Trial Information

Characterization of High Risk Breast Duct Epithelium by Cytology, Breast Duct Endoscopy, and cDNA Gene Expression Profile

Background:

Breast cancer is the most common malignancy in women, occurring in over 180,000 women
annually in the United States.

The vast majority of breast cancers originate in the single layer of epithelial cells that
line the ductal/lobular system of the breast milk ducts. The premalignant changes which
occur in the transformed epithelium are not well understood, however several cytologic or
histologic changes have been identified which are associated with an increased risk for
breast cancer, including ductal or lobular hyperplasia, hyperplasia with atypia, and lobular
or ductal carcinoma in situ.

The identification of cytological or histological abnormalities in breast epithelial cells
is an important component of risk assessment.

Objectives:

The primary objectives are:

To determine the incidence and nature of cytologic changes in ductal epithelial cells from
the high risk breast, in specimens collected by breast duct lavage, and to determine if
these cytologic findings are different from those of normal women not at increased risk for
breast cancer.

To characterize by breast duct endoscopy, high risk breast ductal epithelium and
architecture, and correlate these findings with the cytologic findings from above.

To determine what is the global gene expression pattern of high risk breast epithelial cells
from the high risk breast, and does this differ from that of breast epithelial cells from
normal women not at increased risk for breast cancer. The gene expression profile will be
determined by cDNA microarray and validated by RT-PCR.

To determine by comparative genomic hybridization the gross genomic alterations present in
high risk breast epithelial cells.

To determine the pattern of protein expression for selected proteins in the high risk
epithelial cells using proteomics tissue lysate arrays.

To examine ductal epithelial cell preparations from both normal risk and high risk women for
the presence of mammary stem cells.

Eligibility:

Eligibility for high risk individuals will include:

Women of any age with a unilateral invasive or noninvasive (DCIS) breast cancer of
epithelial origin.

Women without breast cancer, but with a Gail Index greater than 1.67%.

Women known to be BRCA1/2 mutation carriers.

Women with cytologic or histologic evidence of ductal hyperplasia, atypical ductal
hyperplasia, or lobular carcinoma in situ.

Women may be either premenopausal or postmenopausal. Postmenopausal is defined by the
absence of menstrual periods for at least 24 months.

Postmenopausal women who have previously undergone a hysterectomy without oophorectomy must
have a serum FSH level of greater than 40 IU/ml, and a serum estradiol level of less than 40
pg/ml to document postmenopausal status.

Eligibility for normal volunteers will include:

- Women who are premenopausal or postmenopausal with a Gail model risk index less than
1.67%.

- Women who have previously undergone a hysterectomy without oophorectomy must have a
serum FSH level of greater than 40 IU/ml, and a serum estradiol level of less than 40
pg/ml to document postmenopausal status.

- Both breasts must be free of any suspicious areas by physical examination and, for
women over 30 years of age by mammogram. There must be no past history of atypical
hyperplasia, invasive or in situ carcinoma.

Both groups must have acceptable WBC and platelet counts.

Design:

Breast ductal epithelial cells will be collected by breast duct lavage from a.) the breast
in women at increased risk for breast cancer, and b.) the breast of female normal volunteers
who are not at increased risk for breast cancer.

Ductal epithelial cell specimens will be analyzed cytologically for the presence of
hyperplasia, atypia, or in situ changes.

Breast duct endoscopy will be performed in breast cancer patients and in normal volunteers
to determine ductal architectural changes associated with increased risk for breast cancer,
and to provide correlation with cytologic studies.

The gene expression profile of normal and high risk ductal epithelial cells will be studied
by cDNA microarray to determine changes in gene expression associated with increased risk
for breast cancer.

Additional molecular profiling experiments which will be performed as lavage cells are
available include Comparative Genomic Hybridization (CGH), proteomic tissue lysate arrays,
and identification of mammary stem cells.

A total of 70 breast cancer patients and 40 normal volunteers will be studied, divided
approximately evenly between premenopausal and postmenopausal women. After accruing the
first 20 individuals for a given menstrual status, an analysis will be performed. If more
than 5 patients yield less than or equal to 7000 ductal cells/lavage, then the trial will be
re-evaluated prior to proceeding with additional enrollment.