Inhibition of endopeptidase and exopeptidase activity of cathepsin B impairs extracellular matrix degradation and tumour invasion
Abstract
Cathepsin B is a lysosomal cysteine protease that is implicated in a number of physiological processes, including protein turnover in lysosomes. Changes in its expression are associated with a variety of pathological processes, including cancer. Due to the structural feature, termed the occluding loop, cathepsin B differs from other cysteine proteases in possessing both, endopeptidase and exopeptidase activity. Here we investigated the impact of both cathepsin B activities on intracellular and extracellular collagen IV degradation and tumour cell invasion using new selective synthetic inhibitors, 2-{[(8-hydroxy-5-nitroquinoline-7-yl)methyl]amino}-acetonitrile (1), 8-(4-methylpiperidin-1-yl)-5-nitroquinoline (2) and 7-[(4-methylpiperidin-1yl)methyl]-5-nitroquinolin-8-ol (3). All three compounds (5 μm) reduced extracellular degradation of collagen IV by MCF-10A neoT cells by 45–70% as determined by spectrofluorimetry and they (50 μm) attenuated intracellular collagen IV degradation by 40-60% as measured with flow cytometry. Furthermore, all three compounds (5 μm) impaired MCF-10A neoT cell invasion by 40–80% as assessed by measuring electrical impedance in real time. Compounds 1 and 3 (5 μm), but not compound 2, significantly reduced the growth of MMTV-PyMT multicellular tumour spheroids. Collectively, these data suggest that the efficient strategy to impair harmful cathepsin B activity in tumour progression may include simultaneous and potent inhibition of cathepsin B endopeptidase and exopeptidase activities.
Acknowledgments
This work was supported by the Slovenian Research Agency (grant number J4-5529 to J.K.).
References
Aggarwal, N. and Sloane, B.F. (2014). Cathepsin B: multiple roles in cancer. Proteomics Clin. Appl. 8, 427–437.10.1002/prca.201300105Suche in Google Scholar
Almeida, P.C., Nantes, I.L., Chagas, J.R., Rizzi, C.C., Faljoni-Alario, A, Carmona, E., Juliano, L., Nader, H.B., and Tersariol, I.L. (2001). Cathepsin B activity regulation. Heparin-like glycosaminogylcans protect human cathepsin B from alkaline pH-induced inactivation. J. Biol. Chem. 276, 944–951.10.1074/jbc.M003820200Suche in Google Scholar
Berardi, S., Lang, a, Kostoulas, G., Hörler, D., Vilei, E.M., and Baici, A. (2001). Alternative messenger RNA splicing and enzyme forms of cathepsin B in human osteoarthritic cartilage and cultured chondrocytes. Arthritis Rheum. 44, 1819–1831.10.1002/1529-0131(200108)44:8<1819::AID-ART319>3.0.CO;2-4Suche in Google Scholar
Bian, B., Mongrain, S., Cagnol, S., Langlois, M.-J., Boulanger, J., Bernatchez, G., Carrier, J.C., Boudreau, F., and Rivard, N. (2015). Cathepsin B promotes colorectal tumorigenesis, cell invasion, and metastasis. Mol. Carcinog. 204, 525–540.Suche in Google Scholar
Cavallo-Medved, D., Rudy, D., Blum, G., Bogyo, M., Caglic, D., and Sloane, B.F. (2009). Live-cell imaging demonstrates extracellular matrix degradation in association with active cathepsin B in caveolae of endothelial cells during tube formation. Exp. Cell Res. 315, 1234–1246.10.1016/j.yexcr.2009.01.021Suche in Google Scholar
Eisenberg, M.C., Kim, Y., Li, R., Ackerman, W.E., Kniss, D.A., and Friedman, A. (2011). Mechanistic modeling of the effects of myoferlin on tumor cell invasion. Proc. Natl. Acad. Sci. USA 108, 20078–20083.10.1073/pnas.1116327108Suche in Google Scholar
