Non-invasive Detection of Breast Cancer by Breath Analysis (BreathBC)

Non-invasive Detection of Breast Cancer by Mass-spectrometry-based Profiling of Exhaled Breath

The goal of this prospective observational study is to compare the exhaled breath profiles of patient with invasive breast cancer and control subjects in the perspective of a diagnostic application in oncology.

The main question it aims to answer is whether the exhaled breath profile of breast cancer patients can be discriminate from the exhaled breath of control subjects.

Participants to the study are recruited in a first discovery series including about 500 subjects and in a following independent validation series including more than 400 subjects. Breath samples are collected in a dedicated room using sampling bags and following anti-Covid-19 measures. Only authorized personnel could access the sampling room which was equipped with a HEPA Filter air purification system. Sampling bags are made of Nalophan® tubular foil and 2 taps, one in which the study participant inflated with their breath and sealed after use, while the second is used to introduce breath samples into the analyzer. Bags are sterilized with the Sterrad™ system with H2O2 vapors before use. To contain the potentially infectious particulate included in the participant exhaled breath, an electrostatic filter is mounted on the tap of the bag used to inflate it with breath. All participants are supplied with a leaflet explaining aims and technical/clinical information of the study. At each breath sampling, study participants are asked not to smoke, eat, drink (except for water), brush their teeth or use lipstick for at least 2 h before breath collection to minimize the presence of contaminant molecules in exhaled breath.

Breath profiling is performed by mass spectrometry analysis using an untargeted approach. Breath samples are profiled by Secondary Electrospray ionization - High resolution Mass Spectrometry (SESI-HR-MS) using a high resolution instrumentation (LTQ Orbitrap Elite,Thermo Fisher Scientific) equipped with a SuperSESI ionization source (Fossiliontech).

Data analysis is based on our previous work (Martínez-Lozano el al., 2015 J Breath Res 9:031001) and will be implemented with machine learning and other Artificial Intelligence (AI) methods.

Once all the breath profiles will be collected and analysed, researchers will compare data from patient with breast cancer and data from tumor-free subjects to identify methods and patterns able to specifically identify the pathological breath samples and discriminate them from the control breath samples.

Study Overview

• Status: Active, not recruiting
• Conditions: Breast Neoplasm Female
• Intervention / Treatment: Diagnostic test: Sampling of exhaled breath

• Study Type: Observational
• Enrollment (Actual): 1000

Contacts and Locations

Study Locations

• Italy
  • Milan, Italy
    • Fondazione IRCCS Istituto Nazionale Tumori

Participation Criteria

Eligibility Criteria

• Ages Eligible for Study: Adult; Older Adult
• Accepts Healthy Volunteers: Yes

• Sampling Method: Non-Probability Sample

Study Population

Patients are recruited at Breast Surgery unit of Fondazione IRCCS Istituto Nazionale Tumori in Milan.

Controls are recruited during breast screening at Breast Imaging Unit of Fondazione IRCCS Istituto Nazionale Tumori in Milan.

Description

Inclusion Criteria:

1. be >= 18 years
2. Patients

2.a patients at early stage of breast cancer 2.b exahled breath collected and analysed at the baseline before any surgical or pharmacological treatment

3. Controls 3.1 subjects with negative breast imaging (BI-RADS= 1,2) 3.2 exhaled breath collected and analysed on the day of breast screening

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Exclusion Criteria:

1. any cancer in the 5 years before the inclusion
2. major surgery within 4 weeks before the inclusion
3. previous or concomitant long-term treatment with systemic steroids or immunosuppressive/immunomodulating drugs
4. concomitant acute infection, viral hepatitis, HIV infection, COVID19 infection or other serious concomitant medical disorders

5. organ allograft

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Study Plan

How is the study designed?

Number of groups / cohorts

2

Cohorts and Interventions

Group / Cohort	Intervention / Treatment
Tumor; Women carrying a breast cancer	Diagnostic test: Sampling of exhaled breath; Exhaled breath of all participants is collected following our standardized procedure
Control; Women with negative breast imaging	Diagnostic test: Sampling of exhaled breath; Exhaled breath of all participants is collected following our standardized procedure

What is the study measuring?

Primary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Classification of the exhaled breath of breast cancer patients and controls as assessed by the comparison of the global breath profile of volatile organic compounds (VOCs) measured using Mass Spectrometry analysis.	Breath profile of each study participant was determined by Mass Spectrometry analysis (Secondary Electrospray ionization - High resolution Mass Spectrometry, SESI-HR-MS). The risk of cancer is identified in the validation cohort of the present study and is based on the diagnostic model developed in the discovery phase of the same study. The predictive result is compared, in the case of breast cancer patients, with the histopathological proven diagnosis and, in the case of control subjects, with the imaging proven diagnosis.	Breath collection and MS analysis were assessed at the baseline.

Collaborators and Investigators

• Sponsor: Fondazione IRCCS Istituto Nazionale dei Tumori, Milano
• Investigators: Principal Investigator: Rosaria Orlandi, Fondazione IRCCS Istituto Nazionale Tumori

Publications and helpful links

General Publications

• Martinez-Lozano Sinues P, Landoni E, Miceli R, Dibari VF, Dugo M, Agresti R, Tagliabue E, Cristoni S, Orlandi R. Secondary electrospray ionization-mass spectrometry and a novel statistical bioinformatic approach identifies a cancer-related profile in exhaled breath of breast cancer patients: a pilot study. J Breath Res. 2015 Sep 21;9(3):031001. doi: 10.1088/1752-7155/9/3/031001.

Study record dates

Study Major Dates

• Study Start (Actual): June 3, 2021
• Primary Completion (Actual): March 29, 2024
• Study Completion (Estimated): December 31, 2027

Study Registration Dates

• First Submitted: June 4, 2024
• First Submitted That Met QC Criteria: July 8, 2024
• First Posted (Actual): July 12, 2024

Study Record Updates

• Last Update Posted (Actual): May 15, 2026
• Last Update Submitted That Met QC Criteria: May 12, 2026
• Last Verified: December 1, 2025

More Information

Terms related to this study

• Keywords: breath analysis; non invasive detection
• Additional Relevant MeSH Terms: Skin Diseases; Neoplasms; Neoplasms by Site; Breast Diseases; Breast Neoplasms
• Other Study ID Numbers: IstitutoTumori-BreathBC

Plan for Individual participant data (IPD)

• Plan to Share Individual Participant Data (IPD)?: UNDECIDED

Drug and device information, study documents

• Studies a U.S. FDA-regulated drug product: No
• Studies a U.S. FDA-regulated device product: No