Abstract P2-20-01: Patient-derived triple-negative breast cancer organoids as a platform for glucocorticoid receptor- mediated mechanisms of immunotherapy response and resistance
Authors: Shiang Christine , Frerich Candace , Durdana Ishrat , Lewis Cheryl , Bennett Lynda B. , Arteaga Carlos , Conzen Suzanne D. 
Published: March 2023

Background: In early-stage triple-negative breast cancer (TNBC), high glucocorticoid receptor (GR) expression correlates with worse relapse-free survival across TNBC subtypes, induces oncogenic gene expression and tumor cell survival, and potentiates anti-inflammatory or an anti-immunogenic phenotype. Immune checkpoint therapy has increased pathologic complete response rates in the neoadjuvant setting, however response to immunotherapy remains modest overall and the explanatory mechanisms for this phenomenon are incompletely understood. We sought a three-dimensional model of the TNBC immune microenvironment using patient-derived organoids (PDOs) that capture tumor heterogeneity, partially recapitulates tumor microenvironment, and can be established for longer-term in vitro/in-vivo translational study after exposure to immunotherapy. We hypothesize that during chemoimmunotherapy treatment of early-stage triple-negative breast cancer, high glucocorticoid receptor activity mediates an immunosuppressive phenotype and glucocorticoid receptor modulation can restore anti-tumor immunity. Methods and Results: We are successfully growing 5 PDOs out of 8 surgically resectedion samples tumors from patients with early-stage triple-negative breast cancer treated with chemotherapy vs. chemoimmunotherapy as per KEYNOTE-522. Interrogation of complementary TNBC cell line data revealed that GR activation downregulates the expression of immune checkpoint genes (PD-L1, B7-H3, B7-H4), while GR knockdown or treatment with a selective GR modulator restores expression of these immune checkpoint genes. TNBC-intrinsic GR activation increases the proportion of regulatory T cells vs. CD8+ T cells. In a coculture system of TNBC spheroids and T cells, GR activation in TNBC results in decreased activated CD8+ CD137+ T cells and use of the selective GR modulator increases the proportion of activated CD8+ CD137+ T cells. Conclusions and Future Directions: In early-stage triple-negative breast cancer, GR activation downregulates expression of immune checkpoint genes and reduction of GR activity restores expression. In a coculture system that recapitulates T cell repertoire found in triple-negative breast cancer, tumor-intrinsic GR activation results in increased regulatory T cell population and reduction of GR activity increases activated CD8+ CD137+ T cells. Ongoing analyses of TNBC organoid-immune coculture systems modeling high GR states will be used to (i) define cooperativity amongst immune checkpoint proteins in costimulation vs. coinhibition of anti-tumor responses and (ii) define novel pairs of receptor-ligands amongst TNBC:CD8+ vs. TNBC:Treg vs. CD8+:Treg as discovery of potential new therapeutic targets.

Citation Format: Christine Shiang, Candace Frerich, Ishrat Durdana, Cheryl Lewis, Lynda B. Bennett, Carlos Arteaga, Suzanne D. Conzen. Patient-derived triple-negative breast cancer organoids as a platform for glucocorticoid receptor- mediated mechanisms of immunotherapy response and resistance [abstract]. In: Proceedings of the 2022 San Antonio Breast Cancer Symposium; 2022 Dec 6-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2023;83(5 Suppl):Abstract nr P2-20-01.