Abstract 1177: Mechanical strain induces phenotypic changes in breast cancer cells and promotes immunosuppression in the tumor microenvironment

Abstract

Abstract Introduction: Cells within the breast tumor mass are met with a variety of biophysical or mechanical signals that is associated with elevated compression or solid stress within the tumor interior, tension at the tumor periphery, and altered interstitial fluid pressure. Understanding how the mechanical stress within the tumor microenvironment (TME) regulates cancer cell phenotype is of interest, yet the response of breast cancer (BCa) cells to these forces is largely unknown. Our study aims to identify the impact of mechanical stress on BCa cell phenotype by mimicking the tension at the tumor periphery. Materials and Methods: BCa cells (MCF-7 or 4T1.2) were cultured to confluence on collagen coated FlexCell culture plates. These plates were then subjected to 10% uniaxial cyclic/oscillatory strain at 0.3 Hz, or 10% constant strain, or no strain for 48 hours. Strained or control cells were isolated for analysis of proliferation (MTT assay), and migration (8.0 μm pore transwell). Exosomes from conditioned media were isolated via differential centrifugation and purified exosomes were characterized by ImageStream. 5x105 4T1.2 cells or PKH-labeled strained or control cells were injected into the mammary fat pad of BALB/c mice. Tumor volume was measured over 14 days. Tumor-infiltrating immune cells and the uptake of exosomes were analyzed by flow cytometry on day-14 post implantation. Results: We determined significant increases in proliferation and migration of 4T1.2 and MCF-7 cells in vitro following exposure to oscillatory forces. The populations of CD63+, CD63+CD24+ and CD63+PD-L1+ exosomes were increased when 4T1.2 cells were exposed to oscillatory strain compared to unstrained control cells. Further, we investigated how oscillatory forces affect tumor growth and tumor-immune cell interactions in vivo by using a syngeneic, orthotopic mouse model of BCa. Mice implanted with 4T1.2 cells that were pre-exposed to oscillatory forces showed a significant increase in primary tumor growth at 8 and 11 days post tumor challenge. The percentages of tumor-infiltrating monocytic myeloid-derived suppressor cells (M-MDSC) and recruited macrophages were increased in the TME of mice implanted with 4T1.2 cells that were pre-exposed to oscillatory forces, while the granulocytic MDSC subset was not significantly different between the two groups. A marginal decrease in the percentage of CD8+ T cells was noted in the TME of mice implanted with strained 4T1.2 when compared to controls, suggesting immune suppression in the TME. Furthermore, exosome uptakes by M-MDSC and recruited macrophages were increased in the TME of mice implanted with PKH-labeled 4T1.2 cells, exposed to oscillatory strain. Conclusion: Together, these data indicate that exposure to mechanical stress changes BCa cell phenotype to an invasive and protumorigenic phenotype that promotes immunosuppressive effects in the TME. Citation Format: Yong Wang, Paige E. Severino, Kayla Goliwas, Kenneth Hough, Derek Van Vessem, Hong Wang, Andra R. Frost, Selvarangan Ponnazhagen, Joel L. Berry, Jessy S. Deshane. Mechanical strain induces phenotypic changes in breast cancer cells and promotes immunosuppression in the tumor microenvironment [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1177.