Abstract LB-230: Refining treatment recommendations for lymph node-negative breast cancer patients using a novel protein-based prognostic signature: The OncoMasTR assay
Authors: Arman Rahman, Seodhna Lynch, Nebras Alattar, Niamh Niamh, Charles Weige, Romina Silva, Claudia Aura, Fiona Lanigan, Adrian Bracken, Björn Nodin, Karin Jirström, William Gallagher
Journal: Cancer Research (2018)

Abstract Introduction Traditionally, many women with lymph node-negative breast cancer (LN-BC) are unnecessarily treated with chemotherapy. This highlights the importance of accurately predicting patient prognosis. We previously identified a novel prognostic signature encompassing a set of master transcriptional regulators (MTRs), termed OncoMasTR. At the mRNA level, OncoMasTR predicted recurrence risk for LN-BC better than current prognostic signatures and accurately classifies &amp;gt;50% of patients as ‘low-risk', thus potentially reducing overtreatment. Here, we describe ongoing validation of the OncoMasTR biomarker signature at the protein level. Methods Commercially available antibodies targeted against markers were validated by Western blotting and immunohistochemistry (IHC) using CRISPR/Cas9 knockout cell-lines and full-face BC sections. IHC was performed on tissue microarrays (TMAs) containing multiple tumour cores from BC patients. Visiopharm's Oncotopix algorithm was used for image analysis, with stained sections verified manually by a pathologist. Image analysis data was obtained, and cut-off points optimised by maximum Chi-square values were used to differentiate low and high expression of individual markers. This was combined with clinical data to generate Kaplan-Meier survival curves. Multivariate analysis was used to determine the optimal combination of biomarkers. Results Antibodies against eight components of the OncoMasTR signature have been validated. IHC analysis demonstrated highly-specific nuclear staining for seven of the markers (UHRF1, ATAD2, HMGB2, E2F1, TCF19, MYBL2, PTTG1), whilst one showed both nuclear and cytoplasmic staining (p16). Image analysis for UHRF1, ATAD2 and HMGB2, using both manual and automated histological scores, showed that high expression of these biomarkers is linked with reduced survival. Additionally, UHRF1 was confirmed to have the same survival correlation in another independent cohort. Interestingly, high nuclear expression of p16 is linked with prolonged survival whilst high cytoplasmic expression of p16 is associated with poorer survival, aligning with previous reports. Conclusions The OncoMasTR signature offers a more accurate stratification of low-vs-high risk, thus showing promise in improving prognosis prediction for early-stage BC. This will translate to reduced overtreatment, decreased costs, improved survival rates and enhanced quality of life for patients. Citation Format: Arman Rahman, Seodhna Lynch, Nebras Alattar, Niamh Niamh, Charles Weige,, Romina Silva, Claudia Aura, Fiona Lanigan, Adrian Bracken, Björn Nodin, Karin Jirström, William Gallagher. Refining treatment recommendations for lymph node-negative breast cancer patients using a novel protein-based prognostic signature: The OncoMasTR assay [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr LB-230.