Harvest of CTCs From MBC Patients Using the Parsortix™ PC1 System

Approximately 200 evaluable MBC patients (either newly diagnosed or patients with
progressive/recurrent disease who are about to start a new line of therapy for the treatment
of their disease) and 200 evaluable healthy volunteer (HV) subjects (healthy women with no
history of cancer) will be enrolled at a minimum of three (3) US based clinical sites. Each
subject will have information about their age race/ethnicity, height and weight, menopausal
status, smoking status, pregnancy and/or nursing status, and a brief medical history captured
at the time of enrollment. Blood will be drawn from each subject into three different EDTA
tubes (minimum of ~7mL up to a maximum of ~23mL of whole blood) specifically for the purposes
of this study. One of the blood tubes collected will be used for a complete blood count (CBC)
with differential testing, while the other two tubes of blood will be processed on the
Parsortix PC1 system for the capture and harvest of CTCs. The cells harvested from one of the
blood tubes will be deposited onto a glass slide and automated Wright-Giemsa staining will be
done to allow for identification of CTCs based on their cytologic features (e.g. size, shape,
nuclear to cytoplasmic ratio, chromatin structure, etc.) by an expert cytopathologist. The
cells harvested from the remaining blood tube will be used for one of three different
evaluations: Fluorescence in-situ hybridization (FISH) for evaluation of Her-2/neu gene
amplification, quantitative reverse-transcriptase real-time PCR (qRT-PCR) for evaluation of
cancer related gene expression, or whole transcriptome sequencing (RNAseq) for determination
of the expression patterns of breast cancer related genes.

MBC Patient Inclusion Criteria

- Female >=22 years of age;

- Documented evidence of metastatic breast cancer (i.e. primary tumor histopathology of
breast cancer and documented evidence of distant sites of metastasis by imaging,
biopsy, or other means) that is either newly diagnosed or currently progressing /
recurrent (disease progression / recurrence may be determined by any means, including
RECIST v1.1 criteria, physical signs and symptoms, rising tumor markers, physician
determination, etc.);

- If newly diagnosed, have not yet started a new line of therapy of any type (e.g.
hormonal, cytotoxic, targeted, etc.) for the treatment and/or management of their
metastatic breast cancer;

- If progressing or recurrent, any number of prior hormonal therapies,
chemotherapies and/or biological/targeted therapies are allowed;

- Willing and able to provide informed consent and agree to complete all aspects of the
study.

MBC Patient Exclusion Criteria

- Female subjects <=21 years old or male subjects;

- Concurrent other malignancies (except for a second primary breast cancer);

- Less than seven days since last administration of a cytotoxic agent;

- Unwilling or unable to provide informed consent or high risk that subject may not
comply with protocol requirements.

HV Inclusion Criteria

- Females >=22 years of age;

- No known fever or active infections at the time of the blood collection;

- No known current diagnosis of acute inflammatory disease or chronic inflammation;

- No known current and/or prior history of malignancy, excluding skin cancers (squamous
cell or basal cell);

- Willing and able to provide informed consent and agree to complete all aspects of the
study.

HV Exclusion Criteria

- Female subjects <=21 years old or male subjects;

- Known illness at the time of the blood collection;

- Known current and/or prior history of malignancy, excluding skin cancers (squamous
cell or basal cell);

- Unwilling or unable to provide informed consent or high risk that subject may not
comply with protocol requirements (e.g. due to health and/or participation in other
research studies).