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Project Title Molecular Biomarkers for Assessing Breast-Cancer Risk in Lactating Women

Researcher: Kathleen Arcaro, Ph.D., University of Massachusetts, Amherst, Amherst, MA at University of Massachusetts

Study Abstract: Accurate assessment of breast-cancer-risk will benefit most women and analysis of promoter hypermethylation in exfoliated epithelial cells in breast milk provides an ideal opportunity to assess breast-cancer-risk. We propose to examine breast milk samples from lactating women who are scheduled for a breast biopsy. About 10% of such women will have breast cancer; the remaining 90% will have benign lesions. Methylation analysis of epithelial cells in milk samples from each breast, will allow us to compare the promoter methylation in the diseased and the non-diseased breast of each woman. In addition, using the pathology reports and questionnaire data, we can compare methylation patterns between women with cancer and those without malignancy. Importantly, since 90% of our samples will come from women with benign disease, we can examine the relationship between promoter methylation patterns and breast-cancer-risk as defined by type of lesion, family history, and other risk factors.
The goal of this research is to develop a noninvasive molecular tool that can be used to accurately assess a woman’s risk of developing breast cancer. Our hypothesis is that epithelial cells isolated from breast milk can serve as a sensitive and reliable tool for assessing breast-cancer-risk.

We will collect breast milk samples from 250 women scheduled for a breast biopsy. Milk samples will be collected from each breast of each woman prior to her biopsy. Pathology report and questionnaire data (risk factors including reproductive history, health history, family cancer history etc,) will also be obtained. The promoter methylation pattern in a panel of genes known to exhibit promoter hypermethylation in breast cancer will be examined. DNA from the epithelial cells (positive selection with immunomagnetic beads) and from the non-epithelial cells (negative fraction containing blood, immune and other cells) will be isolated and after bisulfite treatment and gene specific amplification will analyzed by pyrosequencing. We will compare the milk epithelial methylation scores between 1) malignant disease and contralateral breasts, 2) benign lesion and contralateral breasts, 3) malignant disease and benign lesion breasts, 4) benign lesions with different risk factors, 5) validate comparisons with contralateral breasts through controls not scheduled for biopsy. Finally, milk fat, skim milk, and RNA will be archived for future studies of pollutant load, proteomics and gene expression, respectively.