Consequences of irradiation on blood-brain tumor barrier model of Diffuse Midline Glioma: characterization of physical and metabolic properties

TL;DR

This study examines how radiation affects the blood-brain tumor barrier in a pediatric brain tumor model, revealing changes in physical and metabolic properties.

Contribution

The study introduces a novel in vitro model to characterize the effects of radiation on the blood-brain tumor barrier in diffuse midline glioma.

Findings

Irradiation preserved endothelial permeability but caused Claudin-5 heterogeneity and reduced expression.

Radiation modulated metabolic properties differently in endothelial cells and pericytes, with pericytes showing compensatory effects.

P-gp/BCRP efflux pump activity remained functional post-irradiation, though less efficient, in both endothelial cells and pericytes.

Abstract

Diffuse midline glioma (DMG) is a rare and aggressive pediatric brain tumor, with a median survival of less than 12 months. Due to its location in the pons, surgical resection is impossible, leaving radiation therapy as the only palliative treatment option. Unfortunately, radiation therapy yields minimal improvement in survival. Thus, characterization of the vascular component of the DMG microenvironment at the cellular and molecular levels following radiotherapy to improve therapeutic strategies.

A human syngeneic blood-brain tumor barrier (BBTB) in vitro model, comprising endothelial cells, pericytes and DMG cells was submitted to a single dose of radiation (2 Gγ to 6 Gγ) and was characterized for its physical and metabolic properties over a period of 7 days post-exposure. The results were then compared to the effects of the same irradiation protocol on a physiologic blood-brain barrier (BBB) model.

Following irradiation, the endothelial permeability of the BBB ECs and the BBTB ECs was preserved for up to 7 days but associated with Claudin-5 heterogeneous distribution at the ECs borders and decrease of expression after irradiation. Nevertheless, irradiation was found to potentiate the effect of TNFα on the physical integrity of the BBB, which was less important for the BBTB. The metabolic properties of the BBB and BBTB were modulated by radiation at the transcriptional level. Interestingly, different regulations were observed in endothelial cells and pericytes. Notably, pericytes have demonstrated compensatory effects. Immunoblots confirmed the decrease of BCRP, MRP4 and MFSD2A in BBTB endothelial cells after irradiation. Despite significant reduced efficiency, P-gp/BCRP efflux pump activity remains functional in endothelial cells and pericytes following irradiation.

Irradiation sensitizes the BBB, but to a lesser extent the BBTB, to the effects of pro-inflammatory cytokines. The observed decrease in P-gp/BCRP activity, as well as the involvement of MFSD2A, MRP4 and Claudin-5 regulation, warrant further investigations.

The online version contains supplementary material available at 10.1186/s12987-026-00778-6.

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