A Comprehensive Treatment-Induced Resistance Atlas of Glioblastoma Reveals a Fibrotic Niche Shielding the Tumor from Immunotherapy

Wang, F.; Huang, R.; Ling, H.; Bai, Y.; Yang, C.; Zhao, G.; Wu, X.; Cao, W.; Lu, Y.; Zhang, Y.; Lu, D.; Qiu, Y.; Zhang, J.; Gao, B.; Ma, C.; Dang, H.; Yang, Y.; Sun, T.; Chen, Z.; Wang, Z.

doi:10.64898/2025.12.25.696471

bioRxiv · 10.64898/2025.12.25.696471

A Comprehensive Treatment-Induced Resistance Atlas of Glioblastoma Reveals a Fibrotic Niche Shielding the Tumor from Immunotherapy

Wang, F.·Huang, R.·Ling, H.·Bai, Y.·Yang, C.·Zhao, G.·Wu, X.·Cao, W.·Lu, Y.·Zhang, Y.·Lu, D.·Qiu, Y.·Zhang, J.·Gao, B.·Ma, C.·Dang, H.·Yang, Y.·Sun, T.·Chen, Z.·Wang, Z.

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Abstract

Therapeutic resistance in IDH-wildtype glioblastoma (GBM) is driven by an intricate interplay between cellular plasticity and protective microenvironmental niches. By constructing the GRIT-Atlas--a massive transcriptomic compendium encompassing nearly one million high-quality single cells across 17 cohorts--and cross-analyzing it with 48 independent Visium spatial transcriptomics sections, we discover a recurrent "Spatial Resistance Triad" comprising mesenchymal-like (MES-like) malignant cells, myeloid-derived suppressor cells (MDSCs), and collagen-secreting cancer-associated fibroblasts (CAFs). This large-scale spatial integration reveals that the triad physically fortifies microvascular proliferation (MVP) and pseudopalisading necrosis (PAN) niches. To substantiate these findings at true single-cell resolution, we deploy high-plex spatial molecular imaging (CosMx SMI) to map over 400,000 single cells across a clinical validation cohort. This high-resolution architecture firmly validates the triads geography and, critically, provides structural gradient evidence unmasking a spatiotemporal continuum wherein PAN emerges as a direct functional consequence of MVP advancement and subsequent vascular collapse. Mechanistically, stromal CAF-derived COL6A1 engages CD44 receptors on MES-like cells to accelerate a potent neural stemness cascade within these protective domains. Utilizing a blood-brain barrier (BBB)-penetrant multi-library compound screen, we identify Lacidipine as a multi-modal stromal disruptor that successfully silences CAF activation and halts matrix secretion. Incorporating Lacidipine into the standard chemo-immunotherapy backbone (TMZ + anti-CSF1R) completely dismantles the protective desmoplastic matrix niche, forcing a profound collapse of intracranial tumor burden and dramatically extending overall survival in orthotopic models. Collectively, our study leverages unprecedented single-cell and spatial scale to provide a definitive blueprint of GBM therapeutic evasion, establishing matrix-targeted intervention as a mandatory prerequisite for successful glioblastoma eradication.

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Wang, F., Huang, R., Ling, H., Bai, Y., Yang, C., Zhao, G., Wu, X., Cao, W., Lu, Y., Zhang, Y., Lu, D., Qiu, Y., Zhang, J., Gao, B., Ma, C., Dang, H., Yang, Y., Sun, T., Chen, Z., Wang, Z.. 2025-12-29. A Comprehensive Treatment-Induced Resistance Atlas of Glioblastoma Reveals a Fibrotic Niche Shielding the Tumor from Immunotherapy. https://doi.org/10.64898/2025.12.25.696471

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