Frizler, M., Lohr, F., Furtmann, N., Kläs, J., and Gütschow, M. (2011). Structural optimization of azadipeptide nitriles strongly increases association rates and allows the development of selective cathepsin inhibitors. J. Med. Chem. 54, 396–400.10.1021/jm101272pSuche in Google Scholar
Halangk, W., Lerch, M.M., Brandt-Nedelev, B., Roth, W., Ruthenbuerger, M., Reinheckel, T., Domschke, W., Lippert, H., Peters, C., and Deussing, J. (2000). Role of cathepsin B in intracellular trypsinogen activation and the onset of acute pancreatitis. J. Clin. Invest. 106, 773–781.10.1172/JCI9411Suche in Google Scholar
Hashimoto, Y., Kakegawa, H., Narita, Y., Hachiya, Y., Hayakawa, T., Kos, J., Turk, V., and Katunuma, N. (2001). Significance of cathepsin B accumulation in synovial fluid of rheumatoid arthritis. Biochem. Biophys. Res. Commun. 283, 334–339.10.1006/bbrc.2001.4787Suche in Google Scholar
Hook, V., Toneff, T., Bogyo, M., Greenbaum, D., Medzihradszky, K.F., Neveu, J., Lane, W., Hook, G., and Reisine, T. (2005). Inhibition of cathepsin B reduces β-amyloid production in regulated secretory vesicles of neuronal chromaffin cells: evidence for cathepsin B as a candidate beta-secretase of Alzheimer’s disease. Biol. Chem. 386, 931–940.10.1515/BC.2005.151Suche in Google Scholar
Illy, C., Quraishi, O., Wang, J., Purisima, E., Vernet, T., and Mort, J.S. (1997). Role of the occluding loop in cathepsin B activity. J. Biol. Chem. 272, 1197–1202.10.1074/jbc.272.2.1197Suche in Google Scholar
Jevnikar, Z., Mirković, B., Fonović, U.P., Zidar, N., Švajger, U., and Kos, J. (2012). Three-dimensional invasion of macrophages is mediated by cysteine cathepsins in protrusive podosomes. Eur. J. Immunol. 42, 3429–3441.10.1002/eji.201242610Suche in Google Scholar
Joyce, J.A. and Hanahan, D. (2004). Multiple roles for cysteine cathepsins in cancer. Cell Cycle 3, 1516–1519.10.4161/cc.3.12.1289Suche in Google Scholar
Kelm, J.M., Timmins, N.E., Brown, C.J., Fussenegger, M., and Nielsen, L.K. (2003). Method for generation of homogeneous multicellular tumor spheroids applicable to a wide variety of cell types. Biotechnol. Bioeng. 83, 173–180.10.1002/bit.10655Suche in Google Scholar
Koblinski, J.E., Ahram, M., and Sloane, B.F. (2000). Unraveling the role of proteases in cancer. Clin. Chim. Acta 291, 113–135.10.1016/S0009-8981(99)00224-7Suche in Google Scholar
Kos, J., Mitrović, A., and Mirković, B. (2014). The current stage of cathepsin B inhibitors as potential anticancer agents. Future Med. Chem. 6, 1355–1371.10.4155/fmc.14.73Suche in Google Scholar
Kostoulas, G., Lang, A., Nagase, H., and Baici, A. (1999). Stimulation of angiogenesis through cathepsin B inactivation of the tissue inhibitors of matrix metalloproteinases. FEBS Lett. 455, 286–290.10.1016/S0014-5793(99)00897-2Suche in Google Scholar
Krupa, J.C., Hasnain, S., Nägler, D.K., Ménard, R., and Mort, J.S. (2002). S2′ substrate specificity and the role of His110 and His111 in the exopeptidase activity of human cathepsin B. Biochem. J. 361, 613–619.10.1042/bj3610613Suche in Google Scholar
Kuhelj, R., Dolinar, M., Pungercar, J., and Turk, V. (1995). The preparation of catalytically active human cathepsin B from its precursor expressed in Escherichia coli in the form of inclusion bodies. Eur. J. Biochem. 229, 533–539.10.1111/j.1432-1033.1995.0533k.xSuche in Google Scholar PubMed
