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Journal of Cancer Research and Clinical Oncology

, Volume 145, Issue 2, pp 383–392 | Cite as

Non-invasive profiling of protease-specific elastin turnover in lung cancer: biomarker potential

  • Jeppe Thorlacius-Ussing
  • Stephanie Nina Kehlet
  • Sarah Rank Rønnow
  • Morten Asser Karsdal
  • Nicholas WillumsenEmail author
Original Article – Cancer Research
  • 54 Downloads

Abstract

Purpose

Elastin is a signature protein of lungs. Increased elastin turnover driven by altered proteolytic activity is an important part of lung tumorigenesis. Elastin-derived fragments have been shown to be pro-tumorigenic, however, little is known regarding the biomarker potential of such elastin fragments. Here, we present an elastin turnover profile by non-invasively quantifying five specific elastin degradation fragments generated by different proteases.

Methods

Elastin fragments were assessed in serum from patients with stage I–IV non-small cell lung cancer (NSCLC) (n = 40) and healthy controls (n = 30) using competitive ELISAs targeting different protease-generated fragments of elastin: ELM12 (generated by matrix metalloproteinase MMP-9 and -12), ELM7 (MMP-7), EL-NE (neutrophil elastase), EL-CG (cathepsin G) and ELP-3 (proteinase 3).

Results

ELM12, ELM7, EL-NE and EL-CG were all significantly elevated in NSCLC patients (n = 40) when compared to healthy controls (n = 30) (ELM12, p = 0.0191; ELM7, p < 0.0001; EL-NE, p < 0.0001; EL-CG, p < 0.0001). ELP-3 showed no significant difference between patients and controls (p = 0.8735). All fragments correlated positively (Spearman, r: 0.69–0.81) when compared pairwise, except ELM12 (Spearman, r: 0.042–0.097). In general, all fragments were detectable across all stages of the disease.

Conclusions

Elastin fragments generated by different proteases are elevated in lung cancer patients compared to healthy controls but differ in their presence. This demonstrates non-invasive biomarker potential of elastin fragments in serum from lung cancer patients and suggests that different pathological mechanisms may be responsible for the elastin turnover, warranting further validation in clinical trials.

Keywords

Elastin MMP Neutrophil elastase Cathepsin g Proteinase 3 Biomarker Lung cancer NSCLC ECM Serum 

Notes

Acknowledgements

We acknowledge the Danish Research Foundation for supporting this study.

Compliance with ethical standards

Conflict of interest

All authors are employed at Nordic Bioscience involved in biomarker discovery and assay development.

Ethical approval

All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards.

Informed consent

Informed consent was obtained from all individual participants included in the study.

