Vectorization of biomacromolecules into cells using extracellular vesicles with enhanced internalization induced by macropinocytosis

Sci Rep. 2016 Oct 17;6:34937. doi: 10.1038/srep34937.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Arginine / chemistry
  • CHO Cells
  • Cell Count
  • Cell Line, Tumor
  • Cell Membrane / metabolism
  • Cell-Penetrating Peptides / metabolism
  • Cricetinae
  • Cricetulus
  • Endocytosis
  • Exosomes / metabolism*
  • Extracellular Vesicles / metabolism
  • Fluorescent Dyes / chemistry
  • HeLa Cells
  • Humans
  • Macromolecular Substances*
  • Microscopy, Electron, Transmission
  • Oligopeptides / chemistry
  • Peptides / chemistry
  • Pinocytosis*
  • Ribosome Inactivating Proteins, Type 1 / chemistry*
  • Ribosomes / chemistry*
  • Saporins
  • Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

Substances

  • Cell-Penetrating Peptides
  • Fluorescent Dyes
  • Macromolecular Substances
  • Oligopeptides
  • Peptides
  • Ribosome Inactivating Proteins, Type 1
  • octaarginine
  • Arginine
  • Saporins