Matrix stress relaxation drives glioblastoma cell response in viscoelastic biomaterials

The extracellular matrix (ECM) of glioblastoma (GBM) is known to modulate cell behavior, yet the specific contributions of matrix biochemical and biomechanical signaling remain poorly understood. To address this, we engineer a tunable hyaluronan-elastin-like protein (HELP) hydrogel to independently control ligand presentation and matrix viscoelasticity. Two peptides mimicking fibronectin (FBN) and tenascin-C (TNC) are incorporated into HELP along with hyaluronan, all of which are highly up-regulated in GBM. Using dynamic covalent chemistry, we develop hydrogels with matched stiffness but distinct stress relaxation profiles. Slow stress-relaxing matrices promote cell clustering and elevated expression of P-selectin, a GBM invasion marker. These matrices also result in increased nascent ECM production, increased lipid droplet storage, and altered cytokine secretion. Our results highlight the benefit of protein-engineered, viscoelastic biomaterials for modeling the tumor microenvironment and reveal matrix viscoelasticity as a critical regulator of GBM cell state, offering a tool for identifying previously unrecognized therapeutic targets.