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Inhibition of TMPRSS2 by HAI-2 reduces prostate cancer cell invasion and metastasis

Oncogene volume 39, pages 5950–5963 (2020)Cite this article

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Abstract

TMPRSS2 is an important membrane-anchored serine protease involved in human prostate cancer progression and metastasis. A serine protease physiologically often comes together with a cognate inhibitor for execution of proteolytically biologic function; however, TMPRSS2’s cognate inhibitor is still elusive. To identify the cognate inhibitor of TMPRSS2, in this study, we applied co-immunoprecipitation and LC/MS/MS analysis and isolated hepatocyte growth factor activator inhibitors (HAIs) to be potential inhibitor candidates for TMPRSS2. Moreover, the recombinant HAI-2 proteins exhibited a better inhibitory effect on TMPRSS2 proteolytic activity than HAI-1, and recombinant HAI-2 proteins had a high affinity to form a complex with TMPRSS2. The immunofluorescence images further showed that TMPRSS2 was co-localized to HAI-2. Both KD1 and KD2 domain of HAI-2 showed comparable inhibitory effects on TMPRSS2 proteolytic activity. In addition, HAI-2 overexpression could suppress the induction effect of TMPRSS2 on pro-HGF activation, extracellular matrix degradation and prostate cancer cell invasion. We further determined that the expression levels of TMPRSS2 were inversely correlated with HAI-2 levels during prostate cancer progression. In orthotopic xenograft animal model, TMPRSS2 overexpression promoted prostate cancer metastasis, and HAI-2 overexpression efficiently blocked TMPRSS2-induced metastasis. In summary, the results together indicate that HAI-2 can function as a cognate inhibitor for TMPRSS2 in human prostate cancer cells and may serve as a potential factor to suppress TMPRSS2-mediated malignancy.

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Fig. 1: TMPRSS2 promoted PCa cell invasion.
Fig. 2: Identification and validation of TMPRSS2-interacting proteins.
Fig. 3: Identification of the functional domain of HAI-2 for the inhibition of TMPRSS2 proteolytic activity.
Fig. 4: Inhibitory role of HAI-2 in TMPRSS2-induced cell invasion and substrate cleavage.
Fig. 5: Correlation of the expression of TMPRSS2 and HAI-2 in PCa specimens.
Fig. 6: Suppression role of HAI-2 in TMPRSS2-induced PCa tumor growth and metastasis.
Fig. 7: Schematic model for the inhibition of TMPRSS2 by HAI-2.

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References

  1. Jemal A, Bray F, Center MM, Ferlay J, Ward E, Forman D. Global cancer statistics. CA Cancer J Clin. 2011;61:69–90.

    PubMed  Google Scholar 

  2. Shen CA-SaMM. Molecular genetics of prostate cancer. Genes Dev. 2000;14:26.

    Google Scholar 

  3. Mierke CT. Physical break-down of the classical view on cancer cell invasion and metastasis. Eur J Cell Biol. 2013;92:89–104.

    CAS  PubMed  Google Scholar 

  4. Lopez-Otin C, Bond JS. Proteases: multifunctional enzymes in life and disease. J Biol Chem. 2008;283:30433–7.

    CAS  PubMed  PubMed Central  Google Scholar 

  5. Sevenich L, Joyce JA. Pericellular proteolysis in cancer. Genes Dev. 2014;28:2331–47.

    PubMed  PubMed Central  Google Scholar 

  6. Bugge TH, Antalis TM, Wu Q. Type II transmembrane serine proteases. J Biol Chem. 2009;284:23177–81.

    CAS  PubMed  PubMed Central  Google Scholar 

  7. Lin CY, Anders J, Johnson M, Dickson RB. Purification and characterization of a complex containing matriptase and a Kunitz-type serine protease inhibitor from human milk. J Biol Chem. 1999;274:18237–42.

    CAS  PubMed  Google Scholar 

  8. Ko CJ, Huang CC, Lin HY, Juan CP, Lan SW, Shyu HY, et al. Androgen-induced TMPRSS2 activates matriptase and promotes extracellular matrix degradation, prostate cancer cell invasion, tumor growth, and metastasis. Cancer Res. 2015;75:2949–60.

