Advertisement

Clinical and Experimental Nephrology

, Volume 21, Issue 6, pp 952–960 | Cite as

Long-term treatment with EGFR inhibitor erlotinib attenuates renal inflammatory cytokines but not nephropathy in Alport syndrome mouse model

  • Kohei Omachi
  • Rui Miyakita
  • Ryosuke Fukuda
  • Yukari Kai
  • Mary Ann Suico
  • Tsubasa Yokota
  • Misato Kamura
  • Tsuyoshi Shuto
  • Hirofumi KaiEmail author
Original article

Abstract

Background

Alport syndrome (AS) is a hereditary kidney disease caused by mutation of type IV collagen. Loss of collagen network induces collapse of glomerular basement membrane (GBM) structure. The previous studies showed that upregulation of some tyrosine kinase receptors signaling accompanied GBM disorder in AS mouse model. EGFR signaling is one of the well-known receptor kinase signaling that is involved in glomerular diseases. However, whether EGFR signaling is relevant to AS progression is still uninvestigated. Here, we determined the involvement of EGFR in AS and the effect of suppressing EGFR signaling by erlotinib treatment on AS progression.

Methods

Phosphorylated EGFR expression was investigated by Western blotting analysis and immunostaining of kidney tissues of Col4a5 mutant mice (a mouse model of X-linked AS). To check the effect of blocking EGFR signaling in AS, we administered erlotinib to AS mice once a day (10 mg/kg/day) orally for 18 weeks. Renal function parameters (proteinuria, serum creatinine, and BUN) and renal histology were assessed, and the gene expressions of inflammatory cytokines were analyzed in renal tissues.

Results

Phosphorylated EGFR expression was upregulated in AS mice kidney tissues. Erlotinib slightly reduced the urinary protein and suppressed the expression of renal injury markers (Lcn2, Lysozyme) and inflammatory cytokines (Il-6, Il-1β and KC). Erlotinib did not improve renal pathology, such as glomerular sclerosis and fibrosis.

Conclusion

These findings suggest that EGFR signaling is upregulated in kidney, but although inhibiting this signaling pathway suppressed renal inflammatory cytokines, it did not ameliorate renal dysfunction in AS mouse model.

Keywords

Alport syndrome Erlotinib Inflammatory cytokines Epithelial growth factor receptor (EGFR) 

Notes

Acknowledgements

This work was supported by the Grants-in-Aid for Science Research from the Ministry of Education, Science, Sports, and Culture of Japan (MEXT) (#22390015 to H.K. and #23590082 to M.S.), and by the Program for Leading Graduate Schools HIGO (Health life science: Interdisciplinary and Glocal Oriented), MEXT, Japan.

Compliance with ethical standards

Conflict of interest

The authors have declared that no conflict of interest exists.

Human and animal rights

All procedures performed in studies involving animals were in accordance with the ethical standards of the Animal Care and Use committee of Kumamoto University (#25-230E). This article does not contain any studies with human participants performed by any of the authors.

