Proteasome inhibition induces apoptosis through simultaneous inactivation of MCL-1/BCL-XL by NOXA independent of CHOP and JNK pathways

• Published in: Toxicology (Amsterdam) 2024-11, Vol.508, p.153906, Article 153906
• Main Authors: Chen, Wenjuan, Sun, Mengning, Sun, Yi, Yang, Qinglan, Gao, Hui, Li, Li, Fu, Rongrong, Dong, Na
• Format: Article
• Language: English
• Subjects: Apoptosis; Apoptosis - drug effects; bcl-X Protein - genetics; bcl-X Protein - metabolism; BCL-XL inactivation; Bortezomib - pharmacology; CHOP; GSDME; HeLa Cells; Humans; JNK; MAP Kinase Signaling System - drug effects; Myeloid Cell Leukemia Sequence 1 Protein - genetics; Myeloid Cell Leukemia Sequence 1 Protein - metabolism; NOXA; Proteasome Inhibitors - pharmacology; Proto-Oncogene Proteins c-bcl-2 - genetics; Proto-Oncogene Proteins c-bcl-2 - metabolism; Transcription Factor CHOP - genetics; Transcription Factor CHOP - metabolism
• ISSN: 0300-483X; 1879-3185; 1879-3185
• DOI: 10.1016/j.tox.2024.153906

Proteasome inhibitors have been employed in the treatment of relapsed multiple myeloma and mantle cell lymphoma. The observed toxicity caused by proteasome inhibitors is a universal phenotype in numerous cancer cells with different sensitivity. In this study, we investigate the conserved mechanisms underlying the toxicity of the proteasome inhibitor bortezomib using gene editing approaches. Our findings utilizing different caspase knocking out cells reveal that bortezomib induces classic intrinsic apoptosis by activating caspase-9 and caspase-3/7, leading to pore-forming protein GSDME cleavage and subsequent lytic cell death or called secondary necrosis, a phenotype also observed in many apoptosis triggers like TNFα plus CHX, DTT and tunicamycin treatment in HeLa cells. Furthermore, through knocking out of nearly all BH3-only proteins including BIM, BAD, BID, BMF and PUMA, we demonstrate that NOXA is the sole BH3-only protein responsible for bortezomib-induced apoptosis. Of note, NOXA is well known for selectively binding to MCL-1 and A1, but our studies utilizing different BH3 mimetics as well as immunoprecipitation assays indicate that, except for the constitutive interaction of NOXA with MCL-1, the accumulation of NOXA after bortezomib treatment allows it to interact with BCL-XL, then simultaneous relieving suppression on apoptosis by both anti-apoptotic proteins BCL-XL and MCL-1. In addition, though bortezomib-induced significant ER stress and JNK activation were observed in the study, further genetic depletion experiments prove that bortezomib-induced apoptosis occurs independently of ER stress-related apoptosis factor CHOP and JNK. In summary, these results provide a solid conclusion about the critical role of NOXA in inactivation of BCL-XL except MCL-1 in bortezomib-induced apoptosis. •Bortezomib induces apoptosis with "blowing bubbles" phenotype mediated by GSDME.•NOXA is the sole BH3-protein responsible for bortezomib-induced apoptosis.•NOXA relieves both BCL-XL and MCL-1 suppression after bortezomib exposure.•CHOP and JNK are not required for bortezomib-induced apoptosis.