Abstract PR02: β-Catenin overexpression underlies the aggressive disease course in African American triple-negative breast cancer patients who lack androgen receptor
Authors: Mittal Karuna , Bhattarai Shristi , Klimov Sergey , Krishnamurthi Uma , Li Xiaoxian , Wetherilt Ceyda Sonmez , Aleskandaran Mohammad A. , Green Andrew A. , Rakha Emad A. , Ellis Ian O. , Cantuaria Guilherme , Wei Guanhao , Osan Remus Mihai , Gupta Meenakshi V. , Manne Upender , Rida Padmashree C.G , Aneja Ritu 
Published: February 2019

Background: Androgen receptor (AR) has emerged as a new target for treating TNBC. AR is expressed in 10-43% of TNBCs. Although there are conflicting reports in the literature about the effect of AR status on TNBC prognosis, agents targeting AR signaling (enzalutamide) are already being evaluated in AR-positive TNBCs in early-stage clinical trials. However, no study so far has evaluated the association/correlation of AR status with ethnicity in TNBCs and downstream effects of AR loss in TNBCs. Given the association of AR loss with poor prognosis in breast cancer and that the African American (AA) with TNBC suffers aggressive disease course when compared to European American (EA) TNBCs, we hypothesized that AR loss might be an underlying cause of aggressive disease course in AR-negative TNBCs. Thus, in this project we aimed to study if loss or gain of AR in AA and EA TNBCs regulates the expression of β-catenin and leads to more aggressive disease course by activating downstream canonical Wnt-beta catenin signaling.

Methods: We evaluated AR expression immunohistochemically in 424 formalin-fixed, paraffin-embedded samples from TNBC patients for whom complete clinicopathologic and overall survival (OS) data were available. Samples with &lt;1% nuclear-stained cells were considered quadruple-negative. We also performed gene set enrichment analysis for the AR low and high group in AA and EA TNBCs using the publicly available TCGA gene dataset. Finally, we compared levels of 187 proteins with suspected involvement in breast tumorigenesis in AR-high vs AR-low TNBCs (using median AR expression as a cutpoint, as the IHC-derived AR status of these samples was unknown).

Results: IHC staining of AR indicated that 79.5% of AA TNBCs (n=214) and 70% of EA TNBCs (n=210) were AR negative. Loss of AR was associated with poor overall survival in adjuvant-treated high Ki67 (&gt;14%) (HR=1.72; p=0.095) AA TNBC (n=98) when compared to EA TNBCs (n=80). These data were validated by our in silico findings, which suggested that EA TNBCs (n=81) exhibited higher levels of AR mRNA compared to AA TNBCs (n=41) (p&lt;0.05). Similarly, AR protein expression was higher in EA TNBCs (n=75) than AA TNBCs (n=32) (p&lt;0.05). We further observed that β-catenin protein levels are higher in AA AR-low TNBCs compared with AA AR-high TNBCs (median AR expression used as cut point; p&lt;0.05). This was strengthened by our observation in a cohort of 142 TNBCs wherein AAs with AR-negative TNBC showed a preponderance of cells with nuclear β-catenin staining via IHC compared with EA TNBCs that lack AR. Our GSEA analysis results indicated that Wnt/β-catenin signaling was the top-enriched gene ontology in the AR-low subgroup compared to AR-high subgroup of TNBCs. Furthermore, proteomic data revealed that β-catenin and Dvl3 expression was significantly upregulated in the AR-low subgroup when compared with AR-high group, suggesting that Wnt signaling is in an overdrive in the AR-low subgroup, especially in AA TNBCs.

Conclusion: This study suggests that increased expression of β-catenin coupled with AR loss in AAs may underlie the ethnic disparity in outcomes among TNBC patients and strongly supports the prognostic role of AR and β-catenin in this breast cancer subtype.

Citation Format: Karuna Mittal, Shristi Bhattarai, Sergey Klimov, Uma Krishnamurthi, Xiaoxian Li, Ceyda Sonmez Wetherilt, Mohammad A. Aleskandaran, Andrew A. Green, Emad A. Rakha, Ian O. Ellis, Guilherme Cantuaria, Guanhao Wei, Remus Mihai Osan, Meenakshi V. Gupta, Upender Manne, Padmashree C.G Rida, Ritu Aneja. β-Catenin overexpression underlies the aggressive disease course in African American triple-negative breast cancer patients who lack androgen receptor [abstract]. In: Proceedings of the Tenth AACR Conference on the Science of Cancer Health Disparities in Racial/Ethnic Minorities and the Medically Underserved; 2017 Sep 25-28; Atlanta, GA. Philadelphia (PA): AACR; Cancer Epidemiol Biomarkers Prev 2018;27(7 Suppl):Abstract nr PR02.