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.

  • Original Article
  • Published:

Deregulated hepsin protease activity confers oncogenicity by concomitantly augmenting HGF/MET signalling and disrupting epithelial cohesion

Oncogene volume 35, pages 1832–1846 (2016)Cite this article

Subjects

Abstract

Hepsin belongs to a family of cell-surface serine proteases, which have sparked interest as therapeutic targets because of the accessibility of extracellular protease domain for inhibitors. Hepsin is frequently amplified and/or overexpressed in epithelial cancers, but it is not clear how enhanced hepsin expression confers a potential for oncogenicity. We show that hepsin is consistently overexpressed in more than 40% of examined breast cancers, including all major biological subtypes. The effects of doxycycline-induced hepsin overexpression were examined in mammary epithelial organoids, and we found that induced hepsin acutely downmodulates its cognate inhibitor, hepatocyte growth factor (HGF) activator inhibitor type 1 (HAI-1). Hepsin-induced depletion of cellular HAI-1 led to a sharp increase in pericellular serine protease activity. The derepressed hepsin proteolytically activated downstream serine proteases, augmented HGF/MET signalling and caused deterioration of desmosomes and hemidesmosomes; structures important for cell cohesion and cell-basement membrane interaction. Moreover, chronic induction of hepsin considerably shortened the latency of Myc-dependent tumourigenesis in the mouse mammary gland. The serine protease and uPA system inhibitor WX-UK1, identified as a micromolar range hepsin inhibitor, prevented hepsin from augmenting HGF/MET signalling and disrupting desmosomes and hemidesmosomes. The findings suggest that the oncogenic activity of hepsin arises not only from elevated expression level but also from depletion of HAI-1, events which together trigger gain-of-function activity impacting HGF/MET signalling and epithelial cohesion. Thus, hepsin overexpression is a major oncogenic conferrer to a serine protease activity involved in breast cancer dissemination.

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
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6
Figure 7
Figure 8

Similar content being viewed by others

References

  1. Antalis TM, Buzza MS, Hodge KM, Hooper JD, Netzel-Arnett S . The cutting edge: membrane-anchored serine protease activities in the pericellular microenvironment. Biochem J 2010; 428: 325–346.

    Article  CAS  PubMed  Google Scholar 

  2. Miao J, Mu D, Ergel B, Singavarapu R, Duan Z, Powers S et al. Hepsin colocalizes with desmosomes and induces progression of ovarian cancer in a mouse model. Int J Cancer 2008; 123: 2041–2047.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Xing P, Li JG, Jin F, Zhao TT, Liu Q, Dong HT et al. Clinical and biological significance of hepsin overexpression in breast cancer. J Investig Med 2011; 59: 803–810.

    Article  CAS  PubMed  Google Scholar 

  4. Szabo R, Bugge TH . Type II transmembrane serine proteases in development and disease. Int J Biochem Cell Biol 2008; 40: 1297–1316.

    Article  CAS  PubMed  Google Scholar 

  5. Klezovitch O, Chevillet J, Mirosevich J, Roberts RL, Matusik RJ, Vasioukhin V . Hepsin promotes prostate cancer progression and metastasis. Cancer Cell 2004; 6: 185–195.

    Article  CAS  PubMed  Google Scholar 

  6. Li W, Wang BE, Moran P, Lipari T, Ganesan R, Corpuz R et al. Pegylated kunitz domain inhibitor suppresses hepsin-mediated invasive tumor growth and metastasis. Cancer Res 2009; 69: 8395–8402.

    Article  CAS  PubMed  Google Scholar 

  7. Xuan JA, Schneider D, Toy P, Lin R, Newton A, Zhu Y et al. Antibodies neutralizing hepsin protease activity do not impact cell growth but inhibit invasion of prostate and ovarian tumor cells in culture. Cancer Res 2006; 66: 3611–3619.

