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Journal of Occupational Medicine and Toxicology

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Open Access 01-12-2021 | Silicosis | Research

Genome-wide mRNA profiling identifies the NRF2-regulated lymphocyte oxidative stress status in patients with silicosis

Authors: Yingzheng Zhao, Guangcui Xu, Haibin Li, Meiyu Chang, Cheng Xiong, Yingjun Tao, Yi Guan, Yuchun Li, Sanqiao Yao

Published in: Journal of Occupational Medicine and Toxicology | Issue 1/2021

Abstract

Background

The immunomodulatory abnormalities of silicosis are related to the lymphocyte oxidative stress state. The potential effect of antioxidant therapy on silicosis may depend on the variation in nuclear factor erythroid 2-related factor 2 (NRF2)-regulated antioxidant genes in peripheral blood mononuclear cells (PBMCs). As NRF2 is a redox-sensitive transcription factor, its possible roles and underlying mechanism in the treatment of silicosis need to be clarified.

Methods

Ninety-two male patients with silicosis and 87 male healthy volunteers were randomly selected. PBMCs were isolated from fresh blood from patients with silicosis and healthy controls. The lymphocyte oxidative stress state was investigated by evaluating NRF2 expression and NRF2-dependent antioxidative genes in PBMCs from patients with silicosis. Key differentially expressed genes (DEGs) and signaling pathways were identified utilizing RNA sequencing (RNA-Seq) and bioinformatics technology. Gene set enrichment analysis was used to identify the differences in NRF2 signaling networks between patients with silicosis and healthy controls.

Results

The number of monocytes was significantly higher in patients with silicosis than that of healthy controls. Furthermore, RNA-Seq findings were confirmed using quantitative polymerase chain reaction and revealed that NRF2-regulated DEGs were associated with glutathione metabolism, transforming growth factor-β, and the extracellular matrix receptor interaction signaling pathway in PBMCs from patients with silicosis. The top 10 hub genes were identified by PPI analysis: SMAD2, MAPK3, THBS1, SMAD3, ITGB3, integrin alpha-V (ITGAV), von Willebrand factor (VWF), BMP4, CD44, and SMAD7.

Conclusions

These findings suggest that NRF2 signaling regulates the lymphocyte oxidative stress state and may contribute to fibrogenic responses in human PBMCs. Therefore, NRF2 might serve as a novel preventive and therapeutic candidate for silicosis.

Riley L, Urbine D. Chronic silicosis with progressive massive fibrosis. N Engl J Med. 2019;380(23):2256. https://doi.org/10.1056/NEJMicm1809675.CrossRefPubMed

Leung C, Yu I, Chen W. Silicosis. Lancet. 2012;379:2008–18.CrossRefPubMed

The Lancet Respiratory Medicine. The world is failing on silicosis. Lancet Respir Med. 2019;7:283. https://doi.org/10.1016/S2213-2600(19)30078-5.CrossRefPubMed

Han S, Chen H, Harvey MA, Stemn E, Cliff D. Focusing on coal workers’ lung diseases: a comparative analysis of China, Australia, and the United States. Int J Environ Res Public Health. 2018;15(11):2565. https://doi.org/10.3390/ijerph15112565.

Steenland K, Goldsmith DF. Silica exposure and autoimmune diseases. Am J Ind Med. 1995;28(5):603–8. https://doi.org/10.1002/ajim.4700280505.CrossRefPubMed

Haustein UF. Silica-induced lupus erythematosus. Acta Derm Venereol. 1998;78(1):73–4. https://doi.org/10.1080/00015559850135940.CrossRefPubMed

Rosenman KD, Moore-Fuller M, Reilly MJ. Connective tissue disease and silicosis. Am J Ind Med. 1999;35(4):375–81. https://doi.org/10.1002/(SICI)1097-0274(199904)35:4<375::AID-AJIM8>3.0.CO;2-I.CrossRefPubMed

Zhang XB. Cellular reprogramming of human peripheral blood cells. Genomics Proteomics Bioinformatics. 2013;11(5):264–74. https://doi.org/10.1016/j.gpb.2013.09.001.CrossRefPubMedPubMedCentral

Murashima A, Takasaki Y, Ohgaki M, Hashimoto H, Shirai T, Hirose S. Activated peripheral blood mononuclear cells detected by murine monoclonal antibodies to proliferating cell nuclear antigen in active lupus patients. J Clin Immunol. 1990;10(1):28–37. https://doi.org/10.1007/BF00917495.CrossRefPubMed

10.

