Discovery and validation of potential urinary biomarkers for bladder cancer diagnosis using a pseudotargeted GC-MS metabolomics method

Abstract

Bladder cancer (BC) is the second most prevalent malignancy in the urinary system and is associated with significant mortality; thus, there is an urgent need for novel noninvasive diagnostic biomarkers. A urinary pseudotargeted method based on gas chromatography–mass spectrometry was developed and validated for a BC metabolomics study. The method exhibited good repeatability, intraday and interday precision, linearity and metabolome coverage. A total of 76 differential metabolites were defined in the discovery sample set, 58 of which were verified using an independent validation urine set. The verified differential metabolites revealed that energy metabolism, anabolic metabolism and cell redox states were disordered in BC. Based on a binary logistic regression analysis, a four-biomarker panel was defined for the diagnosis of BC. The area under the receiving operator characteristic curve was 0.885 with 88.0% sensitivity and 85.7% specificity in the discovery set and 0.804 with 78.0% sensitivity and 70.3% specificity in the validation set. The combinatorial biomarker panel was also useful for the early diagnosis of BC. This approach can be used to discriminate non-muscle invasive and low-grade BCs from healthy controls with satisfactory sensitivity and specificity. The results show that the developed urinary metabolomics method can be employed to effectively screen noninvasive biomarkers.

GC–MS analysis of organic acids in rat urine: A protocol of direct ultrasound‐assisted derivatization

“…Gas chromatography mass spectrometry (GC–MS) is a platform of choice for qualitative and quantitative analysis of OAs in laboratory settings (Bouatra et al, ), because of its advantages including high sensitivity, peak resolution and reproducibility. This technology has been applied to determine changes in global metabolite profiling in urine from patients with prostate (Lima et al, ), bladder (Zhou et al, ) and renal cancers (Monteiro et al, ). Several important observations regarding altered levels of OAs have recently been obtained from GC–MS.…”

5-Hydroxyhexanoic Acid Predicts Early Renal Functional Decline in Type 2 Diabetes Patients with Microalbuminuria

“…They found that the level of 5-hydroxyhexanoic acid was significantly increased in patients with BD [40]. Similarly, another two studies based on urinary metabolite analysis reported that 5-hydroxyhexanoic acid was identified as a “good” classifier for post-stroke depression, and levels of 5-hydroxyhexanoic acid were significantly decreased in the bladder cancer group [41, 42]. In an animal study, the concentration of 5-hydroxyhexanoic acid was significantly decreased in Ginseng polysaccharide-treated diabetic rats.…”

Liquid Biopsy Biomarkers in Bladder Cancer: A Current Need for Patient Diagnosis and Monitoring

“…The use of metabolomic analysis in BC has been primarily focused on the distinction between normal-appearing urothelium and BC. Zhou et al found a urinary four-biomarker panel (5-hydroxyvaleric acid, cholesterol, 3-phosphoglyceric acid and glycolic acid) including important metabolic characteristics (e.g., organic acid metabolism, steroid hormone biosynthesis, glycolysis and glyoxylate metabolism) and defined this panel as a combinatorial biomarker for the differentiation between BC patients and healthy controls (AUC = 0.804 with 78.0% sensitivity and 70.3% specificity in the validation set) [ 219 , 220 ]. Besides, Huang and colleagues reported the elevation of component I and decrease of carnitine C9:1 in BC urine samples, compared to healthy controls, as a promising biomarker panel for the identification of BC patients (92.6% sensitivity and 96.9% specificity; AUC = 0.963) [ 221 ].…”