A major focus of breast cancer research has involved the catechol estrogen pathway. Estrogen DNA Adduct (EDA) is a novel biomarker that is hypothesized to play a role in initiating breast cancer. Pilot studies have shown elevated levels of EDA in women at high risk for breast cancer and with breast cancer. The overall purpose of this project was to determine whether the EDA biomarker has potential for clinical use for breast cancer prevention. The first step toward this goal was to examine the extent to which the biomarker varies over time in the same individual. The specific aim was to determine whether the variation between urine samples taken at two points in time is significantly different from samples taken at a single point in time. We utilized a convenience sample from the DEEM (Diet, Exercise and Estrogen Metabolites) study of 6 urine samples from 3 women at 2 separate clinic visits, conducted 3 months apart. Triple quadrupole mass spectrometry was used to process 37 estrogen metabolites in the urine samples. The fragmentation pattern of each metabolite was analyzed and compared to a panel of known estrogen metabolites in which the elution times were known. The correlation coefficient between the 2 clinical visits was 0.49. The results from this analysis indicate moderate correlation between 2 time points. We will await the results from the larger study to decide whether a one point in time urine collection is adequate for the overall project. The implications of the overall project are important because determining breast cancer risk from urine samples would be a sensitive and non-invasive method that could allow healthcare providers to better target the women that should receive careful monitoring, testing, and interventions.

It is known that exercise improves cancer survival, however, there is an unknown molecular link between physical activity and cancer occurrence. The purpose of this study was to determine the feasibility of studying biologic mechanisms related to exercise and cancer survival in the LIVESTRONG YMCA program. Eligible subjects were males and females who were diagnosed with prostate or breast cancer in the last three years, between 35-80 years of age, willing to provide urine samples at four time points, and enrollment in the 12-week LIVESTRONG YMCA program. Through a partnership with the University of Washington (UW) Mass Spectrometry Center and the UW Office of Nursing Research, two specific biomarkers were analyzed from the urine specimens. One of these markers is Estrogen DNA Adduct (EDA), an estrogen metabolite that binds to DNA and creates an increased potential of DNA mutation and cancer development. The other is 8-OH-dG, a biomarker related to oxidation. To determine the feasibility, we administered questionnaires to better determine patient satisfaction and calculate the percentage of participation and retention. One questionnaire surveyed the acceptability of our biospecimen collection procedures to participants, anticipated acceptability of additional biospecimen collection (blood and tissue samples) for future projects, and willingness to have a DEXA scan (for compensation). In order to determine the feasibility of conducting the study on a larger scale, we evaluated the percentage of participants enrolled out of those recruited, percentage of participants who provided urine samples out of those enrolled, and the percentage of participants who completed the program. Continuing to invest in our therapeutic, diagnostic and genetic understanding of cancer means that in the future, patients will have options based on scientific evidence.

 Breast cancer is the most common type of cancer in women, and 40,996 women died of cancer in 2010. I am working with Dr. Keryn Reding from the School of Nursing on examining estrogen DNA adducts (EDA) for their clinical utility as a biomarker for breast cancer. EDA is formed when estrone or estrodial are oxidized to form quinones. These compounds then attach to DNA, causing mutations that lead to cancerous cell growth. The EDA are then excised and released into the blood stream and excreted through urine. I will analyze urine samples in a triple quadrupole mass spectrometer in order to see if the ratio of EDA to parent estrogens is higher in women with breast cancer than women who did not develop breast cancer. My analysis will investigate the inter-variability of EDA in premenopausal and postmenopausal women to see if this biomarker can be accurate and reliable for women of all ages. We expect results to show that premenopausal women will have less EDA due to their younger age and lower risk of developing breast cancer. By May when I present at the URP, I will have results from the mass spectrometer and I will use the McNemar's test for statistical significance. We are investigating a hypothesis that a high EDA ratio will be associated with breast cancer risk, and thus the EDA ratio may have clinical applications for breast cancer prevention and screening. Because it is a non-invasive method of measuring the risk of developing breast cancer, it could be an ideal complement to current screening approaches, such as mammograms or biopsies.