Hu S, Tao D, He P. Immunophenotyping of lymphocyte T and B in the peripheral blood of systemic lupus erythematosus. J Tongji Med Univ. 2001;21:108–9.CrossRefPubMed

11.

Otsuki T, Sakaguchi H, Tomokuni A, Aikoh T, Matsuki T, Isozaki Y, et al. Detection of alternatively spliced variant messages of Fas gene and mutational screening of Fas and Fas ligand coding regions in peripheral blood mononuclear cells derived from silicosis patients. Immunol Lett. 2000;72(2):137–43. https://doi.org/10.1016/S0165-2478(00)00177-2.CrossRefPubMed

12.

Otsuki T, Sakaguchi H, Tomokuni A, Aikoh T, Matsuki T, Kawakami Y, et al. Soluble Fas mRNA is dominantly expressed in cases with silicosis. Immunology. 1998;94(2):258–62. https://doi.org/10.1046/j.1365-2567.1998.00509.x.CrossRefPubMedPubMedCentral

13.

Otsuki T, Tomokuni A, Sakaguchi H, Aikoh T, Matsuki T, Isozaki Y, et al. Over-expression of the decoy receptor 3 (DcR3) gene in peripheral blood mononuclear cells (PBMC) derived from silicosis patients. Clin Exp Immunol. 2000;119(2):323–7. https://doi.org/10.1046/j.1365-2249.2000.01132.x.CrossRefPubMedPubMedCentral

14.

Liu S, Hao C, Bao L, Zhao D, Zhang H, Hou J, et al. Silica particles mediate phenotypic and functional alteration of dendritic cells and induce Th2 cell polarization. Front Immunol. 2019;10:787. https://doi.org/10.3389/fimmu.2019.00787.CrossRefPubMedPubMedCentral

15.

Joshi GN, Goetjen AM, Knecht DA. Silica particles cause NADPH oxidase-independent ROS generation and transient phagolysosomal leakage. Mol Biol Cell. 2015;26(18):3150–64. https://doi.org/10.1091/mbc.e15-03-0126.CrossRefPubMedPubMedCentral

16.

Harijith A, Ebenezer DL, Natarajan V. Reactive oxygen species at the crossroads of inflammasome and inflammation. Front Physiol. 2014;5:352. https://doi.org/10.3389/fphys.2014.00352.CrossRefPubMedPubMedCentral

17.

Zhang ZQ, Zhang CZ, Shao B, Pang DH, Han GZ, Lin L. Effects of abnormal expression of fusion and fission genes on the morphology and function of lung macrophage mitochondria in SiO-induced silicosis fibrosis in rats in vivo. Toxicol Lett. 2019;312:181–7. https://doi.org/10.1016/j.toxlet.2019.04.029.CrossRefPubMed

18.

Zhang L, He YL, Li QZ, Hao XH, Zhang ZF, Yuan JX, et al. N-acetylcysteine alleviated silica-induced lung fibrosis in rats by down-regulation of ROS and mitochondrial apoptosis signaling. Toxicol Mech Methods. 2014;24(3):212–9. https://doi.org/10.3109/15376516.2013.879974.CrossRefPubMed

19.

Huang H, Chen M, Liu F, Wu H, Wang J, Chen J, et al. N-acetylcysteine tiherapeutically protects against pulmonary fibrosis in a mouse model of silicosis. Biosci Rep. 2019;39(7):BSR20190681. https://doi.org/10.1042/BSR20190681.

20.

Nakashima K, Sato T. Regulatory role of heme oxygenase-1 in silica-induced lung injury. Respir Res. 2018;19:144.CrossRefPubMedPubMedCentral

21.

Yang J, Wang T, Li Y, Yao W, Ji X, Wu Q, et al. Earthworm extract attenuates silica-induced pulmonary fibrosis through Nrf2-dependent mechanisms. Lab Investig. 2016;96(12):1279–300. https://doi.org/10.1038/labinvest.2016.101.CrossRefPubMed

22.

