In a groundbreaking development poised to reshape approaches in cellular biology and therapeutic strategies, recent research uncovers a novel mechanism by which disulfiram—a drug historically used to combat chronic alcoholism—activates autophagy, a critical cellular degradation pathway. The study elucidates how disulfiram achieves this activation through proteasome inhibition coupled with the upregulation of pivotal molecular players c-Fos and beclin-1, thereby enhancing autophagic flux. Remarkably, this autophagic activation synergizes with chloroquine, a drug known for its lysosomal inhibition properties, suggesting promising avenues for combination therapies in disease contexts where autophagy modulation is beneficial.
Autophagy, a vital catabolic process, maintains cellular homeostasis by engulfing and degrading dysfunctional organelles and protein aggregates through lysosomal machinery. Dysregulation of autophagy is implicated in numerous pathologies, including neurodegenerative diseases, cancer, and infectious disorders. Hence, agents capable of modulating autophagic pathways have attracted immense scientific interest. Disulfiram’s newly discovered role as an autophagy activator through proteasome inhibition presents a sophisticated mechanism that bridges two fundamental degradative systems: the ubiquitin-proteasome system and autophagy-lysosome pathway.
Proteasomes, responsible for degrading short-lived and misfolded proteins, are crucial for proteostasis. Their inhibition leads to an accumulation of cellular proteins, which is hypothesized to trigger compensatory autophagy. Disulfiram’s capacity to inhibit proteasome activity was demonstrated to stimulate this compensatory autophagic response. Such a dual-targeted modulation disrupts protein degradation homeostasis, effectively pushing cells toward increased autophagic activity as a rescue mechanism. Central to this interplay is the transcription factor c-Fos, whose upregulation orchestrates downstream autophagy-related gene expression changes.
The research highlights the significant upregulation of beclin-1, a core initiator of autophagosome formation, facilitated by c-Fos activation. Beclin-1 forms part of a lipid kinase complex essential for nucleating autophagic vesicles, thereby launching the autophagic cascade. This molecular axis—disulfiram-induced proteasome inhibition leading to c-Fos-driven beclin-1 upregulation—constructs a compelling narrative for the modulation of autophagy at transcriptional and post-translational levels. Such a mechanism might potentiate robust autophagic flux, surpassing basal cellular activity and achieving therapeutic thresholds.
Intriguingly, the study also explores the synergism between disulfiram and chloroquine. Chloroquine, known primarily as an antimalarial agent, serves as a lysosomal inhibitor that hampers autophagosome-lysosome fusion, thereby arresting autophagy at a late stage. Paradoxically, when combined with disulfiram, which accelerates early-stage autophagy initiation, the net effect results in amplified autophagic stress. This interplay effectively overloads the degradative pathways in cells, potentially magnifying cytotoxic efficacy against pathological cells reliant on autophagy for survival, such as certain cancer cell types.
The implications of this synergistic modulation could revolutionize therapeutic regimens, especially within oncology where autophagy serves dual roles—cytoprotective under stress yet tumor-suppressive upon prolonged activation. Harnessing disulfiram’s proteasome inhibition and autophagy induction in concert with chloroquine’s lysosomal blockade may tip the balance toward enhanced cancer cell death while sparing normal cells. This therapeutic window expands the potential for repurposing widely available drugs with established safety profiles, reducing barriers to clinical translation.
Beyond oncology, this mechanistic insight into autophagy regulation opens new research vistas in neurodegeneration and infectious diseases. For example, neurodegenerative illnesses characterized by toxic protein accumulation could benefit from disulfiram’s dual intervention in degrading pathologic proteins via autophagy induction. Concurrently, infectious disease paradigms that exploit autophagic pathways for pathogen clearance might utilize this drug synergy to optimize host responses or overcome microbial resistance.
Technically, the research employed state-of-the-art proteomic analyses, live-cell imaging, and gene expression assays to dissect the molecular choreography underpinning disulfiram’s effects. Proteasome activity assays confirmed direct inhibition, while chromatin immunoprecipitation revealed c-Fos occupancy at the beclin-1 promoter region, confirming transcriptional regulation. Functional autophagy assays using LC3-II lipidation markers and autophagosome quantification substantiated the enhanced autophagy flux upon treatment.
