1. MINT study Principal Investigator: Charles E. Cox, M.D.
Co Investigators: Stefan Glück, M.D.

2. MINT
MINT I: Multi-Institutional Neo-adjuvant Therapy MammaPrint Project I
Principal Investigator: Charles E. Cox, M.D. USF, Tampa FL
Co Investigator: Stefan Glück, M.D. UM/Sylvester Comprehensive Cancer Center University of Miami, Miller School of Medicine FL
Total of 226 patients; up to 10 institutes in the US
Full genome gene expression profiling Agendia Inc
Central pathology review at USF pathology FL
Timelines; Oct Oct 2013

3. Breast Cancer Symphony Suite
MammaPrint: 70-gene profile prognostic and predictive tumor analysis
Will patient benefit from chemotherapy?
TargetPrint: Gene expression of ER/PR/HER Centralized lab confirmation of receptor status
Will patient benefit from hormonal treatment?
BluePrint: 80-gene molecular subtyping profile Basal, Luminal, and HER2 subtypes
Which therapy works best?
TheraPrint: Gene expression of 56 genes Potential markers for prognosis and therapeutic response
Potential therapy options saved for the future

4. pCR rate of MammaPrint and BluePrint in previous neo-adjuvant studies

5. MINT; Study Objectives
To determine the predictive power of chemosensitivity of the combination of MammaPrint and BluePrint as measured by pCR.
To compare TargetPrint single gene read out of ER, PR and HER2 with local and centralized IHC and/or CISH/FISH assessment of ER, PR and HER2.
To identify and/or validate predictive gene expression profiles of clinical response/resistance to chemotherapy.
To identify possible correlations between the TheraPrint Research Gene Panel outcomes and chemoresponsiveness.
To compare the three BluePrint molecular subtype categories with IHC- based subtype classification.

6. MINT; Eligibility Criteria
Inclusion criteria:
Women with histologically proven invasive breast cancer; T2(≥3.5cm)-T4, N0,M0 or T2-4N1M0
DCIS or LCIS are allowed in addition to invasive cancer at T2 or T3 level.
Age ≥ 18 years.
Measurable disease in two dimensions
Adequate bone marrow reserves, adequate renal function, and hepatic function
Signed informed consent
Exclusion criteria
Patients with inflammatory breast cancer.
Tumor sample shipped to Agendia with ≤ 30% tumor cells or that fails QA or QC criteria.
Patients who have had any prior chemotherapy, radiotherapy, or endocrine therapy for the treatment of breast cancer.
Any serious uncontrolled inter current infections, or other serious uncontrolled concomitant disease.

7. MINT; Nodal staging

8. Study Design Flowchart
Central slide pathology review
Patient information & informed consent
Core Needle
Biopsies
TAC
Surgery including nodal staging
HER2-
Sample placed in RNA Retain, send to Agendia
Full Genome Array* successful TC
HER2-
Central slide pathology review
ddAC/FEC100,
paclitaxel or docetaxel
HER2-
TCH
HER2+
Full Genome Array* not successful
T H/FEC H
HER2+
CRF 1
baseline
CRF 2
surgery
Patient ineligible
* (Including diagnostic commercial testing for Symphony Breast cancer Suite)

9. Neo-adjuvant therapy For HER2 negative patients: TAC chemotherapy
TC chemotherapy
Dose Dense AC or FEC100 followed by paclitaxel or docetaxel chemotherapy
For HER2 positive patients:
TCH chemotherapy
T H followed by FEC H
Dose adjustments
Hematological and non-hematological toxicities should be managed by treating oncologist as per routine clinical practice.

10. Future Research – Remaining Tissue
Agendia will store remaining tissue from patient samples
This tissue can be used for future scientific research
Patients are asked in the patient consent form to provide consent for storage and future research
Please note: If a patient does not wish to allow their tissue to be used for future research, it is the responsibility of the site to communicate this with Agendia

11. Tissue Collection
Tissue should be collected by incisional biopsy (when placing port) or via core needle biopsy.
If the tissue is obtained by incisional biopsy then the tissue sample should be no greater than 3 to 4 mm in thickness and between 8 to 10 mm in diameter.
Core needle biopsies should be obtained with a 14 gauge or larger needle.
If a 14 gauge needle is used: 5 cores
If a 11 gauge needle is used: 4 cores
If a 9 gauge needle is used: 3 cores   

12. Sending Sample to Agendia
Remove large specimen tube from kit and open it.
Place large screw cap with small specimen vial on table and open small specimen vial.
Place a barcode label from the completed requisition form on to the vial.
Place the small specimen vial into the large specimen tube and place the tube into the specimen safety bag.
Place the sealed specimen bag into the shipping kit along with the requisition form. IMPORTANT: ensure that the study sticker is affixed to the requisition form.
Package the kit into the FedEx shipping pack and attach the pre printed label for shipment to Agendia Inc.
Please call Fed Ex for pickup.

