ABCR  Vol.4 No.1 , January 2015
An Autoantibody Based Protein Microarray Blood Test to Enhance the Specificity of a Negative Screening Mammogram
Background: Current screening mammography for breast cancer is associated with misdiagnosis in as many as 30% of cases. Objectives: To develop and clinically evaluate a unique autoantibody based protein microarray blood test to improve the accuracy of breast cancer screening. Materials and Methods: A microarray was constructed from commercial antigens and antigens selected from screened cDNA libraries of breast cancer tissue samples. A training set containing 439 healthy controls and 276 biopsy proven breast cancer cases was used to establish a set of separating models between the two groups. These models were used to assign a diagnosis to 285 blind samples from 120 breast cancer patients and 165 healthy controls. Results: The test identified 82 of the 120 breast cancer patients and 160 of the 165 healthy controls. These results can be translated into a sensitivity of 68.3% [CI: 59% -77%] and a specificity of 97% [CI: 93% -99%], with a PPV for this validation set of 94.3% (CI: 87.10% -98.11%), NPV of 80.81% [CI: 74.62% -86.05%] and an AUC of 89.2% [CI: 78% -87%]. Conclusions: The protein microarray can be utilized to reduce the false negative rate of routine screening mammography. Women with a negative mammography and a negative blood test can be reassured and encouraged to continue routine breast cancer screening. A positive test should alert the physician about the possible presence of a breast cancer not detected by routine screening mammography and drive to perform additional investigation, such as breast ultrasound and MRI.

Cite this paper
Allweis, T. , Strauss, L. , Malyutin, Z. , Kapov-Kagan, A. , Novikov, I. , Bevers, T. , Iacobelli, S. , Sandri, M. , Bitterman, A. , Engelman, P. , Piura, B. , Rosenberg, M. and Yahalom, G. (2015) An Autoantibody Based Protein Microarray Blood Test to Enhance the Specificity of a Negative Screening Mammogram. Advances in Breast Cancer Research, 4, 22-38. doi: 10.4236/abcr.2015.41003.
[1]   IARC (2012) IARC [Online].

[2]   D’Orsi, C.J. and Newell, M.S. (2011) On the Frontline of Screening for Breast Cancer. Seminars in Oncology, 38, 119-127.

[3]   Pisano, E.D., et al. (2005) Diagnostic Performance of Digital versus Film Mammography for Breast-Cancer Screening. The New England Journal of Medicine, 353, 1773-1783.

[4]   Berg, W.A., et al. (2012) Detection of Breast Cancer with Addition of Annual Screening Ultrasound or a Single Screening MRI to Mammography in Women with Elevated Breast Cancer Risk. The Journal of the American Medicine Association, 307, 1394-1404.

[5]   Ho, J.M., Jafferjee, N., Covarrubias, G.M., Ghesani, M. and Handler, B. (2014) Dense Breasts: A Review of Reporting Legislation and Available Supplemental Screening Options. American Journal of Roentgenology, 203, 449-456.

[6]   Crystal, P., Strano, S.D., Shcharynski, S. and Koretz, M.J. (2003) Using Sonography to Screen Women with Mammographically Dense Breasts. American Journal of Roentgenology, 181, 177-182.

[7]   Berg, W.A., et al. (2008) Combined Screening with Ultrasound and Mammography vs. Mammography Alone in Women at Elevated Risk of Breast Cancer. The Journal of the American Medicine Association, 299, 2151-2163.

[8]   Kolb, T.M., Lichy, J. and Newhouse, J.H. (2002) Comparison of the Performance of Screening Mammography, Physical Examination, and Breast US and Evaluation of Factors that Influence Them: An Analysis of 27,825 Patient Evaluation. Radiology, 225, 165-175.

[9]   Kuhl, C.K., et al. (2005) Mammography, Breast Ultrasound, and Magnetic Resonance Imaging for Surveillance of Women at High Familial Risk for Breast Cancer. Journal of Clinical Oncology, 23, 8469-8476.

