Cannabidiol modulates exosomal miRNA networks to enhance Imatinib mesylate response in chronic myelogenous leukemia

Abstract

Background/Objectives Chronic myelogenous leukemia (CML) is a clonal myeloproliferative disease driven by the BCR-ABL1 fusion oncogene. Tyrosine kinase inhibitors (TKIs) such as Imatinib mesylate have dramatically improved patient outcomes, yet resistance remains a major obstacle to long-term efficacy. Exosomes, as carriers of bioactive molecules including miRNAs, are increasingly recognized as mediators of drug resistance. CBD has demonstrated antiproliferative and pro-apoptotic effects in several cancer models, but its potential to modulate Imatinib sensitivity or resistance in CML remains unclear. This study aimed to investigate exosomal miRNA signatures associated with Imatinib sensitivity and resistance in the context of treatment with Cannabidiol (CBD), Imatinib mesylate (IM), and their combination.

Methods Following treatment with CBD, IM, and CBD+IM, exosomal miRNA profiles in Imatinib-sensitive (K-562S) and Imatinib-resistant (K-562 R) cell lines were analyzed. Gene Ontology (GO) enrichment and semantic clustering was performed.

Results CBD activated tumor-suppressive and apoptosis-related miRNAs in K-562S cells, whereas K-562 R cells showed a dual response involving oncogenic miRNAs and metabolic regulators. IM induced suppressive cascades in K-562S but caused loss of canonical tumor suppressors in K-562 R. CBD+IM produced synergistic amplification of apoptotic and differentiation-related pathways in sensitive cells, while resistant cells showed partial restoration of apoptosis but persistent loss of tumor suppressors. HMGB1-associated miRNAs were identified, of which suppressed were miR-615-5p, miR-4435, let-7 g-3p, and the miR-548 family, alongside upregulated miR-3191-3p and miR-33a-5p.

Conclusions Circulating miRNAs are valuable biomarkers for TKI resistance in CML. Targeting HMGB1-associated miRNAs, together with combined CBD and IM treatment, may help re-establish apoptotic regulation and overcome resistance mechanisms.

Keywords

Chronic myelogenous leukemia (CML) / Imatinib resistance / Cannabidiol (CBD) / Exosomal miRNAs / HMGB1 regulation

CRediT authorship contribution statement

Conceptualization, Petar P. Donchev; methodology, Petar P. Donchev; formal analysis, Petar P. Donchev; investigation, Petar P. Donchev; data curation, Petar P. Donchev; writing—original draft preparation, Petar P. Donchev; writing—review and editing, Petar P. Donchev and Prof. Svetla Trifonova Danova; visualization, Petar P. Donchev; supervision, Petar P. Donchev; project administration, Petar P. Donchev; funding acquisition, Petar P. Donchev. All authors have read and agreed to the published version of the manuscript.

Funding

This research was funded by the Bulgarian National Science Fund, grant number KП-06-M61/6.

Data availability

The miRNA sequencing data generated and analyzed in this study are publicly available in the ArrayExpress database (Annotare 2.0, EMBL-EBI) under accession number E-MTAB-16287. Further details can be accessed at https://www.ebi.ac.uk/arrayexpress/.

Declaration of Competing Interest

The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Petar Petrov Donchev reports financial support was provided by Bulgarian National Science Fund. Petar Petrov Donchev reports a relationship with Stephan Angeloff Institute of Microbiology Bulgarian Academy of Sciences that includes: employment. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

Declaration of Generative AI and AI-assisted technologies in the writing process

During the preparation of this work the authors used Microsoft Copilot (current version, 2025) in order to assist with data analysis and interpretation. After using this tool, the authors reviewed and edited the content as needed and take full responsibility for the content of the published article.

Acknowledgments

The authors would like to thank the Department of Immunology for accommodating this research and providing valuable support.

The author extends his sincere gratitude to the Bulgarian National Science Fund for the support provided under grant KП-06-M61/6, which was indispensable for carrying out this study.

We thank Assoc. Prof. Emi Haladjova and Prof. Petar Petrov (Institute of Polymers, Bulgarian Academy of Sciences) for their expert support in the DLS verification of exosomes.

Appendix A. Supplementary material

Supplementary data associated with this article can be found in the online version at doi:10.1016/j.gmg.2026.100094.

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