Fast quantification of extracellular vesicles levels in early breast cancer patients by Single Molecule Detection Array (SiMoA)
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...
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| Published in | Breast cancer research and treatment Vol. 192; no. 1; pp. 65 - 74 |
|---|---|
| Main Authors | , , , , , , , , , , |
| Format | Journal Article |
| Language | English |
| Published |
New York
Springer US
01.02.2022
Springer Springer Nature B.V |
| Subjects | |
| Online Access | Get full text |
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| Abstract | 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/µl
vs
613.0 ng/µl,
p
< 0.0001). ROC curve was used to define the optimal cut-off level of the test at 1034.5 ng/µl 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. |
|---|---|
| AbstractList | 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).PURPOSEPreliminary 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).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.METHODSBy 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.The measured levels of EVs resulted significantly higher in BC patients (median values 1179.1 ng/µl vs 613.0 ng/µl, p < 0.0001). ROC curve was used to define the optimal cut-off level of the test at 1034.5 ng/µl 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.RESULTSThe measured levels of EVs resulted significantly higher in BC patients (median values 1179.1 ng/µl vs 613.0 ng/µl, p < 0.0001). ROC curve was used to define the optimal cut-off level of the test at 1034.5 ng/µl 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.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.CONCLUSIONSiMoA 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. 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). 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. The measured levels of EVs resulted significantly higher in BC patients (median values 1179.1 ng/[micro]l vs 613.0 ng/[micro]l, p < 0.0001). ROC curve was used to define the optimal cut-off level of the test at 1034.5 ng/[micro]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. 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. PurposePreliminary 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).MethodsBy 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.ResultsThe measured levels of EVs resulted significantly higher in BC patients (median values 1179.1 ng/µl vs 613.0 ng/µl, p < 0.0001). ROC curve was used to define the optimal cut-off level of the test at 1034.5 ng/µl 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.ConclusionSiMoA 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. 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/µl vs 613.0 ng/µl, p < 0.0001). ROC curve was used to define the optimal cut-off level of the test at 1034.5 ng/µl 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. 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). 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. The measured levels of EVs resulted significantly higher in BC patients (median values 1179.1 ng/µl vs 613.0 ng/µl, p < 0.0001). ROC curve was used to define the optimal cut-off level of the test at 1034.5 ng/µl 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. 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. 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/[micro]l vs 613.0 ng/[micro]l, p < 0.0001). ROC curve was used to define the optimal cut-off level of the test at 1034.5 ng/[micro]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. |
| Audience | Academic |
| Author | Sproviero, Daisy Albasini, Sara Piccotti, Francesca Mollica, Ludovica Morasso, Carlo Corsi, Fabio Sorrentino, Luca Ricciardi, Alessandra Sottotetti, Federico Cereda, Cristina Truffi, Marta |
| Author_xml | – sequence: 1 givenname: Carlo surname: Morasso fullname: Morasso, Carlo organization: Laboratory of Nanomedicine, Istituti Clinici Scientifici Maugeri IRCCS – sequence: 2 givenname: Alessandra surname: Ricciardi fullname: Ricciardi, Alessandra organization: Laboratory of Nanomedicine, Istituti Clinici Scientifici Maugeri IRCCS – sequence: 3 givenname: Daisy surname: Sproviero fullname: Sproviero, Daisy organization: Genomic and Post-Genomic Center, IRCCS Mondino Foundation – sequence: 4 givenname: Marta surname: Truffi fullname: Truffi, Marta organization: Laboratory of Nanomedicine, Istituti Clinici Scientifici Maugeri IRCCS – sequence: 5 givenname: Sara surname: Albasini fullname: Albasini, Sara organization: Breast Unit, Istituti Clinici Scientifici Maugeri IRCCS – sequence: 6 givenname: Francesca surname: Piccotti fullname: Piccotti, Francesca organization: Laboratory of Nanomedicine, Istituti Clinici Scientifici Maugeri IRCCS – sequence: 7 givenname: Federico surname: Sottotetti fullname: Sottotetti, Federico organization: Medical Oncology Unit, Istituti Clinici Scientifici Maugeri IRCCS – sequence: 8 givenname: Ludovica surname: Mollica fullname: Mollica, Ludovica organization: Medical Oncology Unit, Istituti Clinici Scientifici Maugeri IRCCS – sequence: 9 givenname: Cristina surname: Cereda fullname: Cereda, Cristina organization: Genomic and Post-Genomic Center, IRCCS Mondino Foundation – sequence: 10 givenname: Luca surname: Sorrentino fullname: Sorrentino, Luca organization: Colorectal Surgery Unit, Fondazione IRCCS Istituto Nazionale dei Tumori di Milano – sequence: 11 givenname: Fabio orcidid: 0000-0002-6469-4086 surname: Corsi fullname: Corsi, Fabio email: fabio.corsi@unimi.it, fabio.corsi@icsmaugeri.it organization: Breast Unit, Istituti Clinici Scientifici Maugeri IRCCS, Dipartimento di Scienze Biomediche e Cliniche “L. Sacco”, Università di Milano |
| BackLink | https://www.ncbi.nlm.nih.gov/pubmed/34935096$$D View this record in MEDLINE/PubMed |
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| Keywords | Extracellular vesicles Immunoassay Biomarker Breast cancer SiMoA Tetraspanins |
| Language | English |
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| PublicationDate | 2022-02-01 |
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| PublicationDate_xml | – month: 02 year: 2022 text: 2022-02-01 day: 01 |
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| PublicationTitle | Breast cancer research and treatment |
| PublicationTitleAbbrev | Breast Cancer Res Treat |
| PublicationTitleAlternate | Breast Cancer Res Treat |
| PublicationYear | 2022 |
| Publisher | Springer US Springer Springer Nature B.V |
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Preliminary reports suggest that extracellular vesicles (EVs) might be a promising biomarker for breast cancer (BC). However, the quantification of... Preliminary reports suggest that extracellular vesicles (EVs) might be a promising biomarker for breast cancer (BC). However, the quantification of plasmatic... Purpose Preliminary reports suggest that extracellular vesicles (EVs) might be a promising biomarker for breast cancer (BC). However, the quantification of... PurposePreliminary reports suggest that extracellular vesicles (EVs) might be a promising biomarker for breast cancer (BC). However, the quantification of... |
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| SubjectTerms | Analysis Biomarkers Breast cancer Breast Neoplasms - diagnosis Cancer patients Cancer research Cancer surgery Care and treatment CD63 antigen CD9 antigen Extracellular Vesicles Female Humans Medical research Medicine Medicine & Public Health Medicine, Experimental Oncology Patients Plasma Preclinical Study ROC Curve Surgery |
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| Title | Fast quantification of extracellular vesicles levels in early breast cancer patients by Single Molecule Detection Array (SiMoA) |
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