Prediction of Response to Neoadjuvant Chemotherapy in Women With Operable Breast Cancer

The objective of this study is to develop a biomarker to predict pathological complete
response in women treated with neoadjuvant chemotherapy for breast cancer. Such a biomarker
would assist physicians in selecting the most effective chemotherapy for the individual
patient. The anticipated biomarker will take into account clinical factors (such as tumor
stage, tumor size, and age), phenotypic characteristics of the tumor (determined by
pathological immunohistochemistry and ex vivo ChemoResponse assay), and genotypic
characteristics of the tumor and patient (determined by genomic profiling via gene
expression analysis of tumor RNA). It is expected that collective consideration of all of
these factors will be more predictive of patient response to therapy than any of them alone.

Approximately 224 evaluable subjects will be recruited from approximately 30 US sites.
Women with measurable operable invasive breast cancer diagnosed by core needle biopsy will
be eligible for this study. Additional tumor specimens will be obtained prior to the start
of chemotherapy via core needle biopsies to be used for the ex vivo ChemoResponse Assay and
tumor genomic analysis (gene expression), respectively.

All subjects will receive neoadjuvant chemotherapy with one of two standard of care regimens
that must consist of the following agents: doxorubicin (A), cyclophosphamide (C), and a
taxane (T) such as docetaxel, paclitaxel, or Abraxane (nanoparticle albumin-bound paclitaxel
[nab-paclitaxel]); or, docetaxel (T) and cyclophosphamide (C). These must be administered
per NCCN guidelines by the treating physician.

Upon completion of chemotherapy treatment, women will undergo lumpectomy, modified radical
mastectomy or other surgical procedure determined appropriate by the investigator and at
that time will be evaluated for pathological response. At the time of lumpectomy, modified
radical mastectomy, or other surgical procedure, additional tumor excess will be sent to
Precision Therapeutics, Inc. (Precision) for exploratory analysis if there is no pathologic
complete response (pCR), if there are sufficient tumor cells to send, and if the patient
agrees to have her excess tumor cells sent to Precision for this purpose.

During the patient's course of participation on the study, the treating physician will
remain blinded to the results of the ChemoResponse Assay and genomic analysis. If it is
determined there is no pCR at the time of lumpectomy, modified radical mastectomy or other
surgical procedure, Precision will make available a subsequent report to the physician
containing additional information about chemotherapy drugs other than ACT that could benefit
the further treatment decisions for the patient.

Inclusion Criteria

Female subjects who satisfy the following conditions will be considered for enrollment
into the study:

1. The subject must consent to be in the research study and must have signed an approved
consent form conforming to institutional guidelines prior to study entry.

2. The diagnosis of breast cancer can be made by FNA or biopsy (other than incisional or
excisional). The tumor specimen must demonstrate a diagnosis of invasive
adenocarcinoma.

3. The primary breast cancer must be operable and measurable "greater than or equal to"
2.0 cm by use of physical exam and/or ultrasound, MRI, CT scan, or mammogram.

4. T1c, T2, T3, or T4 patients clinically staged as M0 (non-inflammatory) are eligible.

5. Patients with a prior diagnosis and treatment for DCIS are eligible.

6. Patients with multi-focal breast cancer are eligible.

7. The tumor must be confined to either the breast or to the breast and ipsilateral
axilla.

8. The subject must be 18 years or older.

9. The interval between initial cytologic or histologic diagnosis of breast cancer and
registration must be no more than 10 weeks.

10. ECOG Performance Status of 0 or 1 (see Appendix A) is required.

11. The subject must receive standard of care chemotherapy regimens consisting of either
doxorubicin (A), cyclophosphamide (C), and a taxane (T) such as docetaxel,
paclitaxel, or nab-paclitaxel administered in any sequence and combination the
treating physician determines or docetaxel (T) plus cyclophosphamide (C).

Exclusion Criteria

Male subjects are not eligible for this study as the incidence of breast cancer in male
subjects is significantly lower than female subjects. Those subjects who are strongly
HER2-positive will be excluded as they will require treatment by biological agents for
which the ChemoResponse Assay has not yet been validated. Subjects with evidence of
distant metastatic disease are excluded as these subjects would not be good candidates for
neoadjuvant therapy. Women who have had an excisional or incisional biopsy prior to entry
would not have sufficient tumor sample to test or to be measured by physical exam for the
study. Women who have nonmalignant comorbid conditions and diseases that would preclude
them from being treated with doxorubicin (A), cyclophosphamide (C), and a taxane (T), and
from completing the study are also excluded. Women with psychiatric or addictive
disorders are excluded to protect those vulnerable subjects who may not be able to
adequately give informed consent.

Women with one or more of the following conditions will be ineligible for this study:

1. Tumor determined to be strongly HER2-positive by immunohistochemistry (3+) or by
fluorescent in situ hybridization (positive for gene amplification)

2. Definitive clinical or radiologic evidence of distant metastatic disease.

3. Excisional or incisional biopsy for this primary breast tumor.

4. Inflammatory breast cancer.

5. Synchronous contra-lateral breast cancer.

6. Multi-centric breast cancer.

7. Participation in the NSABP B-40 study.

8. Prior therapy for invasive breast cancer, including irradiation, chemo-, immuno-,
and/or hormonal therapy.

a. Note: the only exception is hormonal therapy, which may have been given anytime
after diagnosis and before study entry as long as the hormonal therapy is
discontinued at or before registration. After surgery, hormonal therapy may be
re-started, at the discretion of the treating physician.

9. Current therapy with any hormonal agent such as raloxifene, tamoxifen, or other
selective estrogen receptor modulator (SERM), either for osteoporosis or breast
cancer prevention, or sex hormonal therapy such as birth control pills, ovarian
hormonal replacement therapy, etc. These patients are eligible IF these medications
are discontinued prior to registration.

10. Surgical axillary staging procedure prior to study entry.

a. Note: exceptions include FNA of an axillary node and pre-neoadjuvant sentinel
lymph node biopsy for patients with clinically negative axillary nodes.

11. Nonmalignant systemic disease (cardiovascular, renal, hepatic, etc.) that would
preclude the woman from being treated with doxorubicin (A), cyclophosphamide (C), and
a taxane (T), and from completing the study.

12. Psychiatric or addictive disorders that would preclude obtaining informed consent.