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NLRP3 inflammasome blockade reduces adipose tissue inflammation and extracellular matrix remodeling

Abstract

The NLRP3-IL-1β pathway plays an important role in adipose tissue (AT)-induced inflammation and the development of obesity-associated comorbidities. We aimed to determine the impact of NLRP3 on obesity and its associated metabolic alterations as well as its role in adipocyte inflammation and extracellular matrix (ECM) remodeling. Samples obtained from 98 subjects were used in a case−control study. The expression of different components of the inflammasome as well as their main effectors and inflammation- and ECM remodeling-related genes were analyzed. The impact of blocking NLRP3 using siRNA in lipopolysaccharide (LPS)-mediated inflammation and ECM remodeling signaling pathways was evaluated. We demonstrated that obesity (P < 0.01), obesity-associated T2D (P < 0.01) and NAFLD (P < 0.05) increased the expression of different components of the inflammasome as well as the expression and release of IL-1β and IL-18 in AT. We also found that obese patients with T2D exhibited increased (P < 0.05) hepatic gene expression levels of NLRP3, IL1B and IL18. We showed that NLRP3, but not NLRP1, is regulated by inflammation and hypoxia in visceral adipocytes. We revealed that the inhibition of NLRP3 in human visceral adipocytes significantly blocked (P < 0.01) LPS-induced inflammation by downregulating the mRNA levels of CCL2, IL1B, IL6, IL8, S100A8, S100A9, TLR4 and TNF as well as inhibiting (P < 0.01) the secretion of IL1-β into the culture medium. Furthermore, blocking NLRP3 attenuated (P < 0.01) the LPS-induced expression of important molecules involved in AT fibrosis (COL1A1, COL4A3, COL6A3 and MMP2). These novel findings provide evidence that blocking the expression of NLRP3 reduces AT inflammation with significant fibrosis attenuation.

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References

  1. 1.

    GBDO, Collaborators et al. Health effects of overweight and obesity in 195 countries over 25 years. N. Engl. J. Med. 377, 13–27 (2017).

    Google Scholar 

  2. 2.

    James, W. P. T. Obesity: a global public health challenge. Clin. Chem. 64, 24–29 (2018).

    CAS  PubMed  Google Scholar 

  3. 3.

    Lee, Y. S., Wollam, J. & Olefsky, J. M. An integrated view of immunometabolism. Cell 172, 22–40 (2018).

    CAS  PubMed  Google Scholar 

  4. 4.

    Bray, G. A., Frühbeck, G., Ryan, D. H. & Wilding, J. P. Management of obesity. Lancet 387, 1947–1956 (2016).

    PubMed  Google Scholar 

  5. 5.

    Ouchi, N., Parker, J. L., Lugus, J. J. & Walsh, K. Adipokines in inflammation and metabolic disease. Nat. Rev. Immunol. 11, 85–97 (2011).

    CAS  PubMed  PubMed Central  Google Scholar 

  6. 6.

    Reilly, S. M. & Saltiel, A. R. Adapting to obesity with adipose tissue inflammation. Nat. Rev. Endocrinol. 13, 633–643 (2017).

    CAS  PubMed  Google Scholar 

  7. 7.

    Hotamisligil, G. S. Foundations of immunometabolism and implications for metabolic health and disease. Immunity 47, 406–420 (2017).

    CAS  PubMed  PubMed Central  Google Scholar 

  8. 8.

    Wang, Q. & Wu, H. T cells in adipose tissue: critical players in immunometabolism. Front. Immunol. 9, 2509 (2018).

    PubMed  PubMed Central  Google Scholar 

  9. 9.

    Man, K., Kutyavin, V. I. & Chawla, A. Tissue immunometabolism: development, physiology, and pathobiology. Cell Metab. 25, 11–26 (2017).

    CAS  PubMed  Google Scholar 

  10. 10.

    Brestoff, J. R. & Artis, D. Immune regulation of metabolic homeostasis in health and disease. Cell 161, 146–160 (2015).

    CAS  PubMed  PubMed Central  Google Scholar 

  11. 11.

