Abstract P1-04-03: Host myeloid response to tumor and immunotherapy is associated with heterogeneity in outcomes to anti-PDL1
Authors: Hanna Ann , Sun Xiaopeng , Gonzalez-Ericsson Paula I. , Sanchez Violeta M. , Sanders Melinda E. , Balko Justin M. 
Published: March 2022

Abstract
Background: Immune checkpoint inhibitors (ICI) have improved patient overall and progression-free survival in some cancer types but yielded limited success in breast cancer. Phase-III clinical trials in triple negative breast cancer (TNBC) patients, who harbor extensive tumor-infiltrating lymphocytes within tumor stroma, have demonstrated increased progression-free survival (IMpassion130) and pathologic complete response (KEYNOTE-522). Consequently, combinations of ICI and chemotherapy have been FDA-approved for metastatic TNBC patients, and potentially in the early breast cancer setting. Despite FDA-approval, the therapeutic benefit of ICI alone and the most efficacious chemotherapy combinations are poorly characterized. Objective: We sought to model ICI response in vivo to elucidate the mechanisms responsible for immunotherapy efficacy in breast cancer and ascertain the therapeutic benefits of different chemotherapeutic combinations with ICI. Methods: In this study, we used an immunocompetent EMT6 orthotopic mammary tumor model to investigate the efficacy of single-agent immunotherapy and in combination with standard-of-care chemotherapy (paclitaxel [PAC] or doxorubicin [DOX]). We used single-cell RNA sequencing to analyze the cellular landscape of the primary tumor in response to combinatorial therapeutic strategies. Additionally, we serially sampled and analyzed peripheral blood from mice with differential responses by bulk and T-cell receptor (TCR) sequencing to identify systemic genetic alterations and T-cell expansion. Results: Single-agent anti-PD-L1 robustly suppressed primary tumor growth (p =0.0046) and extended survival (p&lt;0.0001) beyond the isotype control group. While either PAC or DOX demonstrated moderate therapeutic efficacy, neither agent potentiated single-agent anti-PD-L1 benefit. Interestingly, despite using a genetically identical tumor model and murine host, anti-PD-L1 induced heterogeneous responses, ranging from complete response to complete intrinsic resistance. The longitudinal analysis of peripheral blood from heterogeneously responding mice uncovered signatures of myeloid cell recruitment corresponding to transient responses ultimately converting to resistance. We also identified specific clonal T cell expansion present only in responders. Single-cell transcriptomic profiling of the tumor microenvironment revealed an increase of T cells and natural killer cells and reduction of regulatory T cells in the combination groups versus chemotherapy alone, although this did not translate into improved benefit. Finally, we performed gene-set enrichment analysis on infiltrating T cells and identified a robust signature of cytotoxic T cell activation characterized by a significant enrichment in inflammatory pathways in both single-agent anti-PD-L1 and in combination with chemotherapy. Conclusions: This study identifies a heterogeneously ICI-responsive in vivo model that emulates TNBC patient response to combinatorial ICI approaches. We describe the efficacy of single-agent ICI in upregulating cytotoxic immune cell infiltration and expansion within the primary tumor, thereby diminishing tumor growth and enhancing survival. Moreover, this study describes differential responses in a genetically similar host, which reflects heterogeneous patient response to ICI. Further characterization may identify systemic biomarkers and tumor antigen-specific T cell clones to accurately predict immunotherapy response in patients and uncover mechanisms for sensitizing tumors refractory to ICI. This study also has potentially significant clinical implications for re-evaluating the benefits of chemotherapy in combination with ICI in TNBC patients.
Citation Format: Ann Hanna, Xiaopeng Sun, Paula I. Gonzalez-Ericsson, Violeta M. Sanchez, Melinda E. Sanders, Justin M. Balko. Host myeloid response to tumor and immunotherapy is associated with heterogeneity in outcomes to anti-PDL1 [abstract]. In: Proceedings of the 2021 San Antonio Breast Cancer Symposium; 2021 Dec 7-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2022;82(4 Suppl):Abstract nr P1-04-03.