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01-08-2019 | Lung Cancer | LUNG CANCER

Mass Spectrometry Analysis of the Exhaled Breath Condensate and Proposal of Dermcidin and S100A9 as Possible Markers for Lung Cancer Prognosis

Authors: Laura Núñez-Naveira, Luis Antonio Mariñas-Pardo, Carmen Montero-Martínez

Published in: Lung | Issue 4/2019

Abstract

Introduction

New sampling techniques to analyse lung diseases, such as exhaled breath condensate (EBC), are a breakthrough in research field since they are less invasive and less traumatic for the patients compared to lung biopsies. Nevertheless, there is an increasing need to optimize not only the sampling protocols but the storage and processing of specimens to get accurate results.

Methods

Exhaled breath condensate was sampled employing the ECoScreen device. Concentrated protein was obtained after ultracentrifugation, lyophilization and reversed-phase chromatography. MALDI-time of flight (TOF)/TOF mass spectrometry (MS) was applied to determine the protein profile in EBC. Commercially available ELISA kits were used to detect the selected biomarker in the EBC after MALDI-MS proteins identification.

Results

The obtained EBC volume after two periods of 10 min doubled the amount obtained after 20 min. One hundred peptides were detected by MALDI-MS, and 18 proteins were identified after reversed-phase chromatography concentration. Dermcidin (P81605), S100A9 (P06702) and Cathepsin G (P08311) were selected to be analysed by ELISA. Dermcidin and S100A9 expression were statistically higher in lung cancer versus healthy volunteers. VEGF concentrations decreased, respectively, by 5.94 and 11.42-fold after 1 and 2 years of frozen EBC preservation in parallel with the declined number of proteins identified by MALDI-MS.

Conclusion

Exhaled breath condensate analysis combined with MS technique may become a valuable method for lung cancer screening and Dermcidin and S100A9 may serve as biomarkers for lung cancer diagnosis or prognosis.

Sidorenko GI, Zborovskii EI, Levina DI (1980) Surface-active properties of the exhaled air condensate (a new method of studying lung function). Ter Arkh 52:65–68PubMed

Shibata A, Katsunuma T, Tomikawa M et al (2006) Increased leukotriene E4 in the exhaled breath condensate of children with mild asthma. Chest 130:1718–1722. https://doi.org/10.1378/chest.130.6.1718 CrossRefPubMed

Vaughan J, Ngamtrakulpanit L, Pajewski TN et al (2003) Exhaled breath condensate pH is a robust and reproducible assay of airway acidity. Eur Respir J 22:889–894CrossRefPubMed

Montuschi P, Corradi M, Ciabattoni G et al (1999) Increased 8-isoprostane, a marker of oxidative stress, in exhaled condensate of asthma patients. Am J Respir Crit Care Med 160:216–220CrossRefPubMed

Robroeks CM, Rijkers GT, Jobsis Q et al (2010) Increased cytokines, chemokines and soluble adhesion molecules in exhaled breath condensate of asthmatic children. Clin Exp Allergy 40:77–84. https://doi.org/10.1111/j.1365-2222.2009.03397.x CrossRefPubMed

Nunez-Naveira L, Marinas-Pardo LA, Amor-Carro O, Montero-Martinez C (2012) Determination of ELISA reproducibility to detect protein markers in exhaled breath condensate. J Breath Res. https://doi.org/10.1088/1752-7155/6/4/046003 CrossRefPubMed

Conrad DH, Goyette J, Thomas PS (2008) Proteomics as a method for early detection of cancer: a review of proteomics, exhaled breath condensate, and lung cancer screening. J Gen Intern Med 23(Suppl 1):78–84. https://doi.org/10.1007/s11606-007-0411-1 CrossRefPubMed

Torre LA, Bray F, Siegel RL et al (2015) Global cancer statistics, 2012. CA Cancer J Clin 65:87–108. https://doi.org/10.3322/caac.21262 CrossRef

Pallis AG, Fennell DA, Szutowicz E et al (2011) Biomarkers of clinical benefit from anti-epidermal growth factor receptor agents in patients with non-small-cell lung cancer. Br J Cancer 105:1–8. https://doi.org/10.1038/bjc.2011.207 CrossRefPubMedPubMedCentral

10.

