Characterization of large extracellular vesicles released by apoptotic and pyroptotic cells

ABSTRACT: Extracellular vesicles (EVs) are emerging key factors maintaining cellular homeostasis, critical mediators of intercellular communication, potential biomarkers and therapeutic tools. While small EVs have been extensively characterized, the molecular signatures of large EVs (including those generated during regulated cell death pathways) remain poorly defined. Here, we investigated the characteristics of large EVs released during apoptosis and pyroptosis by human monocytic cell lines (THP-1 and U937). Apoptosis was induced by staurosporine and blocked with the pan-caspase inhibitor Q-VD-OPh, whereas pyroptosis was triggered by LPS/nigericin and inhibited with a selective NLRP3 inhibitor. We found that both forms of regulated cell death markedly enhanced the release of large EVs. Both apoptotic and pyroptotic large EVs showed increased Annexin V binding and decreased CD9 expression as compared to those released by healthy living cells. Apoptosis- and pyroptosis-derived large EVs exhibited distinct proteomic profiles. Pyroptotic large EVs were shown to contain interacting protein networks of RNA binding proteins and chromatin-associated proteins many of which are known as established damage associated molecular patterns or alarmins. On the other hand, we found that a subpopulation of apoptotic large EVs was characterized by the presence of dsDNA, and active caspase-3/7. Together, our data shed light on the specific protein content of large EVs released by cells undergoing apoptosis and pyroptosis. This study identifies candidate markers of large EVs released by dying cells and may enhance our understanding of the role of EVs in cell death.

INSTRUMENT(S): Orbitrap Fusion Lumos

ORGANISM(S): Homo Sapiens (ncbitaxon:9606)

SUBMITTER: Edit Buzas  

PROVIDER: MSV000100407 | MassIVE | Thu Jan 08 04:16:00 GMT 2026

SECONDARY ACCESSION(S): PXD072811

REPOSITORIES: MassIVE