Abstract
Chronic rhinosinusitis without nasal polyps (CRSsNP) is a CRS phenotype. However, the mechanisms of CRSsNP remains unclear. Differentially expressed genes (DEGs) were obtained from the GSE36830 and GSE198950 datasets through the GEO2R tool. The six hub genes were screened by the protein–protein interaction (PPI) network analysis and Cytoscape software. Then we constructed the mouse models of CRS and verified the expression levels of hub genes by reverse transcription quantitative PCR (RT-qPCR). Hematoxylin–eosin (HE) staining was employed to observe pathological alterations in mouse tissues. Casepase-3 expression was detected by immunohistochemistry (IHC). The levels of TNF-α, IL-12, IL-6, IL-1β, LDH, and IL-18 were evaluated using enzyme-linked immunosorbent assay (ELISA). Pyroptosis-related protein expressions were measured by western blotting. Cell counting kit-8 (CCK-8) and flow cytometry were performed to assess the proliferation and apoptosis of lipopolysaccharide (LPS)-induced NP69 cells. Six hub DEGs were identified. The expression levels of IRF4, IKZF1, and CD79A were obviously increased in CRSsNP, while those of ADH6, ADH1A, and LDHC were significantly decreased. IRF4 knockdown attenuated the pathologic features of CRSsNP. IRF4 knockdown reduced levels of the TNF-α, IL-12, IL-6 IL-1β, LDH, and IL-18 as well as the proteins expression of Casepase-1, GSDMD, and NLRP3 both in vivo and in vitro, implying that inflammation and pyroptosis were inhibited. IRF4 knockdown hinders the development of CRSsNP by inhibiting the inflammatory response and NLRP3/Caspase-1/GSDMD-mediated pyroptosis, which offers novel promising treatment strategies for clinical intervention.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Data Availability
All data in the manuscript is available through the responsible corresponding author.
References
Cao PP, Li HB, Wang BF, Wang SB, You XJ, Cui YH, Wang DY, Desrosiers M, Liu Z (2009) Distinct immunopathologic characteristics of various types of chronic rhinosinusitis in adult Chinese. J Allergy Clin Immunol 124(3):478–484. https://doi.org/10.1016/j.jaci.2009.05.017
Chang L, Wu H, Huang W, Li Y, Chen Y, Li X, Yao Z, Chen X, Lai X, Zheng R et al (2023) IL-21 induces pyroptosis of Treg cells via Akt-mTOR-NLRP3-caspase 1 axis in eosinophilic chronic rhinosinusitis. J Allergy Clin Immunol 152(3):641-655 e14. https://doi.org/10.1016/j.jaci.2023.04.013
Chen X, Gao YD, Yang J (2012) Elevated interferon regulatory factor 4 levels in patients with allergic asthma. J Asthma 49(5):441–449. https://doi.org/10.3109/02770903.2012.674998
Chen JC, Xing QL, Yang HW, Yang F, Luo Y, Kong WJ, Wang YJ (2022) Construction and analysis of a ceRNA network and patterns of immune infiltration in chronic rhinosinusitis with nasal polyps: based on data mining and experimental verification. Sci Rep 12(1):9735. https://doi.org/10.1038/s41598-022-13818-6
Chen CL, Ma J, Lu RY, Wang YT, Zhao JF, Kang YF, Hu JJ, Wang N, Song J, Zhong J et al (2023a) Perturbated glucose metabolism augments epithelial cell proinflammatory function in chronic rhinosinusitis. J Allergy Clin Immunol 151(4):991-1004 e20. https://doi.org/10.1016/j.jaci.2022.09.036
Chen J, Chen S, Gong G, Yang F, Chen J, Wang Y (2023b) Inhibition of IL-4/STAT6/IRF4 signaling reduces the epithelial-mesenchymal transition in eosinophilic chronic rhinosinusitis with nasal polyps. Int Immunopharmacol 121:110554. https://doi.org/10.1016/j.intimp.2023.110554
Cho SH, Kim DW, Gevaert P (2016) Chronic Rhinosinusitis without Nasal Polyps. J Allergy Clin Immunol Pract 4(4):575–582. https://doi.org/10.1016/j.jaip.2016.04.015
Coll RC, Schroder K, Pelegrin P (2022) NLRP3 and pyroptosis blockers for treating inflammatory diseases. Trends Pharmacol Sci 43(8):653–668. https://doi.org/10.1016/j.tips.2022.04.003
