Abstract PO1-26-06: The effect of localized radiotherapy in combination with chemoimmunotherapy in a humanized triple-negative breast cancer mouse model
Authors: Schreiber Anna , Bagby Stacey , Smoots Stephen , Dominguez Adrian , Fisher Christine , Lang Julie , Pitts Todd , Diamond Jennifer 
Published: May 2024

Background: Metastatic triple-negative breast cancer (TNBC) is an aggressive sub-type of breast cancer that is challenging to treat. While the addition of immunotherapy (IT) with pembrolizumab (pembro) to chemotherapy prolongs overall survival in patients with programmed death-ligand 1 positive (PD-L1+) TNBC, few patients obtain long-term benefit. Radiotherapy (RT) can stimulate cellular damage, release of tumor antigens and promote the T cell anti-tumor response including at distant sites (abscopal response) when used with IT. The purpose of this study was to investigate the combination of RT with pembro and paclitaxel (pac) in a TNBC humanized mouse model.

Methods: Human Immune System BRGS (BALB/c, Rag2-/-, IL2RγC-/-, NODSIRPa ) mice were engrafted with TNBC MDA-MB-231 cells in bilateral flanks. Treatments included: vehicle, pac 10 mg/kg intra-peritoneal (IP) weekly, pembro 15 mg/kg IP weekly, and RT 8 Gy x 3 fractions every other day the first week of treatment to one-sided tumors (contralateral tumors received no RT). Mice were treated and analyzed in the following groups: vehicle, pac + pembro, RT + pac, and RT + pac + pembro. Treatments began when tumors reached 100-300 mm3 and tumors were measured twice weekly. At the end of study, sera, lymph nodes (LNs), spleen, and tumor tissue were collected for immunohistochemistry, single cell suspensions, and flow cytometry analysis for immune cell populations.

Results: All treatment arms resulted in a decrease in tumor volume when compared to vehicle with the greatest reduction in the RT + pac and RT + pac + pembro arms. Combination therapy with RT + paclitaxel (p=0.0002) and RT + pac + pembro (p=0.008) resulted in a statistically significant decrease in specific growth rate (SGR) when compared to vehicle. An abscopal effect was not observed in the non-RT tumors compared to RT-tumors in either group. RT + pembro was associated with an increase in hCD45 T cells in LNs and spleens but not in tumors. In mice that received RT, a decrease was noted in HLA-ABC class I and Intermediate Class II major histocompatibility complex (MHC). In addition, RT-tumors had reduced expression of PD-L1 and polio virus receptor (PVR) and increased expression of PD-1. The percentage of interferon gamma (IFNg+), tumor necrosis factor alpha (TNFa+) CD8+ T cells increased with radiation. This percentage was the highest in the triplet therapy with RT+ pembro + pac.

Conclusion: Combination treatment with RT + immunochemotherapy resulted in a significant increase in anti-tumor activity when compared to immunochemotherapy alone, however, an abscopal effect was not observed in contralateral non-RT tumors. RT resulted in down-regulation of immune inhibitory receptors and tumor MHC.

Citation Format: Anna Schreiber, Stacey Bagby, Stephen Smoots, Adrian Dominguez, Christine Fisher, Julie Lang, Todd Pitts, Jennifer Diamond. The effect of localized radiotherapy in combination with chemoimmunotherapy in a humanized triple-negative breast cancer mouse model [abstract]. In: Proceedings of the 2023 San Antonio Breast Cancer Symposium; 2023 Dec 5-9; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2024;84(9 Suppl):Abstract nr PO1-26-06.