Alcohol Exposure Impacts the Composition of HeLa-Derived Extracellular Vesicles

Abstract

Extracellular vesicles are nanosized vesicles that are under intense investigation for their role in intercellular communication. Extracellular vesicles have begun to be examined for their role in disease protection and their role as disease biomarkers and/or vaccine agents. The goal of this study was to investigate the effects of alcohol exposure on the biogenesis and composition of extracellular vesicles derived from the cervical cancer line, HeLa. The HeLa cells were cultured in exosome-free media and were either mock-treated (control) or treated with 50 mM or 100 mM of alcohol for 24 h and 48 h. Our results demonstrated that alcohol significantly impacts HeLa cell viability and exosome biogenesis/composition. Importantly, our studies demonstrate the critical role of alcohol on HeLa cells, as well as HeLa-derived extracellular vesicle biogenesis and composition. Specifically, these results indicate that alcohol alters extracellular vesicles’ packaging of heat shock proteins and apoptotic proteins. Extracellular vesicles serve as communicators for HeLa cells, as well as biomarkers for the initiation and progression of disease.

Effects of Cocaine on Human Glial-Derived Extracellular Vesicles

“…The size and quantity of isolated EVs were measured using NTA and TEM. We found that cocaine did not alter the size of EVs and EVs are visible and easily recognizable ( Figure 1B and Supplementary Figures 1B,D–F ), which ranged from 30 to 150 nm in diameter across conditions, consistent with previous studies ( Bell et al, 2019 ; Crenshaw et al, 2019 ; Jones et al, 2019 ; Kumar et al, 2020 ). However, 100 μM cocaine slightly reduced the number of EV particles per mL compared with the 1 μM cocaine and control conditions, although this difference was not significant ( Figure 1C and Supplementary Figures 1C,D–F ).…”

“…Medium only was used as a control. After incubating cells with cocaine, EVs released into the medium were isolated and purified using a standard ultracentrifugation procedure ( Bell et al, 2019 ; Crenshaw et al, 2019 ; Jones et al, 2019 ). The size and quantity of isolated EVs were measured using NTA and TEM.…”

Image_4.JPEG

“…The size and quantity of isolated EVs were measured using NTA and TEM. We found that cocaine did not alter the size of EVs and EVs are visible and easily recognizable ( Figure 1B and Supplementary Figures 1B,D-F), which ranged from 30 to 150 nm in diameter across conditions, consistent with previous studies (Bell et al, 2019;Crenshaw et al, 2019;Jones et al, 2019;Kumar et al, 2020). However, 100 µM cocaine slightly reduced the number of EV particles per mL compared with the 1 µM cocaine and control conditions, although this difference was not significant ( Figure 1C and Supplementary Figures 1C,D-F).…”

Mesenchymal Stem Cell-Derived Exosomes: Applications in Regenerative Medicine

“…The two constitutive forms of heat shock proteins (i.e., HSP70 and HSP90) are found in exosomes which perform the function of antigen presentation and aid in loading antigenic peptides onto MHC class I molecules (Table 3). Notably, the MHC class I molecules are present in most isolated exosomes [141][142][143][144].…”