CK1α agonists attenuate medulloblastoma stemness and relapse risk

While outcomes for most children with medulloblastoma (MB) are relatively favorable, those in the Sonic Hedgehog (SHH) subgroup with Tumor Protein P53 ( TP53 ) mutations–known as the SHHα subtype–face a much poorer prognosis. SHHα patients relapse more frequently and rapidly, underscoring the need for therapies that prevent recurrence. We recently identified a Gli-driven Sox2⁺ cell population that promotes relapse in SHH MB. However, few Gli-targeting strategies have shown clinical promise to date. One translational Gli inhibitor is pyrvinium, an FDA-approved compound known to destabilize Gli through increasing Casein Kinase 1α (CK1α) activity. In this study, we tested whether pyrvinium and a brain-permeable derivative, SSTC3, affect stemness and relapse risk in mouse and human-derived SHHα MB models. We found that pyrvinium suppresses the Gli-driven proliferation of Sox2⁺ cells. Unlike other SHH/Gli-targeting approaches, pyrvinium also impaired MB self-renewal by depleting Cluster of Differentiation 15 (CD15)⁺ cells. Mechanistic studies revealed that CD15⁺ cell self-renewal is WNT-dependent and driven by the loss of p53/ microRNA-34a –mediated repression of WNT signaling. Remarkably, pyrvinium and SSTC3 reduced Sox2⁺, CD15⁺, and dual Sox2/CD15-labeled populations in mouse and patient-derived SHHα models. Consistent with their ability to diminish tumor stemness, pyrvinium also impaired primary and secondary tumor engraftment. Together, these findings highlight the translational potential of the CK1α agonist pyrvinium and its derivatives for patients with very high-risk SHHα MB.