Martinez FJ, de Andrade JA, Anstrom KJ, King TE Jr, Raghu G. Randomized trial of acetylcysteine in idiopathic pulmonary fibrosis. N Engl J Med. 2014;370(22):2093–101. https://doi.org/10.1056/NEJMoa1401739.CrossRefPubMedPubMedCentral

23.

Moser MA, Chun OK. Vitamin C and heart health: a review based on findings from epidemiologic studies. Int J Mol Sci. 2016;17(8):1328. https://doi.org/10.3390/ijms17081328.

24.

Castranova V. Generation of oxygen radicals and mechanisms of injury prevention. Environ Health Perspect. 1994;102(Suppl 10):65–8. https://doi.org/10.1289/ehp.94102s1065.CrossRefPubMedPubMedCentral

25.

Lazzarino G, Listorti I, Bilotta G, Capozzolo T, Amorini AM. Water- and fat-soluble antioxidants in human seminal plasma and serum of fertile males. Antioxidants. 2019;8:4. https://doi.org/10.3390/antiox8040096.CrossRef

26.

Serrano I, Romero-Puertas MC, Sandalio LM, Olmedilla A. The role of reactive oxygen species and nitric oxide in programmed cell death associated with self-incompatibility. J Exp Bot. 2015;66(10):2869–76. https://doi.org/10.1093/jxb/erv083.CrossRefPubMed

27.

Palabiyik SS, Girgin G, Tutkun E, Yilmaz OH, Baydar T. Immunomodulation and oxidative stress in denim sandblasting workers: changes caused by silica exposure. Arh Hig Rada Toksikol. 2013;64(3):431–7. https://doi.org/10.2478/10004-1254-64-2013-2312.CrossRefPubMed

28.

Marchitti SA, Chen Y, Thompson DC, Vasiliou V. Ultraviolet radiation: cellular antioxidant response and the role of ocular aldehyde dehydrogenase enzymes. Eye Contact Lens. 2011;37(4):206–13. https://doi.org/10.1097/ICL.0b013e3182212642.CrossRefPubMedPubMedCentral

29.

Yamamoto M, Kensler TW, Motohashi H. The KEAP1-NRF2 system: a thiol-based sensor-effector apparatus for maintaining redox homeostasis. Physiol Rev. 2018;98(3):1169–203. https://doi.org/10.1152/physrev.00023.2017.CrossRefPubMed

30.

Re L, Martinez-Sanchez G, Bordicchia M, Malcangi G, Pocognoli A, Morales-Segura MA, et al. Is ozone pre-conditioning effect linked to Nrf2/EpRE activation pathway in vivo? A preliminary result. Eur J Pharmacol. 2014;742:158–62. https://doi.org/10.1016/j.ejphar.2014.08.029.CrossRefPubMed

31.

Kropat C, Mueller D, Boettler U, Zimmermann K, Heiss EH, Dirsch VM, et al. Modulation of Nrf2-dependent gene transcription by bilberry anthocyanins in vivo. Mol Nutr Food Res. 2013;57(3):545–50. https://doi.org/10.1002/mnfr.201200504.CrossRefPubMed

32.

Zhao Y, Xu G, Li H, Chang M, Guan Y, Li Y, et al. Overexpression of endogenous lipoic acid synthase attenuates pulmonary fibrosis induced by crystalline silica in mice. Toxicol Lett. 2020;323:57–66. https://doi.org/10.1016/j.toxlet.2020.01.023.CrossRefPubMed

33.

Ranu H, Wilde M, Madden B. Pulmonary function tests. Ulster Med J. 2011;80(2):84–90.PubMedPubMedCentral

34.

Miller MR, Crapo R, Hankinson J, Brusasco V, Burgos F, Casaburi R, et al. General considerations for lung function testing. Eur Respir J. 2005;26(1):153–61. https://doi.org/10.1183/09031936.05.00034505.CrossRefPubMed

35.

