Affordable Access

deepdyve-link
Publisher Website

Engineered Exosomes as Vehicles for Biologically Active Proteins.

Authors
  • Sterzenbach, Ulrich1
  • Putz, Ulrich1
  • Low, Ley-Hian2
  • Silke, John3
  • Tan, Seong-Seng1
  • Howitt, Jason4
  • 1 The Florey Institute of Neuroscience and Mental Health, The University of Melbourne, Parkville, Victoria 3052, Australia. , (Australia)
  • 2 University of California San Francisco and Veterans Affairs Medical Center, San Francisco, CA 94121, USA.
  • 3 Walter and Eliza Hall Institute, The University of Melbourne, Parkville, Victoria 3052, Australia. , (Australia)
  • 4 The Florey Institute of Neuroscience and Mental Health, The University of Melbourne, Parkville, Victoria 3052, Australia. Electronic address: [email protected] , (Australia)
Type
Published Article
Journal
Molecular Therapy
Publisher
Elsevier
Publication Date
Jun 07, 2017
Volume
25
Issue
6
Pages
1269–1278
Identifiers
DOI: 10.1016/j.ymthe.2017.03.030
PMID: 28412169
Source
Medline
Keywords
License
Unknown

Abstract

Exosomes represent an attractive vehicle for the delivery of biomolecules. However, mechanisms for loading functional molecules into exosomes are relatively unexplored. Here we report the use of the evolutionarily conserved late-domain (L-domain) pathway as a mechanism for loading exogenous proteins into exosomes. We demonstrate that labeling of a target protein, Cre recombinase, with a WW tag leads to recognition by the L-domain-containing protein Ndfip1, resulting in ubiquitination and loading into exosomes. Our results show that Ndfip1 expression acts as a molecular switch for exosomal packaging of WW-Cre that can be suppressed using the exosome inhibitor GW4869. When taken up by floxed reporter cells, exosomes containing WW-Cre were capable of inducing DNA recombination, indicating functional delivery of the protein to recipient cells. Engineered exosomes were administered to the brain of transgenic reporter mice using the nasal route to test for intracellular protein delivery in vivo. This resulted in the transport of engineered exosomes predominantly to recipient neurons in a number of brain regions, including the olfactory bulb, cortex, striatum, hippocampus, and cerebellum. The ability to engineer exosomes to deliver biologically active proteins across the blood-brain barrier represents an important step for the development of therapeutics to treat brain diseases.

Report this publication

Statistics

Seen <100 times