References

  1. Atkinson JJ, Lutey BA, Suzuki Y et al (2011) The role of matrix metalloproteinase-9 in cigarette smoke-induced emphysema. Am J Respir Crit Care Med 183:876–884.  https://doi.org/10.1164/rccm.201005-0718OC CrossRefGoogle Scholar
  2. Balkwill FR, Capasso M, Hagemann T (2012) The tumor microenvironment at a glance. J Cell Sci 125:5591–5596.  https://doi.org/10.1242/jcs.116392 CrossRefGoogle Scholar
  3. Baud S, Duca L, Bochicchio B et al (2013) Elastin peptides in aging and pathological conditions. Biomol Concepts 4:65–76.  https://doi.org/10.1515/bmc-2011-0062 CrossRefGoogle Scholar
  4. Bihlet AR, Karsdal MA, Sand JMB et al (2017) Biomarkers of extracellular matrix turnover are associated with emphysema and eosinophilic-bronchitis in COPD. Respir Res 18:22.  https://doi.org/10.1186/s12931-017-0509-x CrossRefGoogle Scholar
  5. Blood CH, Sasse J, Brodt P, Zetter BR (1988) Identification of a tumor cell receptor for VGVAPG, an elastin-derived chemotactic peptide. J Cell Biol 107:1987–1993.  https://doi.org/10.1083/jcb.107.5.1987 CrossRefGoogle Scholar
  6. Brown GT, Murray GI (2015) Current mechanistic insights into the roles of matrix metalloproteinases in tumour invasion and metastasis. J Pathol 237:273–281.  https://doi.org/10.1002/path.4586 CrossRefGoogle Scholar
  7. Devy J, Duca L, Cantarelli B et al (2010) Elastin-derived peptides enhance melanoma growth in vivo by upregulating the activation of Mcol-A (MMP-1) collagenase. Br J Cancer 103:1562–1570.  https://doi.org/10.1038/sj.bjc.6605926 CrossRefGoogle Scholar
  8. El Rayes T, Catena R, Lee S et al (2015) Lung inflammation promotes metastasis through neutrophil protease-mediated degradation of Tsp-1. Proc Natl Acad Sci USA 112:16000–16005.  https://doi.org/10.1073/pnas.1507294112 CrossRefGoogle Scholar
  9. Ella E, Harel Y, Abraham M et al (2018) Matrix metalloproteinase 12 promotes tumor propagation in the lung. J Thorac Cardiovasc Surg 155:2164–2175.e1.  https://doi.org/10.1016/j.jtcvs.2017.11.110 CrossRefGoogle Scholar
  10. Gminski J, Mykala-Ciesla J, Machalski M, Drozdz M (1993) Elastin metabolism parameters in sera of patients with lung cancer. Neoplasma 40:41–44Google Scholar
  11. Gong L, Wu D, Zou J et al (2016) Prognostic impact of serum and tissue MMP-9 in non-small cell lung cancer: a systematic review and meta-analysis. Oncotarget 7:18458–18468.  https://doi.org/10.18632/oncotarget.7607 Google Scholar
  12. Guarino C, Legowska M, Epinette C et al (2014) New selective peptidyl di(chlorophenyl) phosphonate esters for visualizing and blocking neutrophil proteinase 3 in human diseases. J Biol Chem 289:31777–31791.  https://doi.org/10.1074/jbc.M114.591339 CrossRefGoogle Scholar
  13. Gudmann NS, Manon-Jensen T, Sand JMB et al (2018) Lung tissue destruction by proteinase 3 and cathepsin G mediated elastin degradation is elevated in chronic obstructive pulmonary disease. Biochem Biophys Res Commun.  https://doi.org/10.1016/j.bbrc.2018.07.038 Google Scholar
  14. Guyot N, Wartelle J, Malleret L et al (2014) Unopposed cathepsin G, neutrophil elastase, and proteinase 3 cause severe lung damage and emphysema. Am J Pathol 184:2197–2210.  https://doi.org/10.1016/j.ajpath.2014.04.015 CrossRefGoogle Scholar
  15. Halaoui R, McCaffrey L (2015) Rewiring cell polarity signaling in cancer. Oncogene 34:939–950.  https://doi.org/10.1038/onc.2014.59 CrossRefGoogle Scholar
  16. Han J-C, Li X-D, Du J et al (2015) Elevated matrix metalloproteinase-7 expression promotes metastasis in human lung carcinoma. World J Surg Oncol 13:5.  https://doi.org/10.1186/1477-7819-13-5 CrossRefGoogle Scholar
  17. Hautamaki RD, Kobayashi DK, Senior RM, Shapiro SD (1997) Requirement for macrophage elastase for cigarette smoke-induced emphysema in mice. Science 277:2002–2004CrossRefGoogle Scholar
  18. Heinz A, Jung MC, Duca L et al (2010) Degradation of tropoelastin by matrix metalloproteinases—cleavage site specificities and release of matrikines. FEBS J 277:1939–1956.  https://doi.org/10.1111/j.1742-4658.2010.07616.x CrossRefGoogle Scholar
  19. Heinz A, Taddese S, Sippl W et al (2011) Insights into the degradation of human elastin by matrilysin-1. Biochimie 93:187–194.  https://doi.org/10.1016/j.biochi.2010.09.011 CrossRefGoogle Scholar
  20. Heinz A, Jung MC, Jahreis G et al (2012) The action of neutrophil serine proteases on elastin and its precursor. Biochimie 94:192–202.  https://doi.org/10.1016/j.biochi.2011.10.006 CrossRefGoogle Scholar
