Abstract 7226: Synergy between ONC201 and temozolomide on ATF4 integrated stress response in glioblastoma

Abstract

Abstract Glioblastoma multiforme (GBM) is the most common primary brain tumor in adults, with a median survival of 15 months. The standard of care involves surgical resection followed by chemoradiotherapy. Even with the full course of treatment, GBM is uncurable, and patients eventually relapse. Although temozolomide (TMZ) is the first-choice chemotherapeutic for GBM, tumor drug resistance limits the effects of the treatment. There is a growing need for new therapies with enhanced efficacy on GBM tumors. Dordaviprone (ONC201) is a TNF-related apoptosis inducing ligand (TRAIL)-inducing compound that is increasingly being studied for GBM treatment. This imipridone small molecule induces apoptosis by activating the integrated stress response, which involves transcription factors ATF4 and CHOP. Subsequently, these transcription factors upregulate proapoptotic protein TRAIL and its receptor, Death Receptor 5 (DR5). This study aims to investigate the combinatory effects of chemotherapeutics TMZ and ONC201 on GBM. Preliminary studies examined the cell viability of GBM cell line U251MG after dual treatment of 0 to 10uM of ONC201 and 0 to 200uM of TMZ using the CellTiter-Glo assay. Synergy was observed between the two drugs at higher concentrations of TMZ. This synergistic interaction was further characterized using western blots. When 0, 2, and 4uM of ONC201 were administered alone or in combination with 0, 60, and 120uM of TMZ, greater ATF4 expression was observed with the dual treatments than with the monotherapies. Subsequent western blots were conducted using three GBM cell lines (U251MG, T98G, and SNB19-GFP) at three time points (8, 24, and 48 hours). All cell lines were treated with 0 and 5uM of ONC201 alone or in combination with 0 and 200uM of TMZ. For all three cell lines, ATF4 expression increased in the combined presence of TMZ and ONC201. However, the effects of TMZ on ONC201-induced ATF4 expression were most pronounced in U251MG and least conclusive in T98G. Additionally, the time points in which this increase in ATF4 expression occurred varied among cell lines. CHOP expression was also probed and was seen to increase with the combination treatment in U251MG. These observations suggest a potential synergistic relationship between chemotherapeutics ONC201 and TMZ in the ATF4 integrated stress response pathway in GBM. By exploring the effects of this dual treatment on upstream and downstream factors in the pathway, we can identify, characterize, and potentially target the root cause of this synergy. Citation Format: Josephine Chen, Tyler J. Roady, Lanlan Zhou, Wafik S. El-Deiry. Synergy between ONC201 and temozolomide on ATF4 integrated stress response in glioblastoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 7226.