PROFILE OF INFLAMMATORY CYTOKINES AND EXTRACELLULAR VESICLES AS BIOMARKERS OF RESPONSE TO RADIOTHERAPY IN PATIENTS WITH HEAD AND NECK CANCER

• Grant number: 23/16407-4
• Support Opportunities: Scholarships in Brazil - Master
• Start date: September 01, 2024
• End date: July 31, 2025
• Field of knowledge: Biological Sciences - Biochemistry - Molecular Biology
• Principal Investigator: Wanessa Fernanda Altei
• Grantee: Igor Sampaio Fagundes
• Host Institution: Hospital do Câncer de Barretos. Fundação Pio XII (FP). Barretos , SP, Brazil

Abstract

Radiotherapy (RT), alone or in combination with other cytotoxic agents, plays a fundamental role in the treatment of head and neck squamous cell carcinoma (HNSCC). Although this treatment has been established as a major cornerstone of oncological treatment for HNSCC, with good response rates and disease control, patients respond differently to this therapy, with some not responding at all. Therefore, an in-depth analysis of the molecular alterations related to this therapeutic regimen may help in identifying factors capable of predicting clinical responses. Given the association between RT and the immune system, the detection of markers derived from damage carried by extracellular vesicles (EVs), through techniques such as liquid biopsy, is essential for the development of new molecular biomarkers. After RT, tumor cells alter the production and content of their EVs, as observed in cases of increased expression of HSP70 (Heat Shock Protein-70) and therapeutic failures associated with PD-L1. In this study, we will investigate the profile of EVs and inflammatory cytokines in patients exposed to RT associated with chemotherapy (CT) at the Barretos Cancer Hospital (BCH). Using samples from these patients available in the institution's biobank, EVs present in radiation-free plasma samples, 5 months post-RT, and 1 year after treatment completion will be isolated. The isolation of these EVs will be performed by size exclusion chromatography (SEC), and their characterization will be carried out by transmission electron microscopy (TEM), nanoparticle tracking analysis (NTA), and protein quantification, in addition to evaluating the profile of biomarkers such as ALIX, HSP70, CD63, VEGFR, TGF-², and PD-L1 present in these vesicles through EV immunolabeling and Western Blotting. The cytokines present in the patients' plasma will be evaluated using the Cytometric Beads Array (CBA) technique. Thus, we may obtain important data that could translate into the prediction of recurrences, metastases, and treatment responses.