Abstract A030: Epigenetic immune modulation by entinostat in breast cancer: Correlative analysis of ENCORE 301 trial
Authors: Ordentlich Peter , Lee Min-Jung , Tomita Yusuke , Lee Sunmin , Tomita Saori , Cruickshank Scott , Chumsri Saranya , Trepel Jane B. 
Published: May 2016

Background: Entinostat, a class I HDAC inhibitor (HDACi), has shown promising activity in ENCORE 301, a randomized, placebo-controlled, phase II trial of entinostat + exemestane (EE) vs. exemestane + placebo (EP) in advanced hormone receptor-positive breast cancer progressed on nonsteroidal aromatase inhibitors. ENCORE 301 met the primary progression free survival endpoint and showed a median 8.3-month improvement in the overall survival (OS) exploratory endpoint for the EE arm. Emerging preclinical work suggests that entinostat has immunomodulatory effects and can eradicate modestly immunogenic mouse tumors in combination with immune checkpoint blockade agents via reduction of circulating myeloid-derived suppressor cells (MDSC). Based on these data, we conducted an analysis of immune subsets in blood samples from ENCORE 301 breast cancer patients.

Method: Blood was collected from a subset of 49 patients (27 EE and 22 EP) representative of the 130 patients enrolled in ENCORE 301 on cycle 1 day 1 (C1D1; pre-treatment), C1D2, C1D8, and C1D15 for biomarker analysis. Of these, 34 patient samples (20 EE and 14 EP) were analyzed for circulating immune subsets. The percent change in subsets at C1D15 vs. baseline was assessed based on the following surface markers: Lin-MDSC (lin; CD3, CD19, CD56)-HLA-DR-CD11b+CD33+), granulocytic MDSC (CD14-CD11b+CD33+), monocytic MDSC (Lin-HLA-DR-CD11b+CD33+CD14+), immature MDSC (Lin-HLA-DR-CD11b+CD33+CD14-), CD8+ T-cells (CD4-CD8+), Foxp3-CD4+ T-cells (CD8-CD4+Foxp3-), and Tregs (CD4+CD8-CD25hiFoxp3+). Monocytes were analyzed for three populations: CD14+, CD14+HLA-DRhi, and CD14+HLA-DRlow/negative. In addition, PD-1, CTLA-4, and TIM-3 were measured on T-cell subsets, and CD40 was measured on MDSCs.

Results: In line with preclinical data, we observed a significant reduction in granulocytic MDSC (-14.67% vs. +20.56%, p 0.029) and monocytic MDSC (-62.3% vs. +1.97%, p 0.002) in EE. Entinostat did not alter immature MDSC levels (-20.9% vs. -15.0%, p 0.93) suggesting a downstream effect of entinostat on MDSC subsets. Interestingly, CD40, a costimulatory receptor required for MDSC-mediated immune suppression was significantly down-regulated in all MDSC subsets except granulocytic MDSC where a downward trend was observed. Entinostat did not significantly impact the ratio of CD8+ T-cells per CD4+ T-cells or per Tregs or alter expression of CTLA4, PD-1, or TIM3 on T-cell subsets. Reduced expression of HLA-DR on monocytes has been associated with poor prognosis in cancer. Consistent with entinostat-mediated immunomodulatory effects, a significant increase in the number of HLA-DR+ monocytes (34.1% vs. -11.38%, p 0.0004) and level of HLA-DR expression on monocytes (16.3% vs. -4.7%; p 0.015) was observed.

Conclusion: Data with entinostat combined with exemestane in ENCORE 301 provide the first evidence of HDACi-mediated reduction of immunosuppressive MDSCs and increased immunocompetent CD14+HLA-DRhi monocytes in patients. These findings may explain the improved OS seen with EE in ENCORE 301 and provide strong rationale for planned combination studies of entinostat with immune checkpoint blockade agents.

Citation Format: Peter Ordentlich, Min-Jung Lee, Yusuke Tomita, Sunmin Lee, Saori Tomita, Scott Cruickshank, Saranya Chumsri, Jane B. Trepel. Epigenetic immune modulation by entinostat in breast cancer: Correlative analysis of ENCORE 301 trial. [abstract]. In: Proceedings of the CRI-CIMT-EATI-AACR Inaugural International Cancer Immunotherapy Conference: Translating Science into Survival; September 16-19, 2015; New York, NY. Philadelphia (PA): AACR; Cancer Immunol Res 2016;4(1 Suppl):Abstract nr A030.