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Egr-1 deficiency protects from renal inflammation and fibrosis

Abstract

NF-κB and TGFβ play critical roles in renal inflammation and fibrosis, and their regulation in the kidney is thus of great interest. Early growth response-1 (Egr-1), a transcription factor belonging to the immediate early gene family, has been found to regulate inflammation and fibrosis in non-kidney tissues, but its role in renal failure has not been clear. In this study, wild-type and Egr1 −/− mice were fed with an adenine-enriched diet to induce tubulointerstitial nephritis (TIN), and primary tubular epithelial cells (PTECs) were treated with pro-inflammatory and pro-fibrotic cytokines. Kidney tissues from patients with or without renal failure were stained for Egr-1. Our results showed that Egr-1 expression was upregulated in the kidney with TIN, and the tubular epithelial cell is the primary site for Egr-1 upregulation and nuclear translocation. Egr1 −/− mice were protected from renal failure, reflected by low levels of serum urea and creatinine. The protective effect was related to an attenuation of tubular injury, immune cell infiltration, NF-κB activity, and cytokine/chemokine expressions in the kidney. Renal fibrotic area and TGFβ signaling were also reduced in Egr1 −/− mice. In vitro study showed that Egr-1 deficiency attenuated the ordinary responses of PTECs to TNFα and TGFβ. Importantly, Egr-1 is of clinical significance since the activity of Egr-1 in renal tubular cells was upregulated in renal failure patients. Our study highlights the integrative role of Egr-1 in renal inflammation and fibrosis. Thus, Egr-1 may serve as a therapeutic target for human kidney diseases.

Key messages

  • Renal failure activates Egr-1 in human and mouse tubular cells.

  • Egr-1 deficiency attenuates NF-κB and TGFβ-mediated renal inflammation/fibrosis.

  • Egr1 −/− PTECs respond weakly to pro-inflammatory or pro-fibrotic stimulation.

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References

  1. 1.

    Sanz AB, Sanchez-Nino MD, Ramos AM, Moreno JA, Santamaria B, Ruiz-Ortega M, Egido J, Ortiz A (2010) NF-kappaB in renal inflammation. J Am Soc Nephrol 21:1254–1262

    CAS  Article  PubMed  Google Scholar 

  2. 2.

    Lorenz G, Darisipudi MN, Anders HJ (2014) Canonical and non-canonical effects of the NLRP3 inflammasome in kidney inflammation and fibrosis. Nephrol Dial Transplant 29:41–48

    CAS  Article  PubMed  Google Scholar 

  3. 3.

    Correa-Costa M, Braga TT, Semedo P, Hayashida CY, Bechara LR, Elias RM, Barreto CR, Silva-Cunha C, Hyane MI, Goncalves GM et al (2011) Pivotal role of Toll-like receptors 2 and 4, its adaptor molecule MyD88, and inflammasome complex in experimental tubule-interstitial nephritis. PLoS One 6:e29004

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  4. 4.

    Carew R, Wang B, Kantharidis P (2012) The role of EMT in renal fibrosis. Cell Tissue Res 347:103–116

    CAS  Article  PubMed  Google Scholar 

  5. 5.

    Garcia-Sanchez O, Lopez-Hernandez FJ, Lopez-Novoa JM (2010) An integrative view on the role of TGF-beta in the progressive tubular deletion associated with chronic kidney disease. Kidney Int 77:950–955

    CAS  Article  PubMed  Google Scholar 

  6. 6.

    Allen K, Jaeschke H, Copple BL (2011) Bile acids induce inflammatory genes in hepatocytes: a novel mechanism of inflammation during obstructive cholestasis. Am J Pathol 178:175–186

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  7. 7.

    Yan SF, Fujita T, Lu J, Okada K, Shan Zou Y, Mackman N, Pinsky DJ, Stern DM (2000) Egr-1, a master switch coordinating upregulation of divergent gene families underlying ischemic stress. Nat Med 6:1355–1361

    CAS  Article  PubMed  Google Scholar 

  8. 8.

    Harja E, Bucciarelli LG, Lu Y, Stern DM, Zou YS, Schmidt AM, Yan SF (2004) Early growth response-1 promotes atherogenesis: mice deficient in early growth response-1 and apolipoprotein E display decreased atherosclerosis and vascular inflammation. Circ Res 94:333–339

    CAS  Article  PubMed  Google Scholar 

  9. 9.