  21. Hinek A, Jung S, Rutka JT (1999) Cell surface aggregation of elastin receptor molecules caused by suramin amplified signals leading to proliferation of human glioma cells. Acta Neuropathol 97:399–407CrossRefGoogle Scholar
  22. Hofmann H-S, Hansen G, Richter G et al (2005) Matrix metalloproteinase-12 expression correlates with local recurrence and metastatic disease in non-small cell lung cancer patients. Clin Cancer Res 11:1086–1092Google Scholar
  23. Houghton AM (2010) The paradox of tumor-associated neutrophils: fueling tumor growth with cytotoxic substances. Cell Cycle 9:1732–1737.  https://doi.org/10.4161/cc.9.9.11297 CrossRefGoogle Scholar
  24. Houghton AM (2013) Mechanistic links between COPD and lung cancer. Nat Rev Cancer 13:233–245.  https://doi.org/10.1038/nrc3477 CrossRefGoogle Scholar
  25. Houghton AM, Grisolano JL, Baumann ML et al (2006) Macrophage elastase (Matrix Metalloproteinase-12) suppresses growth of lung metastases. Cancer Res 66:6149–6155.  https://doi.org/10.1158/0008-5472.CAN-04-0297 CrossRefGoogle Scholar
  26. Jung S, Rutka JT, Hinek A (1998) Tropoelastin and elastin degradation products promote proliferation of human astrocytoma cell lines. J Neuropathol Exp Neurol 57:439–448.  https://doi.org/10.1097/00005072-199805000-00007 CrossRefGoogle Scholar
  27. Karsdal MA, Genovese F, Madsen EA et al (2016) Collagen and tissue turnover as a function of age: implications for fibrosis. J Heptol 64:103–109.  https://doi.org/10.1016/j.jhep.2015.08.014 CrossRefGoogle Scholar
  28. Kettunen E, Anttila S, Seppänen JK et al (2004) Differentially expressed genes in nonsmall cell lung cancer: expression profiling of cancer-related genes in squamous cell lung cancer. Cancer Genet Cytogenet 149:98–106.  https://doi.org/10.1016/S0165-4608(03)00300-5 CrossRefGoogle Scholar
  29. Korkmaz B, Moreau T, Gauthier F (2008) Neutrophil elastase, proteinase 3 and cathepsin G: physicochemical properties, activity and physiopathological functions. Biochimie 90:227–242.  https://doi.org/10.1016/j.biochi.2007.10.009 CrossRefGoogle Scholar
  30. Korkmaz B, Lesner A, Guarino C et al (2016) Inhibitors and antibody fragments as potential anti-inflammatory therapeutics targeting neutrophil proteinase 3 in human disease. Pharmacol Rev 68:603–630.  https://doi.org/10.1124/pr.115.012104 CrossRefGoogle Scholar
  31. Kristensen JH, Karsdal MA, Sand JM et al (2015a) Serological assessment of neutrophil elastase activity on elastin during lung ECM remodeling. BMC Pulm Med 15:53.  https://doi.org/10.1186/s12890-015-0048-5 CrossRefGoogle Scholar
  32. Kristensen JH, Larsen L, Dasgupta B et al (2015b) Levels of circulating MMP-7 degraded elastin are elevated in pulmonary disorders. Clin Biochem 48:1083–1088.  https://doi.org/10.1016/j.clinbiochem.2015.07.009 CrossRefGoogle Scholar
  33. Landskron G, De la Fuente M, Thuwajit P et al (2014) Chronic inflammation and cytokines in the tumor microenvironment. J Immunol Res 2014:149185.  https://doi.org/10.1155/2014/149185 CrossRefGoogle Scholar
  34. Leinonen T, Pirinen R, Böhm J et al (2006a) Expression of matrix metalloproteinases 7 and 9 in non-small cell lung cancer. Lung Cancer 51:313–321.  https://doi.org/10.1016/j.lungcan.2005.11.002 CrossRefGoogle Scholar
  35. Leinonen T, Pirinen R, Böhm J et al (2006b) Expression of matrix metalloproteinases 7 and 9 in non-small cell lung cancer. Relation to clinicopathological factors, beta-catenin and prognosis. Lung Cancer 51:313–321.  https://doi.org/10.1016/j.lungcan.2005.11.002 CrossRefGoogle Scholar
  36. Liang Y, Guo S, Zhou Q (2014) Prognostic value of matrix metalloproteinase-7 expression in patients with non-small cell lung cancer. Tumour Biol 35:3717–3724.  https://doi.org/10.1007/s13277-013-1491-7 CrossRefGoogle Scholar
  37. McGarry Houghton A, Houghton AM, McGarry Houghton A (2015) Matrix metalloproteinases in destructive lung disease. Matrix Biol 44–46:167–174.  https://doi.org/10.1016/j.matbio.2015.02.002 CrossRefGoogle Scholar
  38. Mithieux SM, Weiss AS (2005) Elastin. In: Advances in protein chemistry. Academic Press, Cambridge, pp 437–461Google Scholar
  39. Noël A, Gutiérrez-Fernández A, Sounni NE et al (2012) New and paradoxical roles of matrix metalloproteinases in the tumor microenvironment. Front Pharmacol 3:140.  https://doi.org/10.3389/fphar.2012.00140 Google Scholar
  40. Peters HL, Tripathi SC, Kerros C et al (2017) Serine proteases enhance immunogenic antigen presentation on lung cancer cells. Cancer Immunol Res 5:319–329.  https://doi.org/10.1158/2326-6066.CIR-16-0141 CrossRefGoogle Scholar
