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

ABSTRACT 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 m ⁶ A 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.