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Human cerebral assembloids as a model to study the interaction of glioblastoma with resident glial cells

Kalpazidou, S.; Moysidou-Tsiorva, P.; Özbaykent, B.; Balcioglu, E.; Richter, M.; Shelton, E. R.; Frischmann, M.; Spalevic, J.; Harter, P. N.; Delbridge, C.; Wagner, A.; Goldberg, M.; Negwer, C.; Meyer, B.; Schlegel, J.; Serwane, F.; Schneider, J.; Bühler, D.; Glass, R.; Ninkovic, M.; Pravata, V.; Wimmer, R.; Miinalainen, I.; Cappello, S.; Stricker, S.; Danese, A.; Ninkovic, J.

2026-10-01 neuroscience
10.64898/2026.09.29.755236 bioRxiv
Show abstract

Glioblastoma (GB) is a highly invasive brain cancer characterized by extensive peritumoral glial reactivity. Although GB infiltration and recurrence remain major therapeutic challenges, the contribution of GB-induced gliosis to tumor progression is poorly understood. To investigate glia-GB interactions, we developed human iPSC-derived cerebral forebrain assembloids containing innate microglia and macroglia and transplanted them with GB cells. The model recapitulates key tumor features observed in patients, including distinct tumor regions, reactive gliosis, and altered tissue stiffness. Single-cell RNA sequencing revealed an unexpected enrichment of extracellular vesicles (EVs) in peritumoral reactive glia, which we validated in patient samples. EV inhibition impaired formation of the peritumoral niche, whereas EVs from GB-containing assembloids induced glial reactivity in GB-free assembloids. Furthermore, the modularity of our system enables incorporation of additional niche components, such as monocytes, allowing investigation of the monocyte-glia-tumor axis and providing a powerful platform to dissect mechanisms regulating the GB microenvironment.

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