MDB-76. ONC201 INHIBITS THE CHARACTERISTICS OF CANCER STEM CELLS IN GROUP 3 MEDULLOBLASTOMA

Abstract

Abstract BACKGROUND Medulloblastoma (MB), the most frequent childhood brain tumor, originates in the cerebellum. Despite treatment advances, recurrence affects 40%, with a mortality rate of 30%. Cancer stem cells drive tumor initiation, maintenance, and therapy resistance, in several tumors including MB (MBSC). In response to stress, stem cells activate mechanisms like the unfolded protein response (UPR) to maintain homeostasis and survive adverse conditions. The UPR restores endoplasmic reticulum balance, supporting cell function and survival. Recent studies have shown that the UPR plays an important role in tumorigenesis, however, its function in MBSC remains unknown. METHODS To understand the involvement of the UPR in stem cell maintenance, we have investigated the biological effects of its pharmacological inhibition on Group 3 (G3) MBSC. We selected three G3 MB cell lines (D283-Med, D341-Med and Med411), for in vitro studies. Cell line selection was based on their fidelity to the expected methylation class, corresponding to the subgroup of the parental tumors, and were grown in “stem-like” conditions as neurospheres. We evaluated drugs targeting the UPR stress program, identifying ONC201 as the most effective against MBSC. ONC201 is an imipridone compound that activates p53-independent apoptosis causing changes in gene expression similar to those caused by UPR. RESULTS Our findings revealed that ONC201 modulates protein synthesis via ATF4, a pivotal UPR molecule, with stronger induction observed in MB cells cultured under “stem-like” conditions. This ATF4 upregulation triggers the activation of pro-apoptotic programs. Additionaly, ONC201 was effective on reducing the stem-like features of MBSC, including self-renewal, stem cell markers expression, migration, and invasion potential. Notably, ONC201 demonstrates synergy with vincristine and methotrexate chemotherapy. CONCLUSIONS These results propose ONC201 as a promising agent for targeting G3 medulloblastoma by compromising the stem cell compartment. Further investigation is warranted to confirm its efficacy and potential clinical application.