Abstract P4-06-23: Feasibility of sygeneic mice models of breast cancer for research of immune checkpoint blockades

Abstract

Background: With the increasing success of immune checkpoint blockades for cancer treatment, we increasingly need well-characterized preclinical models. Syngeneic mice models (with a fully competent immune system) have advantages that they are easily established and cost less, though they do not reflect genetic complexity of human tumors. We evaluated feasibility of syngeneic mice models of breast cancer by analyzing efficacy of immune checkpoint blockade and dynamic change of tumor immune microenvironment. Methods: We used syngeneic mice model of JC, 4T1, and EMT6 cells, which are all murine triple negative breast cancer in BALB/c mice. At the time when subcutaneous tumors reach at 50˜100mm^3, each mice models were divided into 2 groups for treatment versus no-treatment control. In the treatment group, mice version of anti-PD-1 antibody was intraperitoneally injected (q 3 days, x 6). Anti-tumor efficacy was monitored by measuring tumor volume. 9Tumor response9 was defined as a case with tumor volume less than that of control group by a standard error at a determined time point. Immune microenvironment was evaluated by measuring serum cytokines (IL-2, IL-6, IL-10, IFNγ, and TNFα) with legendplex and immune cells (CD3, CD4, CD8, CD56, and FOXP3) of peripheral blood with FACS before injection of PD-1 blockade, after 1st injection, and when euthanized. Tumor-infiltrating immune cells were evaluated with FACS, when euthanized. Results: The tumor response rate to PD-1 blockade was highest in the 4T1 model (54.5%, 6/11) compared to JC model (40%, 4/10) or EMT6 model (36.4%, 4/11). Bleeding 3 times and tumor obtainment when euthanized in each mouse were feasible for profiling of cytokines and immune cells. Although before treatment with PD-1 blockade, CD3+T cells in peripheral blood were slightly lower in 4T1 model (18.3±8.1%) than JC model (24.6±4.7%) or EMT6 model (27.9±6.3%), after injection of one dose of PD-1 blockade, CD3+T cells increased 1.5 times in 4T1 model (18.3% to 27.3%), whereas those CD3+T cells decreased slightly in JC model and EMT6 model. Dynamic changes were not observed in other subsets of peripheral immune cells in all 3 models. Serum TNFα (with statistical significance) and IFNγ (with borderline significance) were higher in responders than in non-responders or no-treatment control. Conclusions: Syngeneic mice models of breast cancer were feasible to investigate immune checkpoint blockades and monitor dynamic change of immune microenvironment. In this regard, such models may be used to evaluate immune checkpoint blockade-based combination therapy as well. Citation Format: Moon YW, Park N, Hur J, Pandey K, Cho YB, Kim SK, Lee SA, Son GW, Jo JM, An H-J. Feasibility of sygeneic mice models of breast cancer for research of immune checkpoint blockades [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 P4-06-23.