 The spatial organization of diffuse midline glioma, H3K27M mutant, DMG, and glioblastoma, GBM, remains to be investigated. Here, the authors integrate short read and long read spatial profiling of DMG and GBM to identify regulatory programs and cellular ecosystems in distinct glioma niches. This article was authored by Yang Mingren, Zongyao Huang, Lingling Zhu, and others.