An oxo-mechanical regulation of cellular morphology in 3D

How do combinatorial mechano-chemical cues drive cellular behavior? Oxygen availability and ECM properties are two of the most prevalent, yet dramatically varying microenvironmental cues across most physiological settings. While we understand the biological effects of these two entities in isolation, little is known about how they simultaneously alter cellular behavior. Our present work subjects cells to a combination of oxygen partial pressures and ECM densities - an oxo-mechanical cue - and investigates their morphology at the single cell level by combining morphometric measurements, bulk transcriptome analyses, as well as chemical modulation of intracellular mechanics and oxygen-driven signaling. At lower ECM densities, acute oxygen deprivation significantly alters cell morphology, whereas, at higher ECM densities, the effect of oxygen deprivation on cellular morphology is negligible. We independently show that a cell's perception and response to varying oxygen availability depends on both substrate and intracellular mechanics; while the cell's engagement with mechanically diverse substrates is influenced by oxygen-driven signalling processes. Together, our work identifies an oxo-mechanical regulation of cellular behavior in 3D ECM-like systems.