Abstract
NLRP3-inflammasome-driven inflammation is involved in the pathogenesis of a variety of diseases. Identification of endogenous inflammasome activators is essential for the development of new anti-inflammatory treatment strategies. Here, we identified that apolipoprotein C3 (ApoC3) activates the NLRP3 inflammasome in human monocytes by inducing an alternative NLRP3 inflammasome via caspase-8 and dimerization of Toll-like receptors 2 and 4. Alternative inflammasome activation in human monocytes is mediated by the Toll-like receptor adapter protein SCIMP. This triggers Lyn/Syk-dependent calcium entry and the production of reactive oxygen species, leading to activation of caspase-8. In humanized mouse models, ApoC3 activated human monocytes in vivo to impede endothelial regeneration and promote kidney injury in an NLRP3- and caspase-8-dependent manner. These data provide new insights into the regulation of the NLRP3 inflammasome and the pathophysiological role of triglyceride-rich lipoproteins containing ApoC3. Targeting ApoC3 might prevent organ damage and provide an anti-inflammatory treatment for vascular and kidney diseases.
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References
Hansson, G. K. & Hermansson, A. The immune system in atherosclerosis. Nat. Immunol. 12, 204–212 (2011).
Broderick, L., De Nardo, D., Franklin, B. S., Hoffman, H. M. & Latz, E. The inflammasomes and autoinflammatory syndromes. Annu. Rev. Pathol. 10, 395–424 (2015).
Leemans, J. C., Kors, L., Anders, H. J. & Florquin, S. Pattern recognition receptors and the inflammasome in kidney disease. Nat. Rev. Nephrol. 10, 398–414 (2014).
Lamkanfi, M. & Dixit, V. M. Mechanisms and functions of inflammasomes. Cell 157, 1013–1022 (2014).
Cai, X. et al. Prion-like polymerization underlies signal transduction in antiviral immune defense and inflammasome activation. Cell 156, 1207–1222 (2014).
Man, S. M. & Kanneganti, T. D. Converging roles of caspases in inflammasome activation, cell death and innate immunity. Nat. Rev. Immunol. 16, 7–21 (2016).
Latz, E., Xiao, T. S. & Stutz, A. Activation and regulation of the inflammasomes. Nat. Rev. Immunol. 13, 397–411 (2013).
Yang, Y., Wang, H., Kouadir, M., Song, H. & Shi, F. Recent advances in the mechanisms of NLRP3 inflammasome activation and its inhibitors. Cell Death Dis. 10, 128 (2019).
Bauernfeind, F. G. et al. Cutting edge: NF-κB activating pattern recognition and cytokine receptors license NLRP3 inflammasome activation by regulating NLRP3 expression. J. Immunol. 183, 787–791 (2009).
Ridker, P. M. et al. Antiinflammatory therapy with canakinumab for atherosclerotic disease. N. Engl. J. Med. 377, 1119–1131 (2017).
Speer, T. et al. Abnormal high-density lipoprotein induces endothelial dysfunction via activation of Toll-like receptor-2. Immunity 38, 754–768 (2013).
Lepedda, A. J. et al. Proteomic analysis of plasma-purified VLDL, LDL, and HDL fractions from atherosclerotic patients undergoing carotid endarterectomy: identification of serum amyloid A as a potential marker. Oxid. Med Cell Longev. 2013, 385214 (2013).
Shridas, P., De Beer, M. C. & Webb, N. R. High-density lipoprotein inhibits serum amyloid A-mediated reactive oxygen species generation and NLRP3 inflammasome activation. J. Biol. Chem. 293, 13257–13269 (2018).
Westerterp, M. et al. Cholesterol accumulation in dendritic cells links the inflammasome to acquired immunity. Cell Metab. 25, 1294–1304.e6 (2017).
Juntti-Berggren, L. et al. Apolipoprotein CIII promotes Ca2+-dependent β cell death in type 1 diabetes. Proc. Natl Acad. Sci. USA 101, 10090–10094 (2004).
Zheng, C. et al. Statins suppress apolipoprotein CIII-induced vascular endothelial cell activation and monocyte adhesion. Eur. Heart J. 34, 615–624 (2013).
Gaidt, M. M. et al. Human monocytes engage an alternative inflammasome pathway. Immunity 44, 833–846 (2016).
Tseng, H. H., Vong, C. T., Kwan, Y. W., Lee, S. M. & Hoi, M. P. TRPM2 regulates TXNIP-mediated NLRP3 inflammasome activation via interaction with p47 phox under high glucose in human monocytic cells. Sci. Rep. 6, 35016 (2016).
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).
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).
Gross, O. et al. Syk kinase signalling couples to the Nlrp3 inflammasome for anti-fungal host defence. Nature 459, 433–436 (2009).
