Application of novel breast biospecimen cell type adjustment identifies shared DNA methylation alterations in breast tissue and milk with breast cancer risk factors.

BACKGROUND: DNA methylation patterning is cell-type-specific and altered DNA methylation is well established to occur early in breast carcinogenesis, affecting non-cancerous, histopathologically normal breast tissue. Previous work assessing risk factor-associated alterations to DNA methylation in breast tissue has been limited, with even less published research in breast milk, a non-invasively obtained biospecimen containing sloughed mammary epithelial cells that may identify early alterations indicative of cancer risk.

METHODS: Here, we present a novel library for the estimation of the cellular composition of breast tissue and milk and subsequent assessment of cell type-independent alterations to DNA methylation associated with established breast cancer risk factors in solid breast tissue (n = 95) and breast milk (n = 48) samples using genome-scale DNA methylation measures from the Illumina HumanMethylation450 array.

RESULTS: We identified 772 hypermethylated CpGs (P < 0.01) associated with age consistent between breast tissue and breast milk samples. Age-associated hypermethylated CpG loci were significantly enriched for CpG island shore regions known to be important for regulating gene expression. Among the overlapping hypermethylated loci mapping to genes, a differentially methylated region was identified in the promoter region of SFRP2, a gene observed to undergo promoter hypermethylation in breast cancer.

CONCLUSIONS: Our findings suggest the potential to identify epigenetic biomarkers of breast cancer risk in noninvasively obtained, tissue-specific breast milk specimens.

IMPACT: This work demonstrates the potential of using breast milk as a non-invasive biomarker of breast cancer risk, improving our ability to detect early stage disease and lowering the overall disease burden.