    Article  CAS  PubMed  Google Scholar 

  8. Nandana S, Ellwood-Yen K, Sawyers C, Wills M, Weidow B, Case T et al. Hepsin cooperates with MYC in the progression of adenocarcinoma in a prostate cancer mouse model. Prostate 2010; 70: 591–600.

    CAS  PubMed  PubMed Central  Google Scholar 

  9. Partanen JI, Tervonen TA, Myllynen M, Lind E, Imai M, Katajisto P et al. Tumor suppressor function of Liver kinase B1 (Lkb1) is linked to regulation of epithelial integrity. Proc Natl Acad Sci USA 2012; 109: E388–E397.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Herter S, Piper DE, Aaron W, Gabriele T, Cutler G, Cao P et al. Hepatocyte growth factor is a preferred in vitro substrate for human hepsin, a membrane-anchored serine protease implicated in prostate and ovarian cancers. Biochem J 2005; 390: 125–136.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Somoza JR, Ho JD, Luong C, Ghate M, Sprengeler PA, Mortara K et al. The structure of the extracellular region of human hepsin reveals a serine protease domain and a novel scavenger receptor cysteine-rich (SRCR) domain. Structure 2003; 11: 1123–1131.

    Article  CAS  PubMed  Google Scholar 

  12. Kirchhofer D, Peek M, Lipari MT, Billeci K, Fan B, Moran P . Hepsin activates pro-hepatocyte growth factor and is inhibited by hepatocyte growth factor activator inhibitor-1B (HAI-1B) and HAI-2. FEBS Lett 2005; 579: 1945–1950.

    Article  CAS  PubMed  Google Scholar 

  13. Ganesan R, Kolumam GA, Lin SJ, Xie MH, Santell L, Wu TD et al. Proteolytic activation of pro-macrophage-stimulating protein by hepsin. Mol Cancer Res 2011; 9: 1175–1186.

    Article  CAS  PubMed  Google Scholar 

  14. Hsu YC, Huang HP, Yu IS, Su KY, Lin SR, Lin WC et al. Serine protease hepsin regulates hepatocyte size and hemodynamic retention of tumor cells by hepatocyte growth factor signaling in mice. Hepatology 2012; 56: 1913–1923.

    Article  CAS  PubMed  Google Scholar 

  15. Chen M, Chen LM, Lin CY, Chai KX . Hepsin activates prostasin and cleaves the extracellular domain of the epidermal growth factor receptor. Mol Cell Biochem 2010; 337: 259–266.

    Article  CAS  PubMed  Google Scholar 

  16. Buzza MS, Martin EW, Driesbaugh KH, Desilets A, Leduc R, Antalis TM . Prostasin is required for matriptase activation in intestinal epithelial cells to regulate closure of the paracellular pathway. J Biol Chem 2013; 288: 10328–10337.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Chen YW, Wang JK, Chou FP, Chen CY, Rorke EA, Chen LM et al. Regulation of the matriptase-prostasin cell surface proteolytic cascade by hepatocyte growth factor activator inhibitor-1 during epidermal differentiation. J Biol Chem 2010; 285: 31755–31762.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. Moran P, Li W, Fan B, Vij R, Eigenbrot C, Kirchhofer D . Pro-urokinase-type plasminogen activator is a substrate for hepsin. J Biol Chem 2006; 281: 30439–30446.

    Article  CAS  PubMed  Google Scholar 

  19. Moser TL, Enghild JJ, Pizzo SV, Stack MS . The extracellular matrix proteins laminin and fibronectin contain binding domains for human plasminogen and tissue plasminogen activator. J Biol Chem 1993; 268: 18917–18923.

