Excess phosphate promotes SARS‑CoV‑2 N protein‑induced NLRP3 inflammasome activation via the SCAP‑SREBP2 signaling pathway

Liu,Mi-Hua; Lin,Xiao-Long; Xiao,Le-Le

doi:10.3892/mmr.2024.13173

Excess phosphate promotes SARS‑CoV‑2 N protein‑induced NLRP3 inflammasome activation via the SCAP‑SREBP2 signaling pathway

Authors:
- Mi-Hua Liu
- Xiao-Long Lin
- Le-Le Xiao
View Affiliations

Affiliations: Department of Clinical Laboratory, The Affiliated Ganzhou Hospital of Nanchang University, Ganzhou, Jiangxi 341000, P.R. China, Department of Pathology, Huizhou Third People's Hospital, Guangzhou Medical University, Huizhou, Guangdong 516002, P.R. China, Department of Nursing, The Affiliated Ganzhou Hospital of Nanchang University, Ganzhou, Jiangxi 341000, P.R. China
Published online on: January 25, 2024 https://doi.org/10.3892/mmr.2024.13173
Article Number: 48
Copyright: © Liu et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Hyperphosphatemia or severe acute respiratory syndrome coronavirus 2 (SARS‑CoV‑2) infection can promote cardiovascular adverse events in patients with chronic kidney disease. Hyperphosphatemia is associated with elevated inflammation and sterol regulatory element binding protein 2 (SREBP2) activation, but the underlying mechanisms in SARS‑CoV‑2 that are related to cardiovascular disease remain unclear. The present study aimed to elucidate the role of excess inorganic phosphate (PI) in SARS‑CoV‑2 N protein‑induced NLRP3 inflammasome activation and the underlying mechanisms in vascular smooth muscle cells (VSMCs). The expression levels of SARS‑CoV‑2 N protein, SREBP cleavage‑activating protein (SCAP), mature N‑terminal SREBP2, NLRP3, procaspase‑1, cleaved caspase‑1, IL‑1β and IL‑18 were examined by western blotting. The expression levels of SREBP2, HMG‑CoA reductase, HMGCS1, low density lipoprotein receptor, proprotein convertase subtilisin/kexin type 9 (PCSK9), SREBP1c, fatty acid synthase, stearyl coenzyme A desaturase 1, acetyl‑CoA carboxylase α and ATP‑citrate lyase were determined by reverse transcription‑quantitative PCR. The translocation of SCAP or NLRP3 from the endoplasmic reticulum to the Golgi was detected by confocal microscopy. The results showed that excess PI promoted SCAP‑SREBP and NLRP3 complex translocation to the Golgi, potentially leading to NLRP3 inflammasome activation and lipogenic gene expression. Furthermore, PI amplified SARS‑CoV‑2 N protein‑induced inflammation via the SCAP‑SREBP pathway, which facilitates NLRP3 inﬂammasome assembly and activation. Inhibition of phosphate uptake with phosphonoformate sodium alleviated NLRP3 inflammasome activation and reduced SREBP‑mediated lipogenic gene expression in VSMCs stimulated with PI and with SARS‑CoV‑2 N protein overexpression. Inhibition of SREBP2 or small interfering RNA‑induced silencing of SREBP2 effectively suppressed the effect of PI and SARS‑CoV‑2 N protein on NLRP3 inflammasome activation and lipogenic gene expression. In conclusion, the present study identified that PI amplified SARS‑CoV‑2 N protein‑induced NLRP3 inflammasome activation and lipogenic gene expression via the SCAP‑SREBP signaling pathway.

