Study of early stage non-small-cell lung cancer using Orbitrap-based global serum metabolomics.
OBJECTIVE
The aim of the project was to apply ultra-high-performance liquid chromatography-quadrupole-Orbitrap-high-resolution mass spectrometry for serum metabolite profiling of non-small-cell lung cancer (NSCLC). This Orbitrap-based methodology has been applied for a study of NSCLC potential markers for the first time.


METHODS
After extraction using protein precipitation, sera were separated on the ACE Excel 2 C18-PFP (100 × 2.1 mm, 2.0 µm) column using gradient elution and analyzed within the range of 70-1000 m/z. Only patients with early stage disease (stages IA-IIB) were included in the study, providing opportunity to find biomarkers for early lung cancer detection. The resulting metabolite profiles were subjected to univariate and multivariate statistical tests.


RESULTS
36 features were found significantly changed between NSCLC group and controls after FDR adjustment and 19 were identified using various metabolite databases (in-house library, HMDB, mzCloud). The study revealed a number of NSCLC biomarker candidates which belong to such compound classes as acylcarnitines, organic acids, and amino acids. Multivariate ROC curve built using 12 identified metabolites was characterized by AUC = 0.836 (0.722-0.946). There were no significant differences in the serum metabolite profiles between two most common histological types of lung cancer-adenocarcinoma and squamous cell carcinoma.


CONCLUSIONS
Through identification of novel potential tumor markers, Orbitrap-based global metabolic profiling is a useful strategy in cancer research. Our study can accelerate development of new diagnostic and therapeutic strategies in NSCLC. The metabolites involved in discrimination between NSCLC patients and the control subjects should be further explored using a targeted approach.
Department of Inorganic and Analytical Chemistry, Poznan University of Medical Sciences, Grunwaldzka 6 Street, 60-780, Poznan, Poland.
Agnieszka
A
Klupczynska
Department of Inorganic and Analytical Chemistry, Poznan University of Medical Sciences, Grunwaldzka 6 Street, 60-780, Poznan, Poland.
Paweł
P
Dereziński
Department of Pathology, Immunology and Laboratory Medicine, College of Medicine, University of Florida, 1395 Center Drive, Gainesville, FL, 32610, USA.
Timothy J
TJ
Garrett
Department of Pathology, Immunology and Laboratory Medicine, College of Medicine, University of Florida, 1395 Center Drive, Gainesville, FL, 32610, USA.
Vanessa Y
VY
Rubio
Department of Thoracic Surgery, Poznan University of Medical Sciences, Szamarzewskiego 62 Street, 60-569, Poznan, Poland.
Wojciech
W
Dyszkiewicz
Department of Thoracic Surgery, Poznan University of Medical Sciences, Szamarzewskiego 62 Street, 60-569, Poznan, Poland.
Mariusz
M
Kasprzyk
Department of Inorganic and Analytical Chemistry, Poznan University of Medical Sciences, Grunwaldzka 6 Street, 60-780, Poznan, Poland. zkokot@ump.edu.pl.
Zenon J
ZJ
Kokot
D000368:Aged; D000369:Aged, 80 and over; D002289:Carcinoma, Non-Small-Cell Lung / Q000097:blood*; D002851:Chromatography, High Pressure Liquid; D005260:Female; D006801:Humans; D008175:Lung Neoplasms / Q000097:blood*; D008297:Male; D013058:Mass Spectrometry; D055432:Metabolomics
Global metabolomics; Lung cancer; Mass spectrometry; Metabolite profiling; Orbitrap