Effective extracellular vesicle-based intracellular delivery of an artificially encapsulated protein

Extracellular vesicles (EVs, exosomes) are approximately 30- to 200-nm-long vesicles that have received increased attention due to their role in cell-to-cell communication. Although EVs are highly anticipated to be a next-generation intracellular delivery tool because of their pharmaceutical advantages, including non-immunogenicity, their cellular uptake efficacy is low because of the repulsion of EVs and negatively charged cell membranes and size limitations in endocytosis.

Here, researchers from Osaka Prefecture University demonstrate a methodology for achieving enhanced cellular EV uptake using arginine-rich cell-penetrating peptides (CPPs) to induce active macropinocytosis. The induction of macropinocytosis via a simple modification to the exosomal membrane using stearylated octaarginine, which is a representative CPP, significantly enhanced the cellular EV uptake efficacy. Consequently, effective EV-based intracellular delivery of an artificially encapsulated ribosome-inactivating protein, saporin, in EVs was attained.

Schematic representation of the intracellular delivery of EVs

Objective therapeutic molecules are encapsulated in EVs by electroporation. EVs are then modified with stearyl-r8 peptide on EV membranes, resulting in the active induction of macropinocytosis and effective cellular uptake.