A blueprint for local and distal invasion programs in glioblastoma.

Glioblastoma invasion into brain parenchyma presents significant challenges for treatment but remains poorly understood. In this study, we combine single-cell RNA sequencing, spatial transcriptomics, and multiplexed imaging of orthotopic xenograft models to investigate glioblastoma invasion. We first screen 20 patient-derived gliomasphere models for their distal (i.e., extending to the contralateral hemisphere) and local invasive potential in mice. We show that models with distal invasion potential are enriched with oligodendrocyte progenitor-like cells, while models with only local invasion potential are enriched with mesenchymal-like cells. These patterns reflect predominantly peri-axonal vs peri-vascular invasion routes, respectively. Next, we analyze the transcriptomes of invading cells within models (compared to tumor core) and identify programs associated with distal and local invasion. Thus, we decouple transcriptional features associated with invasion potential from those associated with the process of invasion. We validate our findings by spatial transcriptomics and multiplexed imaging, further describing the spatial niche of invasive cells. Taken together, our results provide a blueprint for the invasive potential of glioblastoma cell states and of the programs associated with invasion across different scales.