BIG Molecular Screening Feasibility Study
BIG Molecular Screening Feasibility Study
This pilot study examines the feasibility and turnaround time of performing and obtaining
 data from a few key molecular assays. These assays will be performed using different
 laboratories and technologies from core biopsies taken from patients diagnosed with invasive
 recurrent or metastatic breast cancer. All results will be uploaded, stored and assessed
 using the IT Molecular Screening Prototype Platform (MSPP). The MSPP will also be evaluated
 for ease of use to screen patients for participation in future molecularly defined clinical
 trials in breast cancer.
This study will accept patients with metastatic/recurrent breast cancer disease. The core
 biopsies must be taken from a metastatic lesion. To note, these patients can be biopsied at
 any phase of their metastatic disease (at diagnosis, at progression etc).

 -  The patient will sign a specific Informed Consent Form (ICF).

 -  The Investigator will access the MSPP, register the patient and enter basic patient
 clinical data necessary for the verification of the eligibility criteria.

 -  The patient will prospectively undergo invasive recurrent or metastatic lesions (1 site
 easily accessible, such as skin, lymph node or liver) core biopsies including the
 collection of tumor samples consisting of 2 Formalin Fixed Paraffin Embedded (FFPE)
 Tissues and 1 (2 recommended) fresh frozen samples embedded in Optimal Cutting
 Temperature (OCT) compound or stored in RNAlater. One whole blood sample (1x10mL) will
 also be collected.

 -  The Investigator will record the biological samples via the MSPP bio-tracking system

 -  The Investigator is responsible for the immediate dispatch of the samples to the
 designated central laboratories.

 -  The assays will be performed at the central laboratories. It should be noted that:

 -  Two FFPE samples will be sent to IEO, Milan, Italy. One FFPE sample will be stored.
 The second FFPE will be used to perform pathological tests. The tests include ER,
 HER2, Ki67 and PTEN status evaluation by immunohistochemistry (IHC) and FISH (for
 HER2 only); and of PIK3CA hot spot somatic mutations identification by Sanger DNA
 sequencing. Unstained sections (10x5µm) and extracted DNA, taken from the FFPE
 tissue core used for the testing, will be sent to IPG and Sanger, respectively, by
 the central laboratory.

 -  The assays performed at IPG and Sanger will consist of targeted breast cancer genes
 mutations identification by Ion Proton or HiSeq 2000 DNA sequencing respectively.
 To note, targeted genes screen will also be coupled with identification of other
 substitutions, short indels and copy number variants (CNVs).

 -  One fresh frozen sample embedded in OCT or stored in RNAlater will be sent to
 IJBordet, Brussels, Belgium, together with the blood sample, for Affymetrix gene
 expression profiling and for chromosomal and SNP-analysis using the Cytoscan
 platform (Affymetrix). The blood sample will be stored.

 -  The central laboratories will upload the processed data that is generated as a result of
 the central testing onto the MSPP.

 -  An alert, by e-mail, will be sent to the Investigator when the central results are
 available. The Investigator will log on to MSPP and obtain the results.

 -  The residual biological samples and derivatives will be stored in the BIG study
 Repository for 15 years or the maximum allowed by local regulations whichever is the
 shortest. If needed, remaining material will be used for future research as high
 throughput genetic analysis.

 It should be noted that the results obtained from the BIG Molecular Screening Feasibility
 Study will NOT be used for treatment decision-making. Patients should receive anti-cancer
 therapy as per the patient's treating physicians decision and in accordance with local
 institutional guidelines. There is no planned follow-up period. The trial will end after all
 the information from the 30 accrued patients is entered into the MSPP, within a maximum of 2
 months after the recruitment of the last patient.
Inclusion Criteria:

 1. Written informed consent for all study procedures according to local regulatory
 requirements prior to enrollment into the study.

 2. Age ≥ 18 years.

 3. Histologically proven metastatic or locally recurrent invasive breast cancer.

 4. Tumor tissue (FFPE and frozen) from recurrent or metastatic lesions available for
 research purposes.

 Exclusion Criteria:

 1. Eastern Cooperative Oncology Group (ECOG) Performance Status >2.

 2. The biopsy procedure is estimated to be too risky for the patient.

 3. Any bevacizumab treatment administered less than 3 weeks before new biopsy procedure.

 4. No appropriate wash-out period for patients on anticoagulation therapy.