Noninvasive Urinary Metabolomic Profiling Identifies Diagnostic and Prognostic Markers in Lung Cancer

Abstract

Lung cancer remains the most common cause of cancer deaths worldwide, yet there is currently a lack of diagnostic noninvasive biomarkers that could guide treatment decisions. Small molecules (<1500 Da) were measured in urine collected from 469 lung cancer patients and 536 population controls using unbiased liquid chromatography-mass spectrometry. Clinical putative diagnostic and prognostic biomarkers were validated by quantitation and normalized to creatinine levels at two different time points and further validated in an independent sample set, which comprises 80 cases and 78 population controls, with similar demographic and clinical characteristics when compared to the training set. Creatine riboside (IUPAC name: 2-{2-[(2R,3R,4S,5R)-3,4-dihydroxy-5-(hydroxymethyl)-oxolan-2-yl]-1-methylcarbamimidamido}acetic acid), a novel molecule identified in this study, and N-acetylneuraminic acid (NANA), were each significantly (P <0.00001) elevated in non–small cell lung cancer (NSCLC) and associated with worse prognosis (hazard ratio (HR) =1.81 [P =0.0002], and 1.54 [P =0.025], respectively). Creatine riboside was the strongest classifier of lung cancer status in all and stage I–II cases, important for early detection, and also associated with worse prognosis in stage I–II lung cancer (HR =1.71, P =0.048). All measurements were highly reproducible with intraclass correlation coefficients ranging from 0.82 – 0.99. Both metabolites were significantly (P <0.03) enriched in tumor tissue compared to adjacent non-tumor tissue (N =48), thus revealing their direct association with tumor metabolism. Creatine riboside and NANA may be robust urinary clinical metabolomic markers that are elevated in tumor tissue and associated with early lung cancer diagnosis and worse prognosis.

&lt;italic&gt;Mup&lt;/italic&gt;-knockout mice generated through CRISPR/Cas9-mediated deletion for use in urinary protein analysis

“…Metabolomics analyses have revealed more than 2000 metabolites in mouse urine [20]. Small volatile organic compounds have been studied as potential biomarkers of lung cancer in a recent study [21]. Urinary miRNA molecules have also been evaluated as biomarkers [22].…”

Comparison of standardization approaches applied to metabolomics data

“…The positive ionization mode (ESI+) of MTBLS28 provided LC/MS based metabolomics profiles of 1,005 samples (469 lung cancer patients and 536 healthy individuals) [8]. The samples were further divided into training group (900 samples including 400 lung cancer patients and 500 healthy individuals) and independent testing group (105 samples) by random sampling.…”

Nuclear magnetic resonance spectroscopy as a new approach for improvement of early diagnosis and risk stratification of prostate cancer

“…NMR technology has evolved into a precise, generally applicable and, at least in high throughput, cost-efficient method (Sokolenko et al, 2013;Mathé et al, 2014). The idea of using a complex mixture of small urine compounds to assay for PCa is the basis of metabolomics in its holistic approach.…”