Abstract
Cyclosporine A (CsA) successfully prevents allograft rejection, but nephrotoxicity is still a dose-limiting adverse effect. TLR4 activation promotes kidney damage but whether this innate immunity receptor mediates CsA nephrotoxicity is unknown. The in vivo role of TLR4 during CsA nephrotoxicity was studied in mice co-treated with CsA and the TLR4 inhibitor TAK242 and also in TLR4−/− mice. CsA-induced renal TLR4 expression in wild-type mice. Pharmacological or genetic targeting of TLR4 reduced the activation of proinflammatory signaling, including JNK/c-jun, JAK2/STAT3, IRE1α and NF-κB and the expression of Fn14. Expression of proinflammatory factors and cytokines was also decreased, and kidney monocyte and lymphocyte influx was prevented. TLR4 inhibition also reduced tubular damage and drastically prevented the development of kidney fibrosis. In vivo and in vitro CsA promoted secretion of the TLR ligand HMGB1 by tubular cells upstream of TLR4 activation, and prevention of HMGB1 secretion significantly reduced CsA-induced synthesis of MCP-1, suggesting that HMGB1 may be one of the mediators of CsA-induced TLR4 activation. These results suggest that TLR4 is a potential pharmacological target in CsA nephrotoxicity.
Similar content being viewed by others
References
Berzal S, Alique M, Ruiz-Ortega M, Egido J, Ortiz A, Ramos AM (2012) GSK3, snail, and adhesion molecule regulation by cyclosporine A in renal tubular cells. Toxicol Sci 127(2):425–437
Borthwick LA, Wynn TA, Fisher AJ (2013) Cytokine mediated tissue fibrosis. Biochim Biophys Acta 1832(7):1049–1060
Campbell MT, Hile KL, Zhang H, Asanuma H, Vanderbrink BA, Rink RR et al (2011) Toll-like receptor 4: a novel signaling pathway during renal fibrogenesis. J Surg Res 168(1):e61–e69
Cao Q, Harris DC, Wang Y (2015) Macrophages in kidney injury, inflammation, and fibrosis. Physiology 30(3):183–194
Carlisle RE, Heffernan A, Brimble E, Liu L, Jerome D, Collins CA et al (2012) TDAG51 mediates epithelial-to-mesenchymal transition in human proximal tubular epithelium. Am J Physiol Renal Physiol 303(3):F467–F481
Choi YM, Cho HY, Anwar MA, Kim HK, Kwon JW, Choi S (2014) ATF3 attenuates cyclosporin A-induced nephrotoxicity by downregulating CHOP in HK-2 cells. Biochem Biophys Res Commun 448(2):182–188
de Borst MH, Prakash J, Sandovici M, Klok PA, Hamming I, Kok RJ et al (2009) c-Jun NH2-terminal kinase is crucially involved in renal tubulo-interstitial inflammation. J Pharmacol Exp Ther 331(3):896–905
Dong B, Zhou H, Han C, Yao J, Xu L, Zhang M et al (2014) Ischemia/reperfusion-induced CHOP expression promotes apoptosis and impairs renal function recovery: the role of acidosis and GPR4. PLoS ONE 9(10):e110944
Du S, Hiramatsu N, Hayakawa K, Kasai A, Okamura M, Huang T et al (2009) Suppression of NF-kappaB by cyclosporin a and tacrolimus (FK506) via induction of the C/EBP family: implication for unfolded protein response. J Immunol 182(11):7201–7211
El-Achkar TM, Huang X, Plotkin Z, Sandoval RM, Rhodes GJ, Dagher PC (2006) Sepsis induces changes in the expression and distribution of Toll-like receptor 4 in the rat kidney. Am J Physiol Renal Physiol 290(5):F1034–F1043
Fujioka S, Niu J, Schmidt C, Sclabas GM, Peng B, Uwagawa T et al (2004) NF-kappaB and AP-1 connection: mechanism of NF-kappaB-dependent regulation of AP-1 activity. Mol Cell Biol 24(17):7806–7819
