Engineered Extracellular Vesicles Prepared by Membrane Extrusion Are Good Substitutes for Natural Extracellular Vesicles

Extracellular vesicles (EV) as drug delivery nanocarriers are under intense investigation. Although clinical-grade EVs have been produced on a large-scale, low yield and high production costs of natural EVs (nEV) limit the relevant industrial translation. Recent studies show that mechanical extrusion of cells can generate nEV-like engineered EVs (eEV) which can also be used as drug nanocarriers. Moreover, in comparison with nEVs, eEVs have similar physicochemical properties. Nevertheless, a comprehensive comparison of cargo between nEVs and eEVs has not been investigated yet. Therefore, the aim of this study is to profile and compare eEVs to nEVs. Our results show that no significant difference was found in size, morphology, and classical markers between nEVs and eEVs derived from MDA-MB-231 cells. Protein sequencing data reveals the similarity of membrane proteins between the two groups was ~71%, while it was ~21% when pertaining to total protein cargo. Notably, a high similarity of membrane proteins was also found between nEVs and eEVs derived from eight additional cancer cell lines. Moreover, analysis of the top 1000 small RNA with RNA sequencing showed a ~65% similarity between the two groups. Altogether, we infer from the high similarity of membrane proteins and small RNA cargo that eEVs can be a good substitute for nEVs. In brief, our findings support previous studies which discovered eEVs own comparable performance of nEVs and could pave the way for clinical implementation of eEV-based therapeutics in the future.