Untargeted ultra-high-resolution mass spectrometry metabolomic profiling of blood serum in bladder cancer

Abstract

Bladder cancer (BC) is a common urological cancer of high mortality and recurrence rates. Currently, cystoscopy is performed as standard examination for the diagnosis and subsequent monitoring for recurrence of the patients. Frequent expensive and invasive procedures may deterrent patients from regular follow-up screening, therefore it is important to look for new non-invasive methods to aid in the detection of recurrent and/or primary BC. In this study, ultra-high-performance liquid chromatography coupled with ultra-high-resolution mass spectrometry was employed for non-targeted metabolomic profiling of 200 human serum samples to identify biochemical signatures that differentiate BC from non-cancer controls (NCs). Univariate and multivariate statistical analyses with external validation revealed twenty-seven metabolites that differentiate between BC patients from NCs. Abundances of these metabolites displayed statistically significant differences in two independent training and validation sets. Twenty-three serum metabolites were also found to be distinguishing between low- and high-grade of BC patients and controls. Thirty-seven serum metabolites were found to differentiate between different stages of BC. The results suggest that measurement of serum metabolites may provide more facile and less invasive diagnostic methodology for detection of bladder cancer and recurrent disease management.

Unmasking the Metabolite Signature of Bladder Cancer: A Systematic Review

“…The majority of experimental studies that performed an LC-MS-based analysis included the use of cold organic solvents, such as acetonitrile [ 18 , 22 , 30 , 31 ] or methanol [ 17 , 20 , 23 , 32 ], for deproteinization prior to separation by centrifugation. In addition, metabolite extraction may also include the use of a mixture of solvents, such as acetone and methanol or acetonitrile and methanol [ 22 , 33 , 34 ]. Meanwhile, two studies that performed a GC-MS-based analysis used polydimethylsiloxane/divinylbenzene for the solid-phase microextraction of metabolites [ 35 , 36 ].…”

“…In a different study, Nizioł et al performed a serum metabolomics analysis of 100 BCa patients and 100 healthy controls. As a result, the molecular signatures revealed 27 metabolites that discriminated between BCa patients and healthy controls, 23 metabolites that differentiated between LG- and HG-BCa from controls and 39 metabolites that could distinguish BCa in distinct stages [ 33 ]. These metabolites were essentially lipids and lipid-like molecules.…”

“…↑: increase; ↓: decrease. BCa: bladder cancer; HG: high grade; LG: low grade; NMIBC: non-muscle-invasive bladder cancer; MIBC: muscle-invasive bladder cancer [ 17 , 18 , 19 , 23 , 30 , 33 , 34 , 38 , 39 , 46 , 54 , 60 , 72 , 106 ].…”

Untargeted urinary metabolomics for bladder cancer biomarker screening with ultrahigh-resolution mass spectrometry

“…Metabolite separation was achieved with a gradient of mobile phases using a Waters UPLC column ACQUITY BEH (C18 silica, 1.7 μm particles, 50 × 2.1 mm) with a compatible column guard was used for all analyses. Further details are described in our previous publication 35 and supplementary information 1 (section S1 ).…”

Untargeted urinary metabolomics for bladder cancer biomarker screening with ultrahigh-resolution mass spectrometry

“…Metabolite separation was achieved with a gradient of mobile phases using a Waters UPLC column ACQUITY BEH (C18 silica, 1.7 µm particles, 50 x 2.1 mm) with a compatible column guard was used for all analyses. Further details are described in our previous publication 35 and supplementary information 1 (section S1).…”