Prof.ssa Canonico, Dott. Tiboni, Dott.ssa Montanari, Prof Casettari, Prof Guescini, Prof.ssa Rossi, Prof Magnani

Abstract

Extracellular vesicles (EVs) are promising natural nanocarriers for the delivery of therapeutic  agents. As with any other kind of cell, red blood cells (RBCs) produce a limited number of EVs under  physiological and pathological conditions. Thus, RBC-derived extracellular vesicles (RBCEVs) have  been recently suggested as next-generation delivery systems for therapeutic purposes. In this  paper, we show that thanks to their unique biological and physicochemical features, RBCs can  be efciently pre-loaded with several kinds of molecules and further used to generate RBCEVs. A  physical vesiculation method, based on “soft extrusion”, was developed, producing an extremely  high yield of cargo-loaded RBCEV mimetics. The RBCEVs population has been deeply characterized  according to the new guidelines MISEV2023, showing great homogeneity in terms of size, biological  features, membrane architecture and cargo. In vitro preliminary results demonstrated that RBCEVs  are abundantly internalized by cells and exert peculiar biological efects. Indeed, efcient loading and  delivery of miR-210 by RBCEVs to HUVEC has been proven, as well as the inhibition of a known mRNA  target. Of note, the bench-scale process can be scaled-up and translated into clinics. In conclusion, this  investigation could open the way to a new biomimetic platform for RNA-based therapies and/or other  therapeutic cargoes useful in several diseases.

Keyword

Extracellular vesicles, RBC-derived extracellular vesicles, EV engineering, Drug delivery

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