Abstract P2-03-03: BRG1-SOX4 mediates a novel and essential signaling network that activates PI3K/Akt signaling in TNBC
Authors: Gatza ML , Angus SP , Khella CA , Tong K , Verzi M , Mehta GA 
Published: May 2019

Background:

Triple negative (TNBC) breast cancer, which is largely synonymous with the basal-like molecular subtype, is an aggressive malignancy that accounts for nearly 1 in 4 breast cancer related deaths and disproportionately affects younger women and women of African American decent. Given the lack of drug-able targets expressed by TNBC tumors, few therapeutic options exist beyond currently utilized cytotoxic therapies, and the overall prognosis for these patients remains poor. While TNBC tumors are characterized by high PI3K signaling, clinical trials targeting this pathway have had limited success. Therefore, identifying mechanisms driving key oncogenic pathways, including PI3K, is paramount to understanding the transformation process and enabling the development of rational, personalized therapeutic regimens.

Methods:

We utilized a PI3K gene expression signature as a conceptual framework to analyze genome-wide mRNA expression and DNA copy number data from human breast tumors to identify genetic drivers of PI3K/Akt signaling. Kinome profiling was used to identify changes in the drug-able kinome regulated by these genes and in vitro studies were used to delineate mechanisms by which identified genes mediate oncogenic PI3K signaling in TNBC.

Results:

Integrative proteogenomic analyses of orthogonal genome-wide data from ˜3,000 human tumors from the TCGA and METABRIC studies identified amplification and overexpression of the oncogenic transcription factor SOX4 as well as the SWI/SNF ATPase BRG1 in tumors with high PI3K activity. These alterations were predominantly expressed in TNBC or basal-like breast tumors. Chromatin immunoprecipitation followed by DNA sequencing (ChIPseq) as well as shRNA-based studies confirmed that BRG1 regulates SOX4 expression in TNBC cell lines. Analyses of data from a genome-wide RNA interference (RNAi) screen in 27 breast cancer cell lines further indicated that SOX4 is essential in cell lines with high PI3K activity and this was confirmed by colony formation and cell proliferation assays. Importantly, in vitro analyses confirmed that both BRG1 and SOX4 regulate Akt phosphorylation and down-stream signaling. Profiling of the drug-able kinome in SOX4 depleted cell lines compared to control cells using Multiplexed kinase Inhibitor Beads couple with quantitative Mass Spectrometry (MIB/MS) identified 21 drug-able kinases regulated by SOX4 activity including TGFBR2 which has been previously shown to regulate PI3K activity. RNA sequencing (RNAseq) and RT-PCR analyses confirmed that SOX4 mediates TGFBR2 mRNA levels and co-immunoprecipitation experiments in conjunction with ChIP assays demonstrated that BRG1 and SOX4 form a complex at the TGFBR2 promoter and enhancer region to regulate TGFBR2 expression.

Conclusions:

In this study, we demonstrated that BRG1-SOX4 constitutes a novel and essential signaling pathway which promotes PI3K/Akt activity through TGFβ signaling in TNBC/basal-like breast cancer and leads to activation of additional, drug-able kinases. Given the essentiality of BRG1 and SOX4, our data suggest that targeting this interaction and/or the down-stream components of this pathway may represent a novel therapeutic strategy in TNBC.

Citation Format: Gatza ML, Angus SP, Khella CA, Tong K, Verzi M, Mehta GA. BRG1-SOX4 mediates a novel and essential signaling network that activates PI3K/Akt signaling in TNBC [abstract]. In: Proceedings of the 2018 San Antonio Breast Cancer Symposium; 2018 Dec 4-8; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2019;79(4 Suppl):Abstract nr P2-03-03.