(540b) Engineering Confined Microenvironments Using Granular Hydrogel Scaffolds to Investigate the Role of Dynein in Cancer Cell Migration

The challenges posed by cancer metastasis significantly contribute to patient fatalities associated with solid tumors. In vitro cancer models enable studying cell migration within confined microenvironments that mimic native tumor tissues. Granular hydrogel scaffolds (GHS), composed of packed hydrogel microparticles (microgels), provide interconnected, micron-sized void spaces with tunable local stiffness that mimic mechano-structural properties of the confining conditions of solid tissue microenvironments in vivo. Here, we developed GHS using gelatin methacryloyl (GelMA) microgels with varying local stiffnesses, partially mimicking the extracellular matrix (ECM) of native tumor microenvironment. To investigate the impact of dynein on confined three-dimensional (3D) cell migration, metastatic and non-metastatic breast cancer cells were seeded in GHS and treated with dynein and/or myosin inhibitors. The average depth of cell migratory infiltration in all treated samples was significantly lower than in untreated cells.These findings suggest that both actomyosin and dynein contractilities are required for 3D confined cell migration. In conclusion, engineered in vitro GHS models provide a suitable, biologically relevant platform for studying the mechanisms of cell motility in confined spaces, which may pave the way for developing therapeutics aimed at overcoming cancer metastasis.