Gilchrist M, Thorsson V, Li B, Rust AG, Korb M, Roach JC et al (2006) Systems biology approaches identify ATF3 as a negative regulator of Toll-like receptor 4. Nature 441(7090):173–178
Gluba A, Banach M, Hannam S, Mikhailidis DP, Sakowicz A, Rysz J (2010) The role of Toll-like receptors in renal diseases. Nat Rev Nephrol 6(4):224–235
González-Guerrero C, Ocaña-Salceda C, Berzal S, Carrasco S, Fernández-Fernández B, Cannata-Ortiz P et al (2013) Calcineurin inhibitors recruit protein kinases JAK2 and JNK, TLR signaling and the UPR to activate NF-κB-mediated inflammatory responses in kidney tubular cells. Toxicol Appl Pharmacol 272(3):825–841
Grivennikov SI, Karin M (2010) Dangerous liaisons: sTAT3 and NF-kappaB collaboration and crosstalk in cancer. Cytokine Growth Factor Rev 21(1):11–19
Haverty TP, Kelly CJ, Hines WH, Amenta PS, Watanabe M, Harper RA et al (1988) Characterization of a renal tubular epithelial cell line which secretes the autologous target antigen of autoimmune experimental interstitial nephritis. J Cell Biol 107(4):1359–1368
Issa N, Kukla A, Ibrahim HN (2013) Calcineurin inhibitor nephrotoxicity: a review and perspective of the evidence. Am J Nephrol 37(6):602–612
Justo P, Lorz C, Sanz A, Egido J, Ortiz A (2003) Intracellular mechanisms of cyclosporin A-induced tubular cell apoptosis. J Am Soc Nephrol 14(12):3072–3080
Justo P, Sanz AB, Sanchez-Niño MD, Winkles JA, Lorz C, Egido J et al (2006) Cytokine cooperation in renal tubular cell injury: the role of TWEAK. Kidney Int 70(10):1750–1758
Kanellis J, Ma FY, Kandane-Rathnayake R, Dowling JP, Polkinghorne KR, Bennett BL et al (2010) JNK signalling in human and experimental renal ischaemia/reperfusion injury. Nephrol Dial Transplant 25(9):2898–2908
Kitching AR, Holdsworth SR (2011) The emergence of TH17 cells as effectors of renal injury. J Am Soc Nephrol 22(2):235–238
Koike K, Ueda S, Yamagishi S, Yasukawa H, Kaida Y, Yokoro M et al (2014) Protective role of JAK/STAT signaling against renal fibrosis in mice with unilateral ureteral obstruction. Clin Immunol 150(1):78–87
Lee S, Huen S, Nishio H, Nishio S, Lee HK, Choi BS et al (2011) Distinct macrophage phenotypes contribute to kidney injury and repair. J Am Soc Nephrol 22(2):317–326
Lee JJ, Kim DH, Kim DG, Lee HJ, Min W, Rhee MH et al (2013) Toll-like receptor 4-linked Janus kinase 2 signaling contributes to internalization of Brucella abortus by macrophages. Infect Immun 81(7):2448–2458
Li HF, Cheng CF, Liao WJ, Lin H, Yang RB (2010) ATF3-mediated epigenetic regulation protects against acute kidney injury. J Am Soc Nephrol 21(6):1003–1013
Lim BJ, Hong SW, Jeong HJ (2009) Renal tubular expression of Toll-like receptor 4 in cyclosporine nephrotoxicity. APMIS 117(8):583–591
Lim SW, Li C, Ahn KO, Kim J, Moon IS, Ahn C, Lee JR, Yang CW (2005) Cyclosporine-Induced Renal Injury Induces Toll-like Receptor and Maturation of Dendritic cells. Transplantation 80(5):691–699
Loiarro M, Ruggiero V, Sette C (2010) Targeting TLR/IL-1R signalling in human diseases. Mediat Inflamm 2010:674363
Ma LJ, Fogo AB (2009) PAI-1 and kidney fibrosis. Front Biosci (Landmark Ed) 14:2028–2041
Martinon F, Chen X, Lee AH, Glimcher LH (2010) TLR activation of the transcription factor XBP1 regulates innate immune responses in macrophages. Nat Immunol 11(5):411–418
