Akademik Veri Yönetim Sistemi
Extracellular Vesicles Conferences, Ankara, Türkiye, 20 Eylül 2024, cilt.89, ss.32, (Özet Bildiri)
The Effects of Exosome-Encapsulated RO5963 Inhibitor on Human Chronic Myeloid Leukemia Cell Line
Chronic myeloid leukemia (CML) is a clonal disease originating from primitive pluripotent stem cells,
characterized by myeloid hyperplasia in the bone marrow and a high leukocyte count composed of
myeloid cells in peripheral blood. Overexpression of MDM2/MDMX negatively regulates p53
activation, accelerating disease progression and conferring apoptotic resistance in CML. Exosomes, the
smallest extracellular vesicles, have been discovered to not only carry prognostic markers but also
function as nanoparticle-like carriers of bioactive compounds. This study aims to determine the effects
of the exosome-treated dual MDM2/MDMX inhibitor RO5963 on cell viability in a chronic myeloid
leukemia cell line. The IC50 value of RO5963 in K562 cells was determined for a 24-hour incubation
period. Exosomes were isolated from serum using the "Exofast Exosome Isolation Reagent" (Cat. No:
D030). Characterization of isolated exosomes was performed using flow cytometry with CD9 and CD81
staining. Characterized exosomes were treated with RO5963 using incubation and sonication (3
minutes) methods. Cell viability of RO5963 treated with and without exosomes was evaluated using
the WST-1 method, validation was performed using UV-Vis spectrophotometry. The IC50 value of
RO5963 in K562 cells (24-hour incubation) was found to be 105 µM. Flow cytometry confirmed
successful exosome isolation with positive staining for CD9 and CD81. Cytotoxicity tests showed cell
viability at 24 hours as follows: Sonication-Exosome-encapsulated-RO5963 group (Exo-RO-S) at
63.42%, non-encapsulated free RO-5963 group (Exo-RO-F) at 66.02%, and inkubation-exosome-
encapsulated-RO5963 group (Exo-RO-I) at 86.81%. UV-Vis analysis at 387 nm showed results
consistent with the WST-1 findings for the Exo-RO-S, Exo-RO-F and Exo-RO-I groups. It has been
reported in many studies that encapsulation with exosome increases drug effectiveness. For example,
Hua et al. determined that exosome-encapsulated miRNA-FAM and 7-coumarin increased apoptosis
compared to use alone. Değirmenci et al. found that exosomes-encapsulated lapatinib increased the
effectiveness of the drug by suppressing the proliferation of breast cancer cells. Bagheri et al. reported
that exosome-encapsulated adriamycin target molecule showed higher efficacy than free doxorubicin
in the treatment of colorectal cancer. Consistent with these data, in this study, exosomes-encapsulated
with RO-5963 by sonication showed higher cytotoxic activity than free RO-5963 group. The findings
suggest that RO5963 dual inhibitor treated with exosomes via sonication could be a potential therapeutic
agent for CML. Exosome-based nanoparticles may represent an effective and alternative treatment
strategy. Further immunological analyses are needed to elucidate the molecular mechanism and
apoptotic effects of the exosome-RO5963 complex in CML.
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International journal of pharmaceutics, 651, 123788. https://doi.org/10.1016/j.ijpharm.2024.123788
Keyword: Exosome, RO5963, Chronic Myeloid Leukemia, MDM2, MDMX