Journal
BREAST CANCER RESEARCH AND TREATMENT
Volume 192, Issue 1, Pages 65-74Publisher
SPRINGER
DOI: 10.1007/s10549-021-06474-3
Keywords
Breast cancer; Extracellular vesicles; Biomarker; SiMoA; Immunoassay; Tetraspanins
Categories
Funding
- Ricerca Corrente Istituti Clinici Scientifici Maugeri IRCCS scheme - Italian Ministry of Health
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The study developed a new SiMoA detection method for directly measuring EVs levels in plasma without prior processing. The research found significantly higher levels of EVs in BC patients, which decreased significantly after cancer surgery. There was no correlation between EVs concentration and clinical features of BC.
Purpose Preliminary reports suggest that extracellular vesicles (EVs) might be a promising biomarker for breast cancer (BC). However, the quantification of plasmatic levels of EVs is a complex task. To overcome these limitations, we developed a new, fast, and easy to use assay for the quantification of EVs directly in plasma based on the use of Single-Molecule Array (SiMoA). Methods By using SiMoA to identify CD9+/CD63+ EVs, we analyzed plasma samples of 181 subjects (95 BC and 86 healthy controls, HC). A calibration curve, made of a serial dilution of lyophilized standards from human plasma, was used in each run to ensure the obtainment of quantitative results from the assay. In a subgroup of patients, EVs concentrations were estimated in plasma before and after 30 days from cancer surgery. Additional information on the size of EVs were also acquired using a Nanosight system to obtain a clearer understanding of the mechanism underlying the releases of EVs associated with the presence of cancer. Results The measured levels of EVs resulted significantly higher in BC patients (median values 1179.1 ng/mu l vs 613.0 ng/mu l, p < 0.0001). ROC curve was used to define the optimal cut-off level of the test at 1034.5 ng/mu l with an AUC of 0.75 [95% CI 0.68-0.82]. EVs plasmatic concentrations significantly decreased after cancer surgery compared to baseline values (p = 0.014). No correlation was found between EVs concentration and clinical features of BC. Conclusion SiMoA assay allows plasmatic EVs levels detection directly without any prior processing. EVs levels are significantly higher in BC patients and significantly decreases after cancer surgery.
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