View Figures

View References

1	Tchidjou HK, Caron F, Ferec A, Braun K, Hery L, Castelain S and Romeo B: Severe hyperphosphatasemia and severe acute respiratory syndrome coronavirus 2 infection in children. Blood Coagul Fibrinolysis. 31:575–577. 2020. View Article : Google Scholar : PubMed/NCBI
2	Martin M, Valls J, Betriu A, Fernandez E and Valdivielso JM: Association of serum phosphorus with subclinical atherosclerosis in chronic kidney disease. Sex makes a difference. Atherosclerosis. 241:264–270. 2015. View Article : Google Scholar : PubMed/NCBI
3	Vervloet MG, Sezer S, Massy ZA, Johansson L, Cozzolino M and Fouque D; ERA-EDTA Working Group on Chronic Kidney Disease-Mineral and Bone Disorders and the European Renal Nutrition Working Group, : The role of phosphate in kidney disease. Nat Rev Nephrol. 13:27–38. 2017. View Article : Google Scholar : PubMed/NCBI
4	Rubio-Aliaga I and Krapf R: Phosphate intake, hyperphosphatemia, and kidney function. Pflugers Arch. 474:935–947. 2022. View Article : Google Scholar : PubMed/NCBI
5	Sabaghian T, Honarvar M, Safavi-Naini SAA, Sadeghi Fadaki AS, Pourhoseingholi MA and Hatamabadi H: Effect of electrolyte imbalance on mortality and late acute kidney injury in hospitalized COVID-19 patients. Iran J Kidney Dis. 16:228–237. 2022.PubMed/NCBI
6	Rothan HA and Byrareddy SN: The epidemiology and pathogenesis of coronavirus disease (COVID-19) outbreak. J Autoimmun. 109:1024332020. View Article : Google Scholar : PubMed/NCBI
7	Hu X, Chen D, Wu L, He G and Ye W: Declined serum high density lipoprotein cholesterol is associated with the severity of COVID-19 infection. Clin Chim Acta. 510:105–110. 2020. View Article : Google Scholar : PubMed/NCBI
8	Wei X, Zeng W, Su J, Wan H, Yu X, Cao X, Tan W and Wang H: Hypolipidemia is associated with the severity of COVID-19. J Clin Lipidol. 14:297–304. 2020. View Article : Google Scholar : PubMed/NCBI
9	Lee W, Ahn JH, Park HH, Kim HN, Kim H, Yoo Y, Shin H, Hong KS, Jang JG, Park CG, et al: COVID-19-activated SREBP2 disturbs cholesterol biosynthesis and leads to cytokine storm. Signal Transduct Target Ther. 5:1862020. View Article : Google Scholar : PubMed/NCBI
10	Kim H, Lee HS, Ahn JH, Hong KS, Jang JG, An J, Mun YH, Yoo SY, Choi YJ, Yun MY, et al: Lung-selective 25-hydroxycholesterol nanotherapeutics as a suppressor of COVID-19-associated cytokine storm. Nano Today. 38:1011492021. View Article : Google Scholar : PubMed/NCBI
11	Okute Y, Shoji T, Shimomura N, Tsujimoto Y, Nagata Y, Uedono H, Nakatani S, Morioka T, Mori K, Fukumoto S, et al: Serum phosphate as an independent factor associated with cholesterol metabolism in patients undergoing hemodialysis: A cross-sectional analysis of the DREAM cohort. Nephrol Dial Transplant. 38:1002–1008. 2023. View Article : Google Scholar : PubMed/NCBI
12	Ellam T, Wilkie M, Chamberlain J, Crossman D, Eastell R, Francis S and Chico TJ: Dietary phosphate modulates atherogenesis and insulin resistance in apolipoprotein E knockout mice-brief report. Arterioscler Thromb Vasc Biol. 31:1988–1990. 2011. View Article : Google Scholar : PubMed/NCBI
13	Zhou C, He Q, Gan H, Zeng T, Liu Q, Moorhead JF, Varghese Z, Ouyang N and Ruan XZ: Hyperphosphatemia in chronic kidney disease exacerbates atherosclerosis via a mannosidases-mediated complex-type conversion of SCAP N-glycans. Kidney Int. 99:1342–1353. 2021. View Article : Google Scholar : PubMed/NCBI
