A Pilot Study on the Potential of RNA-Associated to Urinary Vesicles as a Suitable Non-Invasive Source for Diagnostic Purposes in Bladder Cancer

Abstract

Bladder cancer is one of the most common cancers and, together with prostate carcinoma, accounts for the majority of the malignancies of the genitourinary tract. Since prognosis ameliorates with early detection, it will be beneficial to have a repertoire of diagnostic markers that could complement the current diagnosis protocols. Recently, cell-secreted extracellular vesicles have received great interest as a source of low invasive disease biomarkers because they are found in many body fluids, including urine. The current work describes a pilot study to generate an array-based catalogue of mRNA associated to urinary vesicles, and also a comparison with samples obtained from bladder cancer patients. After an analysis of presence/absence of transcripts in bladder cancer EVs, a list of genes was selected for further validation using PCR technique. We found four genes differentially expressed in cancer samples. LASS2 and GALNT1 were present in cancer patients, while ARHGEF39 and FOXO3 were found only in non-cancer urinary vesicles. Previous studies have pointed to the involvement of those genes in tumour progression and metastasis.

Time-dependent release of extracellular vesicle subpopulations in tumor CABA I cells

“…Due to their role in pathological processes, EVs have been widely studied in the past few years (21)(22)(23)(24). Moreover, there is an increasing expectation in their potential use as clinical targets and as diagnostic and prognostic biomarkers since vesicles occur in bloodstream and other biological fluids (e.g., urine, semen, amniotic fluid, saliva, synovial and bronchoalveolar fluid, breast milk, spinal fluid, ascites, malignant and pleural effusion), particularly the ones that are exposed to primary tumors (2,19,(25)(26)(27)(28).…”

Extracellular Micro-RNAs in Health and Disease: Basic Science, Biogenesis and Release

“…This means that, in case of crowd and overflow, the cell endosomal pathway is unable to cope with the abrupt increase in the need for degradation through the endocytic pathway, hence activation of an alternative pathway to eliminate and secrete these contents outside the cell. Other observations have been supporting the hypothesis of active sorting; high EVs miR-NA level than that contained in the parental cells which may be explained by the observation that certain miRNAs can be lost during extraction from samples [66]. The differences in the blood collection protocols willimpact the comparison between extracellular versus intracellular miRNA profiles expressed in different cell types [67]- [71].…”

Prostasomes as a source of diagnostic biomarkers for prostate cancer

“…The presence of prostasomes in EV fractions isolated from urine was confirmed by detection of prostate-specific proteins, including prostate-specific membrane antigen (PSMA), prostatic acid phosphatase, and prostate transglutaminase (67,(70)(71)(72)(73). One advantage of collecting EVs from urine, as compared with blood, is that such isolates are more enriched in prostasomes relative to other constituents, although tissues within the urogenital system other than the prostate, including the kidney (74) and bladder (75), also contribute EVs to urine. Moreover, urine also contains intact PCa cells and PCa cell-derived apoptotic bodies (74).…”