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Cathepsin B/NLRP3/GSDMD axis-mediated macrophage pyroptosis induces inflammation and fibrosis in systemic sclerosis

Published:December 24, 2022DOI:https://doi.org/10.1016/j.jdermsci.2022.12.006

      Highlights

      • Macrophage pyroptosis was increased in SSc patients and BLM-induced SSc mouse model.
      • Inhibition of pyroptosis reduced inflammation and fibrosis of SSc mouse model.
      • Inhibiting NLRP3/GSDMD-mediated macrophage pyroptosis inactivates fibroblasts.

      Abstract

      Background

      Pyroptosis is a newly discovered type of programmed cell death associated with inflammatory and fibrotic diseases. Macrophages play an important role in inducing early immune inflammation in systemic sclerosis (SSc).

      Objective

      To investigate the effect of macrophages pyroptosis on fibrosis of SSc.

      Methods

      Pyroptosis/inflammatory markers in serum and skin of SSc patients were detected. Bleomycin (BLM) was subcutaneously injected to establish SSc mouse model. The levels of pyroptosis markers, dermal thickness and collagen deposition in skin were assessed before and after the administration of pyroptosis inhibitors, including MCC950, Disulfiram and necrosulfonamide (NSA). Human-derived monocyte-macrophage cell line (THP-1) or mouse bone marrow-derived macrophages (BMDMs) were primed with lipopolysaccharide (LPS) and stimulated by silicon dioxide (SiO2) to induce cell pyroptosis. Fibroblasts from patients with SSc were co-cultured with pyroptotic THP-1 cells, and the collagen production was assessed.

      Results

      Pyroptotic/inflammatory proteins, including NLRP3, cleaved-Caspase (CASP)1, GSDMD-N terminal and IL-18 were increased in the serum, and ASC aggregation and GSDMD were elevated in macrophages in the skin of SSc patients. SSc mice showed increased pyroptosis markers, dermal thickness and collagen deposition in skins, which were alleviated by MCC950, Disulfiram and NSA. Pyroptosis of THP-1 cells and BMDMs was induced by LPS/SiO2, and it was reduced by the inhibitors of Cathepsin B, NLRP3, CASP1 and GSDMD. Co-culture with pyroptotic THP-1 cells increased the fibrotic proteins in fibroblasts, which were alleviated by pyroptosis inhibitors.

      Conclusions

      SSc patients and BLM-induced mouse model presented increased pyroptosis. LPS/SiO2-induced macrophage pyroptosis promoted fibrosis of SSc through Cathepsin B/NLRP3/GSDMD pathway.

      Abbreviations:

      ASC (Apoptosis-associated speck-like protein containing a caspase recruitment domain), α-SMA (α-Smooth muscle actin), BLM (Bleomycin), BMDMs (Mouse bone marrow-derived macrophages), CASP1 (Caspase-1), DAPI (4′,6-diamidino-2-phenylindole), ELISA (Enzyme-linked immunosorbent assay), FBS (Fetal bovine serum), GSDMD (Gasdermin D), GSDMD-N (GSDMD N-terminal), HCs (Healthy controls), H&E (Hematoxylin and eosin), IL-1β (Interleukin-1β), IL-18 (Interleukin-18), LPS (Lipopolysaccharide), LDH (Lactate dehydrogenase), M-CSF (Macrophage colony-stimulating factor), NLRs (Nod-like receptors), NLRP3 (NLRs family pyrin domain containing 3), NSA (Necrosulfonamide), PMA (Phorbol 12-myristate 13-acetate), PI (Propidium iodide), SiO2 (Silicon dioxide), SSc (Systemic sclerosis), THP-1 (Human acute monocytic leukemia cell line)