    Lamkanfi, M. & Dixit, V. M. Inflammasomes and their roles in health and disease. Annu. Rev. Cell Dev. Biol. 28, 137–161 (2012).

    CAS  PubMed  Google Scholar 

  12. 12.

    Osborn, O. & Olefsky, J. M. The cellular and signaling networks linking the immune system and metabolism in disease. Nat. Med. 18, 363–374 (2012).

    CAS  PubMed  Google Scholar 

  13. 13.

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

    CAS  PubMed  Google Scholar 

  14. 14.

    Schroder, K. & Tschopp, J. The inflammasomes. Cell 140, 821–832 (2010).

    CAS  PubMed  Google Scholar 

  15. 15.

    Lamkanfi, M. & Dixit, V. M. Mechanisms and functions of inflammasomes. Cell 157, 1013–1022 (2014).

    CAS  PubMed  Google Scholar 

  16. 16.

    De Nardo, D. & Latz, E. NLRP3 inflammasomes link inflammation and metabolic disease. Trends Immunol. 32, 373–379 (2011).

    PubMed  PubMed Central  Google Scholar 

  17. 17.

    Dinarello, C. A. Interleukin-1 in the pathogenesis and treatment of inflammatory diseases. Blood 117, 3720–3732 (2011).

    CAS  PubMed  PubMed Central  Google Scholar 

  18. 18.

    Donath, M. Y. & Shoelson, S. E. Type 2 diabetes as an inflammatory disease. Nat. Rev. Immunol. 11, 98–107 (2011).

    CAS  PubMed  Google Scholar 

  19. 19.

    Netea, M. G. et al. Deficiency of interleukin-18 in mice leads to hyperphagia, obesity and insulin resistance. Nat. Med. 12, 650–656 (2006).

    CAS  PubMed  Google Scholar 

  20. 20.

    Murphy, A. J. et al. IL-18 production from the NLRP1 inflammasome prevents obesity and metabolic syndrome. Cell Metab. 23, 155–164 (2016).

    CAS  PubMed  Google Scholar 

  21. 21.

    Stienstra, R. et al. The inflammasome-mediated caspase-1 activation controls adipocyte differentiation and insulin sensitivity. Cell Metab. 12, 593–605 (2010).

    CAS  PubMed  PubMed Central  Google Scholar 

  22. 22.

    Stienstra, R., Tack, C. J., Kanneganti, T. D., Joosten, L. A. & Netea, M. G. The inflammasome puts obesity in the danger zone. Cell Metab. 15, 10–18 (2012).

    CAS  PubMed  Google Scholar 

  23. 23.

    Netea, M. G. & Joosten, L. A. The NLRP1-IL18 connection: a stab in the back of obesity-induced inflammation. Cell Metab. 23, 6–7 (2016).

    CAS  PubMed  Google Scholar 

  24. 24.

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

    CAS  PubMed  PubMed Central  Google Scholar 

  25. 25.

    Sun, K., Tordjman, J., Clement, K. & Scherer, P. E. Fibrosis and adipose tissue dysfunction. Cell Metab. 18, 470–477 (2013).

    CAS  PubMed  PubMed Central  Google Scholar 

  26. 26.

    Virtue, S. & Vidal-Puig, A. Adipose tissue expandability, lipotoxicity and the metabolic syndrome-an allostatic perspective. Biochim. Biophys. Acta 1801, 338–349 (2010).

    CAS  PubMed  Google Scholar 

  27. 27.

    Khan, T. et al. Metabolic dysregulation and adipose tissue fibrosis: role of collagen VI. Mol. Cell Biol. 29, 1575–1591 (2009).

    CAS  PubMed  Google Scholar 

  28. 28.

    Kleiner, D. E. & Brunt, E. M. Nonalcoholic fatty liver disease: pathologic patterns and biopsy evaluation in clinical research. Semin. Liver Dis. 32, 3–13 (2012).

    CAS  PubMed  Google Scholar 

  29. 29.

    Ye, J. Emerging role of adipose tissue hypoxia in obesity and insulin resistance. Int. J. Obes. (Lond.) 33, 54–66 (2009).