Linebaugh, B.E., Sameni, M., Day, N.A., Sloane, B.F., and Keppler, D. (1999). Exocytosis of active cathepsin B: Enzyme activity at pH 7.0, inhibition and molecular mass. Eur. J. Biochem. 264, 100–109.10.1046/j.1432-1327.1999.00582.xSuche in Google Scholar PubMed
Mirković, B., Premzl, A., Hodnik, V., Doljak, B., Jevnikar, Z., Anderluh, G., and Kos, J. (2009). Regulation of cathepsin B activity by 2A2 monoclonal antibody. FEBS J. 276, 4739–4751.10.1111/j.1742-4658.2009.07171.xSuche in Google Scholar PubMed
Mirković, B., Renko, M., Turk, S., Sosič, I., Jevnikar, Z., Obermajer, N., Turk, D., Gobec, S., and Kos, J. (2011). Novel mechanism of cathepsin B inhibition by antibiotic nitroxoline and related compounds. ChemMedChem 6, 1351–1356.10.1002/cmdc.201100098Suche in Google Scholar
Mirković, B., Markelc, B., Butinar, M., Mitrović, A., Sosič, I., Gobec, S., Vasiljeva, O., Turk, B., Čemažar, M., Serša, G., and Kos, J. (2015). Nitroxoline impairs tumor progression in vitro and in vivo by regulating cathepsin B activity. Oncotarget 6, 19027–19042.10.18632/oncotarget.3699Suche in Google Scholar
Mohamed, M.M. and Sloane, B.F. (2006). Cysteine cathepsins: multifunctional enzymes in cancer. Nat. Rev. Cancer 6, 764–775.10.1038/nrc1949Suche in Google Scholar
Mueller-Klieser, W. (2000). Tumor biology and experimental therapeutics. Crit. Rev. Oncol. Hematol. 36, 123–139.10.1016/S1040-8428(00)00082-2Suche in Google Scholar
Murata, M., Miyashita, S., Yokoo, C., Tamai, M., Hanada, K., Hatayama, K., Towatari, T., Nikawa, T., and Katunuma, N. (1991). Novel epoxysuccinyl peptides: selective inhibitors of cathepsin B, in vitro. FEBS Lett. 280, 307–310.10.1016/0014-5793(91)80318-WSuche in Google Scholar
Musil, D., Zucic, D., Turk, D., Engh, R.A, Mayr, I., Huber, R., Popovic, T., Turk, V., Towatari, T., and Katunuma, N. (1991). The refined 2.15 Å X-ray crystal structure of human liver cathepsin B: the structural basis for its specificity. EMBO J. 10, 2321–2330.10.1002/j.1460-2075.1991.tb07771.xSuche in Google Scholar
Nägler, D.K., Storer, A.C., Portaro, F.C. V., Carmona, E., Juliano, L., and Ménard, R. (1997). Major increase in endopeptidase activity of human cathepsin B upon removal of occluding loop contacts. Biochemistry 36, 12608–12615.10.1021/bi971264+Suche in Google Scholar
Premzl, A., Zavašnik-Bergant, V., Turk, V., and Kos, J. (2003). Intracellular and extracellular cathepsin B facilitate invasion of MCF-10A neoT cells through reconstituted extracellular matrix in vitro. Exp. Cell Res. 283, 206–214.10.1016/S0014-4827(02)00055-1Suche in Google Scholar
Premzl, A., Turk, V., and Kos, J. (2006). Intracellular proteolytic activity of cathepsin B is associated with capillary-like tube formation by endothelial cells in vitro. J. Cell. Biochem. 97, 1230–1240.10.1002/jcb.20720Suche in Google Scholar PubMed
Rawlings, N.D., Waller, M., Barrett, A.J., and Bateman, A. (2014). MEROPS: The database of proteolytic enzymes, their substrates and inhibitors. Nucleic Acids Res. 42, D503–D509.10.1093/nar/gkt953Suche in Google Scholar PubMed PubMed Central
Roshy, S., Sloane, B.F., and Moin, K. (2003). Pericellular cathepsin B and malignant progression. Cancer Metastasis Rev. 22, 271–286.10.1023/A:1023007717757Suche in Google Scholar
Shoji, A., Kabeya, M., Ishida, Y., Yanagida, A., Shibusawa, Y., and Sugawara, M. (2014). Evaluation of cathepsin B activity for degrading collagen IV using a surface plasmon resonance method and circular dichroism spectroscopy. J. Pharm. Biomed. Anal. 95, 47–53.10.1016/j.jpba.2014.02.009Suche in Google Scholar
