Methodologies to evaluate the radiation-induced changes on extracellular vesicles.

The extracellular vesicles (EVs) are carriers that actively transfer functional biomolecules between cells affecting the intercellular communication (Pitt, Kroemer, & Zitvogel, 2016). EV signaling has consequences on the targeted cell behavior impacting multiple processes from health to disease, including cancer (Yates et al., 2022). Radiation treatment (RT) is one of the gold standard and effective treatments for cancer, as curative or palliative (Chandra, Keane, Voncken, & Thomas, 2021). RT induces different release of EVs and their cargo is altered. In addition, the uptake of EVs secreted by irradiated cells is affected. Hence, a deep investigation is required to better understand how RT influence the cell-to-cell communication thought signals shuttle by EVs. Here, detailed methods to study the RT effects on EV size and secretion, EV protein expression, EV uptake will be described. Alterations and adaptions might make the protocols applicable to different cell lines, and with different types of RT and dose exposures.