Researcher(s)
- Max Polonsky, Chemical Engineering, University of Delaware
Faculty Mentor(s)
- April Kloxin, Chemical and Biomolecular Engineering & Materials Science and Engineering, University of Delaware
Abstract
More than 380,000 people in the US are expected to be diagnosed with breast cancer in 2026. Triple negative breast cancer (TNBC) accounts for ~15-20% of these cases and is known for being highly aggressive, invasive, and for its early recurrence rates. Additionally, once TNBC metastasizes, the survival rate is only ~14.9%. One main driver of this clinical risk is dormancy, a reversible state of growth arrest that allows cancer cells to adapt and survive harsh environments and evade treatment. In this work, I aimed to investigate how various microenvironmental stressors affect morphology, growth dynamics, and cellular fate in a model TNBC cell line (MDA-MB-231). Initially, I analyzed images of these cells in a three-dimensional (3D) culture. However, we chose to start with two-dimensional (2D) culture as it is high-throughput and more resource efficient. To establish a baseline for how these cells grow in 2D, we first tracked morphological changes and cell counts under growth, exhausted (no media replacement), and death conditions. Driven by literature on nutrient starvation, we further introduced a serum deprivation condition, measuring metabolic activity and proliferation using metabolic assays and immunostaining. Finally, to establish an optimal cytokine dosing in our 2D models for this TNBC cell line, an experiment was conducted to identify the most viable concentration of the inflammatory cytokine TNF-⍺, evaluating cellular responses across a range of concentrations (10-200 ng/mL). Our findings indicate that microenvironmental stressors, specifically serum deprivation, induces slower growth or death of the cells, reflected with reduced metabolic activity and morphological changes, consistent with literature reports. Future experiments will translate 2D findings into 3D models to more accurately replicate the tumor microenvironment, with future opportunities for testing novel therapeutics in high-throughput 2D cultures by comparing their effect to the conditions we have tested in this study.