Quantitative multiplexed analysis of ErbB family coexpression for primary breast cancer prognosis in a large retrospective cohort

• Published in: Cancer 2009-06, Vol.115 (11), p.2400-2409
• Main Authors: Giltnane, Jennifer M., Moeder, Christopher B., Camp, Robert L., Rimm, David L.
• Format: Article
• Language: English
• Subjects: automated quantitative analysis; Biological and medical sciences; Biomarkers, Tumor - analysis; Breast Neoplasms - metabolism; Breast Neoplasms - mortality; Cell Line, Tumor; epidermal growth factor receptor; Female; Gynecology. Andrology. Obstetrics; HER‐2; HER‐3; HER‐4; Humans; immunohistochemistry; Mammary gland diseases; Medical sciences; Middle Aged; Prognosis; Receptor, Epidermal Growth Factor - analysis; Receptor, ErbB-2 - analysis; Receptor, ErbB-3 - analysis; Receptor, ErbB-4; Receptors, Estrogen - analysis; Receptors, Progesterone - analysis; survival; Tissue Array Analysis; Tumors
• ISSN: 0008-543X; 1097-0142
• DOI: 10.1002/cncr.24277

BACKGROUND: Assessment of outcome using a single prognostic or predictive marker is the current basis of targeted therapy, but is inherently limited by its simplicity. Multiplexing has provided better classification, but has only been done quantitatively using RNA or DNA. Automated quantitative analysis is a new technology that allows quantitative in situ assessment of protein expression. The authors hypothesized that multiplexed quantitative measurement of ErbB receptor family proteins may allow better prediction of outcome. METHODS: The authors quantitatively assessed the expression of 6 proteins in 4 subcellular compartments in 676 patients using breast carcinoma tissue microarrays. Then using Cox proportional hazards modeling and unsupervised hierarchical clustering, they assessed the prognostic value of the expression singly and multiplexed. RESULTS: Epidermal growth factor receptor (EGFR), HER‐2, and HER‐3 expression were associated with decreased survival. Multivariate analysis showed high HER‐2 and HER‐3 expression maintained independence as prognostic markers. Hierarchical clustering of expression data defined a small class enriched for HER‐2 expression with 40% 10‐year survival, compared with 55% using conventional methods. Clustering also revealed a similarly poor‐prognostic subgroup coexpressing EGFR and HER‐3 (but low for estrogen receptor, progesterone receptor, and HER‐2) with a 42% 10‐year survival. CONCLUSIONS: This work shows that the combined analysis of protein expression improved prognostic classification, and that multiplexed models were superior to any single‐marker–based method for prediction of 10‐year survival. These methods illustrate a protein‐based, multiplexed approach that could more accurately identify patients for targeted therapies. Cancer 2009. © 2009 American Cancer Society. Automated fluorescence‐based quantitative analysis of breast cancer tissue microarrays allows for multiplexing and clustering of protein expression data. Analysis of the 4 ErbB family proteins along with estrogen and progesterone receptor allows definition of a subset of HER‐2–positive cases with worse outcome.