Engineered 3D hydrogel model reveals divergence of adhesion-migration balance in Glioblastoma under simulated microgravity

Glioblastoma is an aggressive brain cancer whose cells can switch between different modes of invasion in response to their surroundings, making the disease difficult to predict and treat. How physical forces influence this adaptability remains poorly understood. Here, we used simulated microgravity together with engineered hydrogels that independently control adhesion, degradability, and mechanical properties to test how gravity affects glioblastoma invasion. Microgravity strongly reduced invasion and shifted cells from elongated, protrusive behavior to a more cohesive state. Proteomic analysis showed reduced invasive signaling together with increased cell-matrix and cell-cell adhesion, consistent with a redistribution of contractile forces toward the cell edge. Under normal gravity, blocking CD44, integrin β1, or N-cadherin reduced matrix-dependent invasion. In contrast, under microgravity, inhibiting these same adhesion pathways restored invasion, indicating that microgravity traps cells in an overly adhesive, cohesive state that limits movement rather than motility itself. These findings show that gravity is an important regulator of cancer cell plasticity and reveal a mechanically induced vulnerability in glioblastoma invasion. More broadly, combining defined biomaterials with gravitational modulation provides a new way to study how physical forces shape tumor behavior.