      Keywords

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      References

        • Maehara T.
        • Kaneko N.
        • Perugino C.A.
        • Mattoo H.
        • Kers J.
        • Allard-Chamard H.
        • et al.
        Cytotoxic CD4+ T lymphocytes may induce endothelial cell apoptosis in systemic sclerosis.
        J. Clin. Invest. 2020; 130: 2451-2464
        • Skaug B.
        • Khanna D.
        • Swindell W.R.
        • Hinchcliff M.E.
        • Frech T.M.
        • Steen V.D.
        • et al.
        Global skin gene expression analysis of early diffuse cutaneous systemic sclerosis shows a prominent innate and adaptive inflammatory profile.
        Ann. Rheum. Dis. 2020; 79: 379-386
        • Matsushita T.
        • Kobayashi T.
        • Mizumaki K.
        • Kano M.
        • Sawada T.
        • Tennichi M.
        • et al.
        BAFF inhibition attenuates fibrosis in scleroderma by modulating the regulatory and effector B cell balance.
        Sci. Adv. 2018; 4: eaas9944
        • Laurent P.
        • Sisirak V.
        • Lazaro E.
        • Richez C.
        • Duffau P.
        • Blanco P.
        • et al.
        Innate immunity in systemic sclerosis fibrosis: recent advances.
        Front Immunol. 2018; 9: 1702
        • Xu D.
        • Mu R.
        • Wei X.
        The roles of IL-1 family cytokines in the pathogenesis of systemic sclerosis.
        Front Immunol. 2019; 10: 2025
        • 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: 660-665
        • Broz P.
        • Dixit V.M.
        Inflammasomes: mechanism of assembly, regulation and signalling.
        Nat. Rev. Immunol. 2016; 16: 407-420
        • Martinez-Godinez M.A.
        • Cruz-Dominguez M.P.
        • Jara L.J.
        • Dominguez-Lopez A.
        • Jarillo-Luna R.A.
        • Vera-Lastra O.
        • et al.
        Expression of NLRP3 inflammasome, cytokines and vascular mediators in the skin of systemic sclerosis patients.
        Isr. Med Assoc. J. 2015; 17: 5-10
        • Lu A.
        • Magupalli V.G.
        • Ruan J.
        • Yin Q.
        • Atianand M.K.
        • Vos M.R.
        • et al.
        Unified polymerization mechanism for the assembly of ASC-dependent inflammasomes.
        Cell. 2014; 156: 1193-1206
        • Latz E.
        • Xiao T.S.
        • Stutz A.
        Activation and regulation of the inflammasomes.
        Nat. Rev. Immunol. 2013; 13: 397-411
        • Lescoat A.
        • Ballerie A.
        • Lecureur V.
        • Belhomme N.
        • Cazalets C.
        • Jouneau S.
        • et al.
        The neglected association of crystalline silica exposure and systemic sclerosis.
        Rheumatology. 2020; 59: 3587-3588
        • Patel S.
        • Morrisroe K.
        • Proudman S.
        • Hansen D.
        • Sahhar J.
        • Sim M.R.
        • et al.
        Occupational silica exposure in an Australian systemic sclerosis cohort.
        Rheumatology. 2020; 59: 3900-3905
        • Zhang L.
        • Yan J.W.
        • Wang Y.J.
        • Wan Y.N.
        • Wang B.X.
        • Tao J.H.
        • et al.
        Association of interleukin 1 family with systemic sclerosis.
        Inflammation. 2014; 37: 1213-1220
        • Kumar A.
        • Gupta P.
        • Rana M.
        • Chandra T.
        • Dikshit M.
        • Barthwal M.K.
        Role of pyruvate kinase M2 in oxidized LDL-induced macrophage foam cell formation and inflammation.
        J. Lipid Res. 2020; 61: 351-364
        • Zhu L.
        • Gao D.
        • Yang J.
        • Li M.
        Characterization of the phenotype of high collagen-producing fibroblast clones in systemic sclerosis, using a new modified limiting-dilution method.
        Clin. Exp. Dermatol. 2012; 37: 395-403
        • Liu C.
        • Zhou X.
        • Lu J.
        • Zhu L.
        • Li M.
        Autophagy mediates 2-methoxyestradiol-inhibited scleroderma collagen synthesis and endothelial-to-mesenchymal transition induced by hypoxia.
        Rheumatology. 2019; 58: 1966-1975