References

  1. 1.
    Hudson BG, Tryggvason K. Alport“s syndrome, Goodpasture”s syndrome, and type IV collagen. N Engl J Med. 2003;348:2543–56.CrossRefPubMedGoogle Scholar
  2. 2.
    Suleiman H, Zhang L, Roth R, Heuser JE, Miner JH. Nanoscale protein architecture of the kidney glomerular basement membrane. eLife. 2013;2:e01149.PubMedPubMedCentralGoogle Scholar
  3. 3.
    Zallocchi M, Johnson BM, Meehan DT. α1β1 integrin/Rac1-dependent mesangial invasion of glomerular capillaries in Alport syndrome. Am J Pathol. 2013;183:1269–80.CrossRefPubMedPubMedCentralGoogle Scholar
  4. 4.
    Randles MJ, Collinson S, Starborg T, Mironov A, Krendel M, Königshausen E, et al. Three-dimensional electron microscopy reveals the evolution of glomerular barrier injury. Sci Rep Nat Publ Group; 2016;6:35068.Google Scholar
  5. 5.
    Gross O, Beirowski B, Koepke ML, Kuck J. Preemptive ramipril therapy delays renal failure and reduces renal fibrosis in COL4A3-knockout mice with Alport syndrome1. Kidney Int. 2003;63:438–46.CrossRefPubMedGoogle Scholar
  6. 6.
    Gross O, Licht C, Anders HJ, Hoppe B, Beck B, nshoff BTO, et al. Early angiotensin-converting enzyme inhibition in Alport syndrome delays renal failure and improves life expectancy. Kidney Int. 2011;81:494–501.CrossRefPubMedGoogle Scholar
  7. 7.
    Stock J, Kuenanz J, Glonke N, Sonntag J, Frese J, Tönshoff B, et al. Prospective study on the potential of RAAS blockade to halt renal disease in Alport syndrome patients with heterozygous mutations. Pediatr Nephrol. 2017;32:131–37.CrossRefPubMedGoogle Scholar
  8. 8.
    Gross O, Girgert R, Beirowski B, Kretzler M, Kang HG, Kruegel J, et al. Loss of collagen-receptor DDR1 delays renal fibrosis in hereditary type IV collagen disease. Matrix Biol. 2010;29:346–56.CrossRefPubMedGoogle Scholar
  9. 9.
    Rubel D, Kruegel J, Martin M, Leibnitz A, Girgert R, Miosge N, et al. Collagen receptors integrin alpha2beta1 and discoidin domain receptor 1 regulate maturation of the glomerular basement membrane and loss of integrin alpha2beta1 delays kidney fibrosis in COL4A3 knockout mice. Matrix Biol. 2014;34:13–21.CrossRefPubMedGoogle Scholar
  10. 10.
    Fukuda R, Suico MA, Kai Y, Omachi K, Motomura K, Koga T, et al. Podocyte p53 Limits the severity of experimental Alport syndrome. J Am Soc Nephrol. 2016;27:144–57.CrossRefPubMedGoogle Scholar
  11. 11.
    Sharma SV, Bell DW, Settleman J, Haber DA. Epidermal growth factor receptor mutations in lung cancer. Nat Rev Cancer. 2007;7:169–81.CrossRefPubMedGoogle Scholar
  12. 12.
    Lynch TJ, Bell DW, Sordella R, Gurubhagavatula S, Okimoto RA, Brannigan BW, et al. Activating mutations in the epidermal growth factor receptor underlying responsiveness of non–small-cell lung cancer to gefitinib. N Engl J Med. 2004;350:2129–39.CrossRefPubMedGoogle Scholar
  13. 13.
    Hynes NE, Lane HA. ERBB receptors and cancer: the complexity of targeted inhibitors. Nat Rev Cancer. 2005;5:341–54.CrossRefPubMedGoogle Scholar
  14. 14.
    Saghir Akhtar IFB. The Role of Epidermal growth factor receptor in diabetes-induced cardiac dysfunction. BioImpacts BI. 2013;3:5.PubMedPubMedCentralGoogle Scholar
  15. 15.
    Smith NJ, Chan H-W, Qian H, Bourne AM, Hannan KM, Warner FJ, et al. Determination of the exact molecular requirements for Type 1 Angiotensin receptor epidermal growth factor receptor transactivation and cardiomyocyte hypertrophy. Hypertension. 2011;57:973–80.CrossRefPubMedGoogle Scholar
  16. 16.
    Bollée G, Flamant M, Schordan S, Fligny C, Rumpel E, Milon M, et al. Epidermal growth factor receptor promotes glomerular injury and renal failure in rapidly progressive crescentic glomerulonephritis. Nat Med. 2011;17:1242–50.CrossRefPubMedPubMedCentralGoogle Scholar
  17. 17.
    Torres VE, Sweeney WE, Wang X, Qian Q. EGF receptor tyrosine kinase inhibition attenuates the development of PKD in Han: SPRD rats. Kidney Int. 2003;64:1573–79.CrossRefPubMedGoogle Scholar
  18. 18.