    CAS  Google Scholar 

  30. 30.

    Zhang, S. Y. et al. Adipocyte-derived lysophosphatidylcholine activates adipocyte and adipose tissue macrophage nod-like receptor protein 3 inflammasomes mediating homocysteine-induced insulin resistance. EBioMedicine 31, 202–216 (2018).

    PubMed  PubMed Central  Google Scholar 

  31. 31.

    Ahechu, P. et al. NLRP3 inflammasome: a possible link between obesity-associated low-grade chronic inflammation and colorectal cancer development. Front. Immunol. 9, 2918 (2018).

    CAS  PubMed  PubMed Central  Google Scholar 

  32. 32.

    Mangan, M. S. J. et al. Targeting the NLRP3 inflammasome in inflammatory diseases. Nat. Rev. Drug Discov. 17, 688 (2018).

    CAS  PubMed  Google Scholar 

  33. 33.

    Zhou, R., Tardivel, A., Thorens, B., Choi, I. & Tschopp, J. Thioredoxin-interacting protein links oxidative stress to inflammasome activation. Nat. Immunol. 11, 136–140 (2010).

    CAS  PubMed  Google Scholar 

  34. 34.

    Esser, N. et al. Obesity phenotype is related to NLRP3 inflammasome activity and immunological profile of visceral adipose tissue. Diabetologia 56, 2487–2497 (2013).

    CAS  PubMed  Google Scholar 

  35. 35.

    Moschen, A. R. et al. Adipose and liver expression of interleukin (IL)-1 family members in morbid obesity and effects of weight loss. Mol. Med. 17, 840–845 (2011).

    CAS  PubMed  PubMed Central  Google Scholar 

  36. 36.

    Jourdan, T. et al. Activation of the Nlrp3 inflammasome in infiltrating macrophages by endocannabinoids mediates beta cell loss in type 2 diabetes. Nat. Med. 19, 1132–1140 (2013).

    CAS  PubMed  PubMed Central  Google Scholar 

  37. 37.

    Lamkanfi, M. et al. Glyburide inhibits the cryopyrin/Nalp3 inflammasome. J. Cell Biol. 187, 61–70 (2009).

    CAS  PubMed  PubMed Central  Google Scholar 

  38. 38.

    Kursawe, R. et al. A role of the inflammasome in the low storage capacity of the abdominal subcutaneous adipose tissue in obese adolescents. Diabetes 65, 610–618 (2016).

    CAS  PubMed  Google Scholar 

  39. 39.

    Mridha, A. R. et al. NLRP3 inflammasome blockade reduces liver inflammation and fibrosis in experimental NASH in mice. J. Hepatol. 66, 1037–1046 (2017).

    CAS  PubMed  PubMed Central  Google Scholar 

  40. 40.

    Csak, T. et al. Fatty acid and endotoxin activate inflammasomes in mouse hepatocytes that release danger signals to stimulate immune cells. Hepatology 54, 133–144 (2011).

    CAS  PubMed  PubMed Central  Google Scholar 

  41. 41.

    Kim, W. R., Flamm, S. L., Di Bisceglie, A. M. & Bodenheimer, H. C. Public Policy Committee of the American Association for the Study of Liver D. Serum activity of alanine aminotransferase (ALT) as an indicator of health and disease. Hepatology 47, 1363–1370 (2008).

    CAS  PubMed  Google Scholar 

  42. 42.

    Weisberg, S. P. et al. Obesity is associated with macrophage accumulation in adipose tissue. J. Clin. Invest. 112, 1796–1808 (2003).

    CAS  PubMed  PubMed Central  Google Scholar 

  43. 43.

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

    CAS  PubMed  PubMed Central  Google Scholar 

  44. 44.

    Lee, H. M. et al. Upregulated NLRP3 inflammasome activation in patients with type 2 diabetes. Diabetes 62, 194–204 (2013).

    CAS  PubMed  Google Scholar 

  45. 45.

