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Abstract
Introduction. The efficacy of systemic cytostatic therapy for oncological diseases, including ovarian cancer, is limited by toxic effects on healthy tissues. The development of targeted drug delivery technologies for anticancer agents represents an urgent priority. A promising strategy involves creating innovative delivery systems based on extracellular nanovesicles (ENVs) modified with DNA aptamers.
Aim. To develop a vesicular membrane modification technology using DNA aptamers and evaluate the efficacy of this system for targeted doxorubicin delivery to ovarian cancer cells in vitro.
Materials and Methods. The study utilized ovarian cancer peritoneal metastasis tissue samples (n=3) and Skov-3 and Ovcar-3 cell lines. ENVs were isolated from healthy donor plasma and characterized using nanoparticle tracking analysis (NTA) and flow cytometry. Doxorubicin (DOX) loading was achieved through an optimized sonoporation protocol, followed by purification of DOX-ENV complexes using size-exclusion chromatography. Vesicular surface modification was accomplished via hydrophobic interaction between the membrane and cholesterol-conjugated DNA aptamers. System cytotoxicity was assessed using MTT assay.
Results. Initial candidate aptamer selection was performed through literature analysis and binding affinity assessment to ovarian cancer cells. The vesicular delivery system development in the second stage of the study involved: ENV isolation from donor plasma via ultracentrifugation; doxorubicin loading through sonoporation; purification of DOX-ENV complexes; and surface modification to create DOX-ENV-APT complexes. The optimized system demonstrated significantly enhanced cytotoxicity compared to non-targeted complexes (survival rates: Skov-3: 1% vs 57%; Ovcar-3: 0.03% vs 0.3%).
Conclusion. Aptamer-mediated surface modification significantly enhanced the targeted delivery efficiency of cytostatic drugs to ovarian cancer cells in vitro. DNA aptamer-functionalized extracellular vesicles represent a promising platform for improving the therapeutic index of systemic chemotherapy.
References
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