14	Li D, Liu M, Li Z, Zheng G, Chen A, Zhao L, Yang P, Wei L, Chen Y and Ruan XZ: Sterol-resistant SCAP overexpression in vascular smooth muscle cells accelerates atherosclerosis by increasing local vascular inflammation through activation of the NLRP3 inflammasome in mice. Aging Dis. 12:747–763. 2021. View Article : Google Scholar : PubMed/NCBI
15	Pan P, Shen M, Yu Z, Ge W, Chen K, Tian M, Xiao F, Wang Z, Wang J, Jia Y, et al: SARS-CoV-2 N protein promotes NLRP3 inflammasome activation to induce hyperinflammation. Nat Commun. 12:46642021. View Article : Google Scholar : PubMed/NCBI
16	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(−Delta Delta C(T)) method. Methods. 25:402–408. 2001. View Article : Google Scholar : PubMed/NCBI
17	Ganji R, Paulo JA, Xi Y, Kline I, Zhu J, Clemen CS, Weihl CC, Purdy JG, Gygi SP and Raman M: The p97-UBXD8 complex regulates ER-mitochondria contact sites by altering membrane lipid saturation and composition. Nat Commun. 14:6382023. View Article : Google Scholar : PubMed/NCBI
18	Zeng H, Qin H, Liao M, Zheng E, Luo X, Xiao A, Li Y, Chen L, Wei L, Zhao L, et al: CD36 promotes de novo lipogenesis in hepatocytes through INSIG2-dependent SREBP1 processing. Mol Metab. 57:1014282022. View Article : Google Scholar : PubMed/NCBI
19	Kong Y, Wu M, Wan X, Sun M, Zhang Y, Wu Z, Li C, Liang X, Gao L, Ma C and Yue X: Lipophagy-mediated cholesterol synthesis inhibition is required for the survival of hepatocellular carcinoma under glutamine deprivation. Redox Biol. 63:1027322023. View Article : Google Scholar : PubMed/NCBI
20	Liu MH, Lin XL and Xiao LL: SARS-CoV-2 nucleocapsid protein promotes TMAO-induced NLRP3 inflammasome activation by SCAP-SREBP signaling pathway. Tissue Cell. 86:1022762023. View Article : Google Scholar : PubMed/NCBI
21	Jono S, McKee MD, Murry CE, Shioi A, Nishizawa Y, Mori K, Morii H and Giachelli CM: Phosphate regulation of vascular smooth muscle cell calcification. Circ Res. 87:E10–E17. 2000. View Article : Google Scholar : PubMed/NCBI
22	Guo C, Chi Z, Jiang D, Xu T, Yu W, Wang Z, Chen S, Zhang L, Liu Q, Guo X, et al: Cholesterol homeostatic regulator SCAP-SREBP2 integrates NLRP3 inflammasome activation and cholesterol biosynthetic signaling in macrophages. Immunity. 49:842–856.e7. 2018. View Article : Google Scholar : PubMed/NCBI
23	Bepouka B, Mayasi N, Mandina M, Longokolo M, Odio O, Mangala D, Mbula M, Kayembe JM and Situakibanza H: Risk factors for mortality in COVID-19 patients in sub-Saharan Africa: A systematic review and meta-analysis. PLoS One. 17:e02760082022. View Article : Google Scholar : PubMed/NCBI
24	Gibertoni D, Reno C, Rucci P, Fantini MP, Buscaroli A, Mosconi G, Rigotti A, Giudicissi A, Mambelli E, Righini M, et al: COVID-19 incidence and mortality in non-dialysis chronic kidney disease patients. PLoS One. 16:e02545252021. View Article : Google Scholar : PubMed/NCBI
25	Chung EYM, Palmer SC, Natale P, Krishnan A, Cooper TE, Saglimbene VM, Ruospo M, Au E, Jayanti S, Liang A, et al: Incidence and outcomes of COVID-19 in people with CKD: A systematic review and meta-analysis. Am J Kidney Dis. 78:804–815. 2021. View Article : Google Scholar : PubMed/NCBI
