Inflammasomes are mediators of inflammation, and constitutively activated NLRP3 inflammasomes have been linked to interleukin-1β (IL-1β)-mediated tumorigenesis in human melanoma. Whereas NLRP3 regulation of caspase-1 activation requires the adaptor protein ASC (apoptosis-associated speck-like protein containing a CARD (caspase recruitment domain)), caspase-1 activation by another danger-signaling sensor NLRP1 does not require ASC because NLRP1 contains a C-terminal CARD domain that facilitates direct caspase-1 activation via CARD–CARD interaction. We hypothesized that NLRP1 has additional biological activities besides IL-1β maturation and investigated its role in melanoma tumorigenesis. NLRP1 expression in melanoma was confirmed by analysis of 216 melanoma tumors and 13 human melanoma cell lines. Unlike monocytic THP-1 cells with prominent nuclear localization of NLRP1, melanoma cells expressed NLRP1 mainly in the cytoplasm. Knocking down NLRP1 revealed a tumor-promoting property of NLRP1 both in vitro and in vivo. Mechanistic studies showed that caspase-1 activity, IL-1β production, IL-1β secretion and nuclear factor-kB activity were reduced by knocking down of NLRP1 in human metastatic melanoma cell lines 1205Lu and HS294T, indicating that NLRP1 inflammasomes are active in metastatic melanoma. However, unlike previous reports showing that NLRP1 enhances pyroptosis in macrophages, NLRP1 in melanoma behaved differently in the context of cell death. Knocking down NLRP1 increased caspase-2, -9 and -3/7 activities and promoted apoptosis in human melanoma cells. Immunoprecipitation revealed interaction of NLRP1 with CARD-containing caspase-2 and -9, whereas NLRP3 lacking a CARD motif did not interact with the caspases. Consistent with these findings, NLRP1 activation but not NLRP3 activation reduced caspase-2, -9 and -3/7 activities and provided protection against apoptosis in human melanoma cells, suggesting a suppressive role of NLRP1 in caspase-3/7 activation and apoptosis via interaction with caspase-2 and -9. In summary, we showed that NLRP1 promotes melanoma growth by enhancing inflammasome activation and suppressing apoptotic pathways. Our study demonstrates a tumor-promoting role of NLRP1 in cancer cells.
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Diakos CI, Charles KA, McMillan DC, Clarke SJ . Cancer-related inflammation and treatment effectiveness. Lancet Oncol 2014; 15: e493–e503.
Solinas G, Marchesi F, Garlanda C, Mantovani A, Allavena P . Inflammation-mediated promotion of invasion and metastasis. Cancer Metastasis Rev 2010; 29: 243–248.
Multhoff G, Molls M, Radons J . Chronic inflammation in cancer development. Front Immunol 2012; 2: 98.
Watari K, Shibata T, Kawahara A, Sata K, Nabeshima H, Shinoda A et al. Tumor-derived interleukin-1 promotes lymphangiogenesis and lymph node metastasis through M2-type macrophages. PLoS ONE 2014; 9: e99568.
Song X, Krelin Y, Dvorkin T, Bjorkdahl O, Segal S, Dinarello CA et al. CD11b+/Gr-1+ immature myeloid cells mediate suppression of T cells in mice bearing tumors of IL-1beta-secreting cells. J Immunol 2005; 175: 8200–8208.
Voronov E, Shouval DS, Krelin Y, Cagnano E, Benharroch D, Iwakura Y et al. IL-1 is required for tumor invasiveness and angiogenesis. Proc Natl Acad Sci USA 2003; 100: 2645–2650.
Dunn JH, Ellis LZ, Fujita M . Inflammasomes as molecular mediators of inflammation and cancer: potential role in melanoma. Cancer Lett 2012; 314: 24–33.
Okamoto M, Liu W, Luo Y, Tanaka A, Cai X, Norris DA et al. Constitutively active inflammasome in human melanoma cells mediating autoinflammation via caspase-1 processing and secretion of interleukin-1beta. J Biol Chem 2010; 285: 6477–6488.
Davis BK, Wen H, Ting JP . The inflammasome NLRs in immunity, inflammation, and associated diseases. Annu Rev Immunol 2011; 29: 707–735.
Kolb R, Liu GH, Janowski AM, Sutterwala FS, Zhang W . Inflammasomes in cancer: a double-edged sword. Protein Cell 2014; 5: 12–20.
