PROTEOMIC ANALYSIS FOR CANCER DIAGNOSIS: DETECTING AND STAGING BREAST CANCER PRIOR TO OVERT CLINICAL SYMPTOMS

Seth M. Miller^1,2, Dr. M. P. Nandakumar¹, M. Megan Fitzpatrick², Suzanne O. Rosenberg², Brian P. Bradley² and Mark R. Marten¹

Departments of Chemical and Biochemical Engineering¹ and Biological Sciences², University of Maryland, Baltimore County, 1000 Hilltop Circle, Baltimore, MD 21250

Methods are needed to detect small malignant primary tumors, to determine if precancerous states exist [e.g. ductal carcinoma in situ (DCIS), atypical mammary hyperplasia], and to establish whether micrometastases exist. We hypothesize that the different stages of breast cancer (e.g. hyperplasia, DCIS, and mammary adenocarcinoma) may result in production of tumor encoded or host-produced proteins characteristic of these steps. We propose that by using a proteomic approach we will be able to analyze serum from individuals developing breast cancer and stage the disease or even predict the patient’s outcome. To test this theory, transgenic NeuT mice containing a mutated her-2/neu gene, a gene that causes breast cancer, will be used. Her-2/neu transgenic NeuT mice develop breast cancer beginning with atypical mammary hyperplasia (3 weeks of age), progressing to DCIS (13-17 weeks), and then to overt adenocarcinoma (~week 20). Serum samples will be obtained from these mice at approximately 2-week fine points and correlated with evident tumor onset. Serum proteins will be analyzed using two-dimensional gel electrophoresis. Specific proteins diagnostic of the different stages of breast cancer will be sought. If such biomarkers are identified, they will be distinguished by Matrix Assisted Laser Desorption/Ionization- Time of Flight Mass Spectrometry (MALDI-TOF MS).