Serum Deprivation of Mesenchymal Stem Cells Improves Exosome Activity and Alters Lipid and Protein Composition
Reka Agnes Haraszti,
Rachael Miller,
Michelle L. Dubuke,
Hannah E. Rockwell,
Andrew H. Coles,
Ellen Sapp,
Marie-Cecile Didiot,
Dimas Echeverria,
Matteo Stoppato,
Yves Y. Sere,
John Leszyk,
Julia F. Alterman,
Bruno M.D.C. Godinho,
Matthew R. Hassler,
Justice McDaniel,
Niven R. Narain,
Rachel Wollacott,
Yang Wang,
Scott A. Shaffer,
Michael A. Kiebish,
Marian DiFiglia,
Neil Aronin,
Anastasia Khvorova
Affiliations
Reka Agnes Haraszti
RNA Therapeutics Institute, University of Massachusetts Medical School, Worcester, MA, USA; Corresponding author
Rachael Miller
RNA Therapeutics Institute, University of Massachusetts Medical School, Worcester, MA, USA; Department of Medicine, University of Massachusetts Medical School, Worcester, MA, USA
Michelle L. Dubuke
Mass Spectrometry Facility, Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School, Shrewsbury, MA, USA
Hannah E. Rockwell
BERG LLC, Framingham, MA, USA
Andrew H. Coles
RNA Therapeutics Institute, University of Massachusetts Medical School, Worcester, MA, USA
Ellen Sapp
Mass General Institute for Neurodegenerative Disease, Boston, MA, USA
Marie-Cecile Didiot
RNA Therapeutics Institute, University of Massachusetts Medical School, Worcester, MA, USA
Dimas Echeverria
RNA Therapeutics Institute, University of Massachusetts Medical School, Worcester, MA, USA
Matteo Stoppato
MassBiologics, Boston, MA, USA
Yves Y. Sere
MassBiologics, Boston, MA, USA
John Leszyk
Mass Spectrometry Facility, Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School, Shrewsbury, MA, USA
Julia F. Alterman
RNA Therapeutics Institute, University of Massachusetts Medical School, Worcester, MA, USA
Bruno M.D.C. Godinho
RNA Therapeutics Institute, University of Massachusetts Medical School, Worcester, MA, USA
Matthew R. Hassler
RNA Therapeutics Institute, University of Massachusetts Medical School, Worcester, MA, USA
Justice McDaniel
BERG LLC, Framingham, MA, USA
Niven R. Narain
BERG LLC, Framingham, MA, USA
Rachel Wollacott
MassBiologics, Boston, MA, USA
Yang Wang
MassBiologics, Boston, MA, USA
Scott A. Shaffer
Mass Spectrometry Facility, Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School, Shrewsbury, MA, USA
Michael A. Kiebish
BERG LLC, Framingham, MA, USA
Marian DiFiglia
Mass General Institute for Neurodegenerative Disease, Boston, MA, USA
Neil Aronin
RNA Therapeutics Institute, University of Massachusetts Medical School, Worcester, MA, USA; Department of Medicine, University of Massachusetts Medical School, Worcester, MA, USA; Corresponding author
Anastasia Khvorova
RNA Therapeutics Institute, University of Massachusetts Medical School, Worcester, MA, USA; Program in Molecular Medicine, University of Massachusetts Medical School, Worcester, MA, USA; Corresponding author
Summary: Exosomes can serve as delivery vehicles for advanced therapeutics. The components necessary and sufficient to support exosomal delivery have not been established. Here we connect biochemical composition and activity of exosomes to optimize exosome-mediated delivery of small interfering RNAs (siRNAs). This information is used to create effective artificial exosomes. We show that serum-deprived mesenchymal stem cells produce exosomes up to 22-fold more effective at delivering siRNAs to neurons than exosomes derived from control cells. Proteinase treatment of exosomes stops siRNA transfer, indicating that surface proteins on exosomes are involved in trafficking. Proteomic and lipidomic analyses show that exosomes derived in serum-deprived conditions are enriched in six protein pathways and one lipid class, dilysocardiolipin. Inspired by these findings, we engineer an “artificial exosome,” in which the incorporation of one lipid (dilysocardiolipin) and three proteins (Rab7, Desmoplakin, and AHSG) into conventional neutral liposomes produces vesicles that mimic cargo delivering activity of natural exosomes. : Biochemistry; Biological Sciences; Lipidomics; Molecular Biology; Proteomics Subject Areas: Biochemistry, Biological Sciences, Lipidomics, Molecular Biology, Proteomics
