Comparative analyses of volatile organic compounds (VOCs) from patients, tumors and transformed cell lines for the validation of lung cancer-derived breath markers

Breath analysis for the purpose of non-invasive diagnosis of lung cancer has yielded numerous candidate compounds with still questionable clinical relevance. To arrive at suitable volatile organic compounds our approach combined the analysis of different sources: isolated tumor samples compared to healthy lung tissues, and exhaled breath from lung cancer patients and healthy controls. Candidate compounds were further compared to substances previously identified in the comparison of transformed and normal lung epithelial cell lines. For human studies, a breath sampling device was developed enabling automated and CO2-controlled collection of the end-tidal air. All samples were first preconcentrated on multibed sorption tubes and analyzed with gas chromatography mass spectrometry (GC-MS). Significantly (p < 0.05) higher concentrations in all three types of cancer samples studied were observed for ethanol and n-octane. Additional metabolites (inter alia 2-methylpentane, n-hexane) significantly released by lung cancer cells were observed at higher levels in cancer lung tissues and breath samples (compared to respective healthy controls) with statistical significance (p < 0.05) only in breath samples. The results obtained confirmed the cancer-related origin of volatile metabolites, e.g. ethanol and octane that were both detected at significantly (p < 0.05) elevated concentrations in all three kinds of cancer samples studied. This work is an important step towards identification of volatile breath markers of lung cancer through the demonstration of cancer-related origin of certain volatile metabolites.

[1]  M. Fiegl,et al.  Noninvasive detection of lung cancer by analysis of exhaled breath , 2009, BMC Cancer.

[2]  Anton Amann,et al.  TD-GC-MS Analysis of Volatile Metabolites of Human Lung Cancer and Normal Cells In vitro , 2010, Cancer Epidemiology, Biomarkers & Prevention.

[3]  C. Kneepkens,et al.  The potential of the hydrocarbon breath test as a measure of lipid peroxidation. , 1994, Free radical biology & medicine.

[4]  E. Weitzberg,et al.  Increased amount of nitric oxide in exhaled air of asthmatics. , 1993, The European respiratory journal.

[5]  Frank Ullrich,et al.  Identification of the most intense volatile flavour compounds formed during autoxidation of linoleic acid , 1987 .

[6]  Bogusław Buszewski,et al.  Identification of volatile organic compounds secreted from cancer tissues and bacterial cultures. , 2008, Journal of chromatography. B, Analytical technologies in the biomedical and life sciences.

[7]  Derrick Crump,et al.  A review of the emission of VOCs from polymeric materials used in buildings , 1998 .

[8]  W. Miekisch,et al.  A novel visually CO2 controlled alveolar breath sampling technique. , 2006, Technology and health care : official journal of the European Society for Engineering and Medicine.

[9]  Anton Amann,et al.  Lung cancer detection by proton transfer reaction mass-spectrometric analysis of human breath gas , 2007 .

[10]  J. Troppmair,et al.  Characterization of volatile metabolites taken up by or released from Streptococcus pneumoniae and Haemophilus influenzae by using GC-MS. , 2012, Microbiology.

[11]  D. Berry,et al.  Benefits and harms of CT screening for lung cancer: a systematic review. , 2012, JAMA.

[12]  D. Naidich,et al.  Screening for Lung Cancer Diagnosis and Management of Lung Cancer, 3rd ed: American College of Chest Physicians Evidence-Based Clinical Practice Guidelines , 2013 .

[13]  Julian King,et al.  Stability of selected volatile breath constituents in Tedlar, Kynar and Flexfilm sampling bags. , 2013, The Analyst.

[14]  A. Jemal,et al.  Cancer statistics, 2014 , 2014, CA: a cancer journal for clinicians.

[15]  Armando Santoro,et al.  A randomized study of lung cancer screening with spiral computed tomography: three-year results from the DANTE trial. , 2009, American journal of respiratory and critical care medicine.

[16]  Y. Ung,et al.  Positron Emission Tomography in Staging Early Lung Cancer , 2009, Annals of Internal Medicine.

[17]  E. V. van Beek,et al.  The 7th lung cancer TNM classification and staging system: Review of the changes and implications. , 2012, World journal of radiology.

[18]  Julian King,et al.  Blood and breath levels of selected volatile organic compounds in healthy volunteers. , 2013, The Analyst.

[19]  Ulrike Tisch,et al.  A nanomaterial-based breath test for short-term follow-up after lung tumor resection. , 2013, Nanomedicine : nanotechnology, biology, and medicine.

[20]  Anton Amann,et al.  Analysis of volatile organic compounds (VOCs) in the headspace of NCI-H1666 lung cancer cells. , 2011, Cancer biomarkers : section A of Disease markers.

