mRNA-LNP vaccination induces microenvironment remodeling and generation of tissue-resident memory T cells in glioblastoma

Snelling, S. M.; Manoharan, V. T.; Pyczek, J.; Lun, X.; Wang, J.; Liu, J.; Gold, M. J.; Nechanitzky, R.; Ellestad, K.; Patel, A.; Olsen, K.; Wu, Y.; Karpov, Y.; Menon, H.; Tan, Y.; Krishnan, R.; Mahoney, D. J.; Morrissy, S.; Martin-Orozco, N.; Chan, J. A.

doi:10.64898/2026.11.11.762754

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bioRxiv · 10.64898/2026.11.11.762754

mRNA-LNP vaccination induces microenvironment remodeling and generation of tissue-resident memory T cells in glioblastoma

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Abstract

Glioblastoma remains highly refractory to immunotherapy, in part because of a profoundly immunosuppressive tumour microenvironment that limits effective T-cell responses. Although mRNA vaccines can induce anti-tumour immunity, how they reshape the glioblastoma immune ecosystem to support durable tumour control is not well understood. Here we use an immunocompetent EGFRvIII-driven syngeneic glioblastoma model and antigen-specific mRNA lipid nanoparticle (mRNA-LNP) vaccination to define the cellular and spatial mechanisms underlying therapeutic response. In our model, vaccination induced durable EGFRvIII-specific humoral and cellular immunity, resulting in complete tumour eradication and long-term protection against intracranial rechallenge. Integrative analyses incorporating flow cytometry, tumour interstitial fluid profiling, multiplex immunostaining, spatial transcriptomics and ligand-receptor modelling demonstrated extensive remodelling of the tumour microenvironment following vaccination. Early tumour control was associated with expansion of pro-inflammatory and antigen-presenting myeloid populations, increased cytotoxic CD8 T-cell infiltration and activation, enhanced interferon-responsive signalling, and the formation of immune-supportive spatial niches enriched for antigen presentation and chemokine-mediated cellular communication. Vaccination promoted inflammatory chemokine relay networks centred on the CXCL16-CXCR6 axis, a pathway linked to tissue residency programming in the central nervous system. Functional depletion studies identified CD4 and CD8 T cells as essential mediators of therapeutic efficacy, while durable protection was maintained by tumour-specific brain-resident memory CD8 T cells that persisted despite peripheral lymphocyte depletion. Together, these findings identify T-cell-driven microenvironmental remodelling as a critical mechanism linking mRNA vaccination to the establishment of long-term brain-resident anti-tumour immunity in glioblastoma.

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Snelling, S. M., Manoharan, V. T., Pyczek, J., Lun, X., Wang, J., Liu, J., Gold, M. J., Nechanitzky, R., Ellestad, K., Patel, A., Olsen, K., Wu, Y., Karpov, Y., Menon, H., Tan, Y., Krishnan, R., Mahoney, D. J., Morrissy, S., Martin-Orozco, N., Chan, J. A.. 2026-11-13. mRNA-LNP vaccination induces microenvironment remodeling and generation of tissue-resident memory T cells in glioblastoma. https://doi.org/10.64898/2026.11.11.762754

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