SPATIOTEMPORAL GLIAL DYNAMICS IN GLIOBLASTOMA

Glioblastoma (GBM) is the most aggressive primary malignant tumor of the central nervous system (CNS) and remains invariably fatal despite advances in surgical and pharmacological treatments. Beyond tumor-intrinsic alterations, GBM progression is increasingly recognized as a disease-driven process arising from altered neuroglial interactions and loss of CNS homeostasis.

Here, we investigated the spatiotemporal dynamics of glial responses during GBM progression, focusing on astrocyte and microglia activation and on regulators of glial communication and neuroinflammatory signaling, including GFAP, Iba1, Connexin 43 (Cx43), and the purinergic receptor P2X4R. Acute and organotypic brain slice cultures from C57BL/6J mice were used to preserve native tissue architecture and multicellular interactions. GL261 glioblastoma cells were injected into the cortical parenchyma, and analyses were performed at early (3 days) and progressive (7 days) stages after tumor implantation. Tumor proliferation was assessed by Ki67 immunoreactivity combined with histological analysis and confocal immunofluorescence.

GBM cells exhibited coordinated invasive behavior, migrating along organized cellular substrates and extending beyond the primary tumor mass. Early tumor infiltration was associated with localized astrocytic reactivity, whereas progressive stages showed sustained astrocyte activation and pronounced microglial morphological remodeling, indicative of an injury-associated inflammatory response. GBM progression induced region- and time-dependent modulation of Cx43 expression, together with dynamic changes in P2X4R levels consistent with adaptive regulation of purinergic signaling.

Overall, these findings identify GBM as a spatiotemporally evolving CNS pathology characterized by coordinated glial remodeling and disruption of neuroglial network homeostasis.