Program Director/Principal Investigator (Last, First, Middle): Darie, Costel C. Project summary The R15 AREA is designed to support small scale research projects at primarily undergraduate institutions that provide bachelors and advanced degrees and that do not have major support from NIH. Our R15 plan will expose undergraduate students to the interdisciplinary concepts required to understand biomedical research, and conduct research in the field of proteomics-based analysis of biological fluids, as applied to breast cancer (BC) biomarker discovery. The project type and the location of our university is a perfect match for the goals of R15 AREA grant. BC in young women (reproductive age, pre-menopausal) is associated with increased mortality, and current methods of detecting BC in this group of women have known limitations. Tools for accurately assessing personal BC risk in young women are needed to identify those women who would benefit the most from earlier intervention. Breast milk provides a noninvasive way to examine the health of the breast. We will apply quantitative proteomics to a unique collection of breast milk samples to determine if there is a group of proteins (proteomic signature) that can be used to detect early breast cancer and predict which women are at increased risk of developing BC. We hypothesize that a set of proteins exist in breast milk that can be used to identify women with BC and women at increased risk of developing BC at a young age. We also hypothesize that the set of proteins, detected in the breast milk, can also be detected in the blood and be used to detect BC in non-lactating women. We will identify a proteomic BC signature in milk using archived samples from 20 women who were diagnosed with Invasive Ductal Carcinoma (IDC) of the breast, with a focus on samples provided before diagnosis of cancer and from an age- and parity-matched comparison group of 20 women without BC (Aim 1). We will also quantify the protein biomarker candidates identified in our preliminary studies. Our draft protein signature includes downregulated caseins, bile salt stimulated lipase, xanthine dehydrogenase/oxidase, lactoferrins, fatty acid synthase and upregulated Zn-alpha2-glycoprotein and anti- chymotrypsin. In Aim 2, we will identify a proteomic BC signature in sera from 50 donors with IDC BC and 50 matched controls. We will also assess the applicability of the milk proteomic BC-signature to serum using serum samples described earlier. In Aim 3, we will use bioinformatics to structurally and functionally characterize the proteins that are found dysregulated in milk and serum proteomics (this Aim will be conducted only by undergraduate students). The unique aspects of our study include proteomics analysis of breast milk for assessing BC risk. Translation of the proteomic signature from a local microenvironment (breasts) to a systemic environment (blood) will then allow its use in the detection of BC in non- lactating women. Our pro...