High-Throughput Evaluation of Breast Cancer Markers

High-Throughput Evaluation of Breast Cancer Markers
amphiregulin, CA15-3, CD14, epidermal growth factor (EGF), EGF receptor, E-selectin, basic fibroblast growth factor, heparin-binding EGF, Her2, hepatocyte growth factor, intracellular adhesion molecule, insulin-like growth factor 1, interleukins 1alpha and 18, matrix metalloproteases 1, 2 and 9, platelet-derived growth factor, prostate-selective antigen, RANTES, transforming growth factor alpha, tissue necrosis factor alpha, urokinase plasminogen activator, vascular endothelial growth factor
No design specified.
Breast and Gynecologic Cancers Research Group

The objective of this study is to use a combination of AMT proteomics and ELISA microarray testing to discover and validate novel biomarkers for breast cancer.

Aim #1. Identify new candidate biomarkers of breast cancer by using a semi-quantitative proteomic analysis of plasma. This analysis will employ capillary liquid chromatography (cLC) interfaced with Fourier transform ion cyclotron resonance (FTICR) mass spectrometry (MS), which is an exceptionally sensitive method for measuring peptides. We are independently developing a database of proteins secreted into nipple aspirate fluid (NAF) from women with or without breast cancer. This database will allow us to identify proteins in plasma that are produced by the breast, thereby allowing selection of proteins that are more likely to be specific markers for breast disease. Because some proteins may only be useful for detecting a single breast cancer subtype, we will analyze 20 samples/group from four groups of women: 1) healthy women, 2) women with ductal carcinoma in situ (DCIS; stage 0), 3) early ductal cancer (stages 1+2), and 4) early lobular cancer (stages 1+2). Aim #2. Develop an enzyme-linked immunosorbent assay (ELISA) microarray chip that can simultaneously assay ~50 proteins that are potential markers for non-invasive breast cancer. These proteins will be selected based on the results of Aim #1, guidance from the EDRN Steering Committee, published reports and other defined criteria. Where possible, commercial antigen and antibody reagents will be used for these analyses. However, it is anticipated that a major effort will be required for the development and validation of reagents for many of the ELISA assays. Aim #3. Determine whether the quantitative analysis of 25 proteins by ELISA microarray can distinguish between plasma samples from women with or without breast cancer. A set of 1000 plasma samples, approximately equally divided between women with and without breast cancer, will be obtained from the Army’s CBCP. This is an exceptionally high-quality set of samples that was collected under well-defined clinical protocols with an extensive set of epidemiological data. The epidemiological information will be used to evaluate the influence of factors other than breast cancer on individual protein levels, with the goal of accounting for predictable variability in marker levels. Profile analysis of the potential cancer markers will then be undertaken. For this analysis, the samples will be evenly split into a “training” set and a “validation” set of samples, with each set having equal numbers of samples from the control and cancer groups.
High mass accuracy LC-MS proteomics and ELISA microarrays.

There are currently no biomarkers annotated for this protocol.

No datasets are currently associated with this protocol.

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