Abstract P3-06-08: Imipridone compounds inhibit breast cancer mTORC1 signaling through integrated stress response-mediated upregulation of endogenous mTORC1 inhibitor sestrin2

Abstract

Abstract Breast cancer is a major cause of cancer-related death and there is a need for novel therapies with increased efficacy and decreased toxicity. The small molecule ONC201 was initially identified as a TRAIL pathway inducer. The compound has entered early phase clinical trials and is being tested in a range of solid tumors and hematological malignancies. Our previously published data demonstrate that ONC201 has potent anti-proliferative and pro-apoptotic effects in a broad range of breast cancer subtypes through TRAIL-dependent and TRAIL-independent mechanisms. Analogs of ONC201 with a shared pharmacophore have been developed and form the novel “imipridone” class. When compared with ONC201, imipridones ONC206, ONC212, and ONC213 showed increased potency of anti-proliferative or pro-apoptotic effects in breast cancer cells. We were interested in further defining the previously unstudied anti-proliferative effects of the imipridone compounds. Single agent efficacy of potent imipridone ONC212 in a xenograft model of TNBC was observed in the absence of apoptosis induction. This indicates that the anti-proliferative actions of the compound are sufficient for an in vivo anti-tumor effect. Our lab has previously shown that ONC201 activates an ATF4-dependent integrated stress response (ISR), essential for apoptosis induction in colon cancer cells. In contrast, in breast cancer cells, although ATF4 knockdown did not block cell death induced by ONC201 it did partially abrogate the anti-proliferative effects of the compound. The mammalian target of rapamycin complex 1 (mTORC1) is a well-known regulator of cellular growth and proliferation. mTORC1 signaling is inactivated in breast cancer cells following treatment with ONC201 and its analogs ONC212 and ONC213, regardless of whether the cells undergo apoptosis. Knockdown of ATF4 abrogated ONC201-mediated inhibition of p70 S6 kinase and ribosomal protein S6 phosphorylation, linking ISR induction to mTORC1 inhibition. We hypothesized that sestrin2, an endogenous mTORC1 inhibitor known to be upregulated following cellular stress, might represent a link between induction of ATF4 and inhibition of mTORC1. Treatment with ONC201 and its analogs ONC212 and ONC213 in multiple breast cancer cell lines resulted in sestrin2 upregulation. This was blocked by ATF4 knockdown. Furthermore, knockdown of sestrin2 abrogated the effects of ONC201 on mTORC1 signaling in breast cancers from multiple molecular subtypes. Previous mechanistic studies have focused exclusively on the relevance of ATF4 in the pro-apoptotic effects of ONC201. The novel findings described here help to elucidate the mechanism behind the potent and understudied anti-proliferative effects of the imipridones. Our findings also strengthen the preclinical rationale for testing of imipridone compounds against breast cancers regardless of molecular subtype. Citation Format: Ralff MD, Kline CLB, El-Deiry WS. Imipridone compounds inhibit breast cancer mTORC1 signaling through integrated stress response-mediated upregulation of endogenous mTORC1 inhibitor sestrin2 [abstract]. In: Proceedings of the 2017 San Antonio Breast Cancer Symposium; 2017 Dec 5-9; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2018;78(4 Suppl):Abstract nr P3-06-08.