Extracellular Vesicle-Associated Angiopoietin-2 and Cell Migration-Inducing Protein in Lung Cancer Progression and Brain Metastases.

BACKGROUND: Angiopoietin-2 (ANGPT2) and cell migration-inducing protein (CEMIP) are key regulators of angiogenesis, extracellular matrix remodeling, and metastatic progression in various cancers, including lung cancer (LC). The presence of these biomarkers in extracellular vesicles (EVs) may offer valuable insights into the molecular mechanisms underlying LC progression and metastasis. Extracellular vesicles play a critical role in LC by enhancing intercellular communication and supporting processes such as angiogenesis, immune evasion, and metastasis, transferring key molecules like vascular endothelial growth factor (VEGF) and pro-angiogenic microRNAs (miRNAs). METHODS: This study aimed to investigate the presence and quantification of ANGPT2 and CEMIP in the cargo of EVs isolated from plasma samples obtained from the peripheral venous blood of patients with localized lung cancer (LLC group), lung cancer with brain metastases (LCM group), and healthy controls (HC group). EVs were isolated using the density gradient ultracentrifugation method, and their characterization was performed through scanning and transmission electron microscopy as well as flow cytometry. Western blot analysis was used to identify ANGPT2 and CEMIP in EV cargo, and band intensity from western blot images was quantified using specialized software. RESULTS: The expression of ANGPT2 and CEMIP in EV cargo was significantly higher in the oncologic groups (LLC and LCM combined) compared to the HC group. Notably, EV CEMIP levels were, on average, 59% higher in patients with brain metastases than in those with localized lung cancer. Following surgical resection, postoperative EV ANGPT2 and EV CEMIP levels decreased by 36% and 8.5%, respectively, in the LLC group, and by 40% and 4.6%, respectively, in the LCM group. CONCLUSION: These findings emphasize the potential of ANGPT2 and CEMIP as biomarkers for LC progression and prognosis. To our knowledge, no previous study has evaluated the presence and quantification of ANGPT2 and CEMIP in EV cargo from lung cancer patients. To further validate their role in cancer progression, functional studies should explore the mechanistic effects of EV-associated ANGPT2 and CEMIP on angiogenesis, immune modulation, cell migration, extracellular matrix remodeling, and tumor progression in lung cancer models.