Consortium TGO. Expansion of the gene ontology knowledgebase and resources. Nucleic Acids Res. 2017;45(D1):D331–8. https://doi.org/10.1093/nar/gkw1108.CrossRef

36.

Kanehisa M, Araki M, Goto S, Hattori M, Hirakawa M, Itoh M, et al. KEGG for linking genomes to life and the environment. Nucleic Acids Res. 2008;36(Database issue):D480–4. https://doi.org/10.1093/nar/gkm882.CrossRefPubMed

37.

Subramanian A, Tamayo P, Mootha VK, Mukherjee S, Ebert BL, Gillette MA, et al. Gene set enrichment analysis: a knowledge-based approach for interpreting genome-wide expression profiles. Proc Natl Acad Sci U S A. 2005;102(43):15545–50. https://doi.org/10.1073/pnas.0506580102.CrossRefPubMedPubMedCentral

38.

Bader GD, Hogue CW. An automated method for finding molecular complexes in large protein interaction networks. BMC Bioinformatics. 2003;4(1):2. https://doi.org/10.1186/1471-2105-4-2.CrossRefPubMedPubMedCentral

39.

Azuaje FJ. Selecting biologically informative genes in co-expression networks with a centrality score. Biol Direct. 2014;9(1):12. https://doi.org/10.1186/1745-6150-9-12.CrossRefPubMedPubMedCentral

40.

Abdelaziz R, Elkashef W, Said E. Tadalafil reduces airway hyperactivity and protects against lung and respiratory airways dysfunction in a rat model of silicosis. Int Immunopharmacol. 2016;40:530–41. https://doi.org/10.1016/j.intimp.2016.10.007.CrossRefPubMed

41.

Lu Y, Sun Y, Liu Z, Lu Y, Zhu X, Lan B, et al. Activation of NRF2 ameliorates oxidative stress and cystogenesis in autosomal dominant polycystic kidney disease. Sci Transl Med. 2020;12(554):eaba3613. https://doi.org/10.1126/scitranslmed.aba3613.

42.

Zhu Z, Yang G, Wang Y, Yang J, Gao A, Niu P, et al. Suppression of thioredoxin system contributes to silica-induced oxidative stress and pulmonary fibrogenesis in rats. Toxicol Lett. 2013;222(3):289–94. https://doi.org/10.1016/j.toxlet.2013.08.004.CrossRefPubMed

43.

Barbarin V, Arras M, Misson P, Delos M, McGarry B, Phan SH, et al. Characterization of the effect of interleukin-10 on silica-induced lung fibrosis in mice. Am J Respir Cell Mol Biol. 2004;31(1):78–85. https://doi.org/10.1165/rcmb.2003-0299OC.CrossRefPubMed

44.

Nardi J, Nascimento S, Göethel G, Gauer B, Sauer E, Fão N, et al. Inflammatory and oxidative stress parameters as potential early biomarkers for silicosis. Clin Chim Acta. 2018;484:305–13. https://doi.org/10.1016/j.cca.2018.05.045.CrossRefPubMed

45.

Liu H, Cheng Y, Yang J, Wang W, Fang S, Zhang W, et al. BBC3 in macrophages promoted pulmonary fibrosis development through inducing autophagy during silicosis. Cell Death Dis. 2017;8(3):e2657. https://doi.org/10.1038/cddis.2017.78.CrossRefPubMedPubMedCentral

46.

Faria ACD, Carvalho ARS, Guimarães ARM, Lopes AJ, Melo PL. Association of respiratory integer and fractional-order models with structural abnormalities in silicosis. Comput Methods Prog Biomed. 2019;172:53–63. https://doi.org/10.1016/j.cmpb.2019.02.003.CrossRef

47.

Cai W, Xu H, Zhang B, Gao X, Li S, Wei Z, et al. Differential expression of lncRNAs during silicosis and the role of LOC103691771 in myofibroblast differentiation induced by TGF-β1. Biomed Pharmacother. 2020;125:109980. https://doi.org/10.1016/j.biopha.2020.109980.CrossRefPubMed

48.

Yao W, Yang P, Qi Y, Jin L, Zhao A, Ding M, et al. Transcriptome analysis reveals a protective role of liver X receptor alpha against silica particle-induced experimental silicosis. Sci Total Environ. 2020;747:141531. https://doi.org/10.1016/j.scitotenv.2020.141531.CrossRefPubMed

49.