Importantly, the study draws important distinctions regarding dosage and temporal dynamics. Disulfiram doses sufficient for proteasome inhibition triggered sustained autophagy induction without acute cytotoxicity in normal cell lines, underscoring the therapeutic relevance and safety margin. Meanwhile, the timing of chloroquine administration was critical to ensure proper synergy—administered subsequent to disulfiram-induced autophagy initiation for maximum lysosomal disruption.
This precision in pharmacodynamic modulation underscores the sophistication of therapeutic strategy emerging from the research. It advocates for personalized medicine approaches that integrate drug timing, dosage, and cellular context, improving efficacy while minimizing unintended adverse effects. Such frameworks could be pivotal in managing complex diseases responsive to autophagy modulation.
The discovery also rekindles interest in disulfiram’s versatile pharmacology beyond its classical enzyme inhibition of aldehyde dehydrogenase. Its chemical structure and redox properties enable interactions with diverse molecular targets, facilitating proteasome inhibition and induction of stress-responsive transcription factors like c-Fos. This multi-targeted nature paves the way for combination therapies exploiting synergistic mechanisms intrinsic to disulfiram.
Moreover, the potential to repurpose an FDA-approved drug with a well-characterized safety profile accelerates the translational trajectory of these findings. Clinical trials adapting disulfiram for oncological or neurodegenerative conditions could be expedited, leveraging existing pharmacokinetic data and manufacturing pipelines. Concurrently, chloroquine offers cost-effective adjunct treatment, enhancing accessibility in resource-limited settings.
The study’s implications extend to fundamental biology as well, providing deeper understanding of how cells reciprocally regulate proteasomal and autophagic systems in stress and homeostasis. This crosstalk ensures cellular adaptability to proteotoxic insults, with transcription factors like c-Fos acting as molecular switches. Understanding such regulatory networks enriches the conceptual framework of cellular maintenance and disease pathophysiology.
Finally, this work prompts further research to delineate the full spectrum of molecular players involved in disulfiram-mediated autophagy activation. Questions remain regarding potential off-target effects, long-term consequences of combined drug regimens, and varying responses across different cell types and disease states. Expanding this knowledge base will be instrumental for refining therapeutic applications and exploring novel drug combinations.
In essence, the unveiling of disulfiram as a potent activator of autophagy through proteasome inhibition and c-Fos/beclin-1 upregulation represents a paradigm shift in drug repurposing and autophagy modulation. Coupled with chloroquine, this synergy holds immense promise for innovative treatments targeting diseases characterized by dysregulated protein homeostasis. The scientific community eagerly anticipates subsequent translational research that will harness this mechanistic insight to generate effective, durable clinical interventions.
Subject of Research:
Disulfiram’s role in autophagy activation via proteasome inhibition and c-Fos/beclin-1 upregulation, and its synergistic effects with chloroquine.
Article Title:
Disulfiram activates autophagy via proteasome inhibition and c-Fos/beclin-1 upregulation, synergizing with chloroquine.
Article References:
Wang, K., Wang, Z., Peng, W. et al. Disulfiram activates autophagy via proteasome inhibition and c-Fos/beclin-1 upregulation, synergizing with chloroquine. Cell Death Discov. (2025). https://doi.org/10.1038/s41420-025-02899-7
Image Credits: AI Generated
DOI: https://doi.org/10.1038/s41420-025-02899-7
Tags: autophagic flux enhancementcellular homeostasis and autophagychloroquine synergy with disulfiramcombination therapies for neurodegenerative diseasesdisulfiram and autophagy activationdisulfiram in chronic alcoholism treatmentimplications of autophagy dysregulation inlysosomal inhibition and drug developmentmolecular mechanisms of autophagy modulationproteasome inhibition and cellular healthrole of c-Fos and beclin-1 in autophagytherapeutic strategies for cancer treatment