Gou X, Xu W, Liu Y, Peng Y, Xu W, Yin Y, Zhang X (2022) IL-6 prevents lung macrophage death and lung inflammation injury by inhibiting GSDME- and GSDMD-mediated pyroptosis during pneumococcal pneumosepsis. Microbiol Spectr 10(2):e0204921. https://doi.org/10.1128/spectrum.02049-21
Hoshino A, Boutboul D, Zhang Y, Kuehn HS, Hadjadj J, Ozdemir N, Celkan T, Walz C, Picard C, Lenoir C et al (2022) Gain-of-function IKZF1 variants in humans cause immune dysregulation associated with abnormal T/B cell late differentiation. Sci Immunol 7(69):eabi7160. https://doi.org/10.1126/sciimmunol.abi7160
Huber M, Lohoff M (2014) IRF4 at the crossroads of effector T-cell fate decision. Eur J Immunol 44(7):1886–1895. https://doi.org/10.1002/eji.201344279
Huse K, Bai B, Hilden V, Bollum L, Våtsveen T, Munthe L, Smeland E, Irish J, Wälchli S, Myklebust JJJoi, (2022) Mechanism of CD79A and CD79B support for IgM+ B cell fitness through B cell receptor surface expression. J Immunol 209(10):2042–2053. https://doi.org/10.4049/jimmunol.2200144
Jo M, Yang H, Park J, Shin J, Park IJPo, (2023) Glycolytic reprogramming is involved in tissue remodeling on chronic rhinosinusitis. PLoS ONE 18(2):e0281640. https://doi.org/10.1371/journal.pone.0281640
Kanehisa M, Goto S (2000) KEGG: kyoto encyclopedia of genes and genomes. Nucleic Acids Res 28(1):27–30. https://doi.org/10.1093/nar/28.1.27
Kato A, Schleimer RP, Bleier BS (2022) Mechanisms and pathogenesis of chronic rhinosinusitis. J Allergy Clin Immunol 149(5):1491–1503. https://doi.org/10.1016/j.jaci.2022.02.016
Kovacs SB, Miao EA (2017) Gasdermins: effectors of pyroptosis. Trends Cell Biol 27(9):673–684. https://doi.org/10.1016/j.tcb.2017.05.005
Kratchmarov R, Nish SA, Lin WW, Adams WC, Chen YH, Yen B, Rothman NJ, Klein U, Reiner SL (2017) IRF4 couples anabolic metabolism to Th1 cell fate determination. Immunohorizons 1(7):156–161. https://doi.org/10.4049/immunohorizons.1700012
Kuehn HS, Nunes-Santos CJ, Rosenzweig SD (2021) IKZF1Germline mutations and their impact on immunity: IKAROS-associated diseases and pathophysiology. Exp Rev Clin Immunol 17(4):407–416. https://doi.org/10.1080/1744666x.2021.1901582
Lai H, Rogers DF (2010) New pharmacotherapy for airway mucus hypersecretion in asthma and COPD: targeting intracellular signaling pathways. J Aerosol Med Pulm Drug Deliv 23(4):219–231. https://doi.org/10.1089/jamp.2009.0802
Li Y, Chang LH, Huang WQ, Bao HW, Li X, Chen XH, Wu HT, Yao ZZ, Huang ZZ, Weinberg SE et al (2022) IL-17A mediates pyroptosis via the ERK pathway and contributes to steroid resistance in CRSwNP. J Allergy Clin Immunol 150(2):337–351. https://doi.org/10.1016/j.jaci.2022.02.031
Liu Z, Liu H, Yu D, Gao J, Ruan B, Long R (2021) Downregulation of miR-29b-3p promotes α-tubulin deacetylation by targeting the interaction of matrix metalloproteinase-9 with integrin β1 in nasal polyps. Int J Mol Med. https://doi.org/10.3892/ijmm.2021.4959
Liu Z, Fu Y, Huang W, Li C, Wei X, Zhan J, Zheng J (2023a) LINC01094 promotes human nasal epithelial cell epithelial-to-mesenchymal transition and pyroptosis via upregulating HMGB1. Rhinology. https://doi.org/10.4193/Rhin23.184
Liu Z, Wang C, Lin CJLs, (2023b) Pyroptosis as a double-edged sword: The pathogenic and therapeutic roles in inflammatory diseases and cancers. Life Sci 318:121498. https://doi.org/10.1016/j.lfs.2023.121498
Ma Y, Zheng C, Shi LJC (2018) The kinase LRRK2 is differently expressed in chronic rhinosinusitis with and without nasal polyps. Clin Transl Allergy 8:8. https://doi.org/10.1186/s13601-018-0194-y
Maffei R, Fiorcari S, Atene CG, Martinelli S, Mesini N, Pilato F, Lagreca I, Barozzi P, Riva G, Nasillo V et al (2023) The dynamic functions of IRF4 in B cell malignancies. Clin Exp Med 23(4):1171–1180. https://doi.org/10.1007/s10238-022-00968-0
Man K, Gabriel SS, Liao Y, Gloury R, Preston S, Henstridge DC, Pellegrini M, Zehn D, Berberich-Siebelt F, Febbraio MA et al (2017) Transcription factor IRF4 promotes CD8 (+) T cell exhaustion and limits the development of memory-like T cells during chronic infection. Immunity 47(6):1129-1141 e5. https://doi.org/10.1016/j.immuni.2017.11.021