    Yin, Z. et al. Transcriptome analysis of human adipocytes implicates the NOD-like receptor pathway in obesity-induced adipose inflammation. Mol. Cell Endocrinol. 394, 80–87 (2014).

    CAS  PubMed  PubMed Central  Google Scholar 

  46. 46.

    Crewe, C., An, Y. A. & Scherer, P. E. The ominous triad of adipose tissue dysfunction: inflammation, fibrosis, and impaired angiogenesis. J. Clin. Invest. 127, 74–82 (2017).

    PubMed  PubMed Central  Google Scholar 

  47. 47.

    Furuoka, M. et al. TNF-alpha induces caspase-1 activation independently of simultaneously induced NLRP3 in 3T3-L1 cells. J. Cell Physiol. 231, 2761–2767 (2016).

    CAS  PubMed  Google Scholar 

  48. 48.

    Vince, J. E. et al. The mitochondrial apoptotic effectors BAX/BAK activate caspase-3 and -7 to trigger NLRP3 inflammasome and caspase-8 driven IL-1beta activation. Cell Rep. 25, 2339–2353.e2334 (2018).

    CAS  PubMed  Google Scholar 

  49. 49.

    Gurung, P. et al. Toll or interleukin-1 receptor (TIR) domain-containing adaptor inducing interferon-beta (TRIF)-mediated caspase-11 protease production integrates Toll-like receptor 4 (TLR4) protein- and Nlrp3 inflammasome-mediated host defense against enteropathogens. J. Biol. Chem. 287, 34474–34483 (2012).

    CAS  PubMed  PubMed Central  Google Scholar 

  50. 50.

    Wree, A. et al. NLRP3 inflammasome activation results in hepatocyte pyroptosis, liver inflammation, and fibrosis in mice. Hepatology 59, 898–910 (2014).

    CAS  PubMed  PubMed Central  Google Scholar 

  51. 51.

    Gasse, P. et al. Uric acid is a danger signal activating NALP3 inflammasome in lung injury inflammation and fibrosis. Am. J. Respir. Crit. Care Med. 179, 903–913 (2009).

    CAS  PubMed  Google Scholar 

  52. 52.

    Youm, Y. H. et al. Elimination of the NLRP3-ASC inflammasome protects against chronic obesity-induced pancreatic damage. Endocrinology 152, 4039–4045 (2011).

    CAS  PubMed  PubMed Central  Google Scholar 

  53. 53.

    Bakker, P. J. et al. Nlrp3 is a key modulator of diet-induced nephropathy and renal cholesterol accumulation. Kidney Int. 85, 1112–1122 (2014).

    CAS  PubMed  Google Scholar 

  54. 54.

    Watanabe, A. et al. Inflammasome-mediated regulation of hepatic stellate cells. Am. J. Physiol. Gastrointest. Liver Physiol. 296, G1248–G1257 (2009).

    CAS  PubMed  PubMed Central  Google Scholar 

  55. 55.

    Robert, S. et al. Involvement of matrix metalloproteinases (MMPs) and inflammasome pathway in molecular mechanisms of fibrosis. Biosci. Rep. 36, 1–11 (2016).

  56. 56.

    Qu, J., Yuan, Z., Wang, G., Wang, X. & Li, K. The selective NLRP3 inflammasome inhibitor MCC950 alleviates cholestatic liver injury and fibrosis in mice. Int. Immunopharmacol. 70, 147–155 (2019).

    CAS  PubMed  Google Scholar 

  57. 57.

    Sun, S., Xia, S., Ji, Y., Kersten, S. & Qi, L. The ATP-P2X7 signaling axis is dispensable for obesity-associated inflammasome activation in adipose tissue. Diabetes 61, 1471–1478 (2012).

    CAS  PubMed  PubMed Central  Google Scholar 

  58. 58.

    Martinon, F., Mayor, A. & Tschopp, J. The inflammasomes: guardians of the body. Annu. Rev. Immunol. 27, 229–265 (2009).

    CAS  PubMed  Google Scholar 

  59. 59.

    American Diabetes Association. Classification and diagnosis of diabetes: standards of medical care in diabetes-2019. Diabetes Care 42, S13–S28 (2019).

