Abstract 5039: The relationship between interstitial fluid pressure, collective invasion and YAP activation in engineered human breast tumors

Abstract

Abstract The tumor microenvironment exhibits increased interstitial fluid pressure (IFP) due to combined effects of vascular leakage and inefficient drainage. Elevated IFP affects the transition of metastatic cells from the tumor to the healthy tissue. However, the complex architecture of the heterogeneous tumor tissues has made it difficult to correlate IFP with chemical and physical tumor responses that regulate invasion and metastasis. To overcome these limitations, we have engineered 3D breast tumor models for the quantitative characterization of biophysical and biochemical features that delineate its transition from a passive non-invasive to a metastatic state. In brief, we embed MDA-MB-231 human breast cancer cells in a collagen type I cavity molded within polydimethylsiloxane (PDMS) channels. The multicellular aggregates are subject to gradients of hydrostatic pressure through opposing reservoirs of culture media that are located at the base (Pbase) and the tip (Ptip) of the tumor. The pressure differential |Pbase – Ptip| defines the IFP gradient across the tumor. We found that the morphology of the engineered breast tumor models varies in response to the differential pressure gradients. For Pbase = Ptip and Pbase > Ptip, the engineered tumors displayed limited invadopodia formation. In contrast, for Pbase < Ptip, the engineered tumors displayed enhanced invadopodia formation. Moreover, we found that IFP affected cellular motility and invasion. Tumor cells demonstrated the highest average speed and directionality and were most invasive for Pbase < Ptip. Furthermore, we found that IFP-induced collective invasion coincides with the elevated expression of epithelial-mesenchymal transition (EMT) markers (vimentin, E-cadherin, Snail and keratin-8) in the engineered breast tumor models. In contrast with the previously demonstrated tumor suppressive activity of E-cadherin, engineered tumors that overexpressed E-cadherin developed a more invasive phenotype. The IFP-induced invasion through EMT in our engineered breast tumors motivated an investigation into the role of Yes-associated protein (YAP). YAP activation is implicated in EMT and regulated by fluid shear stress. Immunofluorescence staining for nuclear YAP suggests that YAP activation modulates the invasion and cell motility responses to IFP. Traction force microscopy will be used to further examine the relationship between IFP, force transduction into the surrounding microenvironment, and YAP activation. Citation Format: Andreas P. Kourouklis, Allison K. Simi, Alexandra Piotrowski-Daspit, Joe Tien, Celeste M. Nelson. The relationship between interstitial fluid pressure, collective invasion and YAP activation in engineered human breast tumors [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 5039.