    CAS  PubMed  Google Scholar 

  9. Lucas JM, Heinlein C, Kim T, Hernandez SA, Malik MS, True LD, et al. The androgen-regulated protease TMPRSS2 activates a proteolytic cascade involving components of the tumor microenvironment and promotes prostate cancer metastasis. Cancer Discov. 2014;4:1310–25.

    CAS  PubMed  PubMed Central  Google Scholar 

  10. Wilson S, Greer B, Hooper J, Zijlstra A, Walker B, Quigley J, et al. The membrane-anchored serine protease, TMPRSS2, activates PAR-2 in prostate cancer cells. Biochem J. 2005;388:967–72.

    CAS  PubMed  PubMed Central  Google Scholar 

  11. Lam DK, Dang D, Flynn AN, Hardt M, Schmidt BL. TMPRSS2, a novel membrane-anchored mediator in cancer pain. Pain. 2015;156:923–30.

    CAS  PubMed  PubMed Central  Google Scholar 

  12. Garten W, Braden C, Arendt A, Peitsch C, Baron J, Lu Y, et al. Influenza virus activating host proteases: Identification, localization and inhibitors as potential therapeutics. Eur J Cell Biol. 2015;94:375–83.

    CAS  PubMed  Google Scholar 

  13. Hoffmann M, Kleine-Weber H, Schroeder S, Kruger N, Herrler T, Erichsen S, et al. SARS-CoV-2 Cell entry depends on ACE2 and TMPRSS2 and is blocked by a clinically proven protease inhibitor. Cell. 2020;181:271–80. e278.

    CAS  PubMed  PubMed Central  Google Scholar 

  14. Tomlins SA, Rhodes DR, Perner S, Dhanasekaran SM, Mehra R, Sun XW, et al. Recurrent fusion of TMPRSS2 and ETS transcription factor genes in prostate cancer. Science. 2005;310:644–8.

    CAS  PubMed  Google Scholar 

  15. Seth A, Watson DK. ETS transcription factors and their emerging roles in human cancer. Eur J Cancer. 2005;41:2462–78.

    CAS  PubMed  Google Scholar 

  16. Chen YW, Lee MS, Lucht A, Chou FP, Huang W, Havighurst TC, et al. TMPRSS2, a serine protease expressed in the prostate on the apical surface of luminal epithelial cells and released into semen in prostasomes, is misregulated in prostate cancer cells. Am J Pathol. 2010;176:2986–96.

    CAS  PubMed  PubMed Central  Google Scholar 

  17. Lucas JM, True L, Hawley S, Matsumura M, Morrissey C, Vessella R, et al. The androgen-regulated type II serine protease TMPRSS2 is differentially expressed and mislocalized in prostate adenocarcinoma. J Pathol. 2008;215:118–25.

    CAS  PubMed  Google Scholar 

  18. Shimomura T, Denda K, Kitamura A, Kawaguchi T, Kito M, Kondo J, et al. Hepatocyte growth factor activator inhibitor, a novel Kunitz-type serine protease inhibitor. J Biol Chem. 1997;272:6370–6.

    CAS  PubMed  Google Scholar 

  19. Kawaguchi T, Qin L, Shimomura T, Kondo J, Matsumoto K, Denda K, et al. Purification and cloning of hepatocyte growth factor activator inhibitor type 2, a Kunitz-type serine protease inhibitor. J Biol Chem. 1997;272:27558–64.

    CAS  PubMed  Google Scholar 

  20. Kawaguchi M, Kataoka H. Mechanisms of hepatocyte growth factor activation in cancer tissues. Cancers (Basel). 2014;6:1890–904.

    Google Scholar 

  21. Lin CY, Anders J, Johnson M, Sang QA, Dickson RB. Molecular cloning of cDNA for matriptase, a matrix-degrading serine protease with trypsin-like activity. J Biol Chem. 1999;274:18231–6.

    CAS  PubMed  Google Scholar 

  22. 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;390:125–36.

    CAS  PubMed  PubMed Central  Google Scholar 

  23. 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;280:34513–20.

