Abstract 6791: Investigation of sex hormone effects in an organ chip model of endothelial barrier dysfunction in the triple negative breast cancer microenvironment
Authors: Carlino Delia , Martier Ashley , Maurice Yasmin , Kpeli G. Wills , Burow Matthew , Mondrinos Mark 
Published: March 2024

Abstract
Inflammation and pathological vasculature are common features of the tumor microenvironment (TME). Sex hormones potently modulate vascular physiology and immune cell functions, and there are established correlations with the outcomes of triple negative (TNBC) despite the defining feature of receptor negativity. Circulating estrogens and androgens have been hypothesized as drivers of TNBC through the actions exerted by receptor-positive cells in the TME. Here, we present an organ chip model of the TNBC microenvironment for quantifying the impact of configurable TME variables such as the ECM composition and the relative densities and phenotypes of tumor cells, cancer associated fibroblasts, and myeloid cells on endothelial barrier function. After establishing baselines of vascular permeability in the presence and absence of TNBC cells, we tested the effect of estradiol, estrone, and dihydrotestosterone on barrier permeability. Multilayer polydimethylsiloxane (PDMS) organ chips were fabricated using standard soft lithography. The layers of the device were separated with semi-permeable membranes that allow exchange of soluble factors. Once assembled, the upper channel was loaded with female (XX) human umbilical vein endothelial cells. Collagen hydrogels were injected into the tissue layer and the devices were cultured on a rocker to induce endothelial barrier alignment via application of an average shear stress of 5 dynes/cm2. We optimized rocking conditions by assessing endothelial layer alignment, junctional organization, and permeability. TNBC cells and XX mammary carcinoma fibroblasts were added to the hydrogel to model the effects of TME-derived signals on endothelial barrier permeability. 40 kDa FITC- Dextran was chosen to assay permeability as it can show leakage due to active disruption of the barrier. Staining for F-actin and ZO-1 allowed for quantification of junctional organization. Sex hormone modulation of proinflammatory gene expression was assessed by RT-qPCR. The presence of TNBC cells significantly increased transport of 40kDa FITC-Dextran across the endothelial barrier, disrupted intercellular junction quality, and induced the expression of inflammatory adhesion molecules including ICAM-1. Our results demonstrate that sex hormones modulate TNBC-associated changes in endothelial cell bioenergetics, junctional organization, and macromolecular barrier permeability. Ongoing work will investigate the effects of age-matched XX monocyte derived macrophages on barrier function in the model. This organ chip platform provides an assay for measuring altered vascular inflammation and permeability in the TNBC microenvironment and may provide a venue for investigating vascular-normalizing effects of various candidate therapeutics.
Citation Format: Delia Carlino, Ashley Martier, Yasmin Maurice, G. Wills Kpeli, Matthew Burow, Mark Mondrinos. Investigation of sex hormone effects in an organ chip model of endothelial barrier dysfunction in the triple negative breast cancer microenvironment [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 6791.