[10]   Lehman, C.D., et al. (2007) Cancer Yield of Mammography, MR, and US in High-Risk Women. Radiology, 244, 381-388.

[11]   Warner, E., Plewes, D.B., Shumak, R.S., Catzavelos, G.C., Di Prospero, L.S., Yaffe, M.J., et al. (2001) Comparison of Breast Magnetic Resonance Imaging, Mammography, and Ultrasound for Surveillance of Women at High Risk for Hereditary Breast Cancer. Journal of Clinical Oncology, 19, 3524-3531.

[12]   Weigelt, B., Geyer, F.C. and Reis-Filho, J.S. (2010) Histological Types of Breast Cancer: How Special Are They? Molecular Oncology, 4, 192-208.

[13]   Piura, E. and Piura, B. (2010) Autoantibodies to Tummor-Associated Antigens in Breast Carcinoma. Journal of Oncology, 2010, Article ID: 264926.

[14]   Baselga, J., Seidman, A.D., Norton, L. and Rosen, P.P. (1997) HER2 Overexpression and Paclitaxel Sensitivity in Breast Cancer: Therapeutic Implications. Oncology, 11, 43-48.

[15]   Yang, E., Hu, X.F. and Xing, P.X. (2007) Advances of MUC1 as a Target for Breast Cancer Immunotherapy. Histology and Histopathology, 22, 905-922.

[16]   Daniels, T., Zhang, J.Y., Gutierrez, I., Elliot, M.L., Yamada, B., Jo Heeb, M., et al. (2005) Antinuclear Autoantibodies in Prostate Cancer: Immunity to LEDGF/p75, a Survival Protein Highly Expressed in Prostate Tumors and Cleaved during Apoptosis. The Prostate, 62, 14-26.

[17]   Soussi, T. (2000) p53 Antibodies in the Sera of Patients with Various Types of Cancer: A Review. Cancer Research, 60, 1777-1788.

[18]   Rohayem, J., Diestelkoetter, P., Weigle, B., Oehmichen, A., Schmitz, M., Mehlhorn, J., Conrad, K., Rieber, E.P., et al. (2000) Antibody Response to the Tumor-Associated Inhibitor of Apoptosis Protein Survivin in Cancer Patients. Cancer Research, 60, 1815-1817.

[19]   Zhang, J.Y., Casiano, C.A., Peng, X.X., Koziol, J.A., Chan, E.K. and Tan, E.M. (2003) Enhancement of Antibody Detection in Cancer Using Panel of Recombinant Tumor-Associated Antigens. Cancer Epidemiology, Biomarkers & Prevention, 12, 136-143.

[20]   Ganapathy, V., Daniels, T. and Casiano, C.A. (2003) LEDGF/p75: A Novel Nuclear Autoantigen at the Crossroads of Cell Survival and Apoptosis. Autoimmunity Reviews, 2, 290-297.

[21]   Ganapathy, V. and Casiano, C.A. (2004) Autoimmunity to the Nuclear Autoantigen DFS70 (LEDGF): What Exactly Are the Autoantibodies Trying to Tell Us? Arthritis & Rheumatism, 50, 684-688.

[22]   Ludwig, N., Keller, A., Comtesse, N., Rheinheimer, S., Pallasch, C., Fischer, U., et al. (2008) Pattern of Serum Autoantibodies Allows Accurate Distinction between a Tumor and Pathologies of the Same Organ. Clinical Cancer Research, 14, 4767-4774.

[23]   Leidinger, P., Keller, A., Ludwig, N., Rheinheimer, S., Hamacher, J., Huwer, H., et al. (2008) Towards an Early Diagnosis of Lung Cancer: An Autoantibody Signature for Squamous Cell Lung Carcinoma. International Journal of Cancer, 123, 1631-1636.

[24]   Lin, H.S., Talwar, H.S., Tarca, A.L., Ionan, A., Chatterjee, M., Ye, B., et al. (2007) Autoantibody Approach for Serum Based Detection of Head and Neck Cancer. Cancer Epidemiology, Biomarkers & Prevention, 16, 2396-2405.