[21]  Perilous potential: The chance to save lives, or lose them, through low dose computed tomography screening for lung cancer , 2013, Journal of surgical oncology.

[22]  R. Margesin,et al.  Volatile Organic Compounds (VOCs) Released by Pathogenic Microorganisms in vitro : Potential Breath Biomarkers for Early-Stage Diagnosis of Disease , 2013 .

[23]  E. Uhdea,et al.  Impact of reaction products from building materials and furnishings on indoor air quality — A review of recent advances in indoor chemistry , 2007 .

[24]  Anton Amann,et al.  Molecular analysis of volatile metabolites released specifically by staphylococcus aureus and pseudomonas aeruginosa , 2012, BMC Microbiology.

[25]  M. L. R. D. Christenson,et al.  Reduced Lung-Cancer Mortality with Low-Dose Computed Tomographic Screening , 2012 .

[26]  T. H. Risby.  CURRENT STATUS OF CLINICAL BREATH ANALYSIS , 2005 .

[27]  Sheila Weinmann,et al.  Use of diagnostic imaging studies and associated radiation exposure for patients enrolled in large integrated health care systems, 1996-2010. , 2012, JAMA.

[28]  Kevin Gleeson,et al.  Detection of lung cancer with volatile markers in the breath. , 2003, Chest.

[29]  T. Mitsui,et al.  Inadequacy of theoretical basis of breath methylated alkane contour for assessing oxidative stress. , 2003, Clinica chimica acta; international journal of clinical chemistry.

[30]  W. Miekisch,et al.  Breath gas aldehydes as biomarkers of lung cancer , 2009, International journal of cancer.

[31]  J Guttmann,et al.  CO(2)-controlled sampling of alveolar gas in mechanically ventilated patients. , 2001, Journal of applied physiology.

[32]  Andrew K Stewart,et al.  Patterns of surgical care of lung cancer patients. , 2005, The Annals of thoracic surgery.

[33]  Hossam Haick,et al.  Assessment, origin, and implementation of breath volatile cancer markers. , 2014, Chemical Society reviews.

[34]  Tomas Mikoviny,et al.  Release of volatile organic compounds (VOCs) from the lung cancer cell line CALU-1 in vitro , 2008, Cancer Cell International.

[35]  Michael K Gould,et al.  Noninvasive staging of non-small cell lung cancer: ACCP evidenced-based clinical practice guidelines (2nd edition). , 2007, Chest.

[36]  Olaf Tietje,et al.  Prediction of lung cancer using volatile biomarkers in breath , 2005 .

[37]  Panagiotis Artemiadis,et al.  A hybrid BMI-based exoskeleton for paresis: EMG control for assisting arm movements , 2017, Journal of neural engineering.

[38]  M. Roizen.  Reduced Lung-Cancer Mortality with Low-Dose Computed Tomographic Screening , 2012 .

[39]  Gert Mayer,et al.  Blood and breath profiles of volatile organic compounds in patients with end-stage renal disease , 2014, BMC Nephrology.

[40]  A. Dzien,et al.  Dependence of exhaled breath composition on exogenous factors, smoking habits and exposure to air pollutants , 2012, Journal of breath research.

[41]  L. T. Tanoue.  Detection of Lung Cancer by Sensor Array Analyses of Exhaled Breath , 2007 .

[42]  W. Miekisch,et al.  Analysis of exhaled breath for screening of lung cancer patients , 2010 .

[43]  W. Miekisch,et al.  Breath biomarkers for lung cancer detection and assessment of smoking related effects--confounding variables, influence of normalization and statistical algorithms. , 2010, Clinica chimica acta; international journal of clinical chemistry.

[44]  T. Gierczak,et al.  Prevention of water vapour adsorption by carbon molecular sieves in sampling humid gases. , 2001, Journal of chromatography. A.

[45]  E. Martinelli,et al.  Lung cancer identification by the analysis of breath by means of an array of non-selective gas sensors. , 2003, Biosensors & bioelectronics.

[46]  K. Cope.  BREATH ETHANE IN DISEASE: METHODS FOR ANALYSIS BASED ON ROOM AIR CORRECTION , 2005 .

[47]  Ren-Shyan Liu,et al.  Infrared spectrometry based 13C‐octanoic acid breath test in measuring human solid gastric emptying , 2003, Journal of gastroenterology and hepatology.

[48]  Anton Amann,et al.  Improved pre-concentration and detection methods for volatile sulphur breath constituents. , 2009, Journal of chromatography. B, Analytical technologies in the biomedical and life sciences.

[49]  Q. Hasan,et al.  Microsatellite instability analysis and its correlation with hMLH1 repair gene hypermethylation status in esophageal pathologies including cancers. , 2010, Cancer biomarkers : section A of Disease markers.

