Effects of Canine Oral Melanoma-Derived Extracellular Vesicles on Tumor Cell Migration Capacity: An In Vitro Study

• Grant number: 25/10935-4
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: August 01, 2025
• End date: July 31, 2026
• Field of knowledge: Agronomical Sciences - Veterinary Medicine
• Principal Investigator: Cristina de Oliveira Massoco Salles Gomes
• Grantee: Isabele da Costa Silva
• Host Institution: Faculdade de Medicina Veterinária e Zootecnia (FMVZ). Universidade de São Paulo (USP). São Paulo , SP, Brazil

Abstract

Canine oral melanoma is a highly aggressive neoplasm with a high metastatic potential, whose progression may be influenced by tumor-derived extracellular vesicles (TEVs). These nanoparticles serve as key mediators of intercellular communication, modulating the tumor microenvironment and promoting processes such as angiogenesis, immune evasion, and cell migration and invasion. In veterinary medicine, the lack of standardized protocols for the isolation and characterization of these vesicles limits the understanding of their specific role in tumor progression.Therefore, the primary aim of this project is to investigate the effect of TEVs on the migratory potential of melanocytic tumor cells through in vitro cell migration assays.For this purpose, TEVs will be isolated and purified from a canine oral melanoma lymph node metastatic cell line using differential centrifugation, with prior treatment of the cells with doxorubicin to induce stress and enhance vesicle secretion.The vesicles will be characterized by nano-flow cytometry, and the migration assay will be conducted by comparing control groups (without TEVs) and groups treated with different vesicle concentrations (1, 10, and 100 ¿g/mL), with temporal monitoring (0, 6, 12, 24, and 48 hours) through image-based analysis. The central hypothesis is that extracellular vesicles will significantly enhance tumor cell migration, supporting their role in metastatic progression.