    Chen S-J, Ning H, Ishida W, Sodin-Semrl S, Takagawa S, Mori Y, Varga J (2006) The early-immediate gene EGR-1 is induced by transforming growth factor-β and mediates stimulation of collagen gene expression. J Biol Chem 281:21183–21197

    CAS  Article  PubMed  Google Scholar 

  10. 10.

    Lee CG, Cho SJ, Kang MJ, Chapoval SP, Lee PJ, Noble PW, Yehualaeshet T, Lu B, Flavell RA, Milbrandt J et al (2004) Early growth response gene 1-mediated apoptosis is essential for transforming growth factor beta1-induced pulmonary fibrosis. J Exp Med 200:377–389

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  11. 11.

    Yao J, Mackman N, Edgington TS, Fan ST (1997) Lipopolysaccharide induction of the tumor necrosis factor-alpha promoter in human monocytic cells. Regulation by Egr-1, c-Jun, and NF-kappaB transcription factors. J Biol Chem 272:17795–17801

    CAS  Article  PubMed  Google Scholar 

  12. 12.

    Cogswell PC, Mayo MW, Baldwin AS Jr (1997) Involvement of Egr-1/RelA synergy in distinguishing T cell activation from tumor necrosis factor-alpha-induced NF-kappa B1 transcription. J Exp Med 185:491–497

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  13. 13.

    Liu C, Adamson E, Mercola D (1996) Transcription factor EGR-1 suppresses the growth and transformation of human HT-1080 fibrosarcoma cells by induction of transforming growth factor beta 1. Proc Natl Acad Sci 93:11831–11836

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  14. 14.

    Chapman NR, Perkins ND (2000) Inhibition of the RelA(p65) NF-kappaB subunit by Egr-1. J Biol Chem 275:4719–4725

    CAS  Article  PubMed  Google Scholar 

  15. 15.

    Chen G, Goeddel DV (2002) TNF-R1 signaling: a beautiful pathway. Science 296:1634–1635

    CAS  Article  PubMed  Google Scholar 

  16. 16.

    Do KH, Choi HJ, Kim J, Park SH, Kim HH, Oh CG, Moon Y (2012) Ambivalent roles of early growth response 1 in inflammatory signaling following ribosomal insult in human enterocytes. Biochem Pharmacol 84:513–521

    CAS  Article  PubMed  Google Scholar 

  17. 17.

    Pritchard MT, Nagy LE (2010) Hepatic fibrosis is enhanced and accompanied by robust oval cell activation after chronic carbon tetrachloride administration to Egr-1-deficient mice. Am J Pathol 176:2743–2752

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  18. 18.

    Bonventre JV, Sukhatme VP, Bamberger M, Ouellette AJ, Brown D (1991) Localization of the protein product of the immediate early growth response gene, Egr-1, in the kidney after ischemia and reperfusion. Cell Regul 2:251–260

    CAS  PubMed  PubMed Central  Google Scholar 

  19. 19.

    Nakamura H, Isaka Y, Tsujie M, Rupprecht HD, Akagi Y, Ueda N, Imai E, Hori M (2002) Introduction of DNA enzyme for Egr-1 into tubulointerstitial fibroblasts by electroporation reduced interstitial alpha-smooth muscle actin expression and fibrosis in unilateral ureteral obstruction (UUO) rats. Gene Ther 9:495–502

    CAS  Article  PubMed  Google Scholar 

  20. 20.

    Lee SL, Tourtellotte LC, Wesselschmidt RL, Milbrandt J (1995) Growth and differentiation proceeds normally in cells deficient in the immediate early gene NGFI-A. J Biol Chem 270:9971–9977

    CAS  Article  PubMed  Google Scholar 

  21. 21.

    Tang MJ, Tannen RL (1990) Relationship between proliferation and glucose metabolism in primary cultures of rabbit proximal tubules. Am J Physiol 259:C455–C461

    CAS  PubMed  Google Scholar 

  22. 22.

    Chen WC, Lin HH, Tang MJ (2014) Regulation of proximal tubular cell differentiation and proliferation in primary culture by matrix stiffness and ECM components. Am J Physiol Renal Physiol 307:F695–F707

    CAS  Article  PubMed  Google Scholar 

  23. 23.

    Masszi A, Fan L, Rosivall L, McCulloch CA, Rotstein OD, Mucsi I, Kapus A (2004) Integrity of cell-cell contacts is a critical regulator of TGF-beta 1-induced epithelial-to-myofibroblast transition: role for beta-catenin. Am J Pathol 165:1955–1967

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  24. 24.

    Kozera B, Rapacz M (2013) Reference genes in real-time PCR. J Appl Genet 54:391–406

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  25. 25.