        • Gaul S.
        • Leszczynska A.
        • Alegre F.
        • Kaufmann B.
        • Johnson C.D.
        • Adams L.A.
        • et al.
        Hepatocyte pyroptosis and release of inflammasome particles induce stellate cell activation and liver fibrosis.
        J. Hepatol. 2021; 74: 156-167
        • Liang Q.
        • Cai W.
        • Zhao Y.
        • Xu H.
        • Tang H.
        • Chen D.
        • et al.
        Lycorine ameliorates bleomycin-induced pulmonary fibrosis via inhibiting NLRP3 inflammasome activation and pyroptosis.
        Pharm. Res. 2020; 158104884
        • Ludwig-Portugall I.
        • Bartok E.
        • Dhana E.
        • Evers B.D.
        • Primiano M.J.
        • Hall J.P.
        • et al.
        An NLRP3-specific inflammasome inhibitor attenuates crystal-induced kidney fibrosis in mice.
        Kidney Int. 2016; 90: 525-539
        • Bayram H.
        • Ghio A.J.
        Killer jeans and silicosis.
        Am. J. Respir. Crit. Care Med. 2011; 184: 1322-1324
        • Tsugita M.
        • Morimoto N.
        • Tashiro M.
        • Kinoshita K.
        • Nakayama M.
        SR-B1 is a silica receptor that mediates canonical inflammasome activation.
        Cell Rep. 2017; 18: 1298-1311
        • Eder C.
        Mechanisms of interleukin-1beta release.
        Immunobiology. 2009; 214: 543-553
        • Monteleone M.
        • Stow J.L.
        • Schroder K.
        Mechanisms of unconventional secretion of IL-1 family cytokines.
        Cytokine. 2015; 74: 213-218
        • Kovacs S.B.
        • Miao E.A.
        Gasdermins: effectors of pyroptosis.
        Trends Cell Biol. 2017; 27: 673-684
        • Artlett C.M.
        • Sassi-Gaha S.
        • Rieger J.L.
        • Boesteanu A.C.
        • Feghali-Bostwick C.A.
        • Katsikis P.D.
        The inflammasome activating caspase 1 mediates fibrosis and myofibroblast differentiation in systemic sclerosis.
        Arthritis Rheum. 2011; 63: 3563-3574
        • Coll R.C.
        • Robertson A.A.
        • Chae J.J.
        • Higgins S.C.
        • Muñoz-Planillo R.
        • Inserra M.C.
        • et al.
        A small-molecule inhibitor of the NLRP3 inflammasome for the treatment of inflammatory diseases.
        Nat. Med. 2015; 21: 248-255
        • Bai H.
        • Yang B.
        • Yu W.
        • Xiao Y.
        • Yu D.
        • Zhang Q.
        Cathepsin B links oxidative stress to the activation of NLRP3 inflammasome.
        Exp. Cell Res. 2018; 362: 180-187
        • Hu J.J.
        • 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: 736-745
        • Terao C.
        • Kawaguchi T.
        • Dieude P.
        • Varga J.
        • Kuwana M.
        • Hudson M.
        • et al.
        Transethnic meta-analysis identifies GSDMA and PRDM1 as susceptibility genes to systemic sclerosis.
        Ann. Rheum. Dis. 2017; 76: 1150-1158
        • Moreno-Moral A.
        • Bagnati M.
        • Koturan S.
        • Ko J.H.
        • Fonseca C.
        • Harmston N.
        • et al.
        Changes in macrophage transcriptome associate with systemic sclerosis and mediate GSDMA contribution to disease risk.
        Ann. Rheum. Dis. 2018; 77: 596-601
        • Li T.
        • Ortiz-Fernández L.
        • Andrés-León E.
        • Ciudad L.
        • Javierre B.M.
        • López-Isac E.
        • et al.
        Epigenomics and transcriptomics of systemic sclerosis CD4+ T cells reveal long-range dysregulation of key inflammatory pathways mediated by disease-associated susceptibility loci.
        Genome Med. 2020; 12: 81
        • Yang H.
        • Shi Y.
        • Liu H.
        • Lin F.
        • Qiu B.
        • Feng Q.
        • et al.
        Pyroptosis executor gasdermin D plays a key role in scleroderma and bleomycin-induced skin fibrosis.
        Cell Death Discov. 2022; 8: 183
        • Broz P.
        • Pelegrin P.
        • Shao F.
        The gasdermins, a protein family executing cell death and inflammation.
        Nat. Rev. Immunol. 2020; 20: 143-157