Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Letter
  • Published:

Identification of human epididymis protein-4 as a fibroblast-derived mediator of fibrosis

Nature Medicine volume 19, pages 227–231 (2013)Cite this article

Subjects

Abstract

The functional contribution of myofibroblasts in fibrosis is not well understood1,2,3. Using a new genetic mouse model to track and isolate myofibroblasts, we performed gene expression profiling followed by biological validation to identify HE4 (encoding human epididymis protein 4, also known as WAP 4-disulfide core domain-2 or Wfdc2) as the most upregulated gene in fibrosis-associated myofibroblasts. The HE4 gene encodes for a putative serine protease inhibitor that is upregulated in human and mouse fibrotic kidneys and is elevated in the serum of patients with kidney fibrosis. HE4 suppresses the activity of multiple proteases, including serine proteases and matrix metalloproteinases, and specifically inhibits their capacity to degrade type I collagen. In particular, we identified two serine proteases, Prss35 and Prss23, as HE4 targets with functional relevance in kidney fibrosis. Administration of HE4-neutralizing antibodies accelerated collagen I degradation and inhibited fibrosis in three different mouse models of renal disease. Collectively these studies suggest that HE4 is a potential biomarker of renal fibrosis and a new therapeutic target.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1: αSMA+ myofibroblasts accumulate in the interstitium and express HE4 in renal fibrosis.
Figure 2: HE4 is a pan–serine protease and an MMP2 and MMP9 inhibitor that prevents type I collagen degradation.
Figure 3: HE4 neutralization inhibits kidney fibrosis.
Figure 4: HE4 expression is elevated in human fibrotic kidneys, human FAFs and serum of patients with renal fibrosis.

Similar content being viewed by others

References

  1. Zeisberg, M., Strutz, F. & Muller, G.A. Role of fibroblast activation in inducing interstitial fibrosis. J. Nephrol. 13 (suppl. 3), S111–S120 (2000).

    PubMed  Google Scholar 

  2. Eddy, A.A. Molecular insights into renal interstitial fibrosis. J. Am. Soc. Nephrol. 7, 2495–2508 (1996).

    CAS  PubMed  Google Scholar 

  3. Strutz, F. & Muller, G.A. Renal fibrosis and the origin of the renal fibroblast. Nephrol. Dial. Transplant. 21, 3368–3370 (2006).

    Article  Google Scholar 

  4. Okada, H., Strutz, F., Danoff, T.M., Kalluri, R. & Neilson, E.G. Possible mechanisms of renal fibrosis. Contrib. Nephrol. 118, 147–154 (1996).

    Article  CAS  Google Scholar 

  5. Zeisberg, M. & Neilson, E.G. Mechanisms of tubulointerstitial fibrosis. J. Am. Soc. Nephrol. 21, 1819–1834 (2010).

    Article  CAS  Google Scholar 

  6. Meran, S. & Steadman, R. Fibroblasts and myofibroblasts in renal fibrosis. Int. J. Exp. Pathol. 92, 158–167 (2011).

    Article  CAS  Google Scholar 

  7. Barnes, J.L. & Glass, W.F. II. Renal interstitial fibrosis: a critical evaluation of the origin of myofibroblasts. Contrib. Nephrol. 169, 73–93 (2011).

    Article  CAS  Google Scholar 

  8. Grgic, I., Duffield, J.S. & Humphreys, B.D. The origin of interstitial myofibroblasts in chronic kidney disease. Pediatr. Nephrol. 27, 183–193 (2012).

    Article  Google Scholar 

  9. Kirchhoff, C. Molecular characterization of epididymal proteins. Rev. Reprod. 3, 86–95 (1998).

    Article  CAS  Google Scholar 

  10. Kirchhoff, C., Habben, I., Ivell, R. & Krull, N. A major human epididymis-specific cDNA encodes a protein with sequence homology to extracellular proteinase inhibitors. Biol. Reprod. 45, 350–357 (1991).

    Article  CAS  Google Scholar 

  11. Hahm, K. et al. Alphav beta6 integrin regulates renal fibrosis and inflammation in Alport mouse. Am. J. Pathol. 170, 110–125 (2007).