26	Cai R, Zhang J, Zhu Y, Liu L, Liu Y and He Q: Mortality in chronic kidney disease patients with COVID-19: A systematic review and meta-analysis. Int Urol Nephrol. 53:1623–1629. 2021. View Article : Google Scholar : PubMed/NCBI
27	Rao A, Ranka S, Ayers C, Hendren N, Rosenblatt A, Alger HM, Rutan C, Omar W, Khera R, Gupta K, et al: Association of kidney disease with outcomes in COVID-19: Results from the american heart association COVID-19 cardiovascular disease registry. J Am Heart Assoc. 10:e0209102021. View Article : Google Scholar : PubMed/NCBI
28	Lambourg EJ, Gallacher PJ, Hunter RW, Siddiqui M, Miller-Hodges E, Chalmers JD, Pugh D, Dhaun N and Bell S: Cardiovascular outcomes in patients with chronic kidney disease and COVID-19: A multi-regional data-linkage study. Eur Respir J. 60:21031682022. View Article : Google Scholar : PubMed/NCBI
29	Harrison SL, Buckley BJR, Rivera-Caravaca JM, Zhang J and Lip GYH: Cardiovascular risk factors, cardiovascular disease, and COVID-19: An umbrella review of systematic reviews. Eur Heart J Qual Care Clin Outcomes. 7:330–339. 2021.PubMed/NCBI
30	Shi S, Qin M, Shen B, Cai Y, Liu T, Yang F, Gong W, Liu X, Liang J, Zhao Q, et al: Association of cardiac injury with mortality in hospitalized patients with COVID-19 in Wuhan, China. JAMA Cardiol. 5:802–810. 2020. View Article : Google Scholar : PubMed/NCBI
31	Linschoten M, Peters S, van Smeden M, Jewbali LS, Schaap J, Siebelink HM, Smits PC, Tieleman RG, van der Harst P, van Gilst WH, et al: Cardiac complications in patients hospitalised with COVID-19. Eur Heart J Acute Cardiovasc Care. 9:817–823. 2020. View Article : Google Scholar : PubMed/NCBI
32	Podestà MA, Valli F, Galassi A, Cassia MA, Ciceri P, Barbieri L, Carugo S and Cozzolino M: COVID-19 in chronic kidney disease: The impact of old and novel cardiovascular risk factors. Blood Purif. 50:740–749. 2021. View Article : Google Scholar : PubMed/NCBI
33	Malinowska J, Malecka-Gieldowska M, Bankowska D, Borecka K and Ciepiela O: Hypermagnesemia and hyperphosphatemia are highly prevalent in patients with COVID-19 and increase the risk of death. Int J Infect Dis. 122:543–549. 2022. View Article : Google Scholar : PubMed/NCBI
34	Dhingra R, Sullivan LM, Fox CS, Wang TJ, D'Agostino RB Sr, Gaziano JM and Vasan RS: Relations of serum phosphorus and calcium levels to the incidence of cardiovascular disease in the community. Arch Intern Med. 167:879–885. 2007. View Article : Google Scholar : PubMed/NCBI
35	Phan O, Ivanovski O, Nguyen-Khoa T, Mothu N, Angulo J, Westenfeld R, Ketteler M, Meert N, Maizel J, Nikolov IG, et al: Sevelamer prevents uremia-enhanced atherosclerosis progression in apolipoprotein E-deficient mice. Circulation. 112:2875–2882. 2005. View Article : Google Scholar : PubMed/NCBI
36	Nikolov IG, Joki N, Nguyen-Khoa T, Guerrera IC, Maizel J, Benchitrit J, Machado dos Reis L, Edelman A, Lacour B, Jorgetti V, et al: Lanthanum carbonate, like sevelamer-HCl, retards the progression of vascular calcification and atherosclerosis in uremic apolipoprotein E-deficient mice. Nephrol Dial Transplant. 27:505–513. 2012. View Article : Google Scholar : PubMed/NCBI
