Abstract
GSDMD and GSDME, members of the gasdermin protein family, are involved in the formation of plasma membrane channels contributing to cell rupture during a certain type of necrosis called pyroptosis. GSDMD is activated in response to immunological stimulation such as lipopolysaccharides (LPS) treatment while GSDME is mainly involved in drug-induced tumor cell death. Here we show that the expression of the GSDMD gene increases significantly during LPS-induced pyroptosis in RAW264.7 murine macrophage cells. In contrast, GSDME expression is decreased in the same cells. The increasing effect of LPS on GSDMD expression was observed only when the cells were cultured in high glucose (4.5 g/l) medium, suggesting that glucose availability is important for this effect. The effect of LPS on GSDMD expression is abolished by 2-deoxyglucose (2DG), confirming that glycolysis plays crucial roles in the increasing effect of LPS. Small interference RNA-mediated knock down of GSDMD or overexpression of GSDME causes LPS-induced pyroptosis to take place through GSDME rather than through GSDMD. Taken together, LPS regulates GSDMD and GSDME expression in opposite directions through, at least in part, its effect on glycolysis. This transcriptional regulation may contribute to the execution of pyroptosis in a GSDMD-dependent manner.
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References
Broz P, Pelegrin P, Shao F (2020) The gasdermins, a protein family executing cell death and inflammation. Nat Rev Immunol 20:143–157
Ding J, Wang K, Liu W et al (2016) Pore-forming activity and structural autoinhibition of the gasdermin family. Nature 535:111–116
Aglietti RA, Estevez A, Gupta A et al (2016) GsdmD p30 elicited by caspase-11 during pyroptosis forms pores in membranes. Proc Natl Acad Sci USA 113:7858–7863
Liu X, Zhang Z, Ruan J et al (2016) Inflammasome-activated gasdermin D causes pyroptosis by forming membrane pores. Nature 535:153–158
Wang Y, Gao W, Shi X et al (2017) Chemotherapy drugs induce pyroptosis through caspase-3 cleavage of a gasdermin. Nature 547:99–103
Rogers C, Fernandes-Alnemri T, Mayes L, Alnemri D, Cingolani G, Alnemri ES (2017) Cleavage of DFNA5 by caspase-3 during apoptosis mediates progression to secondary necrotic/pyroptotic cell death. Nat Commun 8:14128
Kayagaki N, Stowe IB, Lee BL et al (2015) Caspase-11 cleaves gasdermin D for non-canonical inflammasome signalling. Nature 526:666–671
Shi J, Zhao Y, Wang K et al (2015) Cleavage of GSDMD by inflammatory caspases determines pyroptotic cell death. Nature 526:660–665
Taabazuing CY, Okondo MC, Bachovchin DA (2017) Pyroptosis and apoptosis pathways engage in bidirectional crosstalk in monocytes and macrophages. Cell Chem Biol 24:507-514 e504
Aki T, Funakoshi T, Noritake K, Unuma K, Uemura K (2020) Extracellular glucose is crucially involved in the fate decision of LPS-stimulated RAW264.7 murine macrophage cells. Sci Rep 10:10581
Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685
Aki T, Unuma K, Noritake K, Kurahashi H, Funakoshi T, Uemura K (2018) Interaction of carbon monoxide-releasing ruthenium carbonyl CORM-3 with plasma fibronectin. Toxicol In Vitro 50:201–209
Aki T, Unuma K, Noritake K, Hirayama N, Funakoshi T, Uemura K (2019) Formation of high molecular weight p62 by CORM-3. PLoS ONE 14:e0210474
Kayagaki N, Lee BL, Stowe IB et al (2019) IRF2 transcriptionally induces GSDMD expression for pyroptosis. Sci Signal. https://doi.org/10.1126/scisignal.aax4917
Benaoudia S, Martin A, Puig Gamez M et al (2019) A genome-wide screen identifies IRF2 as a key regulator of caspase-4 in human cells. EMBO Rep 20:e48235
Tsuchiya K, Nakajima S, Hosojima S et al (2019) Caspase-1 initiates apoptosis in the absence of gasdermin D. Nat Commun 10:2091
Zhou B, Abbott DW (2021) Gasdermin E permits interleukin-1 beta release in distinct sublytic and pyroptotic phases. Cell Rep 35:108998
Corcoran SE, O’Neill LA (2016) HIF1alpha and metabolic reprogramming in inflammation. J Clin Invest 126:3699–3707
Thygesen SJ, Stacey KJ (2019) IRF1 and IRF2 regulate the non-canonical inflammasome. EMBO Rep 20:e48891
Liu Z, Gan L, Xu Y et al (2017) Melatonin alleviates inflammasome-induced pyroptosis through inhibiting NF-kappaB/GSDMD signal in mice adipose tissue. J Pineal Res 63:e12414
Masuda Y, Futamura M, Kamino H et al (2006) The potential role of DFNA5, a hearing impairment gene, in p53-mediated cellular response to DNA damage. J Hum Genet 51:652–664
Wang Y, Peng J, Mi X, Yang M (2021) p53-GSDME elevation: a path for CDK7 inhibition to suppress breast cancer cell survival. Front Mol Biosci 8:697457
Jia C, Zhang Z, Tang J et al (2021) Epithelial–mesenchymal transition induces GSDME transcriptional activation for inflammatory pyroptosis. Front Cell Dev Biol 9:781365
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Ministry of Education, Culture, Sports, Science and Technology-Japan (Grant Nos. 18H19670 and 20H03955).
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Aki, T., Funakoshi, T., Unuma, K. et al. Inverse regulation of GSDMD and GSDME gene expression during LPS-induced pyroptosis in RAW264.7 macrophage cells. Apoptosis 27, 14–21 (2022). https://doi.org/10.1007/s10495-022-01708-1
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DOI: https://doi.org/10.1007/s10495-022-01708-1