title: Differential fates of biomolecules delivered to target cells via extracellular vesicles.
abstract: Extracellular vesicles (EVs), specifically exosomes and microvesicles (MVs), are presumed to play key roles in cell-cell communication via transfer of biomolecules between cells. The biogenesis of these two types of EVs differs as they originate from either the endosomal (exosomes) or plasma (MVs) membranes. To elucidate the primary means through which EVs mediate intercellular communication, we characterized their ability to encapsulate and deliver different types of macromolecules from transiently transfected cells. Both EV types encapsulated reporter proteins and mRNA but only MVs transferred the reporter function to recipient cells. De novo reporter protein expression in recipient cells resulted only from plasmid DNA (pDNA) after delivery via MVs. Reporter mRNA was delivered to recipient cells by both EV types, but was rapidly degraded without being translated. MVs also mediated delivery of functional pDNA encoding Cre recombinase in vivo to tissues in transgenic Cre-lox reporter mice. Within the parameters of this study, MVs delivered functional pDNA, but not RNA, whereas exosomes from the same source did not deliver functional nucleic acids. These results have significant implications for understanding the role of EVs in cellular communication and for development of EVs as delivery tools. Moreover, studies using EVs from transiently transfected cells may be confounded by a predominance of pDNA transfer.
authors.affiliation: Departments of Pediatrics.
authors.forename: Masamitsu
authors.initials: M
authors.lastname: Kanada
authors.affiliation: Departments of Pediatrics.
authors.forename: Michael H
authors.initials: MH
authors.lastname: Bachmann
authors.affiliation: Departments of Pediatrics.
authors.forename: Jonathan W
authors.initials: JW
authors.lastname: Hardy
authors.affiliation: Microbiology and Immunology, and.
authors.forename: Daniel Omar
authors.initials: DO
authors.lastname: Frimannson
authors.affiliation: Departments of Pediatrics.
authors.forename: Laura
authors.initials: L
authors.lastname: Bronsart
authors.affiliation: Departments of Pediatrics.
authors.forename: Andrew
authors.initials: A
authors.lastname: Wang
authors.affiliation: Microbiology and Immunology, and.
authors.forename: Matthew D
authors.initials: MD
authors.lastname: Sylvester
authors.affiliation: Departments of Pediatrics.
authors.forename: Tobi L
authors.initials: TL
authors.lastname: Schmidt
authors.affiliation: TransDerm Inc., Santa Cruz, CA 95060.
authors.forename: Roger L
authors.initials: RL
authors.lastname: Kaspar
authors.affiliation: Departments of Pediatrics.
authors.forename: Manish J
authors.identifier: http://orcid.org/0000-0002-4490-5595
authors.initials: MJ
authors.lastname: Butte
authors.affiliation: Microbiology and Immunology, and.
authors.forename: A C
authors.initials: AC
authors.lastname: Matin
authors.affiliation: Departments of Pediatrics, Microbiology and Immunology, and Radiology, Stanford University School of Medicine, Stanford, CA 94305; and ccontag@stanford.edu.
authors.forename: Christopher H
authors.initials: CH
authors.lastname: Contag
mesh_terms: D000818:Animals; D017209:Apoptosis; D001692:Biological Transport / Q000235:genetics; D002450:Cell Communication; D002462:Cell Membrane / Q000378:metabolism; D004247:DNA / Q000737:chemistry*; D055354:Exosomes / Q000737:chemistry*; D005434:Flow Cytometry; D020868:Gene Silencing; D017930:Genes, Reporter / Q000235:genetics; D057809:HEK293 Cells; D006801:Humans; D019426:Integrases / Q000378:metabolism; D008055:Lipids / Q000737:chemistry; D046911:Macromolecular Substances / Q000737:chemistry; D051379:Mice; D008822:Mice, Transgenic; D018625:Microscopy, Atomic Force; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D010718:Phosphatidylserines / Q000737:chemistry; D010957:Plasmids; D011092:Polyethylene Glycols / Q000737:chemistry; D012333:RNA, Messenger / Q000378:metabolism; D060149:Tetraspanin 30 / Q000737:chemistry
chemical_list: D008055:Lipids; D046911:Macromolecular Substances; D010718:Phosphatidylserines; D012333:RNA, Messenger; D060149:Tetraspanin 30; D011092:Polyethylene Glycols; D004247:DNA; C045073:Cre recombinase; D019426:Integrases
keywords: apoptotic body; cell communication; exosome; extracellular vesicle; microvesicle