[25]   Piura, E. and Piura, B. (2011) Autoantibodies to Tailor-Made Panels of Tumor Associated Antigens in Breast Carcinoma. Journal of Oncology, 2011, Article ID: 982425.

[26]   Yahalom, G., Weiss, D., Novikov, I., Bevers, T.B., Radvanyi, L.G., Liu, M., et al. (2013) An Antibody-Based Blood Test Utilizing a Panel of Biomarkers as a New Method for Improved Breast Cancer Diagnosis. Biomarkers in Cancer, 5, 71-80.

[27]   Smith-Bindman, R., Chu, P., Miglioretti, D.L., Quale, C., Rosenberg, R.D., Cutter, G., et al. (2005) Physician Predictors of Mammographic Accuracy. Journal of the National Cancer Institute, 97, 358-367.

[28]   Beam, C.A., Conant, E.F., Sickles, E.A. and Weinstein, S.P. (2003) Evaluation of Proscriptive Health Care Policy Implementation in Screening Mammography. Radiology, 229, 534-540.

[29]   Chinni, S.R., Falchetto, R., Gercel-Taylor, C., Shabanowitz, J., Hunt, D.F. and Taylor, D.D. (1997) Humoral Immune Response to Cathepsin D and Glucose Regulated Protein 78 in Ovarian Cancer Patients. Clinical Cancer Research, 3, 1557-1564.

[30]   Lubin, R., Schlichtholz, B., Bengoufa, D., Zalcman, G., Trédaniel, J., Hirsch, A., et al. (1993) Analysis of p53 Antibodies in Patients with Various Cancers Define B-Cell Epitopes of Human p53: Distribution on Primary Structure and Exposure on Protein Surface. Cancer Research, 53, 5872-5876.

[31]   Hansen, M.H., Nielsen, H. and Ditzel, H.J. (2001) The Tumor-Infiltrating B Cell Response in Medullary Breast Cancer Is Oligoclonal and Directed against the Autoantigen Actin Exposed on the Surface of Apoptotic Cancer Cells. Proceedings of the National Academy of Sciences of the United States of America, 98, 12659-12664.

[32]   Erkanli, A.I., Taylor, D.D., Dean, D., Eksir, F., Egger, D., Geyer, J., et al. (2006) Application of Bayesian Modeling of Autologous Antibody Responses against Ovarian Tumor-Associated Antigens to Cancer Detection. Cancer Research, 66, 1792-1798.

[33]   Natoli, C., Iacobelli, S. and Kohn, L. (1996) The Immune Stimulatory Protein 90K Increases Major Histocompatibility Complex Class I Expression in a Human Breast Cancer Cell Line. Biochemical and Biophysical Research Communications, 225, 617-620.

[34]   Yavelsky, V., Rohkin, S., Shaco-Levy, R., Tzikinovsky, A., Amir, T., Kohn, H., et al. (2008) Native Human Autoantibodies Targeting GIPC1 Identify Differential Expression in Malignant Tumors of the Breast and Ovary. BMC Cancer, 8, 247-258.

[35]   Smyth, P.P., Shering, S.G., Kilbane, M.T., Murray, M.J., McDermott, E.W., Smith, D.F., O’Higgins, N.J., et al. (1998) Serum Thyroid Peroxidase Autoantibodies, Thyroid Volume, and Outcome in Breast Carcinoma. Journal of Clinical Endocrinology and Metabolism, 83, 2711-2716.

[36]   Nicolini, A., Capri, A. and Tarro, G. (2006) Biomolecular Markers of Breast Cancer. Frontiers in Bioscience, 11, 1818-1843.

[37]   Gold, P. and Freeman, S.O. (1965) Specific Carcino Embryonic Antigens of the Human Digestive System. Journal of Experimental Medicine, 122, 467-481.

[38]   Korneeva, I., Bongiovanni, A.M., Girotra, M., Caputo, T.A. and Witkin, S.S. (1999) Serum Antibodies to the 27-kd Heat Shock Protein in Women with Gynecologic Cancers. American Journal of Obstetrics and Gynecology, 183, 18-21.