    Article  CAS  Google Scholar 

  12. Zhang, G. et al. Urokinase receptor deficiency accelerates renal fibrosis in obstructive nephropathy. J. Am. Soc. Nephrol. 14, 1254–1271 (2003).

    Article  CAS  Google Scholar 

  13. Bingle, L., Singleton, V. & Bingle, C.D. The putative ovarian tumour marker gene HE4 (WFDC2), is expressed in normal tissues and undergoes complex alternative splicing to yield multiple protein isoforms. Oncogene 21, 2768–2773 (2002).

    Article  CAS  Google Scholar 

  14. Clauss, A., Lilja, H. & Lundwall, A. A locus on human chromosome 20 contains several genes expressing protease inhibitor domains with homology to whey acidic protein. Biochem. J. 368, 233–242 (2002).

    Article  CAS  Google Scholar 

  15. Clauss, A., Lilja, H. & Lundwall, A. The evolution of a genetic locus encoding small serine proteinase inhibitors. Biochem. Biophys. Res. Commun. 333, 383–389 (2005).

    Article  CAS  Google Scholar 

  16. Bingle, L. et al. WFDC2 (HE4): a potential role in the innate immunity of the oral cavity and respiratory tract and the development of adenocarcinomas of the lung. Respir. Res. 7, 61 (2006).

    Article  Google Scholar 

  17. Greer, K.A. et al. Gene expression analysis in a canine model of X-linked Alport syndrome. Mamm. Genome 17, 976–990 (2006).

    Article  CAS  Google Scholar 

  18. Bielesz, B. et al. Epithelial Notch signaling regulates interstitial fibrosis development in the kidneys of mice and humans. J. Clin. Invest. 120, 4040–4054 (2010).

    Article  CAS  Google Scholar 

  19. Bunnag, S.a. Molecular correlates of renal functions in kidney transplant biopsies. J. Am. Soc. Nephrol. 20, 1149–1160 (2009).

    Article  Google Scholar 

  20. Galgano, M.T., Hampton, G.M. & Frierson, H.F. Jr. Comprehensive analysis of HE4 expression in normal and malignant human tissues. Mod. Pathol. 19, 847–853 (2006).

    Article  CAS  Google Scholar 

  21. Sivashanmugam, P. et al. Characterization of mouse Eppin and a gene cluster of similar protease inhibitors on mouse chromosome 2. Gene 312, 125–134 (2003).

    Article  CAS  Google Scholar 

  22. Bignotti, E. et al. Diagnostic and prognostic impact of serum HE4 detection in endometrial carcinoma patients. Br. J. Cancer 104, 1418–1425 (2011).

    Article  CAS  Google Scholar 

  23. Drapkin, R. et al. Human epididymis protein 4 (HE4) is a secreted glycoprotein that is overexpressed by serous and endometrioid ovarian carcinomas. Cancer Res. 65, 2162–2169 (2005).

    Article  CAS  Google Scholar 

  24. Matsuo, S. et al. Multifunctionality of PAI-1 in fibrogenesis: evidence from obstructive nephropathy in PAI-1-overexpressing mice. Kidney Int. 67, 2221–2238 (2005).

    Article  CAS  Google Scholar 

  25. Oda, T. et al. PAI-1 deficiency attenuates the fibrogenic response to ureteral obstruction. Kidney Int. 60, 587–596 (2001).

    Article  CAS  Google Scholar 

  26. Catania, J.M., Chen, G. & Parrish, A.R. Role of matrix metalloproteinases in renal pathophysiologies. Am. J. Physiol. Renal Physiol. 292, F905–F911 (2007).

    Article  CAS  Google Scholar 

  27. Zeisberg, M. et al. Bone morphogenic protein-7 inhibits progression of chronic renal fibrosis associated with two genetic mouse models. Am J. Physiol. Renal Physiol. 285, F1060–F1067 (2003).

    Article  CAS  Google Scholar 

  28. Bechtel, W. et al. Methylation determines fibroblast activation and fibrogenesis in the kidney. Nat. Med. 16, 544–550 (2010).