Han, C. et al. Integrin CD11b negatively regulates TLR-triggered inflammatory responses by activating Syk and promoting degradation of MyD88 and TRIF via Cbl-b. Nat. Immunol. 11, 734–742 (2010).
Hara, H. et al. Phosphorylation of the adaptor ASC acts as a molecular switch that controls the formation of speck-like aggregates and inflammasome activity. Nat. Immunol. 14, 1247–1255 (2013).
Rolli, V. et al. Amplification of B cell antigen receptor signaling by a Syk/ITAM positive feedback loop. Mol. Cell 10, 1057–1069 (2002).
Luo, L. et al. SCIMP is a transmembrane non-TIR TLR adaptor that promotes proinflammatory cytokine production from macrophages. Nat. Commun. 8, 14133 (2017).
Hahm, E. et al. Bone marrow-derived immature myeloid cells are a main source of circulating suPAR contributing to proteinuric kidney disease. Nat. Med. 23, 100–106 (2017).
Zewinger, S. et al. Relations between lipoprotein(a) concentrations, LPA genetic variants, and the risk of mortality in patients with established coronary heart disease: a molecular and genetic association study. Lancet Diabetes Endocrinol. 5, 534–543 (2017).
Jin, M. S. & Lee, J. O. Structures of the Toll-like receptor family and its ligand complexes. Immunity 29, 182–191 (2008).
Kang, J. Y. et al. Recognition of lipopeptide patterns by Toll-like receptor 2–Toll-like receptor 6 heterodimer. Immunity 31, 873–884 (2009).
Jin, M. S. et al. Crystal structure of the TLR1–TLR2 heterodimer induced by binding of a tri-acylated lipopeptide. Cell 130, 1071–1082 (2007).
Lee, H. K., Dunzendorfer, S. & Tobias, P. S. Cytoplasmic domain-mediated dimerizations of Toll-like receptor 4 observed by β-lactamase enzyme fragment complementation. J. Biol. Chem. 279, 10564–10574 (2004).
Latz, E. et al. Ligand-induced conformational changes allosterically activate Toll-like receptor 9. Nat. Immunol. 8, 772–779 (2007).
Liu, T. et al. Single-cell imaging of caspase-1 dynamics reveals an all-or-none inflammasome signaling response. Cell Rep. 8, 974–982 (2014).
Kralova, J. et al. The transmembrane adaptor protein SCIMP facilitates sustained dectin-1 signaling in dendritic cells. J. Biol. Chem. 291, 16530–16540 (2016).
Amoui, M., Draberova, L., Tolar, P. & Draber, P. Direct interaction of Syk and Lyn protein tyrosine kinases in rat basophilic leukemia cells activated via type I Fc epsilon receptors. Eur. J. Immunol. 27, 321–328 (1997).
Katsnelson, M. A., Rucker, L. G., Russo, H. M. & Dubyak, G. R. K+ efflux agonists induce NLRP3 inflammasome activation independently of Ca2+ signaling. J. Immunol. 194, 3937–3952 (2015).
Gimbrone, M. A. Jr & Garcia-Cardena, G. Endothelial cell dysfunction and the pathobiology of atherosclerosis. Circ. Res. 118, 620–636 (2016).
Pollin, T. I. et al. A null mutation in human APOC3 confers a favorable plasma lipid profile and apparent cardioprotection. Science 322, 1702–1705 (2008).
Jorgensen, A. B., Frikke-Schmidt, R., Nordestgaard, B. G. & Tybjaerg-Hansen, A. Loss-of-function mutations in APOC3 and risk of ischemic vascular disease. N. Engl. J. Med. 371, 32–41 (2014).
TG and HDL Working Group of the Exome Sequencing Projectet al. Loss-of-function mutations in APOC3, triglycerides, and coronary disease. N. Engl. J. Med. 371, 22–31 (2014).
Saleheen, D. et al. Human knockouts and phenotypic analysis in a cohort with a high rate of consanguinity. Nature 544, 235–239 (2017).
Khetarpal, S. A. et al. A human APOC3 missense variant and monoclonal antibody accelerate apoC-III clearance and lower triglyceride-rich lipoprotein levels. Nat. Med. 23, 1086–1094 (2017).
Wyler von Ballmoos, M. C., Haring, B. & Sacks, F. M. The risk of cardiovascular events with increased apolipoprotein CIII: a systematic review and meta-analysis. J. Clin. Lipidol. 9, 498–510 (2015).
Gaudet, D. et al. Antisense inhibition of apolipoprotein C-III in patients with hypertriglyceridemia. N. Engl. J. Med. 373, 438–447 (2015).
Freigang, S. et al. Fatty acid-induced mitochondrial uncoupling elicits inflammasome-independent IL-1α and sterile vascular inflammation in atherosclerosis. Nat. Immunol. 14, 1045–1053 (2013).
