Immunomodulatory effects of tumor Lactate Dehydrogenase C (LDHC) in breast cancer.

Abstract

Background: Immunotherapy has significantly improved outcomes for cancer patients; however, its clinical benefits vary among patients and its efficacy across breast cancer subtypes remains unclear. To enhance immunotherapy efficacy, it is important to gain more insight into tumor-intrinsic immunomodulatory factors that could serve as therapeutic targets. We previously identified Lactate Dehydrogenase C (LDHC) as a promising anti-cancer target due to its role in regulating cancer cell genomic integrity. In this study, we investigated the effects of tumor LDHC expression on immune responses.

Methods: TIMER AND TIDE deconvolution methods were used to investigate the relationship between tumor LDHC expression, immune cell infiltration and T cell dysfunction. Multiplex cytokine assays and flow cytometry were used to assess the effect of LDHC knockdown on the secretion of inflammatory molecules and expression of immune checkpoint molecules in breast cancer cells and cancer cell-immune cell co-cultures. T cell activity was determined by IFN-γ ELISPot assays and 7-AAD flow cytometry.

Results: TIMER and TIDE analyses revealed that tumor LDHC expression is associated with T cell dysfunction in breast cancer and poorer post-immunotherapy survival in melanoma. Silencing LDHC in breast cancer cell lines (MDA-MB-468, BT-549, HCC-1954) enhanced early T cell activation and cytolytic activity. To gain a better understanding of the underlying mechanisms, comparative analysis of the effects of LDHC knockdown in cancer cell monocultures and co-cultures was conducted. Following LDHC knockdown, we observed an increase in the secretion of tumor-derived pro-inflammatory cytokines (IFN-γ, GM-CSF, MCP-1, CXCL1), a decrease in the soluble levels of tumor-derived immunosuppressive factors (IL-6, Gal-9) and reduced tumor cell surface PD-L1 expression. In direct co-cultures, LDHC knockdown reduced the levels of pro-tumorigenic cytokines (IL-1β, IL-4 and IL-6) and increased the secretion of the chemokine CXCL1. In addition, the number of CD8 + T cells expressing PD-1 and CTLA-4 and the cell surface expression of CTLA-4, TIGIT, TIM3, and VISTA were reduced.

Conclusions: Our findings suggest that targeting LDHC could enhance anti-tumor immune responses by modulating cytokine and chemokine secretion in addition to impairing immune checkpoint signaling. Further studies are required to elucidate the molecular mechanisms by which LDHC modulates immune responses in breast cancer.