An EYA3/NF-κB/CCL2 signaling axis suppresses cytotoxic NK cells in the pre-metastatic niche to promote triple negative breast cancer metastasis

Rosenbaum, S. R.; Hughes, C. J.; Fields, K. M.; Purdy, S. C.; Gustafson, A.; Wolin, A.; Hampton, D.; Turner, N.; Ebmeier, C. C.; Costello, J. C.; Ford, H. L.

doi:10.1101/2024.07.31.606072

bioRxiv · 10.1101/2024.07.31.606072

An EYA3/NF-κB/CCL2 signaling axis suppresses cytotoxic NK cells in the pre-metastatic niche to promote triple negative breast cancer metastasis

Rosenbaum, S. R.·Hughes, C. J.·Fields, K. M.·Purdy, S. C.·Gustafson, A.·Wolin, A.·Hampton, D.·Turner, N.·Ebmeier, C. C.·Costello, J. C.·Ford, H. L.

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Abstract

Patients with Triple Negative Breast Cancer (TNBC) exhibit high rates of metastases and poor prognoses. The Eyes absent (EYA) family of proteins are developmental transcriptional cofactors/phosphatases that are re-expressed and/or upregulated in numerous cancers. Herein, we demonstrate that EYA3 correlates with decreased survival in breast cancer, and that it strongly, and specifically, regulates metastasis via a novel mechanism that involves NF-kB signaling and an altered innate immune profile at the pre-metastatic niche (PMN). Remarkably, restoration of NF-kB signaling downstream of Eya3 knockdown (KD) restores metastasis without restoring primary tumor growth, isolating EYA3/NF-kB effects to the metastatic site. We show that secreted CCL2, regulated downstream of EYA3/NF-kB, specifically decreases cytotoxic NK cells in the PMN and that re-expression of Ccl2 in Eya3-KD cells is sufficient to rescue activation/levels of cytotoxic NK cells in vitro and at the PMN, where EYA3-mediated decreases in cytotoxic NK cells are required for metastatic outgrowth. Importantly, analysis of public breast cancer datasets uncovers a significant correlation of EYA3 with NF-kB/CCL2, underscoring the relevance of EYA3/NF-kB/CCL2 to human disease. Our findings suggest that inhibition of EYA3 could be a powerful means to re-activate the innate immune response at the PMN, inhibiting TNBC metastasis. SignificanceEYA3 promotes metastasis of TNBC cells by promoting NF-kB-mediated CCL2 expression and inhibiting cytotoxic NK cells at the pre-metastatic niche, highlighting a potential therapeutic target in this subset of breast cancer.

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Rosenbaum, S. R., Hughes, C. J., Fields, K. M., Purdy, S. C., Gustafson, A., Wolin, A., Hampton, D., Turner, N., Ebmeier, C. C., Costello, J. C., Ford, H. L.. 2024-08-03. An EYA3/NF-κB/CCL2 signaling axis suppresses cytotoxic NK cells in the pre-metastatic niche to promote triple negative breast cancer metastasis. https://doi.org/10.1101/2024.07.31.606072

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