KDIGO. KDIGO 2012 clinical practice guideline for the evaluation and management of chronic kidney disease. Kidney Int Suppl. 3, 1–150 (2013).
Winkelmann, B. R. et al. Rationale and design of the LURIC study—a resource for functional genomics, pharmacogenomics and long-term prognosis of cardiovascular disease. Pharmacogenomics 2, S1–S73 (2001).
Fredenrich, A. et al. Plasma lipoprotein distribution of apoC-III in normolipidemic and hypertriglyceridemic subjects: comparison of the apoC-III to apoE ratio in different lipoprotein fractions. J. Lipid Res. 38, 1421–1432 (1997).
Shroff, R. et al. HDL in children with CKD promotes endothelial dysfunction and an abnormal vascular phenotype. J. Am. Soc. Nephrol. 25, 2658–2668 (2014).
Zewinger, S. et al. Serum amyloid A: high-density lipoproteins interaction and cardiovascular risk. Eur. Heart J. 36, 3007–3016 (2015).
Zewinger, S. et al. HDL cholesterol is not associated with lower mortality in patients with kidney dysfunction. J. Am. Soc. Nephrol. 25, 1073–1082 (2014).
Wessel, D. & Flugge, U. I. A method for the quantitative recovery of protein in dilute solution in the presence of detergents and lipids. Anal. Biochem. 138, 141–143 (1984).
Steyrer, E. & Kostner, G. M. Activation of lecithin-cholesterol acyltransferase by apolipoprotein D: comparison of proteoliposomes containing apolipoprotein D, A-I or C-I. Biochim. Biophys. Acta 958, 484–491 (1988).
Jankowski, V. et al. The enzymatic activity of the VEGFR2 receptor for the biosynthesis of dinucleoside polyphosphates. J. Mol. Med. (Berl.) 91, 1095–1107 (2013).
Acknowledgements
This work was supported by grants from the Deutsche Forschungsgemeinschaft (SFB/TRR 219), Else-Kröener Fresenius Foundation, Deutsche Nierenstiftung and European Uremic Toxin Work Group of the ERA-EDTA. This study was supported by funding from the European Community’s program ‘European Regional Development Fund Interreg’ to ‘EURIPIDS’ and the Deutsche Forschungsgemeinschaft (SFB894 A2, SFB1027 C4 and SFB TRR 219).
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S.Z., D.F., U.L. and T.S. conceived of the study idea. S.Z., J.R., E.H., V.J., G.K., C.K., B.A.N., L.R., D.F., U.L. and T.S. designed the methodology. S.Z., W.M. and T.S. performed the formal analysis. S.Z., V.J., G.K., D.A., D.S., S.J.S., R.K., E.A., M.Klug, S.T., C.K., S.-R.S., G.A., R.B., A.P., T.H., B.A.N. and T.S. performed the investigation. W.M., J.J., C.K., M.Kopf, G.K., M.W.L., M.D.M. and B.A.N. provided resources. S.Z., D.F., U.L. and T.S. wrote the original draft of the manuscript. J.R., E.H., V.J., G.K., D.A., D.S., S.J.S., R.K., E.A., C.K., S.-R.S., S.S., G.S., M.S., U.S., W.J.-D., L.R., G.A., R.B., M.W.L., M.D.M., W.M., M.B., J.J., M.Kopf, E.L. and B.A.N. reviewed and edited the manuscript. S.Z., V.J. and T.S. visualized the results. S.Z., D.F., U.L. and T.S. acquired funding and supervised the research.
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Extended data
Extended Data Fig. 1
ApoC3 induces alternative NLRP3 inflammasome activation in human monocytes.
Extended Data Fig. 2
ApoC3 induces TLR-dependent effector responses.
Extended Data Fig. 3
TLR2 and TLR4 mediate activation of human monocytes in response to ApoC3.
Extended Data Fig. 4
TRPM2 is required to mediate alternative inflammasome activation.
Extended Data Fig. 5
ApoC3 induces phosphorylation of Syk downstream of Trif.
Extended Data Fig. 6
Summary on the mechanisms leading to alternative inflammasome activation in human monocytes by ApoC3.
Extended Data Fig. 7
Characterization of the humanized mouse models.
Extended Data Fig. 8
Histological changes in humanized mice after unilateral ureter ligation.
Supplementary information
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Zewinger, S., Reiser, J., Jankowski, V. et al. Apolipoprotein C3 induces inflammation and organ damage by alternative inflammasome activation. Nat Immunol 21, 30–41 (2020). https://doi.org/10.1038/s41590-019-0548-1
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DOI: https://doi.org/10.1038/s41590-019-0548-1
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