Spatial vascular/BTB remodeling and malignant-state plasticity in glioblastoma

Abstract

Background Glioblastoma (GBM) contains spatially heterogeneous malignant and vascular states, but blood-tumor barrier (BTB) remodeling is often described as a binary functional phenotype. We asked whether anatomically distinct GBM compartments contain separable vascular programs that coexist with malignant-state plasticity. Methods We performed donor-aware cross-sectional analyses of 38 histopathology-annotated spatial transcriptomic sections from 6 donors and a separately analyzed endothelial single-nucleus layer from the same GBM-Space atlas. Complementary external datasets tested patient-paired regional remodeling, anatomical replication, cross-technology source localization, and malignant-state architecture. Results THSD1-FLT4 Recognition increased from leading edge to infiltrative tumor (median adjusted effect +0.02875; 4/4 donors positive). Priming increased across this boundary (+0.14814; 3/4) but decreased from infiltrative to cellular tumor (-0.16409; 0/4), whereas Gate remodeling increased from infiltrative to cellular tumor (+0.21296; 4/4). Remodeled endothelium showed higher PLVAP detection (+0.26409; 12/12 donors) and PLVAP pseudobulk expression (+1.61784 log1pCPM; 11/12), with lower MFSD2A pseudobulk expression (-0.71448; 10/12 negative). External cohorts supported regional vascular/BTB remodeling, while GSE131928 supported broad malignant-state architecture and an exploratory within-tumor pseudotemporal continuum. Conclusions GBM contains spatially partitioned vascular/BTB-associated programs alongside malignant-state plasticity. Recognition-Priming-Gate is a cross-sectional discovery framework, not a validated temporal cascade, and the data do not establish BTB permeability, causal tumor-vascular signaling, or therapeutic-delivery benefit. Key Points BTB-associated remodeling is spatially partitioned across GBM compartments. A PLVAP-high/MFSD2A-low endothelial state marks remodeled vasculature. Malignant-state plasticity forms a parallel, non-causal transcriptional axis. Importance of the Study The blood-tumor barrier in glioblastoma is often discussed as either intact or disrupted, yet therapeutic access is likely shaped by regionally distinct vascular states. This study resolves donor-aware spatial differences across leading-edge, infiltrative, and cellular-tumor compartments and identifies a PLVAP-high/MFSD2A-low endothelial remodeling state associated with the cellular-tumor region. Independent datasets support the broader vascular-remodeling phenotype, while a separate adult cohort supports malignant-state plasticity as a parallel axis. The Recognition-Priming-Gate framework is intentionally cross-sectional and does not claim a temporal cascade or measured barrier opening. By defining where barrier-associated transcriptional states are spatially organized, the study provides a testable map for future functional experiments aimed at transient, spatially targeted modulation of therapeutic delivery.