    CAS  PubMed  Google Scholar 

  24. Lee SL, Dickson RB, Lin CY. Activation of hepatocyte growth factor and urokinase/plasminogen activator by matriptase, an epithelial membrane serine protease. J Biol Chem. 2000;275:36720–5.

    CAS  PubMed  Google Scholar 

  25. Miyazawa K. Hepatocyte growth factor activator (HGFA): a serine protease that links tissue injury to activation of hepatocyte growth factor. FEBS J. 2010;277:2208–14.

    CAS  PubMed  Google Scholar 

  26. Delaria KA, Muller DK, Marlor CW, Brown JE, Das RC, Roczniak SO, et al. Characterization of placental bikunin, a novel human serine protease inhibitor. J Biol Chem. 1997;272:12209–14.

    CAS  PubMed  Google Scholar 

  27. Qin L, Denda K, Shimomura T, Kawaguchi T, Kitamura N. Functional characterization of Kunitz domains in hepatocyte growth factor activator inhibitor type 2. FEBS Lett. 1998;436:111–4.

    CAS  PubMed  Google Scholar 

  28. Tsai CH, Teng CH, Tu YT, Cheng TS, Wu SR, Ko CJ, et al. HAI-2 suppresses the invasive growth and metastasis of prostate cancer through regulation of matriptase. Oncogene. 2014;33:4643–52.

    CAS  PubMed  Google Scholar 

  29. Maurer E, Gutschow M, Stirnberg M. Hepatocyte growth factor activator inhibitor type 2 (HAI-2) modulates hepcidin expression by inhibiting the cell surface protease matriptase-2. Biochem J. 2013;450:583–93.

    CAS  PubMed  Google Scholar 

  30. 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;579:1945–50.

    CAS  PubMed  Google Scholar 

  31. Wu SR, Lin CH, Shih HP, Ko CJ, Lin HY, Lan SW, et al. HAI-2 as a novel inhibitor of plasmin represses lung cancer cell invasion and metastasis. Br J Cancer. 2019;120:499–511.

    CAS  PubMed  PubMed Central  Google Scholar 

  32. Roversi FM, Olalla Saad ST, Machado-Neto JA. Serine peptidase inhibitor Kunitz type 2 (SPINT2) in cancer development and progression. Biomed Pharmacother. 2018;101:278–86.

    CAS  PubMed  Google Scholar 

  33. Lai YJ, Chang HH, Lai H, Xu Y, Shiao F, Huang N, et al. N-Glycan Branching Affects the Subcellular Distribution of and Inhibition of Matriptase by HAI-2/Placental Bikunin. PLoS ONE. 2015;10:e0132163.

    PubMed  PubMed Central  Google Scholar 

  34. Schwede T, Kopp J, Guex N, Peitsch MC. SWISS-MODEL: An automated protein homology-modeling server. Nucleic Acids Res. 2003;31:3381–5.

    CAS  PubMed  PubMed Central  Google Scholar 

  35. Biasini M, Bienert S, Waterhouse A, Arnold K, Studer G, Schmidt T, et al. SWISS-MODEL: modelling protein tertiary and quaternary structure using evolutionary information. Nucleic Acids Res. 2014;42:W252–8.

    CAS  PubMed  PubMed Central  Google Scholar 

  36. Comeau SR, Gatchell DW, Vajda S, Camacho CJ. ClusPro: a fully automated algorithm for protein-protein docking. Nucleic Acids Res. 2004;32:W96–9.

    CAS  PubMed  PubMed Central  Google Scholar 

  37. Comeau SR, Gatchell DW, Vajda S, Camacho CJ. ClusPro: an automated docking and discrimination method for the prediction of protein complexes. Bioinformatics. 2004;20:45–50.

    CAS  PubMed  Google Scholar 

  38. Kozakov D, Brenke R, Comeau SR, Vajda S. PIPER: an FFT-based protein docking program with pairwise potentials. Proteins. 2006;65:392–406.

    CAS  PubMed  Google Scholar 

  39. Kozakov D, Hall DR, Xia B, Porter KA, Padhorny D, Yueh C, et al. The ClusPro web server for protein-protein docking. Nat Protoc. 2017;12:255–78.