    Google Scholar 

  60. 60.

    Gómez-Ambrosi, J. et al. Increased cardiometabolic risk factors and inflammation in adipose tissue in obese subjects classified as metabolically healthy. Diabetes Care 37, 2813–2821 (2014).

    PubMed  Google Scholar 

  61. 61.

    Catalán, V. et al. Increased interleukin-32 levels in obesity promote adipose tissue inflammation and extracellular matrix remodeling: effect of weight loss. Diabetes 65, 3636–3648 (2016).

    PubMed  Google Scholar 

  62. 62.

    Angulo, P., Keach, J. C., Batts, K. P. & Lindor, K. D. Independent predictors of liver fibrosis in patients with nonalcoholic steatohepatitis. Hepatology 30, 1356–1362 (1999).

    CAS  PubMed  Google Scholar 

  63. 63.

    Muruzabal, F. J., Frühbeck, G., Gómez-Ambrosi, J., Archanco, M. & Burrell, M. A. Immunocytochemical detection of leptin in non-mammalian vertebrate stomach. Gen. Comp. Endocrinol. 128, 149–152 (2002).

    CAS  PubMed  Google Scholar 

  64. 64.

    Catalán, V. et al. Validation of endogenous control genes in human adipose tissue: relevance to obesity and obesity-associated type 2 diabetes mellitus. Horm. Metab. Res. 39, 495–500 (2007).

    PubMed  Google Scholar 

  65. 65.

    Divoux, A. et al. Fibrosis in human adipose tissue: composition, distribution, and link with lipid metabolism and fat mass loss. Diabetes 59, 2817–2825 (2010).

    CAS  PubMed  PubMed Central  Google Scholar 

  66. 66.

    Rodríguez, A. et al. Acylated and desacyl ghrelin stimulate lipid accumulation in human visceral adipocytes. Int. J. Obes. (Lond.) 33, 541–552 (2009).

    Google Scholar 

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Acknowledgements

The authors gratefully acknowledge the valuable collaboration of all the members of the Multidisciplinary Obesity Team, Clínica Universidad de Navarra, Pamplona, Spain. This work was supported by Plan Estatal I+D+I from the Spanish Instituto de Salud Carlos III–Subdirección General de Evaluación y Fomento de la investigación–FEDER (grants number PI16/01217, PI17/02183 and PI17/02188), by the Gobierno de Navarra (10/2018) and by CIBEROBN, ISCIII, Spain.

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X.U. collected and analyzed data, wrote the first draft of the manuscript, contributed to discussion, and reviewed the manuscript. J.G.-A., A.R. and S.B. collected and analyzed the data, contributed to the discussion, and reviewed the manuscript. V.V., C.S., and J.S. enrolled patients, collected data, contributed to discussion, and reviewed the manuscript. B.R. and R.M. collected data, contributed to discussion, and reviewed the manuscript. G.F. designed the study, wrote the first draft of the manuscript, contributed to discussion, and reviewed the manuscript. V.C. designed the study, collected and analyzed data, wrote the first draft of the manuscript, contributed to discussion, and reviewed the manuscript. V.C. and G.F. are guarantors for the contents of the article and had full access to all the data in the study and take responsibility for the integrity of the data and the accuracy of the data analysis.

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Correspondence to Gema Frühbeck or Victoria Catalán.

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The authors declare no competing interests.

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The study was approved, from an ethical and scientific standpoint, by the Clínica Universidad de Navarra’s Ethical Committee responsible for research, and written informed consent of participants was obtained.

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Unamuno, X., Gómez-Ambrosi, J., Ramírez, B. et al. NLRP3 inflammasome blockade reduces adipose tissue inflammation and extracellular matrix remodeling. Cell Mol Immunol 18, 1045–1057 (2021). https://doi.org/10.1038/s41423-019-0296-z

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Keywords

  • NLRP3
  • Inflammasone
  • Inflammation
  • Obesity
  • Type 2 diabetes
  • Nonalcoholic fatty liver disease

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