Decoding glioblastoma invasion: single-cell profiling of environmental and cell cycle effects in tumour assembloids

Akhunbay-Fudge et al. develop two single-cell profiling techniques to dissect glioblastoma invasion phenotypes, focusing on the effects of lineage commitment (neural versus endodermal) and cell cycle progression within tumour assembloids. The study employs single-cell mRNA sequencing to compare tumour invasion into neural versus endodermal lineage assembloids and establishes the 'DyPheT' tracking algorithm to correlate cell cycle phases with malignant cell migration in cerebral organoids in real time. These complementary methods reveal glioblastoma-induced changes in environmental cells, identify cell-autonomous gene expression profiles of assembloid invasion in tumour cells, and emphasize a "go-and-grow" invasion paradigm, identifying highly migratory cells active during the S/G2 phase. Overall design: Glioblastoma (GB) cells were introduced into neural and endodermal assembloids and analyzed by scRNA-seq, while real-time tracking (DyPheT) was used to link cell cycle phases with invasion dynamics in cerebral organoids.