Intratumoral gene delivery of 4-1BBL boosts IL-12-triggered anti-glioblastoma immunity

Lunavat, T. R.; Nieland, L.; Looij, S. M. v. d.; Reus, A. J. E. M. d.; Couturier, C. P.; Farran, C. A. E.; Miller, T. E.; Lill, J. K.; Schubel, M.; Xiao, T.; Ianni, E. D.; Woods, E. C.; Sun, Y.; Ramos, D. R.; Solinge, T. S. v.; Mahjoum, S.; Grandell, E.; Li, M.; Mangena, V.; Dunn, G. P.; Jenkins, R. W.; Mempel, T. R.; Breakefield, X. O.; Breyne, K.

doi:10.1101/2025.02.03.636330

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bioRxiv · 10.1101/2025.02.03.636330

Intratumoral gene delivery of 4-1BBL boosts IL-12-triggered anti-glioblastoma immunity

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Abstract

The standard of care in high-grade gliomas has remained unchanged in the past 20 years. Efforts to replicate effective immunotherapies in non-cranial tumors have led to only modest therapeutical improvements in glioblastoma (GB). Here, we demonstrate that intratumoral administration of recombinant interleukin-12 (rIL-12) promotes local cytotoxic CD8POS T cell accumulation and conversion into an effector-like state, resulting in a dose-dependent survival benefit in preclinical GB mouse models. This tumor-reactive CD8 T cell response is further supported by intratumoral rIL-12-sensing dendritic cells (DCs) and is accompanied by the co-stimulatory receptor 4-1BB expression on both cell types. Given that DCs and CD8POS T cells are functionally suppressed in the tumor microenvironments of de novo and recurrent glioma patients, we tested whether anti-tumor response at the rIL-12-inflamed tumor site could be enhanced with 4-1BBL, the ligand of 4-1BB. 4-1BBL was delivered using an adeno-associated virus (AAV) vector targeting GFAP-expressing cells and resulted in prolonged survival of rIL-12 treated GB-bearing mice. This study establishes that tumor antigen-specific CD8 T cell activity can be directed using an AAV-vector-mediated gene therapy approach, effectively enhancing anti-GB immunity.

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Lunavat, T. R., Nieland, L., Looij, S. M. v. d., Reus, A. J. E. M. d., Couturier, C. P., Farran, C. A. E., Miller, T. E., Lill, J. K., Schubel, M., Xiao, T., Ianni, E. D., Woods, E. C., Sun, Y., Ramos, D. R., Solinge, T. S. v., Mahjoum, S., Grandell, E., Li, M., Mangena, V., Dunn, G. P., Jenkins, R. W., Mempel, T. R., Breakefield, X. O., Breyne, K.. 2025-02-05. Intratumoral gene delivery of 4-1BBL boosts IL-12-triggered anti-glioblastoma immunity. https://doi.org/10.1101/2025.02.03.636330

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