    CAS  PubMed  Google Scholar 

  20. Yue J, Zhang K, Chen J . Role of integrins in regulating proteases to mediate extracellular matrix remodeling. Cancer Microenviron 2012; 5: 275–283.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Tripathi M, Nandana S, Yamashita H, Ganesan R, Kirchhofer D, Quaranta V . Laminin-332 is a substrate for hepsin, a protease associated with prostate cancer progression. J Biol Chem 2008; 283: 30576–30584.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. List K, Szabo R, Molinolo A, Sriuranpong V, Redeye V, Murdock T et al. Deregulated matriptase causes ras-independent multistage carcinogenesis and promotes ras-mediated malignant transformation. Genes Dev 2005; 19: 1934–1950.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  23. Benaud C, Dickson RB, Lin CY . Regulation of the activity of matriptase on epithelial cell surfaces by a blood-derived factor. Eur J Biochem 2001; 268: 1439–1447.

    Article  CAS  PubMed  Google Scholar 

  24. Fan B, Wu TD, Li W, Kirchhofer D . Identification of hepatocyte growth factor activator inhibitor-1B as a potential physiological inhibitor of prostasin. J Biol Chem 2005; 280: 34513–34520.

    Article  CAS  PubMed  Google Scholar 

  25. Li Y, Yu Z, Zhao X, Shen SH . Identification and characterization of hepsin/-TM, a non-transmembrane hepsin isoform. Biochim Biophys Acta 2005; 1681: 157–165.

    Article  CAS  PubMed  Google Scholar 

  26. Debnath J, Muthuswamy SK, Brugge JS . Morphogenesis and oncogenesis of MCF-10 A mammary epithelial acini grown in three-dimensional basement membrane cultures. Methods 2003; 30: 256–268.

    Article  CAS  PubMed  Google Scholar 

  27. Partanen JI, Tervonen TA, Klefstrom J . Breaking the epithelial polarity barrier in cancer: the strange case of LKB1/PAR-4. Philos Trans R Soc Lond B Biol Sci 2013; 368: 20130111.

    Article  PubMed  PubMed Central  Google Scholar 

  28. Shimomura T, Denda K, Kawaguchi T, Matsumoto K, Miyazawa K, Kitamura N . Multiple sites of proteolytic cleavage to release soluble forms of hepatocyte growth factor activator inhibitor type 1 from a transmembrane form. J Biochem 1999; 126: 821–828.

    Article  CAS  PubMed  Google Scholar 

  29. Godiksen S, Selzer-Plon J, Pedersen ED, Abell K, Rasmussen HB, Szabo R et al. Hepatocyte growth factor activator inhibitor-1 has a complex subcellular itinerary. Biochem J 2008; 413: 251–259.

    Article  CAS  PubMed  Google Scholar 

  30. Kiyomiya K, Lee MS, Tseng IC, Zuo H, Barndt RJ, Johnson MD et al. Matriptase activation and shedding with HAI-1 is induced by steroid sex hormones in human prostate cancer cells, but not in breast cancer cells. Am J Physiol Cell Physiol 2006; 291: C40–C49.

    Article  CAS  PubMed  Google Scholar 

  31. Domoto T, Takino T, Guo L, Sato H . Cleavage of hepatocyte growth factor activator inhibitor-1 by membrane-type MMP-1 activates matriptase. Cancer Sci 2012; 103: 448–454.

    Article  CAS  PubMed  Google Scholar 

  32. Trusolino L, Comoglio PM . Scatter-factor and semaphorin receptors: cell signalling for invasive growth. Nat Rev Cancer 2002; 2: 289–300.

    Article  CAS  PubMed  Google Scholar 

  33. Gastaldi S, Comoglio PM, Trusolino L . The Met oncogene and basal-like breast cancer: another culprit to watch out for? Breast Cancer Res 2010; 12: 208.

    Article  PubMed  PubMed Central  Google Scholar 

  34. Kaipparettu BA, Kuiatse I, Tak-Yee Chan B, Benny Kaipparettu M, Lee AV, Oesterreich S . Novel egg white-based 3-D cell culture system. Biotechniques 2008; 45: 170–161.

    Article  Google Scholar 

  35. Guerin-Dubiard C, Pasco M, Molle D, Desert C, Croguennec T, Nau F . Proteomic analysis of hen egg white. J Agric Food Chem 2006; 54: 3901–3910.

