P2-11-08: Clinical Value of Combination Assay for Quantitative Determination of Cancer Biomarkers C2P and uPA/PAI-1 for Disease Recurrence Prediction of Early Breast Cancer Patients.
Authors: Schmitt M , Kiechle M , Schwarz-Boeger U , Langer R , Nakayama S , Matsushima T , Ishihara H 
Published: June 2012

Abstract
[Background] C2P is an assay measuring specific activities (SA; kinase activity compensated by its protein expression) of cyclin-dependent kinases CDK1 and CDK2 by examining a small piece of fresh-frozen tumor tissue. We reported previously that the C2P risk score given by CDK1SA and CDK2SA is a potent prognostic factor in node-negative breast cancer patients. Likewise, a uPA/PAI-1 ELISA test (FEMTELLE®, American Diagnostica Inc. Stamford, CT) quantitatively determines uPA (urokinase-type plasminogen activator) and PAI-1 (plasminogen activator inhibitor type-1) antigen levels in tumor tissue extracts, to identify patients with high or low risk of disease recurrence of node-negative breast cancer patients. These cancer biomarkers are recommended by the ASCO at the highest level of evidence (LOE-1) for therapy decision making in node-negative breast cancer patients. From the biological point of view, the above two assays can be placed into two categories: tumor cell proliferation for C2P and tumor cell invasion for uPA/PAI-1. This fact led us to examine the concept of combination of the two assays to better select breast cancer patients at risk. [Results] Fifty-nine cases of frozen primary breast cancer tissues were subjected to the C2P assay in a blinded manner. Twenty-one cases (40%) were categorized into “high risk”, 7 cases (14%) into “intermediate risk”, and 24 cases (46%) into “low risk”. Seven cases were judged as “not informative”. The uPA/PAI-1 results and clinical information (26: recurrent cases, 30: non-recurrent cases, 1: stage IV, 2: unknown) were provided by the TUM tumor bank. uPA/PAI-1 risk categories of 36 cases (61%) were classified “high” and 23 cases (39%) categorized “low”. C2P and uPA/PAI-1 showed statistically significant correlation to histological grading (Pearson correlation coefficient; 0.45 and 0.40, respectively). No significant correlation was observed between C2P and uPA/PAI-1. By Kaplan-Meier analysis for disease-free survival, in cases treated with endocrine therapy only, both C2P and uPA/PAI-1 showed a reproducible trend to the respective claimed performances as prognostic factors. In the combination analysis of the two parameters, where low/low was judged as “low” and the others as “high”, 11 cases (24%) were categorized into “low” and 34 cases (76%) into “high”. The sensitivity and negative predictive value for disease recurrence were 90% (19/20) and 91% (10/11), respectively. Strong statistical significance was observed between the risk categories by the log-rank test; p=0.0089, and also by Cox proportional hazards regression analysis; HR=9.18, p=0.032. By multivariate analysis, also including tumor size and nodal status, the CP2 uPA/PAI-1 combination evolved as a significant, statistically independent parameter (HR: 6.51, p&lt;0.01). [Conclusion] In this investigative study, the significance of the combination concept was strongly suggested for early breast cancer patients treated with endocrine therapy only. Yet, prior to implementation in the clinical setting, the practical performance of the combination assay should be validated by investigating an independent patient cohort.
Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P2-11-08.