13. Central Pathology Review
Central pathology review for RCB score will be done at USF Pathology FL
Representative slides of core needle biopsy, sentinel lymph node biopsies and surgical sample should be send to:
USF Pathology
Attn: MINT Trial
12901 Bruce B. Downs Blvd MDC 11
Tampa, FL 33612
Lisette to review and adjust as needed

14. Procedures for Sending Slides
APPENDIX III Pathology Worksheet for Core/Incisional Biopsy
Please send the following, and complete the form below:
One H&E section of tumor
ER immunohistochemical stain
PR immunohistochemical stain
HER2 immunohistochemical stain or FISH/CISH/SISH report
10 unstained sections on positively-charged glass slides
(If ER, PR, and/or HER2 studies not available, please send an additional 10 unstained sections on positively-charged glass slides)
Questions to be completed on worksheet:
1. Laterality
a. Left breast
b. Right breast
2. Location
a. UOQ
b. LOQ
c. UIQ
d. LIQ
3. Time in formalin
a. ≥ 6 to ≤ 48 hours
b. Other

15. Procedures for Sending Slides
APPENDIX IV Pathology Worksheet for Post-Neoadjuvant Specimens
Please send the following, and complete the form below:
Two representative blocks of tumor, or 15 unstained slides on positively charged glass slides
Copy of final pathology report (including gross description)
1. Specimen
a. Lumpectomy/partial mastectomy
b. Total mastectomy
c. Skin-sparing, nipple-sparing total mastectomy
d. Nipple-sparing total mastectomy
e. Modified radical mastectomy
f. Other
2. Residual tumor
a. Grossly identified
i. Dimensions: ____ x ____ x ____ cm
ii. Percent gross necrosis:
b. Not grossly identified *
Fibrotic tumor bed dimensions: ____ x ____ x ____ cm
Distance from closest margin: ____
*NOTE: If no definitive tumor grossly identified, please submit entire tumor bed sequentially

16. Pathology Guidelines
Note: additional guidelines for your pathology are outlined in Appendix II of the protocol.

17. Clinical Report Form Web based data collection/electronic CRF
CRF1; Baseline information
CRF2; After surgery
Relatively easy to complete compared to drug trial
15-20 minutes for CRF1 and 2

18. Accessing the Database
There is one hyperlink to access all the electronic Case Report Forms (CRFs)
You will receive an institution specific site number and password from your Agendia Clinical Research Manager.
Institution:

19. CRF Overview Administrative page
Detailed Data Entry Instructions will be sent to site.
Samples will appear in the database if they are found to be eligible.

20. CRF 1: Questions Pathology Date of histologic diagnosis
Histopathologic tumor type  
Histological grade
ER, PR and Her2-neu status
Vascular invasion
TNM
Nodal staging
Date nodal staging
If sentinel lymph node
 Number of sentinel nodes removed
 Number of counts
Blue dye
Histology of SLN: 
Concordance of nodal histology to primary tumor
Baseline Patient characteristics
Age at diagnosis
Ethnicity/ origin
Menopausal status 
Date of biopsy
Specimen type
Tumor measurements
Kind of imaging used
Primary tumor size
Size largest metastatic lymph node

21. CRF 2: Questions Sentinel Lymph Node Procedure (SLNP) If yes
Neo-adjuvant treatment
Regimen given 
Was specified number of cycles completed?
Any grade 4 or 5 CTC for Adverse Events observed?
Surgery information
Date breast carcinoma surgery
Type of surgery
Lymph node assessments
Pathology
Nodal status
ypTN
Tumor measurements
What kind of imaging has been used ?
Primary tumor size
Size largest metastatic lymph node
Treatment response
Lymph node assessments
Sentinel Lymph Node Procedure (SLNP) If yes
Number of sentinel nodes examined
Number of positive sentinel nodes
Number of negative sentinel nodes
Blue dye
Concordance of nodal histology to primary tumor
 Axillary Lymph Node Dissection (ALND) If yes
Number of axillary nodes examined
Number of positive axillary nodes
Number of negative axillary nodes
SLNP and ALND if yes:

22. Contacts:
Jessica Gibson, Clinical Research Manager (619)