Skrzydlewska, E., Sulkowska, M., Koda, M., and Sulkowski, S. (2005). Proteolytic-antiproteolytic balance and its regulation in carcinogenesis. World J. Gastroenterol. 11, 1251–1266.10.3748/wjg.v11.i9.1251Suche in Google Scholar
Sosič, I., Mirković, B., Arenz, K., Štefane, B., Kos, J., and Gobec, S. (2013). Development of new cathepsin B inhibitors: Combining bioisosteric replacements and structure-based design to explore the structure-activity relationships of nitroxoline derivatives. J. Med. Chem. 56, 521–533.10.1021/jm301544xSuche in Google Scholar
Timmins, N.E., and Nielsen, L.K. (2007). Generation of multicellular tumor spheroids by the hanging-drop method. Methods Mol. Med. 140, 141–151.10.1007/978-1-59745-443-8_8Suche in Google Scholar
Towatari, T., Nikawa, T., Murata, M., Yokoo, C., Tamai, M., Hanada, K., and Katunuma, N. (1991). Novel epoxysuccinyl peptides: A selective inhibitor of cathepsin B, in vivo. FEBS Lett. 280, 311–315.10.1016/0014-5793(91)80319-XSuche in Google Scholar
Yamamoto, A., Hara, T., Tomoo, K., Ishida, T., Fujii, T., Hata, Y., Murata, M., and Kitamura, K. (1997). Binding mode of CA074, a specific irreversible inhibitor, to bovine cathepsin B as determined by X-ray crystal analysis of the complex. J. Biochem. 121, 974–977.10.1093/oxfordjournals.jbchem.a021682Suche in Google Scholar PubMed
©2016 by De Gruyter
Artikel in diesem Heft
- Frontmatter
- Research Articles/Short Communications
- Genes and Nucleic Acids
- Acute hypothalamo-pituitary-adrenal axis response to LPS-induced endotoxemia: expression pattern of kinin type B1 and B2 receptors
- Protein Structure and Function
- Crystal structure of cleaved vaspin (serpinA12)
- Semi-purification procedures of prions from a prion-infected brain using sucrose has no influence on the nonenzymatic glycation of the disease-associated prion isoform
- Involvement of loop 5 lysine residues and the N-terminal β-hairpin of the ribotoxin hirsutellin A on its insecticidal activity
- Membranes, Lipids, Glycobiology
- De novo ceramide synthesis is involved in acute inflammation during labor
- Cell Biology and Signaling
- A role for NGF and its receptors TrKA and p75NTR in the progression of COPD
- Proteolysis
- Inhibition of endopeptidase and exopeptidase activity of cathepsin B impairs extracellular matrix degradation and tumour invasion
Artikel in diesem Heft
- Frontmatter
- Research Articles/Short Communications
- Genes and Nucleic Acids
- Acute hypothalamo-pituitary-adrenal axis response to LPS-induced endotoxemia: expression pattern of kinin type B1 and B2 receptors
- Protein Structure and Function
- Crystal structure of cleaved vaspin (serpinA12)
- Semi-purification procedures of prions from a prion-infected brain using sucrose has no influence on the nonenzymatic glycation of the disease-associated prion isoform
- Involvement of loop 5 lysine residues and the N-terminal β-hairpin of the ribotoxin hirsutellin A on its insecticidal activity
- Membranes, Lipids, Glycobiology
- De novo ceramide synthesis is involved in acute inflammation during labor
- Cell Biology and Signaling
- A role for NGF and its receptors TrKA and p75NTR in the progression of COPD
- Proteolysis
- Inhibition of endopeptidase and exopeptidase activity of cathepsin B impairs extracellular matrix degradation and tumour invasion