Investigating the role of microbiota in bladder cancer

Background and Aims Once thought to be sterile, the urinary tract is naturally colonized with a specific microbial community (urobiome). Bladder cancer is currently the 10th most diagnosed cancer in the world. Understanding which microbes in the urobiome promote a healthy state and which can be indicators of a pathological state is crucial for designing intervention strategies that reestablish/promote the physiological conditions and/or for identifying more sensitive and specific biomarkers for early detection. Methods Following approval by ethic committee, thirteen urine samples of female patients were obtained during transurethral resection of bladder (TURB) tumor. Ten samples from case controls (stress incontinence) were also collected. Standard culture (Std) and Expanded Quantitative Urine Culture (EQUC) were set in both aerobic and anaerobic conditions. For metagenomics studies bacterial DNA was extracted using DNeasy Blood & Tissue Kit (Qiagen). Results We established a successful workflow i) to ensure appropriate long storage of the specimens ii) to effectively culture bacteria in aerobic and anaerobic conditions iii) to extract bacterial DNA for metagenomics analysis within few hours from collection and storage at +4°C. During Std and EQUC, plates were inspected over a period of 5 days and bacterial colonies isolated and stored for future MALDI-TOF analysis. Conclusions Urinary bacteria were cultured and isolated, along with the bacterial DNA for the metagenome analysis and currently under amplicon sequencing. Understanding the dysbiosis in the urobiome is an active field with ongoing efforts to improve our understanding of disease development and identification of important biomarkers at an early stage/onset of the disease.