Liu W, Luo Y, Dunn JH, Norris DA, Dinarello CA, Fujita M . Dual role of apoptosis-associated speck-like protein containing a CARD (ASC) in tumorigenesis of human melanoma. J Invest Dermatol 2013; 133: 518–527.
Faustin B, Lartigue L, Bruey JM, Luciano F, Sergienko E, Bailly-Maitre B et al. Reconstituted NALP1 inflammasome reveals two-step mechanism of caspase-1 activation. Mol Cell 2007; 25: 713–724.
Hofmann K, Bucher P, Tschopp J . The CARD domain: a new apoptotic signalling motif. Trends Biochem Sci 1997; 22: 155–156.
Bouchier-Hayes L, Martin SJ . CARD games in apoptosis and immunity. EMBO Rep 2002; 3: 616–621.
Chu ZL, Pio F, Xie Z, Welsh K, Krajewska M, Krajewski S et al. A novel enhancer of the Apaf1 apoptosome involved in cytochrome c-dependent caspase activation and apoptosis. J Biol Chem 2001; 276: 9239–9245.
Kummer JA, Broekhuizen R, Everett H, Agostini L, Kuijk L, Martinon F et al. Inflammasome components NALP 1 and 3 show distinct but separate expression profiles in human tissues suggesting a site-specific role in the inflammatory response. J Histochem Cytochem 2007; 55: 443–452.
Zhong FL, Mamaï O, Sborgi L, Boussofara L, Hopkins R, Robinson K et al. Germline NLRP1 Mutations cause skin inflammatory and cancer susceptibility syndromes via inflammasome activation. Cell 2016; 167: 187–202.
Raskin L, Fullen DR, Giordano TJ, Thomas DG, Frohm ML, Cha KB et al. Transcriptome profiling identifies HMGA2 as a biomarker of melanoma progression and prognosis. J Invest Dermatol 2013; 133: 2585–2592.
Riker AI, Enkemann SA, Fodstad O, Liu S, Ren S, Morris C et al. The gene expression profiles of primary and metastatic melanoma yields a transition point of tumor progression and metastasis. BMC Med Genomics 2008; 1: 13.
Kabbarah O, Nogueira C, Feng B, Nazarian RM, Bosenberg M, Wu M et al. Integrative genome comparison of primary and metastatic melanomas. PLoS ONE 2010; 5: e10770.
Chanput W, Mes JJ, Wichers HJ . THP-1 cell line: an in vitro cell model for immune modulation approach. Int Immunopharmacol 2014; 23: 37–45.
Martinon F, Burns K, Tschopp J . The inflammasome: a molecular platform triggering activation of inflammatory caspases and processing of proIL-beta. Mol Cell 2002; 10: 417–426.
Datta-Mitra A, Kundu-Raychaudhuri S, Mitra A, Raychaudhuri SP . Cross talk between neuroregulatory molecule and monocyte: nerve growth factor activates the inflammasome. PLoS ONE 2015; 10: e0121626.
Bruey JM, Bruey-Sedano N, Luciano F, Zhai D, Balpai R, Xu C et al. Bcl-2 and Bcl-XL regulate proinflammatory caspase-1 activation by interaction with NALP1. Cell 2007; 129: 45–56.
Levinsohn JL, Newman ZL, Hellmich KA, Fattah R, Getz MA, Liu S et al. Anthrax lethal factor cleavage of Nlrp1 is required for activation of the inflammasome. PLoS Pathog 2012; 8: e1002638.
Kodiha M, Bański P, Stochaj U . Computer-based fluorescence quantification: a novel approach to study nucleolar biology. BMC Cell Biol 2011; 12: 25.
Bani MR, Rak J, Adachi D, Wiltshire R, Trent JM, Kerbel RS et al. Multiple features of advanced melanoma recapitulated in tumorigenic variants of early stage (radial growth phase) human melanoma cell lines: evidence for a dominant phenotype. Cancer Res 1996; 56: 3075–3086.
Chow MT, Sceneay J, Paget C, Wong CS, Duret H, Tschopp J et al. NLRP3 suppresses NK cell-mediated responses to carcinogen-induced tumors and metastases. Cancer Res 2012; 72: 5721–5732.
Ahmad I, Muneer KM, Tamimi IA, Chang ME, Ata MO, Yusuf N . Thymoquinone suppresses metastasis of melanoma cells by inhibition of NLRP3 inflammasome. Toxicol Appl Pharmacol 2013; 270: 70–76.
