FAK inhibitor treatment systemically regulates the glioblastoma immune environment via blocking monocyte trafficking

Webb, E. R.; Black, A.; Carrasco, G.; Furquan, M.; Hollis, R. L.; Loftus, A. E. P.; Ochoa, J. C.; Enjalbert, R.; Best, T.; Peng, B.; Muir, M.; Laing, F.; Lee, M.; Al Shboul, S.; Wang, T.; Smith, C.; Hupp, T. R.; Rajan, A.; Alfaro, J. A.; Brennan, P.; Liu, Z.; Ginhoux, F.; Bernabeu, M. O.; Serrels, A.; Gentek, R.; Frame, M. C.; Brunton, V. G.

doi:10.1101/2025.11.26.690731

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bioRxiv · 10.1101/2025.11.26.690731

FAK inhibitor treatment systemically regulates the glioblastoma immune environment via blocking monocyte trafficking

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Abstract

Glioblastoma (GBM) is the most common primary malignant brain tumour in adults with dismal survival rates, and current therapies, including most immunotherapies, are not efficacious due to the highly immunosuppressive microenvironment. Studies in other solid cancers report that impairment of the integrin effector pathway involving focal adhesion kinase (FAK) can promote anti-tumour immune responses. Therefore, we set out to address whether, and if so how, suppressing FAK function may influence GBM by using both tumour cell-specific FAK gene deletion and systemic delivery of a clinically relevant FAK kinase inhibitor (FAKi) VS-4718 in an orthotopic murine stem cell model of GBM. We found that treatment with the FAKi, but not tumour cell-specific FAK gene deletion, resulted in GBM clearance and improved survival. This was dependent on adaptive immunity, and tumour-infiltrating T cells in FAKi-treated tumours displayed increased cytotoxic potential and reduced exhaustion. We also found a significant reduction in immuno-suppressive peripherally-derived macrophages and FAKi treatment caused sequestration of inflammatory monocytes within the bone marrow, resulting in impaired monocyte trafficking to tumours as judged by adoptive transfer. This is due to suppression of key adhesion and migration signalling through 4{beta}1 integrin and CX3CR1 in peripheral monocytes. Our work here describes a previously unidentified role for FAK in trafficking of peripheral suppressive macrophages to GBM tumours, reducing T cell exhaustion and promoting anti-tumour immunity. This highlights a new way in which systemic FAK inhibitors can be used to provide a beneficial immune modulatory strategy for the treatment of GBM.

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Webb, E. R., Black, A., Carrasco, G., Furquan, M., Hollis, R. L., Loftus, A. E. P., Ochoa, J. C., Enjalbert, R., Best, T., Peng, B., Muir, M., Laing, F., Lee, M., Al Shboul, S., Wang, T., Smith, C., Hupp, T. R., Rajan, A., Alfaro, J. A., Brennan, P., Liu, Z., Ginhoux, F., Bernabeu, M. O., Serrels, A., Gentek, R., Frame, M. C., Brunton, V. G.. 2025-12-01. FAK inhibitor treatment systemically regulates the glioblastoma immune environment via blocking monocyte trafficking. https://doi.org/10.1101/2025.11.26.690731

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