    Article  CAS  PubMed  Google Scholar 

  36. Peters DE, Szabo R, Friis S, Shylo NA, Uzzun Sales K, Holmbeck K et al. The membrane-anchored serine protease prostasin (CAP1/PRSS8) supports epidermal development and postnatal homeostasis independent of its enzymatic activity. J Biol Chem 2014; 289: 14740–14749.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  37. Heinemann V, Ebert MP, Laubender RP, Bevan P, Mala C, Boeck S . Phase II randomised proof-of-concept study of the urokinase inhibitor upamostat (WX-671) in combination with gemcitabine compared with gemcitabine alone in patients with non-resectable, locally advanced pancreatic cancer. Br J Cancer 2013; 108: 766–770.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  38. Pulaski BA, Ostrand-Rosenberg S . Mouse 4T1 breast tumor model. Curr Protoc Immunol 2001; Chapter 20: Unit 20.2.

  39. Dang CV . MYC on the path to cancer. Cell 2012; 149: 22–35.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  40. Gherardi E, Birchmeier W, Birchmeier C, Vande Woude G . Targeting MET in cancer: rationale and progress. Nat Rev Cancer 2012; 12: 89–103.

    Article  CAS  PubMed  Google Scholar 

  41. Szabo R, Molinolo A, List K, Bugge TH . Matriptase inhibition by hepatocyte growth factor activator inhibitor-1 is essential for placental development. Oncogene 2007; 26: 1546–1556.

    Article  CAS  PubMed  Google Scholar 

  42. Friis S, Uzzun Sales K, Godiksen S, Peters DE, Lin CY, Vogel LK et al. A matriptase-prostasin reciprocal zymogen activation complex with unique features: prostasin as a non-enzymatic co-factor for matriptase activation. J Biol Chem 2013; 288: 19028–19039.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  43. Marchant DJ, Bellac CL, Moraes TJ, Wadsworth SJ, Dufour A, Butler GS et al. A new transcriptional role for matrix metalloproteinase-12 in antiviral immunity. Nat Med 2014; 20: 493–502.

    Article  CAS  PubMed  Google Scholar 

  44. Chevillet JR, Park GJ, Bedalov A, Simon JA, Vasioukhin VI . Identification and characterization of small-molecule inhibitors of hepsin. Mol Cancer Ther 2008; 7: 3343–3351.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  45. Ganesan R, Zhang Y, Landgraf KE, Lin SJ, Moran P, Kirchhofer D . An allosteric anti-hepsin antibody derived from a constrained phage display library. Protein Eng Des Sel 2012; 25: 127–133.

    Article  CAS  PubMed  Google Scholar 

  46. Koschubs T, Dengl S, Durr H, Kaluza K, Georges G, Hartl C et al. Allosteric antibody inhibition of human hepsin protease. Biochem J 2012; 442: 483–494.

    Article  CAS  PubMed  Google Scholar 

  47. Tang X, Mahajan SS, Nguyen LT, Beliveau F, Leduc R, Simon JA et al. Targeted inhibition of cell-surface serine protease Hepsin blocks prostate cancer bone metastasis. Oncotarget 2014; 5: 1352–1362.

    PubMed  PubMed Central  Google Scholar 

  48. Goldhirsch A, Wood WC, Coates AS, Gelber RD, Thurlimann B, Senn HJ . Strategies for subtypes—dealing with the diversity of breast cancer: highlights of the St. Gallen International Expert Consensus on the Primary Therapy of Early Breast Cancer 2011. Ann Oncol 2011; 22: 1736–1747.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  49. Meerbrey KL, Hu G, Kessler JD, Roarty K, Li MZ, Fang JE et al. The pINDUCER lentiviral toolkit for inducible RNA interference in vitro and in vivo. Proc Natl Acad Sci USA 2011; 108: 3665–3670.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  50. Partanen JI, Nieminen AI, Makela TP, Klefstrom J . Suppression of oncogenic properties of c-Myc by LKB1-controlled epithelial organization. Proc Natl Acad Sci USA 2007; 104: 14694–14699.