McCormick JP, Thompson HM, Cho DY, Woodworth BA, Grayson JW (2020) Phenotypes in Chronic Rhinosinusitis. Curr Allergy Asthma Rep 20(7):20. https://doi.org/10.1007/s11882-020-00916-6
McDaniel MM, Kottyan LC, Singh H, Pasare C (2020) Suppression of Inflammasome Activation by IRF8 and IRF4 in cDCs Is Critical for T Cell Priming. Cell Rep 31(5):107604. https://doi.org/10.1016/j.celrep.2020.107604
Nam J, Roh Y, Fahad W, Noh H, Ha J, Yoon J, Kim C, Cho HJBo, (2021) Association between obesity and chronic rhinosinusitis with nasal polyps: a national population-based study. BMJ Open 11(5):e047230. https://doi.org/10.1136/bmjopen-2020-047230
Nihlen U, Greiff LJ, Nyberg P, Persson CG, Andersson M (2005) Alcohol-induced upper airway symptoms: prevalence and co-morbidity. Respir Med 99(6):762–769. https://doi.org/10.1016/j.rmed.2004.11.010
Oakley GM, Curtin K, Orb Q, Schaefer C, Orlandi RR, Alt JA (2015) Familial risk of chronic rhinosinusitis with and without nasal polyposis: genetics or environment. Int Forum Allergy Rhinol 5(4):276–282. https://doi.org/10.1002/alr.21469
Odet F, Gabel S, London RE, Goldberg E, Eddy EM (2013) Glycolysis and mitochondrial respiration in mouse LDHC-null sperm. Biol Reprod 88(4):95. https://doi.org/10.1095/biolreprod.113.108530
Qu X, Li H, Meng LJAcp, (2022) XBP1 regulates the transcription of HIF-1a in BALB/c mice with chronic rhinosinusitis without polyps. Anal Cell Pathol 2022:3066456. https://doi.org/10.1155/2022/3066456
Rao Z, Zhu Y, Yang P, Chen Z, Xia Y, Qiao C, Liu W, Deng H, Li J, Ning P et al (2022) Pyroptosis in inflammatory diseases and cancer. Theranostics 12(9):4310–4329. https://doi.org/10.7150/thno.71086
Schleimer RP (2017) Immunopathogenesis of Chronic Rhinosinusitis and Nasal Polyposis. Annu Rev Pathol 12:331–357. https://doi.org/10.1146/annurev-pathol-052016-100401
Seshadri S, Rosati M, Lin DC, Carter RG, Norton JE, Choi AW, Suh L, Kato A, Chandra RK, Harris KE et al (2013) Regional differences in the expression of innate host defense molecules in sinonasal mucosa. J Allergy Clin Immunol 132(5):1227-1230 e5. https://doi.org/10.1016/j.jaci.2013.05.042
Smirnova MG, Guo L, Birchall JP, Pearson JP (2003) LPS up-regulates mucin and cytokine mRNA expression and stimulates mucin and cytokine secretion in goblet cells. Cell Immunol 221(1):42–49. https://doi.org/10.1016/s0008-8749(03)00059-5
Stevens W, Peters A, Tan B, Klingler A, Poposki J, Hulse K, Grammer L, Welch K, Smith S, Conley D et al (2019) Associations between inflammatory endotypes and clinical presentations in chronic rhinosinusitis. J Allergy Clin Immunol 7(8):2812–28203. https://doi.org/10.1016/j.jaip.2019.05.009
Subramanian A, Tamayo P, Mootha VK, Mukherjee S, Ebert BL, Gillette MA, Paulovich A, Pomeroy SL, Golub TR, Lander ES et al (2005) Gene set enrichment analysis: a knowledge-based approach for interpreting genome-wide expression profiles. Proc Natl Acad Sci USA 102(43):15545–15550. https://doi.org/10.1073/pnas.0506580102
Sumsion JS, Pulsipher A, Alt JA (2020) Differential expression and role of S100 proteins in chronic rhinosinusitis. Curr Opin Allergy Clin Immunol 20(1):14–22. https://doi.org/10.1097/ACI.0000000000000595
Sundararaj S, Casarotto MG (2021) Molecular interactions of IRF4 in B cell development and malignancies. Biophys Rev 13(6):1219–1227. https://doi.org/10.1007/s12551-021-00825-6
Tan BK, Min JY, Hulse KE (2017) Acquired immunity in chronic rhinosinusitis. Curr Allergy Asthma Rep 17(7):49. https://doi.org/10.1007/s11882-017-0715-0
Tan H, Tong X, Gao Z, Xu Y, Tan L, Zhang W, Xiang R, Xu YJE (2022) INPP4AThe hMeDIP-Seq identified as a novel biomarker for eosinophilic chronic rhinosinusitis with nasal polyps. Epigenomics 14(12):757–775. https://doi.org/10.2217/epi-2022-0053