Tarassishin L, Casper D, Lee SC . Aberrant expression of interleukin-1β and inflammasome activation in human malignant gliomas. PLoS ONE 2014; 9: e103432.
Kumar S . Caspase 2 in apoptosis, the DNA damage response and tumor suppression: enigma no more? Nat Rev Cancer 2009; 9: 897–903.
Labbe K, Saleh M Pyroptosis: a caspase-1-dependent programmed cell death and a barrier to infection. In: Couillin I, Pétrilli V, Martinon F (eds). The Inflammasomes, Progress in Inflammation Research. Springer Basel AG: Switzerland, 2011, pp 17–36.
Newman ZL, Printz MP, Liu S, Crown D, Breen L, Miller-Randolph S et al. Susceptibility to anthrax lethal toxin-induced rat death is controlled by a single chromosome 10 locus that includes rNlrp1. PLoS Pathog 2010; 6: e1000906.
Brennan MA, Cookson BT . Salmonella induces macrophage death by caspase-1-dependent necrosis. Mol Microbiol 2000; 38: 31–40.
Cervantes J, Nagata T, Uchijima M, Shibata K, Koide Y . Intracytosolic Listeria monocytogenes induces cell death through caspase-1 activation in murine macrophages. Cell Microbiol 2008; 10: 41–52.
Rayamajhi M, Zhang Y, Miao EA . Detection of pyroptosis by measuring released lactate dehydrogenase activity. Methods Mol Biol 2013; 1040: 85–90.
Chavarría-Smith J, Vance RE . The NLRP1 inflammasomes. Immunol Rev 2015; 265: 22–34.
Frederick Lo C, Ning X, Gonzales C, Ozenberger BA . Induced expression of death domain genes NALP1 and NALP5 following neuronal injury. Biochem Biophys Res Commun 2008; 366: 664–669.
Modlin IM, Kidd M, Latich I, Zikusoka MN, Eick GN, Mane SM et al. Genetic differentiation of appendiceal tumor malignancy: a guide for the perplexed. Ann Surg 2006; 244: 52–60.
Meng XF, Wang XL, Tian XJ, Yang ZH, Chu GP, Zhang J et al. Nod-like receptor protein 1 inflammasome mediates neuron injury under high glucose. Mol Neurobiol 2014; 49: 673–684.
Kent A, Blander JM . Nod-like receptors: key molecular switches in the conundrum of cancer. Front Immunol 2014; 5: 185.
Trask DK, Muller MT . Stabilization of type I topoisomerase-DNA covalent complexes by actinomycin D. Proc Natl Acad Sci USA 1988; 85: 1417–1421.
Pommier Y, Leo E, Zhang H, Marchand C . DNA topoisomerases and their poisoning by anticancer and antibacterial drugs. Chem Biol 2010; 17: 421–433.
Baptiste-Okoh N, Barsotti AM, Prives C . A role for caspase 2 and PIDD in the process of p53-mediated apoptosis. Proc Natl Acad Sci USA 2008; 105: 1937–1942.
Caporali S, Levati L, Graziani G, Muzi A, Atzori MG, Bonmassar E et al. NF-κB is activated in response to temozolomide in an AKT-dependent manner and confers protection against the growth suppressive effect of the drug. J Transl Med 2012; 10: 252.
We thank the University of Colorado Cancer Center (UCCC) Support Grant (P30CA046934), the Skin Diseases Research Cores Grant (P30AR057212), the Flow Cytometry Core (Christine Childs and Karen Helm) for helping with FACS, and Dr Archana Gopalan (University of Colorado Anschutz Medical Campus) for helping with animal experiments. Imaging experiments were performed in the University of Colorado Anschutz Medical Campus Advance Light Microscopy Core (Radu Moldovan) supported in part by NIH/NCATS Colorado CTSI Grant Number UL1 TR001082. ZZ and JMS are recipients of a training grant, 5T32AR007411-32. This work was supported, in whole or in part, by Veterans Affairs Merit Review Award 5I01BX001228 (to MF), NIH/NCI R01CA197919 (to MF), Cancer League of Colorado (to MF) and NIH/NIAMS 2R01AR045584 (to RAS and MF).
The authors declare no conflict of interest.
Supplementary Information accompanies this paper on the Oncogene website
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Zhai, Z., Liu, W., Kaur, M. et al. NLRP1 promotes tumor growth by enhancing inflammasome activation and suppressing apoptosis in metastatic melanoma. Oncogene 36, 3820–3830 (2017). https://doi.org/10.1038/onc.2017.26
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