Christopher Yusuf Akhunbay-Fudge, Bronwyn K. Irving, Alima Ismail, Sabrina Samuel, Emma Smedley, Holly E. Bradford, Steven Bagley, Alexander Baker, Iain M. Hagan, Deena Gendoo, Kevin Critchley, Ryan K. Mathew, Heiko Wurdak
Akhunbay-Fudge et al. develop two complementary single-cell profiling methods to determine glioblastoma (GB) invasion phenotypes, focusing on the influence of host organoid developmental lineage (neural versus endodermal) and cell cycle progression on GB invasion within tumor assembloids. Notably, GB cells invaded both neural and endodermal organoid hosts, whereas non-malignant adult brain cells lacked this capacity. Single-cell mRNA sequencing revealed gene expression changes in invading tumor cells and surrounding environmental assembloid cells. Concurrently, the "DyPheT" automated tracking tool enabled real-time correlation of cell cycle phases with malignant cell migration within cerebral organoids, which can be utilized for treatment response assessment, exemplified by the investigational compound RP-6306. Collectively, these approaches identify an intrinsic (cell autonomous) gene expression signature linked to GB invasion and support a "go-and-grow" paradigm by revealing a highly migratory (and RP-6306-refractory) GB subpopulation active in the G2/M phase of cell cycle.