[50]  R. Damhuis,et al.  Resection rates and postoperative mortality in 7,899 patients with lung cancer. , 1996, The European respiratory journal.

[51]  David Smith,et al.  Breath analysis: the approach towards clinical applications. , 2007, Mini reviews in medicinal chemistry.

[52]  B. de Lacy Costello,et al.  A review of the volatiles from the healthy human body , 2014, Journal of breath research.

[53]  K. Rostásy,et al.  3-Heptanone as a potential new marker for valproic acid therapy , 2009, Journal of breath research.

[54]  Hossam Haick,et al.  Volatile organic compounds of lung cancer and possible biochemical pathways. , 2012, Chemical reviews.

[55]  Tomas Mikoviny,et al.  Release of volatile organic compounds from the lung cancer cell line NCI-H2087 in vitro. , 2009, Anticancer research.

[56]  G. Teschl,et al.  Isoprene and acetone concentration profiles during exercise on an ergometer , 2009, Journal of breath research.

[57]  P. J. Barnes,et al.  Increased nitric oxide in exhaled air of asthmatic patients , 1994, The Lancet.

[58]  Sabine Kischkel,et al.  Impact of sampling procedures on the results of breath analysis , 2008, Journal of breath research.

[59]  Terence H Risby,et al.  Effects of ventilation on the collection of exhaled breath in humans. , 2004, Journal of applied physiology.

[60]  H. Hinterhuber,et al.  Dynamic profiles of volatile organic compounds in exhaled breath as determined by a coupled PTR-MS/GC-MS study , 2010, Physiological measurement.

[61]  T. Gierczak,et al.  Adsorption of water vapour from humid air by selected carbon adsorbents. , 2005, Journal of chromatography. A.

[62]  D. Kleinbaum,et al.  Applied Regression Analysis and Other Multivariate Methods , 1978 .

[63]  M. Phillips,et al.  Effect of oxygen on breath markers of oxidative stress , 2003, European Respiratory Journal.

[64]  G. Bepler,et al.  Screening for lung cancer. , 2000, The New England journal of medicine.

[65]  G. Teschl,et al.  Breath isoprene: muscle dystrophy patients support the concept of a pool of isoprene in the periphery of the human body. , 2012, Biochemical and biophysical research communications.

[66]  Tarek Mekhail,et al.  Lung cancer screening , 2007, Current oncology reports.

[67]  Julian King,et al.  Physiological modeling of isoprene dynamics in exhaled breath. , 2010, Journal of theoretical biology.

[68]  Michael Hill,et al.  Public policy and private provisions: changes in residential care from 1991 to 2001. , 2006, Health & social care in the community.

[69]  O. Miettinen,et al.  Survival of Patients with Stage I Lung Cancer Detected on CT Screening , 2008 .

[70]  C. Liao,et al.  The 13C‐urea breath test to detect Helicobacter pylori infection: a validated simple methodology with 50 mg 13C‐urea , 2002, Alimentary pharmacology & therapeutics.

[71]  A. Amann,et al.  Suitability of different polymer bags for storage of volatile sulphur compounds relevant to breath analysis. , 2009, Journal of chromatography. B, Analytical technologies in the biomedical and life sciences.

[72]  C. Ager,et al.  Optimization of sampling parameters for collection and preconcentration of alveolar air by needle traps , 2012, Journal of Breath Research.

[73]  Julian King,et al.  Release and uptake of volatile organic compounds by human hepatocellular carcinoma cells (HepG2) in vitro , 2013, Cancer Cell International.

[74]  M. Phillips,et al.  Volatile Markers of Breast Cancer in the Breath , 2003, The breast journal.

[75]  Josef Guttmann,et al.  CO2-controlled sampling of alveolar gas in mechanically ventilated patients , 2001 .

[76]  L. Tanoue,et al.  The new lung cancer staging system. , 2009, Chest.

[77]  L. Gaspar,et al.  National survey of non-small cell lung cancer in the United States: epidemiology, pathology and patterns of care. , 2007, Lung cancer.

[78]  Paul Pinsky,et al.  Baseline findings of a randomized feasibility trial of lung cancer screening with spiral CT scan vs chest radiograph: the Lung Screening Study of the National Cancer Institute. , 2004, Chest.

[79]  L. Bianchi,et al.  Exhaled volatile organic compounds in patients with non-small cell lung cancer: cross sectional and nested short-term follow-up study , 2005, Respiratory research.

[80]  Ulrike Tisch,et al.  The scent fingerprint of hepatocarcinoma: in-vitro metastasis prediction with volatile organic compounds (VOCs) , 2012, International journal of nanomedicine.