Characterization of Extracellular Vesicles in Breast Cancer Patients

Characterization of Extracellular Vesicles in Breast Cancer Patients' Plasma by Single Molecule Detection Array (SiMoA) Digital ELISA

Extracellular vesicles (EVs) are lipid bilayer-delimited particles, naturally released from cells and mediators of intercellular cross-talk. In breast cancer (BC), EVs seem to be involved in the tumor microenvironment's shaping, in cancer cells invasion and in the set-up of metastasis.

Clinical studies have provided initial evidence that these vesicles may have a prognostic and predictive value in breast cancer. Considering their ubiquitous presence in body fluids and their minimally invasive assessment through blood sampling, EVs could have a potential as liquid biopsy-derived biomarkers. Their quantification though is a complex task requiring complicated and time-consuming pre-analytical procedures of EVs isolation.

This protocol want to develop a new method for the detection of tumor-derived-EVs associated proteins, based on the use of Single Molecule Array (SiMoA), a digital ELISA technology able to detect and quantify extremely low concentrations of target proteins or particles.

The aim of this study is to evaluate how this new technology can allow the quantification of EVs plasma levels in patients affected by BC, providing useful diagnostic and prognostic information.

Study Overview

• Status: Recruiting
• Conditions: Breast Cancer
• Intervention / Treatment: Diagnostic test: Blood sample

Detailed Description

This is a prospective, observational, monocentric and no profit study. The study involves the analysis of plasma from patients with breast cancer in order to quantify and characterize tumor-derived EVs at specific disease stages.

The enrollment of consecutive patients affected by BC referring to an EUSOMA-accredited Breast Unit is planned.

The patients will be divided into three pre-planned groups, as follows:

Population 1: patients diagnosed with early breast cancer patients (stage I-III) with indication to curative surgery.

Population 2: a control group made of sex- and age-matched healthy volunteers, not affected by cancer or chronic diseases.

Population 3: patients with metastatic breast cancer diagnosis. For each patient a blood sample will be collected and plasma will be isolated. A new SiMoA assay based on the use of anti-CD63 and anti-CD9 antibodies, two well known protein markers of EVs, will be used to capture and quantify EVs directly from plasma without requiring any prior sample processing.

The study will be conducted following the International Conference on Harmonization [ICH] Good Clinical Practice [GCP] guidelines. The Ethical Committee of ICS Maugeri authorized the study as protocol 2490/2020.

• Study Type: Observational
• Enrollment (Estimated): 105

Contacts and Locations

• Study Contact: Name: Fabio Corsi, Professor; Phone Number: 0382592272; Email: fabio.corsi@icsmaugeri.it
• Study Contact Backup: Name: SARA PAOLA ALBASINI, MsC; Phone Number: 3497378405; Email: sara.albasini@icsmaugeri.it

Study Locations

• Italy
  • Lombardy
    • Pavia, Lombardy, Italy, 27100
      • Recruiting
      • Istituti Clinici Scientifici Maugeri SpA
      • Contact: Fabio Corsi, Professor; Phone Number: 0382592272; Email: fabio.corsi@icsmaugeri.it
      • Contact: Sara Albasini, MsC; Phone Number: 3497378405; Email: sara.albasini@icsmaugeri.it

Participation Criteria

Eligibility Criteria

• Ages Eligible for Study: 18 years to 90 years (Adult, Older Adult)
• Accepts Healthy Volunteers: Yes

• Sampling Method: Non-Probability Sample

Study Population

Breast cancer patients

Description

Inclusion Criteria:

• Signing of a specific informed consent for participation to the study
• Female sex

Population 1:

• Diagnosis of early breast cancer;
• Indication for surgery after multidisciplinary discussion.

Population 2:

- Patients having a negative mammography or breast ultrasound within 12 months from the study enrollment.

Population 3:

• Diagnosis of metastatic breast cancer confirmed by cytological/histological examination or by imaging;
• Indication to chemotherapy.

Exclusion Criteria:

Population 1:

• Presence of distant metastases
• Synchronous presence of a different tumor
• Indication to neoadjuvant chemotherapy

Population 2:

• Diagnosis of breast cancer
• Synchronous presence of a different tumor or chronic disease

Population 3:

- Patients who are unfit for systemic chemotherapy treatment

Study Plan

How is the study designed?

Number of groups / cohorts

3

Cohorts and Interventions

Group / Cohort	Intervention / Treatment
Early Breast Cancer patients; Diagnosis of early breast cancer;; Indication for surgery after multidisciplinary discussion.	Diagnostic test: Blood sample; Analysis of plasma in order to quantify and characterize EVs
Metastatic Breast Cancer patients; First diagnosis of metastatic breast cancer confirmed by cytological/histological examination or by imaging;; Indication to chemotherapy.	Diagnostic test: Blood sample; Analysis of plasma in order to quantify and characterize EVs
Healthy patients; - Patients having a negative mammography or breast ultrasound within 12 months from the study enrollment.	Diagnostic test: Blood sample; Analysis of plasma in order to quantify and characterize EVs

What is the study measuring?

Primary Outcome Measures

Outcome Measure	Measure Description	Time Frame
EVs levels in early BC	Determination of plasma EVs levels in patients with EBC by SIMOA digital ELISA and evaluation of the difference of their concentration compared to healthy controls	36 months
EVs levels in metastatic BC	Determination of plasma EVs levels in patients with first diagnosis of metastatic BC by SIMOA digital ELISA and evaluation of the difference of their concentration compared to healthy controls	36 months

Secondary Outcome Measures

Outcome Measure	Measure Description	Time Frame
EVs levels after surgery	Determination of plasma EVs levels in patients with early BC one month after surgery and comparison with pre-surgery EVs levels in order to evaluate any qualitative of quantitative modifications in circulating EVs	36 months
EVs levels after chemotherapy treatment	Determination of plasma EVs levels in patients with first diagnosis of metastatic BC after 4 months of systemic therapy and comparison with pre-therapy EV's plasma levels	36 months

Collaborators and Investigators

• Sponsor: Istituti Clinici Scientifici Maugeri SpA

Study record dates

Study Major Dates

• Study Start (Actual): December 1, 2020
• Primary Completion (Actual): September 30, 2023
• Study Completion (Estimated): September 30, 2026

Study Registration Dates

• First Submitted: March 21, 2023
• First Submitted That Met QC Criteria: April 3, 2023
• First Posted (Actual): April 4, 2023

Study Record Updates

• Last Update Posted (Estimated): November 25, 2025
• Last Update Submitted That Met QC Criteria: November 20, 2025
• Last Verified: May 1, 2025

More Information

Terms related to this study

• Additional Relevant MeSH Terms: Neoplasms by Site; Neoplasms; Skin Diseases; Breast Diseases; Skin and Connective Tissue Diseases; Breast Neoplasms; Investigative Techniques; Specimen Handling; Clinical Laboratory Techniques; Diagnostic Techniques and Procedures; Diagnosis; Punctures; Surgical Procedures, Operative; Blood Specimen Collection
• Other Study ID Numbers: 2490

Plan for Individual participant data (IPD)

• Plan to Share Individual Participant Data (IPD)?: NO

Drug and device information, study documents

• Studies a U.S. FDA-regulated drug product: No
• Studies a U.S. FDA-regulated device product: No
• product manufactured in and exported from the U.S.: No