37	Tanaka S, Yamamoto H, Nakahashi O, Kagawa T, Ishiguro M, Masuda M, Kozai M, Ikeda S, Taketani Y and Takeda E: Dietary phosphate restriction induces hepatic lipid accumulation through dysregulation of cholesterol metabolism in mice. Nutr Res. 33:586–593. 2013. View Article : Google Scholar : PubMed/NCBI
38	Lee JH, Lee SH, Lee EH, Cho JY, Song DK, Lee YJ, Kwon TK, Oh BC, Cho KW, Osborne TF, et al: SCAP deficiency facilitates obesity and insulin resistance through shifting adipose tissue macrophage polarization. J Adv Res. 45:1–13. 2023. View Article : Google Scholar : PubMed/NCBI
39	Abu-Farha M, Thanaraj TA, Qaddoumi MG, Hashem A, Abubaker J and Al-Mulla F: The role of lipid metabolism in COVID-19 virus infection and as a drug target. Int J Mol Sci. 21:35442020. View Article : Google Scholar : PubMed/NCBI
40	Al Heialy S, Hachim MY, Senok A, Gaudet M, Abou Tayoun A, Hamoudi R, Alsheikh-Ali A and Hamid Q: Regulation of angiotensin-converting enzyme 2 in obesity: Implications for COVID-19. Front Physiol. 11:5550392020. View Article : Google Scholar : PubMed/NCBI
41	Yuan S, Chu H, Chan JF, Ye ZW, Wen L, Yan B, Lai PM, Tee KM, Huang J, Chen D, et al: SREBP-dependent lipidomic reprogramming as a broad-spectrum antiviral target. Nat Commun. 10:1202019. View Article : Google Scholar : PubMed/NCBI
42	Yuan S, Chan CC, Chik KK, Tsang JO, Liang R, Cao J, Tang K, Cai JP, Ye ZW, Yin F, et al: Broad-spectrum host-based antivirals targeting the interferon and lipogenesis pathways as potential treatment options for the pandemic coronavirus disease 2019 (COVID-19). Viruses. 12:6282020. View Article : Google Scholar : PubMed/NCBI
43	Orr AW, Hastings NE, Blackman BR and Wamhoff BR: Complex regulation and function of the inflammatory smooth muscle cell phenotype in atherosclerosis. J Vasc Res. 47:168–180. 2010. View Article : Google Scholar : PubMed/NCBI
44	Abbate A, Toldo S, Marchetti C, Kron J, Van Tassell BW and Dinarello CA: Interleukin-1 and the inflammasome as therapeutic targets in cardiovascular disease. Circ Res. 126:1260–1280. 2020. View Article : Google Scholar : PubMed/NCBI
45	Burger F, Baptista D, Roth A, da Silva RF, Montecucco F, Mach F, Brandt KJ and Miteva K: NLRP3 inflammasome activation controls vascular smooth muscle cells phenotypic switch in atherosclerosis. Int J Mol Sci. 23:3402021. View Article : Google Scholar : PubMed/NCBI
46	Xiao H, Lu M, Lin TY, Chen Z, Chen G, Wang WC, Marin T, Shentu TP, Wen L, Gongol B, et al: Sterol regulatory element binding protein 2 activation of NLRP3 inflammasome in endothelium mediates hemodynamic-induced atherosclerosis susceptibility. Circulation. 128:632–642. 2013. View Article : Google Scholar : PubMed/NCBI
47	Coperchini F, Chiovato L, Croce L, Magri F and Rotondi M: The cytokine storm in COVID-19: An overview of the involvement of the chemokine/chemokine-receptor system. Cytokine Growth Factor Rev. 53:25–32. 2020. View Article : Google Scholar : PubMed/NCBI
48	Soares VC, Dias SG, Santos JC, Azevedo-Quintanilha IG, Moreira IBG, Sacramento CQ, Fintelman-Rodrigues N, Temerozo JR, da Silva MAN, Barreto-Vieira DF, et al: Inhibition of the SREBP pathway prevents SARS-CoV-2 replication and inflammasome activation. Life Sci Alliance. 6:e2023020492023. View Article : Google Scholar : PubMed/NCBI