  41. Qu P, Du H, Wang X, Yan C (2009) Matrix metalloproteinase 12 overexpression in lung epithelial cells plays a key role in emphysema to lung bronchioalveolar adenocarcinoma transition. Cancer Res 69:7252–7261.  https://doi.org/10.1158/0008-5472.CAN-09-0577 CrossRefGoogle Scholar
  42. Ra H-J, Parks WC (2007) Control of matrix metalloproteinase catalytic activity. Matrix Biol 26:587–596.  https://doi.org/10.1016/j.matbio.2007.07.001 CrossRefGoogle Scholar
  43. Ricard-Blum S, Vallet SD (2016) Matricryptins network with matricellular receptors at the surface of endothelial and tumor cells. Front Pharmacol 7:11.  https://doi.org/10.3389/fphar.2016.00011 CrossRefGoogle Scholar
  44. Safranek J, Pesta M, Holubec L et al (2009) Expression of MMP-7, MMP-9, TIMP-1 and TIMP-2 mRNA in lung tissue of patients with non-small cell lung cancer (NSCLC) and benign pulmonary disease. Anticancer Res 29:2513–2517Google Scholar
  45. Sand JMB, Knox AJ, Lange P et al (2015) Accelerated extracellular matrix turnover during exacerbations of COPD. Respir Res 16:69.  https://doi.org/10.1186/s12931-015-0225-3 CrossRefGoogle Scholar
  46. Scandolera A, Odoul L, Salesse S et al (2016) The elastin receptor complex: a unique matricellular receptor with high anti-tumoral potential. Front Pharmacol 7:1–10.  https://doi.org/10.3389/fphar.2016.00032 CrossRefGoogle Scholar
  47. Shah SA, Spinale FG, Ikonomidis JS et al (2010) Differential matrix metalloproteinase levels in adenocarcinoma and squamous cell carcinoma of the lung. J Thorac Cardiovasc Surg 139:984–990.  https://doi.org/10.1016/j.jtcvs.2009.12.016 CrossRefGoogle Scholar
  48. Siegel RL, Miller KD, Jemal A (2018) Cancer statistics, 2018. CA Cancer J Clin 68:7–30.  https://doi.org/10.3322/caac.21442 CrossRefGoogle Scholar
  49. Skjøt-Arkil H, Clausen RE, Rasmussen LM et al (2013) Acute myocardial infarction and pulmonary diseases result in two different degradation profiles of elastin as quantified by two novel ELISAs. PLoS One.  https://doi.org/10.1371/journal.pone.0060936 Google Scholar
  50. Starcher B, Sauter E, Ho C (2013) Elastin turnover in malignant solid tumors. Connect Tissue Res 54:313–318.  https://doi.org/10.3109/03008207.2013.820723 CrossRefGoogle Scholar
  51. Stenvold H, Donnem T, Andersen S et al (2012) Overexpression of matrix metalloproteinase-7 and -9 in NSCLC tumor and stromal cells: correlation with a favorable clinical outcome. Lung Cancer 75:235–241.  https://doi.org/10.1016/j.lungcan.2011.06.010 CrossRefGoogle Scholar
  52. Sun Y, Li D, Lv X-H et al (2015) Roles of osteopontin and matrix metalloproteinase-7 in occurrence, progression, and prognosis of nonsmall cell lung cancer. J Res Med Sci 20:1138–1146.  https://doi.org/10.4103/1735-1995.172980 CrossRefGoogle Scholar
  53. Taddese S, Weiss AS, Neubert RHH, Schmelzer CEH (2008) Mapping of macrophage elastase cleavage sites in insoluble human skin elastin. Matrix Biol 27:420–428.  https://doi.org/10.1016/j.matbio.2008.02.001 CrossRefGoogle Scholar
  54. Toupance S, Brassart B, Rabenoelina F et al (2012) Elastin-derived peptides increase invasive capacities of lung cancer cells by post-transcriptional regulation of MMP-2 and uPA. Clin Exp Metastasis 29:511–522.  https://doi.org/10.1007/s10585-012-9467-3 CrossRefGoogle Scholar
  55. Wise SG, Weiss AS (2009) Tropoelastin. Int J Biochem Cell Biol 41:494–497.  https://doi.org/10.1016/j.biocel.2008.03.017 CrossRefGoogle Scholar
  56. Woenckhaus M, Klein-Hitpass L, Grepmeier U et al (2006) Smoking and cancer-related gene expression in bronchial epithelium and non-small-cell lung cancers. J Pathol 210:192–204.  https://doi.org/10.1002/path.2039 CrossRefGoogle Scholar
  57. Yang JJ, Preston GA, Pendergraft WF et al (2001) Internalization of proteinase 3 is concomitant with endothelial cell apoptosis and internalization of myeloperoxidase with generation of intracellular oxidants. Am J Pathol 158:581–592.  https://doi.org/10.1016/S0002-9440(10)64000-X CrossRefGoogle Scholar
  58. Zamay T, Zamay G, Kolovskaya O et al (2017) Current and prospective protein biomarkers of lung cancer. Cancers (Basel) 9:155.  https://doi.org/10.3390/cancers9110155 CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Biomarkers & Research, Nordic BioscienceHerlevDenmark
  2. 2.Department of BiologyUniversity of CopenhagenCopenhagen NDenmark

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