Abstract P2-09-10: Epigenetic modulation—unlocking the potential of checkpoint inhibition in breast cancer
Authors: Roussos Torres ET , Rafie C , Armstrong T , Jaffee EM 
Published: May 2019

Immune checkpoint inhibition (ICI) has revolutionized treatment in cancers that are naturally immunogenic by enabling infiltration of T cells into the tumor microenvironment (TME) and promoting cytotoxic signaling pathways. Tumors possessing complex immunosuppressive TME's such as breast cancer present unique therapeutic obstacles as response rates to ICI remain low. Such tumors often recruit myeloid-derived suppressor cells (MDSCs) whose functioning prohibits both T cell activation and infiltration. To date, most studies focus on use of ICI in triple negative disease. Our work aims to uncover the efficacy of ICI in both early and advanced HER2 positive (HER2+) disease and to advance our understanding of how to improve response rates to these new promising therapies.

We are using a HER-2/neu transgenic mouse model with tumor challenge of syngeneic cell lines to test the efficacy of different combinations of an epigenetic agent, the histone deacetylase inhibitor entinostat (ENT), checkpoint inhibitors anti-PD-1 and anti- CTLA-4, on primary and metastatic disease. We are examining treatment effects on primary tumor growth, metastatic burden, and survival. Characterization of tumor infiltrating lymphocytes and their functional capabilities are being investigated using fluorescence-activated cell sorting, gene expression profiling, and ex vivo suppression assays. Western blots, qPCR and other in vitro assays will be performed on MDSCs to investigate mechanisms behind response.

In the HER2+ mouse model of early stage disease, we show that combining ENT, with ICis significantly improves survival and delays tumor growth. Preliminary data in models of advanced disease, show only ENT + a-PD-1 improves survival and metastatic burden. Conversely, in the metastatic model, ENT + a-CTLA-4 negatively effects survival and metastatic burden. In primary tumors, ENT + ICIs leads to significantly decreased suppression by granulocytic-MDSCs. However, MDSC infiltration and function is not affected in lungs containing macrometastatic disease. Interestingly, we found an increase in activated granzyme-B-producing CD8+ T effector cells in mice treated with combination therapy in both primary and metastatic tumors. Finally, gene expression profiling of MDSCs from primary tumors identified significant changes in immune-related pathways, and identified a common downstream regulator –STAT3. Studies are ongoing to evaluate the mechanistic role of STAT3 in the response observed in primary tumors and to determine if STAT3 is involved in response in the metastatic setting.

In summary, addition of ENT to ICIs significantly affects overall survival in early stage models of HER2+ breast cancer however, only addition of a-PD-1 to ENT is beneficial in models of advanced disease. Additionally, the mechanism of action in early stage disease involves altered infiltration and function of MDSCs, allowing for a more robust adaptive immune response. However, a different mechanism of action is likely responsible for the responses seen in advanced stage disease. These novel findings provide a rationale for combination therapy in patients with HER2+ breast cancer and suggest responses to this combination therapy are linked to stage of disease likely due to different mechanisms of action.

Citation Format: Roussos Torres ET, Rafie C, Armstrong T, Jaffee EM. Epigenetic modulation—unlocking the potential of checkpoint inhibition in breast cancer [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 P2-09-10.