Glioblastoma is spatially organized by neurodevelopmental programs and a glial-like wound healing response

Glioblastoma is the deadliest brain cancer, characterized by great cellular diversity and unique histology. To understand the spatial organization of transcriptional cell states, we mapped the expression of 888 genes in centimeter-scaletissuesectionsfromalargepatientcohort.Wefoundahierarchyofcellularstatesakintonormal brain development, including proliferating and differentiated cells interacting with the stroma. We discovered that mesenchymal-like glioblastoma cells comprised a major glial-like wound-response component and a distinct gliosarcoma-specific malignant fibroblast type. Our analysis highlighted hypoxia, tissue damage, and woundhealingasmajorfactorsinglioblastomaspatialorganization.Tumormicroenvironment variedalongthe hypoxia gradient, inducing the recruitment of monocytes and pro-tumorigenic macrophages, and propagating wound response program activation in malignant glial cells. Our study reveals the dynamic progression of glioblastoma organization independent of its mutational profile, in response to tumor-induced injury. ### Competing Interest Statement SL is a paid scientific advisor to Moleculent, Combigene, and the Oslo University Center of Excellence in Immunotherapy. S.L., A.M.A., J.J., S.C., and L.E.B. are shareholders in EEL Transcriptomics AB, which owns the intellectual property to the EEL method (Spatial RNA localization method, provisional US patent application 63/139,701). All other authors declare that they have no competing interests.