Meng XM, Nikolic-Paterson DJ, Lan HY (2014) Inflammatory processes in renal fibrosis. Nat Rev Nephrol 10(9):493–503
Naesens M, Kuypers DR, Sarwal M (2009) Calcineurin inhibitor nephrotoxicity. Clin J Am Soc Nephrol 4(2):481–508
Neria F, Castilla MA, Sanchez RF, Gonzalez Pacheco FR, Deudero JJ, Calabia O et al (2009) Inhibition of JAK2 protects renal endothelial and epithelial cells from oxidative stress and cyclosporin A toxicity. Kidney Int 75(2):227–234
Noh MR, Kim JI, Han SJ, Lee TJ, Park KM (2015) C/EBP homologous protein (CHOP) gene deficiency attenuates renal ischemia/reperfusion injury in mice. Biochim Biophys Acta 1852(9):1895–1901
Okugawa S, Ota Y, Kitazawa T, Nakayama K, Yanagimoto S, Tsukada K et al (2003) Janus kinase 2 is involved in lipopolysaccharide-induced activation of macrophages. Am J Physiol Cell Physiol 285(2):C399–C408
Ortiz-Muñoz G, Lopez-Parra V, Lopez-Franco O, Fernandez-Vizarra P, Mallavia B, Flores C et al (2010) Suppressors of cytokine signaling abrogate diabetic nephropathy. J Am Soc Nephrol 21(5):763–772
O’Shea JJ, Holland SM, Staudt LM (2013) JAKs and STATs in immunity, immunodeficiency, and cancer. N Engl J Med 368(2):161–170
Ponticelli C (2005) Cyclosporine: from renal transplantation to autoimmune diseases. Ann N Y Acad Sci 1051:551–558
Rayego-Mateos S, Morgado-Pascual JL, Sanz AB, Ramos AM, Eguchi S, Batlle D et al (2013) TWEAK transactivation of the epidermal growth factor receptor mediates renal inflammation. J Pathol 231(4):480–494
Sanchez-Niño MD, Poveda J, Sanz AB, Mezzano S, Carrasco S, Fernandez-Fernandez B et al (2013) Fn14 in podocytes and proteinuric kidney disease. Biochim Biophys Acta 1832(12):2232–2243
Sanz AB, Justo P, Sanchez-Niño MD, Blanco-Colio LM, Winkles JA, Kreztler M et al (2008) The cytokine TWEAK modulates renal tubulointerstitial inflammation. J Am Soc Nephrol 19(4):695–703
Sanz AB, Sanchez-Niño MD, Izquierdo MC, Jakubowski A, Justo P, Blanco-Colio LM et al (2009) Tweak induces proliferation in renal tubular epithelium: a role in uninephrectomy induced renal hyperplasia. J Cell Mol Med 13(9B):3329–3342
Sanz AB, Sanchez-Niño MD, Ramos AM, Moreno JA, Santamaria B, Ruiz-Ortega M et al (2010) NF-kappaB in renal inflammation. J Am Soc Nephrol 21(8):1254–1262
Sawinski D, Trofe-Clark J, Leas B, Uhl S, Tuteja S, Kaczmarek JL et al (2016) Calcineurin inhibitor minimization, conversion, withdrawal, and avoidance strategies in renal transplantation: a systematic review and meta-analysis. Am J Transplant 16(7):2117–2138
Schulze-Osthoff K, Ferrari D, Riehemann K, Wesselborg S (1997) Regulation of NF-kappa B activation by MAP kinase cascades. Immunobiology 198(1–3):35–49
Ucero AC, Benito-Martin A, Fuentes-Calvo I, Santamaria B, Blanco J, Lopez-Novoa JM et al (2013a) TNF-related weak inducer of apoptosis (TWEAK) promotes kidney fibrosis and Ras-dependent proliferation of cultured renal fibroblast. Biochim Biophys Acta 1832(10):1744–1755
Ucero AC, Berzal S, Ocaña-Salceda C, Sancho M, Orzáez M, Messeguer A et al (2013b) A polymeric nanomedicine diminishes inflammatory events in renal tubular cells. PLoS ONE 8(1):e51992
Urano F, Wang X, Bertolotti A, Zhang Y, Chung P, Harding HP et al (2000) Coupling of stress in the ER to activation of JNK protein kinases by transmembrane protein kinase IRE1. Science 287(5453):664–666
