Single-cell and intercellular communication analyses reveal DLL3 as a key regulator in the glioma tumor microenvironment.

Glioma progression is increasingly recognized as a process driven not only by intrinsic genetic alterations in tumor cells but also by dynamic interactions within the tumor microenvironment (TME). However, systematic insight into the remodeling of intercellular communication networks during glioma evolution and their role in tumor progression remains limited. We integrated large-scale single-cell RNA sequencing with computational inference of ligand-receptor interactions to characterize the cellular landscape and communication architecture across disease stages. Single-cell analysis of normal brain and glioma tissues revealed pronounced cellular heterogeneity and stage-dependent remodeling of the TME, involving malignant cells, immune populations, glial cells, and vascular components. Intercellular communication analysis showed extensive reconstructing and strengthening of signaling networks during glioma progression, particularly among glioma cells, myeloid cells, astrocytes, and endothelial cells. Integrative analysis identified six communication-associated genes (DLL3, ADCYAP1R1, LFNG, FGFR3, VEGFA, and EGFR) that were significantly associated with patient prognosis. Functional validation further demonstrated that DLL3 is essential for glioma cell viability and proliferation. DLL3 depletion induced broad transcriptional reprogramming and suppressed cell cycle-related pathways, suggesting that DLL3 may mediate the link between microenvironmental signaling and intrinsic proliferative programs. Together, these findings highlight intercellular communication as a central driver of glioma progression and a potential therapeutic target.