Abstract P2-05-11: Sphingosine-1-phosphate signaling promotes metastatic niches and lung metastasis in obesity-related breast cancer
Authors: Nagahashi M , Tsuchida J , Moro K , Tatsuda K , Koyama Y , Takabe K , Wakai T 
Published: May 2016

Abstract
INTRODUCTION: The link between obesity and elevated breast cancer mortality is well known, however, the underlying mechanisms are poorly understood. Sphingosine-1-phosphate (S1P) is a pleiotropic bioactive lipid mediator produced by sphingosine kinases (SphKs) that plays critical roles in inflammation and cancer progression. Previously, we found that obesity increases levels of S1P not only in breast tumors, but also in the lung. "Metastatic niches" are specialized microenvironments in distant organs primed by factors from cancer cells. We hypothesized that S1P secreted from the primary tumor could promote formation of a "metastatic niche" in the lung, which assists circulating cancer cells to form metastatic lesions. Further, HFD-induced obesity increases S1P secretion from the primary tumor, which could promote the formation of "metastatic niches" in the lung and lung metastasis. The aim of this study is to test these hypotheses.
METHODS: A mouse model utilizing tail vein injection of E0771 syngeneic breast cancer cells was used. Prior to tail vein injections of naive E0771 cells, mice were treated with conditioned media from E0771 breast cancer cells overexpressing SphK1 (K1-CM) or that from E0771 cells cultured with the vector control (CT-CM). Histological analysis, RT-qPCR, and western blot were used.
RESULTS: The lungs after K1-CM treatment demonstrated much more infiltration of macrophages with greater IL-6 secretion than lungs from CT-CM mice in areas without metastasis. Furthermore, SphK1, S1P receptor 1 and IL-6 expression were all significantly higher in the lungs of mice treated with K1-CM than with CT-CM, suggesting that S1P secreted from the primary tumor promotes formation of a metastatic niche in the lung. Next, mice were fed with HFD or ND for 12 weeks before treatment with SphK1-CM, and lungs were examined 7 days after intravenous injection of E0771 cells. Histological analysis demonstrated that there were significantly more lung metastases in mice on HFD than in mice on ND. Importantly, treatment with FTY720, a functional antagonist of S1P receptor 1, significantly reduced the lung metastases in HFD fed animals. Immunofluorescent staining revealed higher expression of IL-6 and greater number of F4/80 positive macrophages in mice fed with HFD compared with mice fed with ND, whereas FTY720 dramatically suppressed both IL-6 and macrophage infiltration in the lung of HFD-fed mice. HFD-induced obesity also increased pERK, pAKT, pStat3, and pp65 in the lung, and FTY720 suppressed these signaling pathways.
CONCLUSION: Our results suggest that S1P plays a role in the formation of "metastatic niches" in the lung and lung metastasis of breast cancer, and obesity promotes this process. S1P will be a promising target for treatment of breast cancer metastasis, especially in condition with obesity. This work was supported by the Japan Society for the Promotion of Science (JSPS) Grant-in-Aid for Scientific Research Grant Number 15H05676 and 15K15471 for M.N and 15H04927 for W.T. M.N. is supported by the Uehara Memorial Foundation, Nakayama Cancer Research Institute, and Tsukada Medical Foundation. K.T. is supported by NIH/NCI grant R01CA160688 and Susan G. Komen Investigator Initiated Research Grant IIR12222224.
Citation Format: Nagahashi M, Tsuchida J, Moro K, Tatsuda K, Koyama Y, Takabe K, Wakai T. Sphingosine-1-phosphate signaling promotes metastatic niches and lung metastasis in obesity-related breast cancer. [abstract]. In: Proceedings of the Thirty-Eighth Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2015 Dec 8-12; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2016;76(4 Suppl):Abstract nr P2-05-11.