    Article  CAS  Google Scholar 

  29. Panicker, L.M., Usha, R., Roy, S. & Mandal, C. Purification and characterization of a serine protease (CESP) from mature coconut endosperm. BMC Res. Notes 2, 81 (2009).

    Article  Google Scholar 

  30. LeBleu, V.S., Sugimoto, H., Miller, C.A., Gattone, V.H. II & Kalluri, R. Lymphocytes are Dispensable for Glomerulonephritis but Required for Renal Interstitial Fibrosis in Matrix Defect Induced Alport Renal Disease. Lab. Invest. 88, 284–292 (2008).

    Article  CAS  Google Scholar 

  31. Lebleu, V.S. et al. Stem Cell Therapies Benefit Alport Syndrome. J. Am. Soc. Nephrol. 20, 2359–2370 (2009).

    Article  CAS  Google Scholar 

  32. Sugimoto, H. et al. Activin-like kinase 3 is important for kidney regeneration and reversal of fibrosis. Nat. Med. 18, 396–404 (2012).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

This study was supported by US National Institutes of Health (NIH) grants DK55001, DK081976, CA125550, CA155370, CA151925 and CA163191, funding from the Harvard Stem Cell Institute, the Metastasis Research Center at the MD Anderson Cancer Center and the Cancer Prevention and Research Institute of Texas (all to R.K.). V.S.L. was funded from the NIH Research Training grant in Gastroenterology (2T32DK007760-11), H.S. was funded by the NIH Research Training grant in Cardiovascular Biology (5T32HL007374-30), and J.T.O. was funded by the NIH Cell and Development Biology Training grant GM07226 and the Department of Defense Breast Cancer Predoctoral Traineeship Award BC083229.

Author information

Authors and Affiliations

  1. Department of Medicine, Division of Matrix Biology, Beth Israel Deaconess Medical Center, Boston, Massachusetts, USA

    Valerie S LeBleu, Yingqi Teng, Joyce T O'Connell, Hikaru Sugimoto & Raghu Kalluri

  2. Department of Medicine, Renal Division, Brigham and Women's Hospital, Boston, Massachusetts, USA

    David Charytan

  3. Department of Nephrology and Rheumatology, Georg-August-University Medical Center, Göttingen, Germany

    Gerhard A Müller & Claudia A Müller

  4. Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, Massachusetts, USA

    Raghu Kalluri

  5. Harvard–Massachusetts Institute of Technology Division of Health Sciences and Technology, Boston, Massachusetts, USA

    Raghu Kalluri

  6. Department of Cancer Biology, Metastasis Research Center, University of Texas MD Anderson Cancer Center, Houston, Texas, USA

    Raghu Kalluri

Authors
  1. Valerie S LeBleu
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Yingqi Teng
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Joyce T O'Connell
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. David Charytan
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Gerhard A Müller
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Claudia A Müller
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Hikaru Sugimoto
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Raghu Kalluri
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

R.K. provided intellectual input and the conceptual framework, designed the study and helped in the writing of the manuscript. V.S.L. designed the study, provided intellectual input, performed experiments, collected data and wrote the manuscript. Y.T., J.T.O. and H.S. performed some experiments and collected data. D.C., G.A.M. and C.A.M. provided human samples for analysis.

Corresponding author

Correspondence to Raghu Kalluri.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Supplementary information

Supplementary Text and Figures

Supplementary Tables 1 and 2, Supplementary Methods and Supplementary Figures 1–5 (PDF 9323 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

LeBleu, V., Teng, Y., O'Connell, J. et al. Identification of human epididymis protein-4 as a fibroblast-derived mediator of fibrosis. Nat Med 19, 227–231 (2013). https://doi.org/10.1038/nm.2989

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nm.2989

This article is cited by

Search

Advanced search

Quick links

Nature Briefing: Translational Research

Sign up for the Nature Briefing: Translational Research newsletter — top stories in biotechnology, drug discovery and pharma.

Get what matters in translational research, free to your inbox weekly. Sign up for Nature Briefing: Translational Research