Dalaveris E, Kerenidi T, Katsabeki-Katsafli A et al (2009) VEGF, TNF-alpha and 8-isoprostane levels in exhaled breath condensate and serum of patients with lung cancer. Lung Cancer 64:219–225. https://doi.org/10.1016/j.lungcan.2008.08.015 CrossRefPubMed

11.

Bloemen K, Van Den Heuvel R, Govarts E et al (2011) A new approach to study exhaled proteins as potential biomarkers for asthma. Clin Exp Allergy 41:346–356. https://doi.org/10.1111/j.1365-2222.2010.03638.x CrossRefPubMed

12.

Kurova VS, Anaev EC, Kononikhin AS et al (2009) Proteomics of exhaled breath: methodological nuances and pitfalls. Clin Chem Lab Med 47:706–712. https://doi.org/10.1515/CCLM.2009.166 CrossRefPubMed

13.

Hoffmann HJ, Tabaksblat LM, Enghild JJ, Dahl R (2008) Human skin keratins are the major proteins in exhaled breath condensate. Eur Respir J 31:380–384. https://doi.org/10.1183/09031936.00059707 CrossRefPubMed

14.

Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685CrossRef

15.

Rabilloud T, Vuillard L, Gilly C, Lawrence JJ (1994) Silver-staining of proteins in polyacrylamide gels: a general overview. Cell Mol Biol 40:57–75PubMed

16.

Dwyer TM (2004) Sampling airway surface liquid: non-volatiles in the exhaled breath condensate. Lung 182:241–250CrossRefPubMed

17.

Banks R, Selby P (2003) Clinical proteomics—insights into pathologies and benefits for patients. Lancet 362:415–416. https://doi.org/10.1016/s0140-6736(03)14096-2 CrossRefPubMed

18.

Mutlu GM, Garey KW, Robbins RA et al (2001) Collection and analysis of exhaled breath condensate in humans. Am J Respir Crit Care Med 164:731–737CrossRefPubMed

19.

Horvath I, Hunt J, Barnes PJ et al (2005) Exhaled breath condensate: methodological recommendations and unresolved questions. Eur Respir J 26:523–548. https://doi.org/10.1183/09031936.05.00029705 CrossRefPubMed

20.

Lacombe M, Marie-Desvergne C, Combes F et al (2018) Proteomic characterization of human exhaled breath condensate. J Breath Res 12:021001. https://doi.org/10.1088/1752-7163/aa9e71 CrossRefPubMed

21.

López-Sánchez LM, Jurado-Gámez B, Feu-Collado N et al (2017) Exhaled breath condensate biomarkers for the early diagnosis of lung cancer using proteomics. Am J Physiol Cell Mol Physiol 313:L664–L676. https://doi.org/10.1152/ajplung.00119.2017 CrossRef

22.

Mateos J, Pintor-Iglesias A, Fernández-Puente P et al (2014) Cryoconservation of peptide extracts from trypsin digestion of proteins for proteomic analysis in a hospital biobank facility. J Proteome Res 13:1930–1937. https://doi.org/10.1021/pr401046u CrossRefPubMed

23.

Jackson AS, Sandrini A, Campbell C et al (2007) Comparison of biomarkers in exhaled breath condensate and bronchoalveolar lavage. Am J Respir Crit Care Med 175:222–227. https://doi.org/10.1164/rccm.200601-107OC CrossRefPubMed

24.

Cruickshank-Quinn C, Armstrong M, Powell R et al (2017) Determining the presence of asthma-related molecules and salivary contamination in exhaled breath condensate. Respir Res 18:57. https://doi.org/10.1186/s12931-017-0538-5 CrossRefPubMedPubMedCentral

25.

Romero PV, Rodríguez B, Martínez S et al (2006) Analysis of oxidative stress in exhaled breath condensate from patients with severe pulmonary infections. Arch Bronconeumol 42:113–119CrossRefPubMed

26.

Rytilä P, Rehn T, Ilumets H et al (2006) Increased oxidative stress in asymptomatic current chronic smokers and GOLD stage 0 COPD. Respir Res 7:69. https://doi.org/10.1186/1465-9921-7-69 CrossRefPubMedPubMedCentral

27.

Nigro E, Imperlini E, Scudiero O et al (2015) Differentially expressed and activated proteins associated with non small cell lung cancer tissues. Respir Res 16:74. https://doi.org/10.1186/s12931-015-0234-2 CrossRefPubMedPubMedCentral

28.

