Glioblastoma (GBM) tumors exhibit potentially actionable genetic alterations against which targeted therapies have been effective in treatment of other cancers. However， these therapies have largely failed in GBM patients. A notable example is kinase inhibitors of EGFR， which display poor clinical efficacy despite overexpression and/or mutation of EGFR in >50% of GBM. In addressing this issue， preclinical models may be limited by the inability to accurately replicate pathophysiologic interactions of GBM cells with unique aspects of the brain extracellular matrix (ECM)， which is relatively enriched in hyaluronic acid (HA) and flexible. In this study， we present a brain-mimetic biomaterial ECM platform for 3D culturing of patient-derived GBM cells， with improved pathophysiologic properties as an experimental model. Compared with orthotopic xenograft assays， the novel biomaterial cultures we developed better preserved the physiology and kinetics of acquired resistance to the EGFR inhibition than gliomasphere cultures. Orthogonal modulation of both HA content and mechanical properties of biomaterial scaffolds was required to achieve this result. Overall， our findings show how specific interactions between GBM cell receptors and scaffold components contribute significantly to resistance to the cytotoxic effects of EGFR inhibition. Three-dimensional culture scaffolds of glioblastoma provide a better physiological representation over current methods of patient-derived cell culture and xenograft models. .