    CAS  PubMed  PubMed Central  Google Scholar 

  40. Kozakov D, Beglov D, Bohnuud T, Mottarella SE, Xia B, Hall DR, et al. How good is automated protein docking? Proteins. 2013;81:2159–66.

    CAS  PubMed  PubMed Central  Google Scholar 

  41. Chou FP, Chen YW, Zhao XF, Xu-Monette ZY, Young KH, Gartenhaus RB, et al. Imbalanced matriptase pericellular proteolysis contributes to the pathogenesis of malignant B-cell lymphomas. Am J Pathol. 2013;183:1306–17.

    CAS  PubMed  PubMed Central  Google Scholar 

  42. Vogel LK, Saebo M, Skjelbred CF, Abell K, Pedersen ED, Vogel U, et al. The ratio of Matriptase/HAI-1 mRNA is higher in colorectal cancer adenomas and carcinomas than corresponding tissue from control individuals. BMC Cancer. 2006;6:176.

    PubMed  PubMed Central  Google Scholar 

  43. Benaud CM, Oberst M, Dickson RB, Lin CY. Deregulated activation of matriptase in breast cancer cells. Clin Exp Metastasis. 2002;19:639–49.

    CAS  PubMed  Google Scholar 

  44. Mestres J, Gregori-Puigjane E, Valverde S, Sole RV. Data completeness–the Achilles heel of drug-target networks. Nat Biotechnol. 2008;26:983–4.

    CAS  PubMed  Google Scholar 

  45. Pisters LL, Troncoso P, Zhau HE, Li W, von Eschenbach AC, Chung LW. c-met proto-oncogene expression in benign and malignant human prostate tissues. J Urol. 1995;154:293–8.

    CAS  PubMed  Google Scholar 

  46. Russo AL, Jedlicka K, Wernick M, McNally D, Kirk M, Sproull M, et al. Urine analysis and protein networking identify met as a marker of metastatic prostate cancer. Clin Cancer Res. 2009;15:4292–8.

    CAS  PubMed  Google Scholar 

  47. Aalinkeel R, Nair MP, Sufrin G, Mahajan SD, Chadha KC, Chawda RP, et al. Gene expression of angiogenic factors correlates with metastatic potential of prostate cancer cells. Cancer Res. 2004;64:5311–21.

    CAS  PubMed  Google Scholar 

  48. Dozmorov MG, Hurst RE, Culkin DJ, Kropp BP, Frank MB, Osban J, et al. Unique patterns of molecular profiling between human prostate cancer LNCaP and PC-3 cells. Prostate. 2009;69:1077–90.

    CAS  PubMed  PubMed Central  Google Scholar 

  49. Debes JD, Tindall DJ. Mechanisms of androgen-refractory prostate cancer. N Engl. J Med (Comment). 2004;351:1488–90.

    CAS  Google Scholar 

  50. Stowell SR, Ju T, Cummings RD. Protein glycosylation in cancer. Annu Rev Pathol. 2015;10:473–510.

    CAS  PubMed  PubMed Central  Google Scholar 

  51. Jin JS, Cheng TF, Tsai WC, Sheu LF, Chiang H, Yu CP. Expression of the serine protease, matriptase, in breast ductal carcinoma of Chinese women: correlation with clinicopathological parameters. Histol Histopathol. 2007;22:305–9.

    CAS  PubMed  Google Scholar 

  52. Kang JY, Dolled-Filhart M, Ocal IT, Singh B, Lin CY, Dickson RB, et al. Tissue microarray analysis of hepatocyte growth factor/Met pathway components reveals a role for Met, matriptase, and hepatocyte growth factor activator inhibitor 1 in the progression of node-negative breast cancer. Cancer Res. 2003;63:1101–5.

    CAS  PubMed  Google Scholar 

  53. Oberst MD, Chen LY, Kiyomiya K, Williams CA, Lee MS, Johnson MD, et al. HAI-1 regulates activation and expression of matriptase, a membrane-bound serine protease. Am J Physiol Cell Physiol. 2005;289:C462–70.

