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LC–MS metabolomics of urine reveals distinct profiles for non-muscle-invasive and muscle-invasive bladder cancer

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Abstract

Purpose

Bladder cancer (BC) is among the most frequent malignancies worldwide. Novel non-invasive markers are needed to diagnose and stage BC with more accuracy than invasive procedures like cystoscopy. To date, no study has identified urine metabolites characteristic of all BC stages. To discover novel urine metabolomic profiles to diagnose and stage non-muscle-invasive (NMIBC) and muscle-invasive (MIBC) patients using mass spectrometry-based metabolomics.

Methods

We prospectively recruited 198 BC patients and 98 age- and sex-matched healthy volunteers without evidence of renal or bladder condition confirmed by ultrasound, from whom we collected a first morning urine sample (before surgery in patients). In a discovery stage, an untargeted metabolomic analysis was conducted in urine samples of a selection of 64 BC patients (19 TaG1, 11 TaG3, 20 T1G3, 12 T2G3, 1 T2G2, 1 T3G3) and 20 controls to identify dysregulated metabolites. Next, after exhaustive multivariate analysis, confirmed dysregulated metabolites were validated in an independent cohort of 134 BC patients (19 TaG1, 62 TaG2, 9 TaG3, 15 T1G2, 16 T1G3, 4 T2G2, 9 T2G3) and 78 controls.

Results

We validated p-cresol glucuronide as potential diagnostic biomarker for BC patients compared to controls (AUC = 0.79). For NMIBC, p-cresol glucuronide was valuable as staging biomarker (AUC = 0.803). And among NMIBCs, p-coumaric acid may be a potential specific staging biomarker for the TaG1 NMIBC; however, future validation experiments should be conducted once the precise version of the standard is commercially available. Remarkably, for MIBC we validated spermine as potential specific staging biomarker (AUC = 0.882).

Conclusion

Ours is the first metabolomics study conducted in urine of a thoroughly characterized cohort comprising all stages of NMIBC, MIBC and healthy controls in which we identified non-invasive diagnostic and staging biomarkers. These may improve BC management, thus reducing the use of current harmful diagnostic techniques.

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Acknowledgements

The MS metabolomics data have been deposited to the Metabolomics Workbench [40] public repository (www.metabolomicsworkbench.org/) with the data set identifier ST001662. We would like to thank Dr. Francisco España for his extraordinary guidance and supervision.

Funding

This research was supported by research grants from Instituto de Salud Carlos III (PI17/00495, PI20/00075, FI21/00171), FEDER una manera de hacer Europa, Generalitat Valenciana (ACIF/2017/138) and Sociedad Española de Trombosis y Hemostasia. All these are nonprofit organizations, and therefore are not involved in experimental/clinical work, data analyses, and preparation of the manuscript.

Author information

Authors and Affiliations

  1. Haemostasis, Thrombosis, Arteriosclerosis and Vascular Biology Research Group, Medical Research Institute Hospital La Fe, Av. Fernando Abril Martorell 106, 46026, Valencia, Spain

    Julia Oto, Álvaro Fernández-Pardo, Emma Plana, Fernando Cana, Raquel Herranz & Pilar Medina

  2. Analytical Unit Platform, Medical Research Institute Hospital La Fe, Valencia, Spain

    Marta Roca

  3. Angiology and Vascular Surgery Service, La Fe University and Polytechnic Hospital, Av. Fernando Abril Martorell 106, 46026, Valencia, Spain

    Emma Plana

  4. Urology Service, La Fe University and Polytechnic Hospital, Av. Fernando Abril Martorell 106, 46026, Valencia, Spain

    Javier Pérez-Ardavín, César David Vera-Donoso & Manuel Martínez-Sarmiento

  5. IIS La Fe-Hospital Universitario y Politécnico La Fe, Torre A, 5ª Planta, Lab. 5-09, Av. Fernando Abril Martorell 106, 46026, Valencia, Spain

    Pilar Medina

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  1. Julia Oto
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Contributions

JO: protocol development, data collection, data analysis and manuscript writing. ÁFP: data analysis. MR: protocol development, data analysis, and manuscript writing. EP: manuscript editing and supervision. FC: protocol development. RH: data analysis. JPA: resources. CDVD: resources and manuscript editing. MMS: resources and manuscript editing. PM: project development, management, data analysis, manuscript writing and funding acquisition. All authors read and approved the final manuscript.

Corresponding author

Correspondence to Pilar Medina.

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The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

Ethical approval and consent to participate

Informed consent was obtained from all participants according to protocols approved by the ethics review board at La Fe University and Polytechnic Hospital. The study was performed according to the declaration of Helsinki, as amended in Edinburgh in 2000.

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Oto, J., Fernández-Pardo, Á., Roca, M. et al. LC–MS metabolomics of urine reveals distinct profiles for non-muscle-invasive and muscle-invasive bladder cancer. World J Urol 40, 2387–2398 (2022). https://doi.org/10.1007/s00345-022-04136-7

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