Brain assembloids reveal efficacy and off target effects in high grade glioma brain cancer

Anti-invasive therapies may limit damage to healthy brain tissue caused by invading high-grade glioma (HGG) cells. However, the actin machinery that drives invasion is shared by cancer and normal cells, and therefore, there is a risk of on-target, off-tumor side effects. We compared omega 3 polyunsaturated fatty acid epoxide analog p-tolyl-ureidopalmitic acid (PTU) effects between HGG multicellular tumor spheroids (MCTSs) embedded in Matrigel and assembloids consisting of HGG MCTS with embryonic stem cell-derived cortical brain organoids. PTU inhibited cofilin and induced cell rounding consistent with a loss of filamentous (F-) actin and efficiently inhibited invasion in 3D Matrigel invasion assays, but effects were cell line specific in assembloids. Immunofluorescence and GeoMx Digital Spatial Profiling of assembloids revealed that PTU disrupts signaling in cancer cells and the "healthy" cortical organoid. This study demonstrates a methodological approach for in vitro testing of efficacy and safety for anti-invasive therapies for HGG.