title: Vectorization of biomacromolecules into cells using extracellular vesicles with enhanced internalization induced by macropinocytosis.
abstract: 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, we 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.
authors.affiliation: Nanoscience and Nanotechnology Research Center, Research Organization for the 21st Century, Osaka Prefecture University, Naka-ku, Sakai, Osaka 599-8570, Japan.
authors.forename: Ikuhiko
authors.initials: I
authors.lastname: Nakase
authors.affiliation: Nanoscience and Nanotechnology Research Center, Research Organization for the 21st Century, Osaka Prefecture University, Naka-ku, Sakai, Osaka 599-8570, Japan.|Graduate School of Science, Osaka Prefecture University, Naka-ku, Sakai, Osaka 599-8531, Japan.
authors.forename: Kosuke
authors.initials: K
authors.lastname: Noguchi
authors.affiliation: Graduate School of Science, Osaka Prefecture University, Naka-ku, Sakai, Osaka 599-8531, Japan.
authors.forename: Ikuo
authors.initials: I
authors.lastname: Fujii
authors.affiliation: Institute for Chemical Research, Kyoto University, Uji, Kyoto 611-0011, Japan.
authors.forename: Shiroh
authors.initials: S
authors.lastname: Futaki
mesh_terms: D000818:Animals; D001120:Arginine / Q000737:chemistry; D016466:CHO Cells; D002452:Cell Count; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000378:metabolism; D057846:Cell-Penetrating Peptides / Q000378:metabolism; D006224:Cricetinae; D003412:Cricetulus; D004705:Endocytosis; D055354:Exosomes / Q000378:metabolism*; D000067128:Extracellular Vesicles / Q000378:metabolism; D005456:Fluorescent Dyes / Q000737:chemistry; D006367:HeLa Cells; D006801:Humans; D046911:Macromolecular Substances; D046529:Microscopy, Electron, Transmission; D009842:Oligopeptides / Q000737:chemistry; D010455:Peptides / Q000737:chemistry; D010873:Pinocytosis; D054789:Ribosome Inactivating Proteins, Type 1 / Q000737:chemistry*; D012270:Ribosomes / Q000737:chemistry*; D000076985:Saporins; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D057846:Cell-Penetrating Peptides; D005456:Fluorescent Dyes; D046911:Macromolecular Substances; D009842:Oligopeptides; D010455:Peptides; D054789:Ribosome Inactivating Proteins, Type 1; C448619:octaarginine; D001120:Arginine; D000076985:Saporins