Pannexin 1 inhibition reduces tumorigenic properties of patient-derived glioblastoma cells through the HIPPO and Wnt signalling pathways

Johnston, D.; Huver, M.; Kanji, R.; Van Kessel, C.; Kelly, J.; Sanchez Pupo, R. E.; O'Donnell, B.; Defamie, N.; Lau, R.; Herrera, C.; Deweyert, A.; Mesnil, M.; Ronald, J.; Hebb, M.; Penuela, S.

doi:10.64898/2026.09.03.749222

bioRxiv · 10.64898/2026.09.03.749222

Pannexin 1 inhibition reduces tumorigenic properties of patient-derived glioblastoma cells through the HIPPO and Wnt signalling pathways

Johnston, D.·Huver, M.·Kanji, R.·Van Kessel, C.·Kelly, J.·Sanchez Pupo, R. E.·O'Donnell, B.·Defamie, N.·Lau, R.·Herrera, C.·Deweyert, A.·Mesnil, M.·Ronald, J.·Hebb, M.·Penuela, S.

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Abstract

Glioblastoma (GBM) is the most common primary brain tumour, with a median survival of 12-18 months, highlighting a need for new treatment targets. We observed that pannexin 1 (PANX1), a channel-forming glycoprotein important in purinergic signalling, is upregulated in GBM compared to normal tissue and expressed throughout patient tumours. Western blot analysis of patient-derived GBM cell lines revealed significantly increased PANX1 expression in these primary lines compared to brain tissue and control glial cells. Bulk RNA-sequencing compared the gene expression of GBM cells devoid of PANX1 via CRISPR/Cas9 deletion (PANX1-KO) compared to controls. Gene Ontology and KEGG gene set analyses revealed PANX1-KO in GBM cells affects cell surface and cell junction components, processes, and pathways, including the HIPPO pathway, in addition to critically downregulating {beta}-catenin mRNA and other components of the Wnt pathway. The deletion of PANX1 resulted in a disruption of the {beta}-catenin protein and a dramatic reduction in migration and cell growth. Pharmacological inhibition of PANX1 in GBM cells with Probenecid (PBN) and Spironolactone (SPIR) demonstrated a significant reduction in live cell numbers and migration via scratch assay. Both blockers dramatically decreased F-actin filament formation, and the cellular localization of beta-catenin became more intracellular compared to controls. Xenografted GBM tumours showed a reduction in tumour cell viability by bioluminescent imaging and reduced hemorrhaging incidence when treated with PBN. These new insights support further investigation of PANX1 as a potential GBM therapeutic target and its role in multiple cancer signaling pathways that regulate this devastating disease.

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Johnston, D., Huver, M., Kanji, R., Van Kessel, C., Kelly, J., Sanchez Pupo, R. E., O'Donnell, B., Defamie, N., Lau, R., Herrera, C., Deweyert, A., Mesnil, M., Ronald, J., Hebb, M., Penuela, S.. 2026-09-07. Pannexin 1 inhibition reduces tumorigenic properties of patient-derived glioblastoma cells through the HIPPO and Wnt signalling pathways. https://doi.org/10.64898/2026.09.03.749222

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