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

iScience (Jun 2019)

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

Journal volume & issue: Vol. 16
pp. 230 – 241

Abstract

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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

Published in iScience

ISSN: 2589-0042 (Online)
Publisher: Elsevier
Country of publisher: United States
LCC subjects: Science
Website: http://www.cell.com/iscience/home

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