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bioRxiv · 10.1101/2025.06.29.661768

Bedaquiline Amplifies Proteasome Inhibitor Efficacy and Overcomes Resistance in Multiple Myeloma

Abstract

Proteasome inhibitors (PIs) are cornerstone therapies for multiple myeloma (MM), yet resistance remains a major barrier to durable responses. To identify druggable vulnerabilities that enhance PIs efficacy, we performed a small-molecule chemical screen in the presence of carfilzomib (CFZ). We identified bedaquiline (BDQ), an FDA-approved antimycobacterial agent, as a potent synergistic partner. BDQ and its fumarate salt (BDQ-F) significantly amplified CFZ-induced cytotoxicity in PI-sensitive and PI-resistant MM cells, in AL amyloidosis and other B-cell malignancies, with minimal toxicity toward normal cells. Mechanistic studies confirmed that BDQ specifically targets the ATP5F1C subunit of mitochondrial ATP synthase. BDQ-CFZ combination triggered extensive apoptosis, exacerbating proteotoxic stress and proteasome-associated pathways. BDQ specifically enhanced CFZs inhibition of the proteasomes chymotrypsin-like activity. Importantly, BDQ synergized with multiple proteasome and ubiquitin-activating enzyme inhibitors, but not with other standard MM agents, underscoring its selective interaction with the UPS pathway. BDQ-CFZ co-treatment markedly reduced MM cell viability and tumor burden in patient-derived cells and zebrafish xenograft models. These findings support the therapeutic repurposing of BDQ to potentiate PIs efficacy and overcome resistance in MM and related B-cell malignancies. KEY POINTSO_LIBedaquiline synergizes with proteasome inhibitors in MM and other hematologic malignancies, sparing normal cells C_LIO_LIBedaquiline targets ATP synthase {gamma} and boosts carfilzomib by enhancing chymotrypsin-like proteasome inhibition C_LI NOVELTYWe identify the antimicrobial bedaquiline and its fumarate salt as potent enhancers of proteasome inhibitor efficacy in multiple myeloma and other B-cell malignancies. By inhibiting ATP synthase {gamma}, bedaquiline amplifies chymotrypsin-like proteasome inhibition and overcomes drug resistance. This study reveals a mitochondria- proteasome vulnerability and proposes a clinically actionable strategy to restore sensitivity and reduce toxicity of proteasome-based therapies. VISUAL ABSTRACT O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=135 SRC="FIGDIR/small/661768v1_ufig1.gif" ALT="Figure 1"> View larger version (42K): org.highwire.dtl.DTLVardef@339391org.highwire.dtl.DTLVardef@1d3271org.highwire.dtl.DTLVardef@daa0faorg.highwire.dtl.DTLVardef@17eb4eb_HPS_FORMAT_FIGEXP M_FIG C_FIG

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BibTeXRIS

Cumerlato, M., Maccagno, M., Labrador, M., Luciano, B., Mereu, E., Porro, M., Bandini, C., Maisano, D., D'Agostino, M., Larocca, A., Gay, F. M., Tamburrini, A., Anselmi, F., Porporato, P. E., Oliviero, S., Bruno, B., Morelli, E., Munshi, N., PIVA, R.. 2025-07-03. Bedaquiline Amplifies Proteasome Inhibitor Efficacy and Overcomes Resistance in Multiple Myeloma. https://doi.org/10.1101/2025.06.29.661768

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