    Bastianelli D, Siciliano C, Puca R, Coccia A, Murdoch C, Bordin A, Mangino G, Pompilio G, Calogero A, De Falco E (2014) Influence of Egr-1 in cardiac tissue-derived mesenchymal stem cells in response to glucose variations. Biomed Res Int 2014:254793

    PubMed  PubMed Central  Google Scholar 

  26. 26.

    Rupprecht HD, Hoffer G, de Heer E, Sterzel RB, Faller G, Schoecklmann HO (1997) Expression of the transcriptional regulator Egr-1 in experimental glomerulonephritis: requirement for mesangial cell proliferation. Kidney Int 51:694–702

    CAS  Article  PubMed  Google Scholar 

  27. 27.

    Joannidis M, Cantley LG, Spokes K, Stuart-Tilley AK, Alper SL, Epstein FH (1997) Modulation of c-fos and egr-1 expression in the isolated perfused kidney by agents that alter tubular work. Kidney Int 52:130–139

    CAS  Article  PubMed  Google Scholar 

  28. 28.

    Sun S, Ning X, Zhai Y, Du R, Lu Y, He L, Li R, Wu W, Sun W, Wang H (2014) Egr-1 mediates chronic hypoxia-induced renal interstitial fibrosis via the PKC/ERK pathway. Am J Nephrol 39:436–448

    CAS  PubMed  Google Scholar 

  29. 29.

    Humphreys BD, Lin SL, Kobayashi A, Hudson TE, Nowlin BT, Bonventre JV, Valerius MT, McMahon AP, Duffield JS (2010) Fate tracing reveals the pericyte and not epithelial origin of myofibroblasts in kidney fibrosis. Am J Pathol 176:85–97

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  30. 30.

    Eddy AA (2014) Overview of the cellular and molecular basis of kidney fibrosis. Kidney Int Suppl 4:2–8

    CAS  Article  Google Scholar 

  31. 31.

    Cao Y, Zhang J, Meng X, Wang D (2013) TNF-alpha induces early growth response gene-1 expression via ERK1/2 activation in endothelial cells. Acta Diabetol 50:27–31

    Article  PubMed  Google Scholar 

  32. 32.

    Shin SY, Lee JM, Lim Y, Lee YH (2013) Transcriptional regulation of the growth-regulated oncogene alpha gene by early growth response protein-1 in response to tumor necrosis factor alpha stimulation. Biochim Biophy Acta 1829:1066–1074

    CAS  Article  Google Scholar 

  33. 33.

    Ramana CV, Cheng GS, Kumar A, Kwon HJ, Enelow RI (2009) Role of alveolar epithelial early growth response-1 (Egr-1) in CD8+ T cell-mediated lung injury. Mol Immunol 47:623–631

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  34. 34.

    Lohoff M, Giaisi M, Kohler R, Casper B, Krammer PH, Li-Weber M (2010) Early growth response protein-1 (Egr-1) is preferentially expressed in T helper type 2 (Th2) cells and is involved in acute transcription of the Th2 cytokine interleukin-4. J Biol Chem 285:1643–1652

    CAS  Article  PubMed  Google Scholar 

  35. 35.

    Singha B, Gatla HR, Manna S, Chang TP, Sanacora S, Poltoratsky V, Vancura A, Vancurova I (2014) Proteasome inhibition increases recruitment of IkappaB kinase beta (IKKbeta), S536P-p65, and transcription factor EGR1 to interleukin-8 (IL-8) promoter, resulting in increased IL-8 production in ovarian cancer cells. J Biol Chem 289:2687–2700

    CAS  Article  PubMed  Google Scholar 

  36. 36.

    Bavendiek U, Libby P, Kilbride M, Reynolds R, Mackman N, Schonbeck U (2002) Induction of tissue factor expression in human endothelial cells by CD40 ligand is mediated via activator protein 1, nuclear factor kappa B, and Egr-1. J Biol Chem 277:25032–25039

    CAS  Article  PubMed  Google Scholar 

  37. 37.

    Parra E, Ferreira J, Ortega A (2011) Overexpression of EGR-1 modulates the activity of NF-kappaB and AP-1 in prostate carcinoma PC-3 and LNCaP cell lines. Int J Oncol 39:345–352

    CAS  PubMed  Google Scholar 

  38. 38.

