Targeting WTAP/ROR1/WNT5A-Mediated Crosstalk Between Glioma Stem Cells and Macrophages to Normalize Tumor Vasculature and Enhance Chemotherapy

Glioma stem cells (GSCs) can trans-differentiate into glioma-derived endothelial cells (GDECs), contributing to glioblastoma progression and therapy resistance. The molecular basis underpinning GSC-GDEC differentiation remain incompletely elucidated. Here, ROR1 is identified as a key regulator of GSC-to-GDEC differentiation under hypoxia. ROR1 expression is elevated in GDECs across patient-derived cells, xenografts, and tumor samples, confirmed by single-cell multiomics, Western blotting, and multiplex staining. Mechanistically, WNT5A, secreted by tumor-associated macrophages (TAMs) under hypoxia, activates ROR1-mediated Wingless-type MMTV integration site family (WNT) signaling in GSCs, promoting GDEC formation. Interestingly, hypoxia-induced WTAP not only enhances ROR1 stability through m6A modification in a HuR-dependent manner, but also contributes to WNT5A+TAMs infiltration. Clinically, high ROR1 and WNT5A expression correlates with poor glioblastoma prognosis, and WNT5A may serve as a circulating biomarker. Therapeutically, targeting ROR1 via endothelial-specific Adno-Associated Virus (AAV) knockdown or the antibody-drug conjugate Zilovertamab vedotin (VLS-101) normalizes vasculature, improves temozolomide delivery, and sensitizes tumors in glioblastoma organoids and xenografts. These findings highlight the ROR1-WNT5A axis as a promising target in glioblastomas treatment.