Wang H, Bloom O, Zhang M, Vishnubhakat JM, Ombrellino M, Che J et al (1999) HMG-1 as a late mediator of endotoxin lethality in mice. Science 285(5425):248–251
Wang S, Schmaderer C, Kiss E, Schmidt C, Bonrouhi M, Porubsky S et al (2010a) Recipient Toll-like receptors contribute to chronic graft dysfunction by both MyD88- and TRIF-dependent signaling. Dis Model Mech 3(1–2):92–103
Wang S, Yang N, Zhang L, Huang B, Tan H, Liang Y et al (2010b) (2007) Jak/STAT signaling is involved in the inflammatory infiltration of the kidneys in MRL/lpr mice. Lupus 19(10):1171–1180
WHO (2015) Annex 1 19th WHO model list of essential medicines (April 2015). http://www.who.int/medicines/publications/essentialmedicines/EML2015_8-May-15.pdf. Accessed 17 June 2016
Wu H, Chen G, Wyburn KR, Yin J, Bertolino P, Eris JM et al (2007) TLR4 activation mediates kidney ischemia/reperfusion injury. J Clin Invest 117(10):2847–2859
Wu H, Wang P, Corpuz TM, Panchapakesan U, Wyburn KR, Chadban SJ (2010) HMGB1 contributes to kidney ischemia reperfusion injury. J Am Soc Nephrol 21(11):1878–1890
Whitmore MM, Iparraguirre A, Kubelka L, Weninger W, Hai T, Williams BR (2007) Negative regulation of TLR-signaling pathways by activating transcription factor-3. J Immunol 179(6):3622–3630
Zafrani L, Ince C (2015) Microcirculation in acute and chronic kidney diseases. Am J Kidney Dis 66(6):1083–1094
Zhang B, Ramesh G, Uematsu S, Akira S, Reeves WB (2008) TLR4 signaling mediates inflammation and tissue injury in nephrotoxicity. J Am Soc Nephrol 19(5):923–932
Zhang X, Dong H, Zhang S, Lu S, Sun J, Qian Y (2015) Enhancement of LPS-induced microglial inflammation response via TLR4 under high glucose conditions. Cell Physiol Biochem 35(4):1571–1581
Zhou TB (2014) Role of high mobility group box 1 and its signaling pathways in renal diseases. J Recept Signal Transduct Res 34(5):348–350
Zhu X, Zhang J, Sun H, Jiang C, Dong Y, Shan Q et al (2014) Ubiquitination of inositol-requiring enzyme 1 (IRE1) by the E3 ligase CHIP mediates the IRE1/TRAF2/JNK pathway. J Biol Chem 289(44):30567–30577
Acknowledgments
This work was supported by grants from the Instituto de Salud Carlos III (Ministerio de Economía y Competitividad, Gobierno de España): FEDER funds ISCIII RETIC REDINREN RD12/0021, PI11/02242, PI13/00047, PI14/00386, PI15/01460; Consejería de Sanidad, Comunidad de Madrid (CIFRA S2010/BMD-2378); Sociedad Española de Nefrología. Salary support: REDINREN to CG-G; Universidad Autónoma de Madrid to JE; Programa Intensificación Actividad Investigadora (ISCIII/Agencia Laín-Entralgo/CM) to AO; Contrato Miguel Servet (ISCIII) to AMR.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Ethical approval
All procedures performed in studies involving animals were in accordance with the ethical standards of the institution or practice at which the studies were conducted.
Conflict of interest
The authors declare that they have no conflict of interest.
Electronic supplementary material
Below is the link to the electronic supplementary material.
Rights and permissions
About this article
Cite this article
González-Guerrero, C., Cannata-Ortiz, P., Guerri, C. et al. TLR4-mediated inflammation is a key pathogenic event leading to kidney damage and fibrosis in cyclosporine nephrotoxicity. Arch Toxicol 91, 1925–1939 (2017). https://doi.org/10.1007/s00204-016-1830-8
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00204-016-1830-8