    CAS  PubMed  Google Scholar 

  54. Pietras K, Ostman A. Hallmarks of cancer: interactions with the tumor stroma. Exp Cell Res. 2010;316:1324–31.

    CAS  PubMed  Google Scholar 

  55. Tao L, Huang G, Song H, Chen Y, Chen L. Cancer associated fibroblasts: An essential role in the tumor microenvironment. Oncol Lett. 2017;14:2611–20.

    PubMed  PubMed Central  Google Scholar 

  56. Kumar D, New J, Vishwakarma V, Joshi R, Enders J, Lin F et al. Cancer-associated fibroblasts drive glycolysis in a targetable signaling loop implicated in head and neck squamous cell carcinoma progression. Cancer Res. 2018;78:3769–82.

  57. Humphrey PA, Halabi S, Picus J, Sanford B, Vogelzang NJ, Small EJ, et al. Prognostic significance of plasma scatter factor/hepatocyte growth factor levels in patients with metastatic hormone- refractory prostate cancer: results from cancer and leukemia group B 150005/9480. Clin Genitourin Cancer. 2006;4:269–74.

    CAS  PubMed  Google Scholar 

  58. Eder T, Weber A, Neuwirt H, Grunbacher G, Ploner C, Klocker H et al. Cancer-associated fibroblasts modify the response of prostate cancer cells to androgen and anti-androgens in three-dimensional spheroid culture. Int J Mol Sci 2016; 17:1458

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Acknowledgements

This study was supported by Ministry of Science and Technology Grants MOST 104-2320-B-002-044-MY3, MOST 105-2911-I-002-521, MOST 106-2320-B-002-046-MY3 and MOST 108-2320-B-002-024-MY3, National Health Research Institutes Grant NHRI-EX106-10401BI and NHRI-EX109-10725BI, and National Taiwan University Grant NTU-CESRP-104R7602C4, NTU105R89612 and NYU107L890504 to M.S. Lee. We appreciate the service of the First Core Laboratory of National Taiwan University College of Medicine. We thank Dr. Chen-Yong Lin at the Georgetown University for his gifts of antibodies. We would like to acknowledge Dr. Chia-Jung Yu (Chang Gung University, Taoyuan, Taiwan; Chang Gung Memorial Hospital, Linkou, Taoyuan, Taiwan) and the Proteomics Core Laboratory of Chang Gung University for providing proteomics technical support.

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  1. These authors contributed equally: Chun-Jung Ko, Ting-Wei Hsu

Authors and Affiliations

  1. Department of Biochemistry and Molecular Biology, College of Medicine, National Taiwan University, Taipei, Taiwan

    Chun-Jung Ko, Ting-Wei Hsu, Shang-Ru Wu, Shao-Wei Lan, Hsin-Ying Lin, Hsin-Hsien Lin, Hsin-Fang Tu, Cheng-Fan Lee, Cheng-Chung Huang & Ming-Shyue Lee

  2. Department of Immunology, The University of Texas MD Anderson Cancer Center, Houston, TX, USA

    Chun-Jung Ko

  3. Graduate Institute of Biomedical Sciences, College of Medicine, Chang Gung University, Taoyuan, Taiwan

    Ting-Feng Hsiao

  4. Molecular Medicine Research Center, Chang Gung University, Taoyuan, Taiwan

    Ting-Feng Hsiao

  5. Department of Bioinformatics and Computational Biology, The University of Texas MD Anderson Cancer Center, Houston, TX, USA

    Mei-Ju May Chen

  6. Agricultural Biotechnology Research Center, Academia Sinica, Taipei, Taiwan

    Pei-Wen Hsiao

  7. Graduate Institute of Medical Genomics and Proteomics, College of Medicine, National Taiwan University, Taipei, Taiwan

    Hsiang-Po Huang

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Ko, CJ., Hsu, TW., Wu, SR. et al. Inhibition of TMPRSS2 by HAI-2 reduces prostate cancer cell invasion and metastasis. Oncogene 39, 5950–5963 (2020). https://doi.org/10.1038/s41388-020-01413-w

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