Vally H, de Klerk N, Thompson PJ (2000) Alcoholic drinks: important triggers for asthma. J Allergy Clin Immunol 105(3):462–467. https://doi.org/10.1067/mai.2000.104548
Vasiliou V, Pappa A, Estey T (2004) Role of human aldehyde dehydrogenases in endobiotic and xenobiotic metabolism. Drug Metab Rev 36(2):279–299. https://doi.org/10.1081/dmr-120034001
Wang PC, Tai CJ, Lin MS, Chu CC, Liang SC (2003) Quality of life in Taiwanese adults with chronic rhino-sinusitis. Qual Life Res 12(4):443–448. https://doi.org/10.1023/a:1023494025748
Wang X, Zhang N, Bo M, Holtappels G, Zheng M, Lou H, Wang H, Zhang L, Bachert C (2016) Diversity of T (H) cytokine profiles in patients with chronic rhinosinusitis: a multicenter study in Europe, Asia, and Oceania. J Allergy Clin Immunol 138(5):1344–1353. https://doi.org/10.1016/j.jaci.2016.05.041
Wu J, Lin S, Chen W, Lian G, Wu W, Chen A, Sagor M, Luo L, Wang H, Xie L (2023) TNF-α contributes to sarcopenia through caspase-8/caspase-3/GSDME-mediated pyroptosis. Cell Death Discov 9(1):76. https://doi.org/10.1038/s41420-023-01365-6
Xia P, Xing XD, Yang CX, Liao XJ, Liu FH, Huang HH, Zhang C, Song JW, Jiao YM, Shi M et al (2022) Activation-induced pyroptosis contributes to the loss of MAIT cells in chronic HIV-1 infected patients. Mil Med Res 9(1):24. https://doi.org/10.1186/s40779-022-00384-1
Yamaya M, Sekizawa K, Suzuki T, Yamada N, Furukawa M, Ishizuka S, Nakayama K, Terajima M, Numazaki Y, Sasaki H (1999) Infection of human respiratory submucosal glands with rhinovirus: effects on cytokine and ICAM-1 production. Am J Physiol 277(2):L362–L371. https://doi.org/10.1152/ajplung.1999.277.2.L362
Yang Y, Sangwung P, Kondo R, Jung Y, McConnell M, Jeong J, Utsumi T, Sessa W, Iwakiri YJJoh, (2021) Alcohol-induced Hsp90 acetylation is a novel driver of liver sinusoidal endothelial dysfunction and alcohol-related liver disease. J Hepatol 75(2):377–386. https://doi.org/10.1016/j.jhep.2021.02.028
Acknowledgements
Not applicable.
Funding
This work was supported by grants from the the Clinical Doctor Research Initial Foundation of Guangzhou Women and Children’s Medical Center (1600088), the National Natural Science Youth Foundation of China (81900915), and the Basic and Applied Basic Research Foundation of Guangdong Province (2021A1515111168).
Author information
Authors and Affiliations
Contributions
Jun Xu: Substantial contributions to conception and design, data acquisition, drafting the article; Jiahui Li, Xiaoya Wang and Yunsong An: data acquisition, drafting the article; Wenlong Liu, Renzhong Luo and Changzhi Sun: data acquisition; reviewing the article; All the authors took part in the experiment. All the authors read and approvaled the manuscript.
Corresponding authors
Ethics declarations
Conflict of interest
The authors declare that they have no conflicts of interest to disclose.
Ethical Approval and Consent to Participate
The animal experiments conformed to the Guide for the Care and Use of Laboratory Animals. Animal study has been approved by the Animal Ethics Committee of Guangzhou Women and Children’s Medical Center, National Children’s Medical Center for South Central Region, Guangzhou Medical University. All experiments were performed in accordance with relevant guidelines and regulations. The manuscript reporting adheres to the ARRIVE guidelines for the reporting of animal experiments.
Consent for Publication
Not applicable.
Additional information
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Supplementary Information
Below is the link to the electronic supplementary material.
Rights and permissions
Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.
About this article
Cite this article
Xu, J., Li, J., Wang, X. et al. IRF4 Knockdown Inhibits the Chronic Rhinosinusitis Without Nasal Polyps Development by Regulating NLRP3/Caspase-1/GSDMD-Mediated Pyroptosis. Biochem Genet (2024). https://doi.org/10.1007/s10528-024-10792-8
Received:
Accepted:
Published:
DOI: https://doi.org/10.1007/s10528-024-10792-8