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.
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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.
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.
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.
Szabo R, Bugge TH . Type II transmembrane serine proteases in development and disease. Int J Biochem Cell Biol 2008; 40: 1297–1316.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
Yue J, Zhang K, Chen J . Role of integrins in regulating proteases to mediate extracellular matrix remodeling. Cancer Microenviron 2012; 5: 275–283.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
Trusolino L, Comoglio PM . Scatter-factor and semaphorin receptors: cell signalling for invasive growth. Nat Rev Cancer 2002; 2: 289–300.
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.
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.
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.
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.
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.
Pulaski BA, Ostrand-Rosenberg S . Mouse 4T1 breast tumor model. Curr Protoc Immunol 2001; Chapter 20: Unit 20.2.
Dang CV . MYC on the path to cancer. Cell 2012; 149: 22–35.
Gherardi E, Birchmeier W, Birchmeier C, Vande Woude G . Targeting MET in cancer: rationale and progress. Nat Rev Cancer 2012; 12: 89–103.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
The authors declare no conflict of interest.
Supplementary Information accompanies this paper on the Oncogene website
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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
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