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Proteomic analysis of urinary and tissue-exudative extracellular vesicles to discover novel bladder cancer biomarkers. | LitMetric

AI Article Synopsis

  • This study explores the use of proteomic analysis on urinary extracellular vesicles (EVs) to identify potential bladder cancer (BCa) biomarkers while addressing the challenge of distinguishing BCa-specific EVs from those derived from normal sources in urine.* -
  • Researchers combined urinary EV analysis with tissue-exudative EVs (Te-EVs) isolated from bladder cancer tissues, resulting in identification of nearly 2,000 proteins in urinary EVs and 1,500 in Te-EVs, with a significant overlap between the two.* -
  • From the analysis, 22 membrane proteins were selected for further validation in a larger sample, leading to the identification of 6 specific proteins that were significantly upregulated in the urine of

Article Abstract

Proteomic analysis of urinary extracellular vesicles (EVs) is a powerful approach to discover potential bladder cancer (BCa) biomarkers, however urine contains numerous EVs derived from the kidney and normal urothelial epithelium, which can obfuscate information related to BCa cell-derived EVs. In this study, we combined proteomic analysis of urinary EVs and tissue-exudative EVs (Te-EVs), which were isolated from culture medium of freshly resected viable BCa tissues. Urinary EVs were isolated from urine samples of 11 individuals (7 BCa patients and 4 healthy individuals), and Te-EVs were isolated from 7 BCa tissues. We performed tandem mass tag (TMT)-labeling liquid chromatography (LC-MS/MS) analysis for both urinary EVs and Te-EVs and identified 1960 proteins in urinary EVs and 1538 proteins in Te-EVs. Most of the proteins identified in Te-EVs were also present in urinary EVs (82.4%), with 55 of these proteins showing upregulated levels in the urine of BCa patients (fold change > 2.0; P < .1). Among them, we selected 22 membrane proteins as BCa biomarker candidates for validation using selected reaction monitoring/multiple reaction monitoring (SRM/MRM) analysis on urine samples from 70 individuals (40 BCa patients and 30 healthy individuals). Six urinary EV proteins (heat-shock protein 90, syndecan-1, myristoylated alanine-rich C-kinase substrate (MARCKS), MARCKS-related protein, tight junction protein ZO-2, and complement decay-accelerating factor) were quantified using SRM/MRM analysis and validated as significantly upregulated in BCa patients (P < .05). In conclusion, the novel strategy that combined proteomic analysis of urinary EVs and Te-EVs enabled selective detection of urinary BCa biomarkers.

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Source
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8088963PMC
http://dx.doi.org/10.1111/cas.14881DOI Listing

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