Background
Immune checkpoint inhibitors (ICIs), such as anti-PD1 therapy, have achieved remarkable success in melanoma and lung cancer but show limited efficacy in breast cancer [1-4]. Tumor infiltration of ICOS-positive (ICOS⁺) regulatory T cells (Tregs) contributes to immune suppression by enhancing PD-1 expression on cytotoxic CD8⁺ T and natural killer (NK) cells. Therefore, blocking the ICOS receptor, which is required for Treg cell activity, could be an effective strategy to reverse the cytotoxic function of immune cells [5, 6]. This study investigates ICOS targeting as a therapeutic strategy in preclinical mammary tumor models.
Methods
Orthotopic syngeneic mammary tumor models (EO771.LMB; hormone receptor-negative) were established in Icos-/- C57BL/6 female mice to understand Icos-driven immunosuppressive mechanisms in breast tumors. The therapeutic efficacy of blocking the Icos receptor was evaluated using monoclonal antibodies in EO771.LMB and 67NR (hormone-receptor positive) models in combination with anti-PD1 therapy. Flow cytometry and single-cell RNA sequencing were used to analyze immune cell modulation. Multiplex immunohistochemistry and spatial transcriptomic analysis of human clinical specimens characterized the cellular and molecular interactions of ICOS+ Tregs in the tumor microenvironment.
Results
Systemic deletion of Icos resulted in a significant reduction in mammary tumor growth, associated with decreased Treg activation and function, and increased infiltration and activity of CD8+ T and NK cells. Moreover, the combination treatment (anti-Icos + anti-PD-1) demonstrated a substantial reduction in mammary tumor burden in both EO771.LMB and 67NR models, indicating synergistic anti-tumor activity through simultaneous depletion of Icos⁺ Treg cells and reinvigoration of exhausted cytotoxic cells. Additionally, combination immunotherapy demonstrated improved metastasis-free survival in both neoadjuvant and adjuvant treatment settings. Spatial-omics data provide a detailed map of ICOS⁺ Treg enrichment in the tumor core, forming immunosuppressive niches with ICOS-Ligand+ cancer cells and exhausted CD8+ T cells.
Conclusions
Pharmacological blockade of the ICOS receptor can suppress Treg-induced immunosuppression, sensitizing breast cancer cells to immunotherapy-driven anti-tumor responses. Hence, an anti-ICOS and anti-PD1 combination may provide a promising therapeutic approach for patients with early-stage or advanced breast cancer. Additionally, integrating ICOS⁺ Tregs into TIL scoring systems would aid in stratifying breast cancer patients for future ICOS-targeted immunotherapy trials.