat-E packaging cells; H. Osada and T. Saito (RIKEN Natural Products Depository) for the authentic chemical compound library of the RIKEN Natural Products Depository; T. Yoshimori and K. Ikegami for critical reading of the manuscript; and members of the Laboratory of Host Defense for assistance. Supported by Japan Society for the Promotion of Science Grant-in-Aid for Specially Promoted Research (S.A.), Japan Society for the Promotion of Science Funding Program for World-Leading Innovative R&D on Science and Technology 'FIRST Program' (S.A.) and Japan Science and Technology Agency Core Research for Evolutional Science and Technology (T.S.).

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T.M. did most of the experiments and analyzed the data; M.T., T.K., H.L. and J.Z. helped with the experiments; T.M. and T.S. designed most of the experiments and wrote the manuscript; and S.A. supervised the overall research project.

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Correspondence to Tatsuya Saitoh or Shizuo Akira.

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The authors declare no competing financial interests.

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Misawa, T., Takahama, M., Kozaki, T. et al. Microtubule-driven spatial arrangement of mitochondria promotes activation of the NLRP3 inflammasome. Nat Immunol 14, 454–460 (2013). https://doi.org/10.1038/ni.2550

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