Su J, Morgani SM, David CJ, Wang Q, Er EE, Huang YH, et al. TGF-β orchestrates fibrogenic and developmental EMTs via the RAS effector RREB1. Nature. 2020;577(7791):566–71. https://doi.org/10.1038/s41586-019-1897-5.CrossRefPubMedPubMedCentral

50.

Li N, Feng F, Wu K, Zhang H, Zhang W, Wang W. Inhibitory effects of astragaloside IV on silica-induced pulmonary fibrosis via inactivating TGF-β1/Smad3 signaling. Biomed Pharmacother. 2019;119:109387. https://doi.org/10.1016/j.biopha.2019.109387.CrossRefPubMed

51.

Mohan V, Das A, Sagi I. Emerging roles of ECM remodeling processes in cancer. Semin Cancer Biol. 2020;62:192–200. https://doi.org/10.1016/j.semcancer.2019.09.004.CrossRefPubMed

52.

Khadilkar RJ, Ho KYL, Venkatesh B, Tanentzapf G. Integrins modulate extracellular matrix organization to control cell signaling during hematopoiesis. Curr Biol. 2020;30:3316–29.e5.CrossRefPubMed

53.

Mitsou I, Multhaupt HAB, Couchman JR. Proteoglycans, ion channels and cell-matrix adhesion. Biochem J. 2017;474(12):1965–79. https://doi.org/10.1042/BCJ20160747.CrossRefPubMed

54.

Chai Y, Ito Y, Han J. TGF-beta signaling and its functional significance in regulating the fate of cranial neural crest cells. Crit Rev Oral Biol Med. 2003;14(2):78–88. https://doi.org/10.1177/154411130301400202.CrossRefPubMed

55.

Aragón E, Wang Q, Zou Y, Morgani SM, Ruiz L, Kaczmarska Z, et al. Structural basis for distinct roles of SMAD2 and SMAD3 in FOXH1 pioneer-directed TGF-β signaling. Genes Dev. 2019;33(21-22):1506–24. https://doi.org/10.1101/gad.330837.119.CrossRefPubMedPubMedCentral

56.

Feng F, Li N, Cheng P, Zhang H, Wang H, Wang Y, et al. Tanshinone IIA attenuates silica-induced pulmonary fibrosis via inhibition of TGF-β1-Smad signaling pathway. Biomed Pharmacother. 2020;121:109586. https://doi.org/10.1016/j.biopha.2019.109586.CrossRefPubMed

57.

Guo J, Yang Z, Jia Q, Bo C, Shao H, Zhang Z. Pirfenidone inhibits epithelial-mesenchymal transition and pulmonary fibrosis in the rat silicosis model. Toxicol Lett. 2019;300:59–66. https://doi.org/10.1016/j.toxlet.2018.10.019.CrossRefPubMed

58.

Bakkebø M, Huse K, Hilden VI, Forfang L, Myklebust JH, Smeland EB, et al. SARA is dispensable for functional TGF-β signaling. FEBS Lett. 2012;586(19):3367–72. https://doi.org/10.1016/j.febslet.2012.07.027.CrossRefPubMed

59.

Ramachandran A, Vizán P, Das D, Chakravarty P, Vogt J, Rogers KW. TGF-β uses a novel mode of receptor activation to phosphorylate SMAD1/5 and induce epithelial-to-mesenchymal transition. Elife. 2018;7:e31756.CrossRefPubMedPubMedCentral

60.

Takebe Y, Tsujigiwa H, Katase N, Siar CH, Takabatake K, Fujii M, et al. Parenchyma-stromal interactions induce fibrosis by secreting CCN2 and promote osteoclastogenesis by stimulating RANKL and CD68 through activated TGF-β/BMP4 in ameloblastoma. J Oral Pathol Med. 2017;46(1):67–75. https://doi.org/10.1111/jop.12467.CrossRefPubMed

61.