    Andersen K, Eltrich N, Lichtnekert J, Anders HJ, Vielhauer V (2014) The NLRP3/ASC inflammasome promotes T-cell-dependent immune complex glomerulonephritis by canonical and noncanonical mechanisms. Kidney Int 86:965–978

    CAS  Article  PubMed  Google Scholar 

  39. 39.

    Shigeoka AA, Mueller JL, Kambo A, Mathison JC, King AJ, Hall WF, Correia Jda S, Ulevitch RJ, Hoffman HM, McKay DB (2010) An inflammasome-independent role for epithelial-expressed Nlrp3 in renal ischemia-reperfusion injury. J Immunol 185:6277–6285

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  40. 40.

    Abnaof K, Mallela N, Walenda G, Meurer SK, Sere K, Lin Q, Smeets B, Hoffmann K, Wagner W, Zenke M et al (2014) TGF-beta stimulation in human and murine cells reveals commonly affected biological processes and pathways at transcription level. BMC Syst Biol 8:55

    Article  PubMed  PubMed Central  Google Scholar 

  41. 41.

    Bhattacharyya S, Chen SJ, Wu M, Warner-Blankenship M, Ning H, Lakos G, Mori Y, Chang E, Nihijima C, Takehara K et al (2008) Smad-independent transforming growth factor-beta regulation of early growth response-1 and sustained expression in fibrosis: implications for scleroderma. Am J Pathol 173:1085–1099

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  42. 42.

    Fichtner-Feigl S, Young CA, Kitani A, Geissler EK, Schlitt HJ, Strober W (2008) IL-13 signaling via IL-13R alpha2 induces major downstream fibrogenic factors mediating fibrosis in chronic TNBS colitis. Gastroenterology 135:2003–2013

    CAS  Article  PubMed  Google Scholar 

  43. 43.

    Wu M, Melichian DS, de la Garza M, Gruner K, Bhattacharyya S, Barr L, Nair A, Shahrara S, Sporn PH, Mustoe TA et al (2009) Essential roles for early growth response transcription factor Egr-1 in tissue fibrosis and wound healing. Am J Pathol 175:1041–1055

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  44. 44.

    Cheong MW, Kuo LH, Cheng YN, Tsai PJ, Ho LC, Tai HC, Chiu WT, Chen SH, Lu PJ, Shan YS et al (2015) Loss of Egr-1 sensitizes pancreatic beta-cells to palmitate-induced ER stress and apoptosis. J Mol Med (Berl) 93:807–818

    CAS  Article  Google Scholar 

  45. 45.

    Waters CM, Hancock DC, Evan GI (1990) Identification and characterisation of the egr-1 gene product as an inducible, short-lived, nuclear phosphoprotein. Oncogene 5:669–674

    CAS  PubMed  Google Scholar 

  46. 46.

    Simon P, Schott K, Williams RW, Schaeffel F (2004) Posttranscriptional regulation of the immediate-early gene EGR1 by light in the mouse retina. Eur J Neurosci 20:3371–3377

    Article  PubMed  Google Scholar 

  47. 47.

    Mou Z, You J, Xiao Q, Wei Y, Yuan J, Liu Y, Brewer G, Ma WJ (2012) HuR posttranscriptionally regulates early growth response-1 (Egr-1) expression at the early stage of T cell activation. FEBS Lett 586:4319–4325

    CAS  Article  PubMed  Google Scholar 

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Acknowledgments

We are grateful for the support from Human Biobank, Research Center of Clinical Medicine, National Cheng Kung University Hospital. We thank Dr. Ming-Jer Tang and his laboratory for providing the protocol and instructions for the primary culture of mouse proximal tubular cells. This study is supported by the research grants of NCKUEDA10201 and NCKUEDA10301 from E-Da Hospital and National Cheng Kung University Hospital, and the research grants of NHRI-EX104-10231SI from National Health Research Institute. It is also a part of the National Cheng Kung University Top-Notch Project.

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Correspondence to Yau-Sheng Tsai.

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The study was conducted according to the study protocol approved by the Institutional Review Board of National Cheng Kung University Hospital (permit number: B-ER-101-008). All Procedures for handling animals were done in accordance to ethical statements approved by the Institutional Animal Care and Use Committee of National Cheng Kung University.

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The authors declare that they have no conflict of interests.

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Ho, LC., Sung, JM., Shen, YT. et al. Egr-1 deficiency protects from renal inflammation and fibrosis. J Mol Med 94, 933–942 (2016). https://doi.org/10.1007/s00109-016-1403-6

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Keywords

  • Early growth response-1
  • Renal inflammation
  • Renal fibrosis
  • Renal failure
  • NLRP3 inflammasome