TAS4464, a NEDD8-activating enzyme inhibitor, activates both intrinsic and extrinsic apoptotic pathways via c-Myc-mediated regulation in acute myeloid leukemia

Abstract

TAS4464, a potent, selective small molecule NEDD8-activating enzyme (NAE) inhibitor, leads to inactivation of cullin-RING E3 ubiquitin ligases (CRLs) and consequent accumulations of its substrate proteins. Here, we investigated the antitumor properties and action mechanism of TAS4464 in acute myeloid leukemia (AML). TAS4464 induced apoptotic cell death in various AML cell lines. TAS4464 treatments resulted in the activation of both the caspase-9-mediated intrinsic apoptotic pathway and caspase-8-mediated extrinsic apoptotic pathway in AML cells; combined treatment with inhibitors of these caspases markedly diminished TAS4464-induced apoptosis. In each apoptotic pathway, TAS4464 induced the mRNA transcription of the intrinsic proapoptotic factor NOXA and decreased that of the extrinsic antiapoptotic factor c-FLIP. RNA-sequencing analysis showed that the signaling pathway of the CRL substrate c-Myc was enriched after TAS4464 treatment. Chromatin immunoprecipitation (ChIP) assay revealed that TAS4464-induced c-Myc bound to the PMAIP1 (encoding NOXA) and CFLAR (encoding c-FLIP) promoter regions, and siRNA-mediated c-Myc knockdown neutralized both TAS4464-mediated NOXA induction and c-FLIP downregulation. TAS4464 activated both caspase-8 and caspase-9 along with an increase in NOXA and a decrease in c-FLIP, resulting in complete tumor remission in a human AML xenograft model. These findings suggest that NAE inhibition leads to anti-AML activity via a novel c-Myc-dependent apoptosis induction mechanism.

Elastic Net Models Based on DNA Copy Number Variations Predicts Clinical Features, Expression Signatures, and Mutations in Lung Adenocarcinoma

“…This also provides possibility for the clinical use of Elastic Net model. For instance, a variety of cell cycle progression and apoptosis signatures, such as c-MYC signature assessed above, may play a predictive role in the efficacy of NEDD8 inhibitors targeting PI3K/c-MYC axis (Ochiiwa et al, 2021), or for MAPK inhibitors by targeting ABL1/2-mediated reactivation of MEK/ERK/c-MYC signaling (Tripathi et al, 2020). The Elastic Net prediction models of c-MYC signature were able to extract out the patients with high proliferation rates, which generally accompanied by MYC amplification, then the use of NEDD8 inhibitors is recommended for these patients.…”

The NF-κB Pharmacopeia: Novel Strategies to Subdue an Intractable Target

“…Accordingly, MLN4924 is currently under investigation in a phase III clinical trial which is evaluating its efficacy and safety in combination with azacytidine, a cytotoxic chemotherapy drug, in AML patients who are not eligible for standard induction therapy (NCT04090736) [ 158 ]. Notably, other E1 inhibitors like TAK-243, TAS-4464 and ML-792 targeting ubiquitin-like modifier-activating enzyme 1 (UBA1), NAE and SUMO E1 activating enzyme, respectively, have been investigated in several cancer models, both in vitro and in vivo, and encouraging results have been reported [ 159 , 160 , 161 , 162 ]. These inhibitors exert their anti-tumor activity by triggering apoptotic cancer cell death, thus reducing tumor growth in xenograft mouse models [ 159 , 160 , 161 , 162 ].…”

Acute Myeloid Leukemia-Related Proteins Modified by Ubiquitin and Ubiquitin-like Proteins

“…TAS4464 is a highly effective and specific inhibitor of NAE and inactivates cullin-RING E3 ubiquitin ligases (CRLs) [ 193 ]. Treatment of TAS4464 in AML cell lines induces apoptosis and activates the intrinsic and extrinsic apoptotic pathways [ 173 ]. Moreover, treatment of HL-60 and THP-1 cell lines with TAS4464 reduces the expression of cellular FADD-like IL-1β-converting enzyme (FLICE)-inhibitory protein (c-FLIP), an anti-apoptotic protein, and increases the expression of phorbol-12-myristate-13-acetate-induced protein 1 (NOXA), a B-cell lymphoma 2 (Bcl-2) family, and pro-apoptotic protein [ 173 ].…”