    Article  PubMed  PubMed Central  Google Scholar 

  51. Goswami R, Mukherjee S, Wohlfahrt G, Ghadiyaram C, Nagaraj J, Chandra BR et al. Discovery of pyridyl bis(oxy)dibenzimidamide derivatives as selective matriptase inhibitors. ACS Med Chem Lett 2013; 4: 1152–1157.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgements

We thank all the members of Klefström laboratory for discussions and critical comments on the manuscript. T Raatikainen and T Välimäki are thanked for technical assistance. Biomedicum Imaging Unit and Biomedicum Functional Genomics Unit are acknowledged for core services and technical support. This study was funded by the Academy of Finland, Sigrid Jusélius Foundation, the Finnish Cancer Society, the Research Funds of the Helsinki University Central Hospital, Jane and Aatos Erkko Foundation, and Helsinki Graduate Program in Biotechnology and Molecular Biology and Innovative Medicines Initiative Joint Undertaking under grant agreement n°115188.

Author information

Author notes

  1. D Belitškin and S M Pant: These authors contributed equally to this work.

Authors and Affiliations

  1. Cancer Cell Circuitry Laboratory, Research Programs Unit, Translational Cancer Biology and Institute of Biomedicine, University of Helsinki, Helsinki, Finland

    T A Tervonen, D Belitškin, S M Pant, J I Englund, E Marques, H Ala-Hongisto, L Nevalaita & J Klefström

  2. Department of Oncology, University of Helsinki & Helsinki University Central Hospital, Helsinki, Finland

    H Sihto & H Joensuu

  3. Department of Pathology, HUSLAB and Haartman Institute, Helsinki University Central Hospital and University of Helsinki, Helsinki, Finland

    P Heikkilä & P E Kovanen

  4. Breast Surgery Unit, Helsinki University Central Hospital, Helsinki, Finland

    M Leidenius

  5. Helsinki Innovation Services Ltd, Tukholmankatu 8A, 00290, University of Helsinki, Helsinki, Finland

    K Hewitson

  6. Aurigene Discovery Technologies Limited, 39-40 KIADB Industrial Area, Electronic City Phase II, Bangalore, India

    M Ramachandra

  7. Orion Corporation, Orion Pharma, Turku, Finland

    A Moilanen

  8. School of Pharmacy, University of Eastern Finland, Kuopio, Finland and Finnish Institute of Molecular Medicine, Helsinki, Finland

    A Poso

Authors
  1. T A Tervonen
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. D Belitškin
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. S M Pant
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. J I Englund
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. E Marques
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. H Ala-Hongisto
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. L Nevalaita
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. H Sihto
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. P Heikkilä
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. M Leidenius
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. K Hewitson
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. M Ramachandra
    View author publications

    You can also search for this author in PubMed Google Scholar

  13. A Moilanen
    View author publications

    You can also search for this author in PubMed Google Scholar

  14. H Joensuu
    View author publications

    You can also search for this author in PubMed Google Scholar

  15. P E Kovanen
    View author publications

    You can also search for this author in PubMed Google Scholar

  16. A Poso
    View author publications

    You can also search for this author in PubMed Google Scholar

  17. J Klefström
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to J Klefström.

Ethics declarations

Competing interests

The authors declare no conflict of interest.

Additional information

Supplementary Information accompanies this paper on the Oncogene website

Supplementary information

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Tervonen, T., Belitškin, D., Pant, S. et al. Deregulated hepsin protease activity confers oncogenicity by concomitantly augmenting HGF/MET signalling and disrupting epithelial cohesion. Oncogene 35, 1832–1846 (2016). https://doi.org/10.1038/onc.2015.248

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/onc.2015.248

This article is cited by

Search

Advanced search

Quick links