Liang D, Wang Y, Zhu Z, Yang G, An G, Li X, et al. BMP-7 attenuated silica-induced pulmonary fibrosis through modulation of the balance between TGF-β/Smad and BMP-7/Smad signaling pathway. Chem Biol Interact. 2016;243:72–81. https://doi.org/10.1016/j.cbi.2015.11.012.CrossRefPubMed

62.

Chen M, Wan B, Zhu S, Zhang F, Jin J, Li X, et al. Geranylgeranyl diphosphate synthase deficiency aggravates lung fibrosis in mice by modulating TGF-β1/BMP-4 signaling. Biol Chem. 2019;400(12):1617–27. https://doi.org/10.1515/hsz-2019-0168.CrossRefPubMed

63.

Maitra S, Das D, Ghosh P, Hajra S, Roy SS, Bhattacharya S. High cAMP attenuation of insulin-stimulated meiotic G2-M1 transition in zebrafish oocytes: interaction between the cAMP-dependent protein kinase (PKA) and the MAPK3/1 pathways. Mol Cell Endocrinol. 2014;393(1-2):109–19. https://doi.org/10.1016/j.mce.2014.06.008.CrossRefPubMed

64.

Chu L, Wang T, Hu Y, Gu Y, Su Z, Jiang H. Activation of Egr-1 in human lung epithelial cells exposed to silica through MAPKs signaling pathways. PLoS One. 2013;8(7):e68943. https://doi.org/10.1371/journal.pone.0068943.CrossRefPubMedPubMedCentral

65.

Akhter N, Madhoun A, Arefanian H, Wilson A, Kochumon S, Thomas R, et al. Oxidative stress induces expression of the toll-like receptors (TLRs) 2 and 4 in the human peripheral blood mononuclear cells: implications for metabolic inflammation. Cell Physiol Biochem. 2019;53(1):1–18. https://doi.org/10.33594/000000117.CrossRefPubMed

66.

Calzada MJ, Roberts DD. Novel integrin antagonists derived from thrombospondins. Curr Pharm Des. 2005;11(7):849-66. https://doi.org/10.2174/1381612053381792.

67.

Mandarapu R, Prakhya BM. Exposure to cypermethrin and mancozeb alters the expression profile of THBS1, SPP1, FEZ1 and GPNMB in human peripheral blood mononuclear cells. J Immunotoxicol. 2016;13(4):463–73. https://doi.org/10.3109/1547691X.2015.1130088.CrossRefPubMed

68.

Chen C, Zhao S, Karnad A, Freeman JW. The biology and role of CD44 in cancer progression: therapeutic implications. J Hematol Oncol. 2018;11(1):64. https://doi.org/10.1186/s13045-018-0605-5.CrossRefPubMedPubMedCentral

69.

Pongcharoen P, Jinawath A, Tohtong R. Silencing of CD44 by siRNA suppressed invasion, migration and adhesion to matrix, but not secretion of MMPs, of cholangiocarcinoma cells. Clin Exp Metastasis. 2011;28(8):827–39. https://doi.org/10.1007/s10585-011-9414-8.CrossRefPubMed

70.

Li S, Li C, Zhang Y, He X, Chen X, Zeng X, et al. Targeting mechanics-induced fibroblast activation through CD44-RhoA-YAP pathway ameliorates crystalline silica-induced silicosis. Theranostics. 2019;9(17):4993–5008. https://doi.org/10.7150/thno.35665.CrossRefPubMedPubMedCentral

Title: Genome-wide mRNA profiling identifies the NRF2-regulated lymphocyte oxidative stress status in patients with silicosis
Authors: Yingzheng Zhao
Guangcui Xu
Haibin Li
Meiyu Chang
Cheng Xiong
Yingjun Tao
Yi Guan
Yuchun Li
Sanqiao Yao
Publication date: 01-12-2021
Publisher: BioMed Central
Keyword: Silicosis
Published in: Journal of Occupational Medicine and Toxicology / Issue 1/2021
Electronic ISSN: 1745-6673
DOI: https://doi.org/10.1186/s12995-021-00332-0

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Genome-wide mRNA profiling identifies the NRF2-regulated lymphocyte oxidative stress status in patients with silicosis

Abstract

Background

Methods

Results

Conclusions

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Abstract

Background

Methods

Results

Conclusions

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