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. 2021 Mar 8;11(3):466.
doi: 10.3390/diagnostics11030466.

Detection and Investigation of Extracellular Vesicles in Serum and Urine Supernatant of Prostate Cancer Patients

Samanta Salvi  1 , Erika Bandini  1 , Silvia Carloni  1 , Valentina Casadio  1 , Michela Battistelli  2 , Sara Salucci  2 , Ilaria Erani  1 , Emanuela Scarpi  3 , Roberta Gunelli  4 , Giacomo Cicchetti  5 , Michele Guescini  2 , Massimiliano Bonafè  6 , Francesco Fabbri  1
Affiliations

Detection and Investigation of Extracellular Vesicles in Serum and Urine Supernatant of Prostate Cancer Patients

Samanta Salvi et al. Diagnostics (Basel). .

Abstract

Prostate Cancer (PCa) is one of the most frequently identified urological cancers. PCa patients are often over-diagnosed due to still not highly specific diagnostic methods. The need for more accurate diagnostic tools to prevent overestimated diagnosis and unnecessary treatment of patients with non-malignant conditions is clear, and new markers and methods are strongly desirable. Extracellular vesicles (EVs) hold great promises as liquid biopsy-based markers. Despite the biological and technical issues present in their detection and study, these particles can be found highly abundantly in the biofluid and encompass a wealth of macromolecules that have been reported to be related to many physiological and pathological processes, including cancer onset, metastasis spreading, and treatment resistance. The present study aims to perform a technical feasibility study to develop a new workflow for investigating EVs from several biological sources. Serum and urinary supernatant EVs of PCa, benign prostatic hyperplasia (BPH) patients, and healthy donors were isolated and investigated by a fast, easily performable, and cost-effective cytofluorimetric approach for a multiplex detection of 37 EV-antigens. We also observed significant alterations in serum and urinary supernatant EVs potentially related to BPH and PCa, suggesting a potential clinical application of this workflow.

Keywords: MACSPlex Exosome kit; extracellular vesicles; prostate cancer; serum; urine.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Representative distribution plot of serum (a) and urinary supernatant (b) EVs and corresponding concentration by Nanosight Instrument in the three groups.
Figure 2
Figure 2
Negative staining of serum and urinary supernatant EVs from the same matched samples. TEM observations showed numerous EVs between 20 and 100 nm. Bar scale = 100 nm.
Figure 3
Figure 3
Range to the min and max of MFI for each EVs marker from serum (a) and urinary supernatant (b). MFI of healthy donors in blue; BPH patients in green; PCa patients in red.
Figure 4
Figure 4
Range of the MFI of the significant markers found in serum EVs (a) and urinary EVs (b). Healthy donors in blue; BPH patients in green; PCa patients in red. * p < 0.05; ** p < 0.0001.
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