Hmmier A, O’Brien ME, Lynch V et al (2017) Proteomic analysis of bronchoalveolar lavage fluid (BALF) from lung cancer patients using label-free mass spectrometry. BBA Clin 7:97–104. https://doi.org/10.1016/j.bbacli.2017.03.001 CrossRefPubMedPubMedCentral

29.

Hao S, Sun S, Xiao X et al (2016) Selective expression of transthyretin in subtypes of lung cancer. J Mol Histol 47:239–247. https://doi.org/10.1007/s10735-016-9666-3 CrossRefPubMed

30.

Chang WC, Huang MS, Yang CJ et al (2010) Dermcidin identification from exhaled air for lung cancer diagnosis. Eur Respir J 35:1182–1185. https://doi.org/10.1183/09031936.00169509 CrossRefPubMed

31.

Maksimowicz T, Chyczewska E, Chyczewski L et al (1997) Activity and tissue localization of cathepsin G in non small cell lung cancer. Rocz Akad Med Bialymst 42(Suppl 1):199–216PubMed

32.

Gregory AD, Hale P, Perlmutter DH, Houghton AM (2012) Clathrin pit-mediated endocytosis of neutrophil elastase and cathepsin G by cancer cells. J Biol Chem 287:35341–35350. https://doi.org/10.1074/jbc.M112.385617 CrossRefPubMedPubMedCentral

33.

Wang Q, Lu J-B, Wu B, Hao L-Y (2010) Expression and clinicopathologic significance of proteolysis-inducing factor in non-small-cell lung cancer: an immunohistochemical analysis. Clin Lung Cancer 11:346–351. https://doi.org/10.3816/CLC.2010.n.044 CrossRefPubMed

34.

Mirza KA, Wyke SM, Tisdale MJ (2011) Attenuation of muscle atrophy by an N-terminal peptide of the receptor for proteolysis-inducing factor (PIF). Br J Cancer 105:83–88. https://doi.org/10.1038/bjc.2011.216 CrossRefPubMedPubMedCentral

35.

Feng P-H, Lee K-Y, Chang Y-L et al (2012) CD14 (+) S100A9 (+) monocytic myeloid-derived suppressor cells and their clinical relevance in non-small cell lung cancer. Am J Respir Crit Care Med 186:1025–1036. https://doi.org/10.1164/rccm.201204-0636OC CrossRefPubMedPubMedCentral

36.

Huang H, Huang Q, Tang T et al (2018) Clinical significance of calcium-binding protein S100A8 and S100A9 expression in non-small cell lung cancer. Thorac Cancer 9:800–804. https://doi.org/10.1111/1759-7714.12649 CrossRefPubMedPubMedCentral

37.

Liu Y, Cui J, Tang Y-L et al (2018) Prognostic roles of mRNA expression of S100 in non-small-cell lung cancer. Biomed Res Int 2018:1–11. https://doi.org/10.1155/2018/9815806 CrossRef

38.

Tian T, Li X, Hua Z et al (2017) S100A1 promotes cell proliferation and migration and is associated with lymph node metastasis in ovarian cancer. Discov Med 23:235–245PubMed

39.

Lim SY, Yuzhalin AE, Gordon-Weeks AN, Muschel RJ (2016) Tumor-infiltrating monocytes/macrophages promote tumor invasion and migration by upregulating S100A8 and S100A9 expression in cancer cells. Oncogene 35:5735–5745. https://doi.org/10.1038/onc.2016.107 CrossRefPubMedPubMedCentral

Title: Mass Spectrometry Analysis of the Exhaled Breath Condensate and Proposal of Dermcidin and S100A9 as Possible Markers for Lung Cancer Prognosis
Authors: Laura Núñez-Naveira
Luis Antonio Mariñas-Pardo
Carmen Montero-Martínez
Publication date: 01-08-2019
Publisher: Springer US
Keywords: Lung Cancer
Lung Cancer
Lung Cancer
Biomarkers
Published in: Lung / Issue 4/2019
Print ISSN: 0341-2040
Electronic ISSN: 1432-1750
DOI: https://doi.org/10.1007/s00408-019-00238-z

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Introduction

Methods

Results

Conclusion

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