Introduction: The etiology of breast cancer remains an area of ongoing investigation. Improving our understanding of factors associated with breast cancer development will strengthen the utility of risk prediction strategies and improve risk stratification. Mammographic breast density remains one of the most influential breast cancer risk factors, with 4 to 6 fold elevated risk observed among women with the highest compared to the lowest levels. These associations have consistently been observed irrespective of the method (visual or automated), used to quantify breast density, showing the robustness of the associations. An understudied breast cancer risk factor is the bilateral asymmetry of mammographic features, with prior studies suggesting that women with higher levels of breast asymmetry may be at elevated breast cancer risk. Increasingly, studies are recognizing the potential of bilateral breast asymmetry defined by mammographic breast density in determining breast cancer risk. Recently, Eriksson and colleagues highlighted a strong influence of breast asymmetry within a model they developed to predict short-term breast cancer risk among women attending breast screening. They observed that the contribution to the model of asymmetry in mammographic breast density, microcalcifications, and masses between the breasts was as substantial as the total number of microcalcifications and masses found within a mammogram, indicating that differences between the breasts may be an important risk marker. To help further understand associations between breast cancer risk factors and asymmetry of breast density, particularly among women at elevated risk for breast cancer, we evaluated risk factor relationships with the bilateral asymmetry of volumetric measures of global and local breast density across the spectrum of premalignant and invasive breast cancer diagnoses. Methods: This study included 882 women enrolled as part of the National Cancer Institute's Breast Radiology Evaluation and Study of Tissues (BREAST)-Stamp Project (2007-2010). The BREAST-Stamp Project is a cross-sectional molecular epidemiologic study that aims to understand how novel breast density measures are related to breast cancer etiology. Women were enrolled if they were referred for diagnostic image-guided breast biopsy following an abnormal mammogram at the University of Vermont Medical Center, and had not previously been diagnosed or treated for cancer, undergone breast surgery within one year or received chemoprevention. Risk factor data were collected at study enrolment via interview and self-administered questionnaires. Breast density measures were estimated using Single X-ray Absorptiometry (SXA), a technique in which an SXA breast density phantom was affixed to the compression paddle of the mammography machine during routine mammography so that it was included in the X-ray field. It served as a reference standard to estimate volumetric breast density. Breast density was assessed using pre-biopsy craniocaudal full-field digital mammograms of both the ipsilateral (affected) and contralateral (unaffected) breast. Firstly, global density from each laterality was determined as percent fibroglandular volume (%FGV). Secondly, localized volumetric density measures were estimated following identification of the biopsy site on the ipsilateral pre-biopsy mammogram by the study radiologist and identifying the corresponding site on the contralateral mammogram. The SXA estimated %FGV in a perilesional volume twice the size of, but excluding, the biopsy target, centered at the biopsy site. Previous estimates of reproducibility for the SXA test phantoms demonstrated a repeatability SD of 2%, with a ±2% accuracy for the entire thickness and density ranges. Breast density asymmetry was defined as an absolute bilateral difference in measures when subtracting the breast density measures of the contralateral breast from the ipsilateral measures. Spearman's correlations (rho) examined associations between breast density measures of the left and right breasts. To determine relationships between breast cancer risk factors (defined as categorical variables) and measures of bilateral breast density asymmetry, analysis of covariance (ANCOVA) models (PROC GLM) were used. Analyses were conducted at the per woman level using SAS. Probability values of <0.05 were considered statistically significant, and all tests were two-tailed. Results:We initially investigated asymmetry within each woman by examining correlations between breast density measures in the left and right breasts. Strong, positive correlations between the ipsilateral and contralateral breast were observed for each breast density measure (rho for global %FGV=0.89, p-value<0.0001; rho for local %FGV=0.79, p-value<0.0001). Breast asymmetry was observed in the majority of the study population. For global %FGV, 76% of women had a bilateral difference ≥2%, with 43% of women having higher %FGV in their ipsilateral affected breast and 33% having higher in their contralateral unaffected breast. For localized %FGV, the majority of women had differences between their two breasts (89%), with 61% of women having higher localized %FGV in their ipsilateral affected breast compared to the remaining 28% who had higher localized breast density in their unaffected breast. We next examined relationships between breast cancer risk factors and breast asymmetry. Overall, no associations were observed between any of the risk factors examined, including age, race, body mass index, education, menopausal status, menopausal hormone therapy use with absolute bilateral differences in global or localized %FGV. Among the study population, most women had a benign breast disease diagnosis, with 33% and 43% being diagnosed with benign non-proliferative and benign proliferative lesions, respectively. Of the study population, 15% were diagnosed with invasive breast cancer. Overall, no differences were observed in bilateral differences in global %FGV according to diagnosis; however, higher mean bilateral differences in localized %FGV were observed for women with invasive compared to other diagnoses including benign and in-situ lesions (p=0.056). Discussion and conclusions: Our findings showed that breast asymmetry, defined by bilateral differences in global and localized mammographic breast density, was mostly unrelated to breast cancer risk factors among women undergoing an image guided breast biopsy. We observed that localized measures, defined according to within-woman bilateral differences in localized %FGV surrounding a suspicious lesion as compared with localized %FVG in a comparable location in the contralateral breast, may be an indication of cancer. This investigation is currently ongoing and efforts are underway to replicate findings using evolving image analysis methods such as Volpara Density Maps, an FDA-approved breast imaging tool which provides volumetric estimates of local glandular tissue distribution. Further, the ascertainment of 10-year follow-up data within this study population is in progress, which will facilitate prospective investigations of the relationship between breast asymmetry and breast cancer risk. Ongoing work will also extend our understanding of whether breast asymmetry is related to individualized breast cancer risk, defined using tailored breast cancer risk assessment tools. In conclusion, further understanding of breast asymmetry is needed to better exploit how bilateral differences in mammographic features may be used to inform breast cancer etiology as well as future breast cancer risk.

Citation Format: Maeve Mullooly, Shaoqi Fan, Ruth M. Pfeiffer, Brian Sprague, Pamela M. Vacek, Donald L. Weaver, John A. Shepherd, Amir Pasha Mahmoudzadeh, Jeff Wang, Serghei Malkov, Jason M. Johnson, Sally D. Herschorn, Gretchen L. Gierach. Investigation of relationships between breast cancer risk factors and bilateral mammographic breast density asymmetry among women undergoing diagnostic image-guided breast biopsies [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr NG15.