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.
The National Cancer Institute's Division of Cancer Prevention has released a new funding opportunity to solicit organ-specific applications for Biomarker Developmental Laboratories (BDLs), one of the four scientific units of the recently funded Early Detection Research Network (EDRN). The EDRN is a national infrastructure funded to discover, develop, and validate biomarkers for risk assessment, detection, and molecular diagnosis and prognosis of early cancer. BDLs are responsible for the discovery, development, characterization, and testing of new, or the refinement of existing, biomarkers and biomarker assays for risk assessment, detection, and molecular diagnosis and prognosis of cancers.
The existing BDLs are primarily focused on ovary and gastrointestinal cancers. The proposed BDLs (to be supported under this funding opportunity) should be focused on one or more of the following cancers: breast, prostate and other genitourinary organs, or lung. In addition, cancers with rapidly rising incidence rates, e.g., endometrial, hepatocellular, kidney, thyroid, oropharyngeal cancers, and/or cancers with unique etiology, e.g., mesothelioma, will be considered.
The newly funded units of the Early Detection Research Network will be announced later in April. Successful applicants have already been notified. Those researchers who were not successful during the last round of applications are encouraged to apply to this opportunity.