Understanding the Role of Glioblastoma-Derived Extracellular Vesicles in Cardiovascular Cell Responses

Introduction Glioblastoma (GBM) is the most aggressive brain cancer, with rapid progression and high recurrence. It is linked to local and peripheral thrombosis, complicating treatment. GBM cells release extracellular vesicles (EVs) carrying oncogenic cargo, contributing to vascular issues, clotting, and metastasis. EV size and subtype further influence these effects. As cancer treatments extend survival, cardiovascular side effects are more evident. While cardio-oncology investigates connection of cancer and its treatment to cardiovascular health, the potential role of cancer-derived EVs in this axis remains unclear. Aim This study aim to explore the effect of GBM-derived EVs of varying sizes on cardiomyocytes and vascular endothelial cells. Methods We isolated EVs from the conditioned media of the human GBM cell line HROG36 using centrifugation and ultracentrifugation. This allowed the separation of small EVs (<200 nm; sEVs) from large EVs (>200 nm; lEVs). Both fractions were characterized by size (NTA) and morphology (cryo-EM). EV internalization in HUVECs and primary Wistar rat cardiomyocytes was examined via fluorescence microscopy and cytochemistry. Following treatment with GBM-derived EVs, HUVEC metabolic activity (PrestoBlue), migration (wound healing), and angiogenesis (tube formation on ECM) were assessed. Additionally, cardiomyocyte metabolic activity and contraction parameters were analyzed post-EV exposure. Results Our findings showed that GBM-derived EVs entered HUVECs and cardiomyocytes within 24 hours. After 72 hours, lEVs significantly increased HUVEC metabolic activity compared to sEVs. Moreover, lEVs notably enhanced HUVEC migration and blood vessel formation, while sEVs had an inhibitory effect. Similarly, lEVs stimulated cardiomyocyte metabolic activity, whereas sEVs did not. However, both EV types prolonged cardiomyocyte contraction time and reduced the beating rate. Conclusions GBM-derived EVs impact cardiovascular cells in a size-dependent manner. lEVs enhance metabolic activity, endothelial migration, and angiogenesis, while sEVs inhibit these processes. Both EV types impair cardiomyocyte function by prolonging contraction time and reducing the beating rate. These findings highlight EV size as a key factor in GBM-related cardiovascular complications, warranting further research in cardio-oncology.