    Dowell J, Minna JD, Kirkpatrick P. Fresh from the pipeline: Erlotinib hydrochloride. Nat Rev Drug Discov. 2005;4:13–4.CrossRefPubMedGoogle Scholar
  19. 19.
    Rheault MN, Kren SM, Thielen BK, Mesa HA, Crosson JT, Thomas W, et al. Mouse model of X-Linked Alport Syndrome. J Am Soc Nephrol. 2004;15:1466–74.CrossRefPubMedGoogle Scholar
  20. 20.
    Koga T, Kai Y, Fukuda R, Morino-Koga S, Suico MA, Koyama K, et al. Mild electrical stimulation and heat shock ameliorates progressive proteinuria and renal inflammation in mouse model of Alport syndrome. PLoS One. 2012;7(8):e43852.CrossRefPubMedPubMedCentralGoogle Scholar
  21. 21.
    Mascia F, Mariani V, Girolomoni G, Pastore S. Blockade of the EGF receptor induces a deranged chemokine expression in keratinocytes leading to enhanced skin inflammation. Am J Pathol. 2003;163:303–12.CrossRefPubMedPubMedCentralGoogle Scholar
  22. 22.
    Huang P, Xu X, Wang L, Zhu B, Wang X, Xia J. The role of EGF-EGFR signalling pathway in hepatocellular carcinoma inflammatory microenvironment. J Cell Mol Med 2014;18:218–30.CrossRefPubMedGoogle Scholar
  23. 23.
    Komposch K, Sibilia M. EGFR signaling in liver diseases. IJMS. 2016;17:30.CrossRefGoogle Scholar
  24. 24.
    Zhang M-Z, Wang Y, Paueksakon P, Harris RC. Epidermal growth factor receptor inhibition slows progression of diabetic nephropathy in association with a decrease in endoplasmic reticulum stress and an increase in autophagy. Diabetes. 2014;63:2063–72.CrossRefPubMedPubMedCentralGoogle Scholar
  25. 25.
    Harvey SJ, Zheng K, Sado Y, Naito I, Ninomiya Y, Jacobs RM, et al. Role of distinct type IV collagen networks in glomerular development and function. Kidney Int. 1998;54(6):1857–66.CrossRefPubMedGoogle Scholar
  26. 26.
    Kalluri R, Shield CF, Todd P, Hudson BG, Neilson EG. Isoform switching of type IV collagen is developmentally arrested in X-linked Alport syndrome leading to increased susceptibility of renal basement membranes to endoproteolysis. J Clin Invest. 1997;99:2470–8.CrossRefPubMedPubMedCentralGoogle Scholar
  27. 27.
    Kruegel J, Rubel D, Gross O. Alport syndrome—insights from basic and clinical research. Nat Rev Nephrol. 2012;9:170–8.CrossRefPubMedGoogle Scholar
  28. 28.
    Moro L, Dolce L, Cabodi S, Bergatto E, Erba EB, Smeriglio M, et al. Integrin-induced epidermal growth factor (EGF) receptor activation requires c-Src and p130Cas and leads to phosphorylation of specific EGF receptor tyrosines. J Biol Chem Am Soc Biochem Mol Biol. 2002;277:9405–14.Google Scholar
  29. 29.
    Balanis N, Carlin CR. Mutual cross-talk between fibronectin integrins and the EGF receptor. Cell Logist. 2014;2:46–51.CrossRefGoogle Scholar
  30. 30.
    Ding J, Lin W. Crosstalk between EGFR and integrin affects invasion and proliferation of gastric cancer cell line, SGC7901. Volume 5. Auckland: OTT. Dove Press; 2012. p. 271–7.Google Scholar
  31. 31.
    Bheda A, Creek KE, Pirisi L. Loss of p53 induces epidermal growth factor receptor promoter activity in normal human keratinocytes. Oncogene. 2008;27:4315–23.CrossRefPubMedPubMedCentralGoogle Scholar
  32. 32.
    Yallowitz AR, Li D, Lobko A, Mott D, Nemajerova A, Marchenko N. Mutant p53 amplifies epidermal growth factor receptor family signaling to promote mammary tumorigenesis. Mol Cancer Res. 2015;13:743–54.CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Japanese Society of Nephrology 2017

Authors and Affiliations

  • Kohei Omachi
    • 1
    • 2
  • Rui Miyakita
    • 1
  • Ryosuke Fukuda
    • 1
  • Yukari Kai
    • 1
  • Mary Ann Suico
    • 1
  • Tsubasa Yokota
    • 1
  • Misato Kamura
    • 1
    • 2
  • Tsuyoshi Shuto
    • 1
  • Hirofumi Kai
    • 1
    Email author
  1. 1.Department of Molecular MedicineGraduate School of Pharmaceutical Sciences, Kumamoto UniversityKumamotoJapan
  2. 2.Program for Leading Graduate Schools “HIGO (Health life science: Interdisciplinary and Glocal Oriented) Program”Kumamoto UniversityKumamoto CityJapan

Personalised recommendations