Pseudotyping exosomes for enhanced protein delivery in mammalian cells

Conary Meyer,1,* Joseph Losacco,1,* Zachary Stickney,1 Lingxuan Li,2 Gerard Marriott,3 Biao Lu1 1Department of Bioengineering, Santa Clara University, Santa Clara, 2Crown College, University of California at Santa Cruz, Santa Cruz, 3Department of Bioengineering, University of California at Berkeley...

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Autores principales:	Meyer C, Losacco J, Stickney Z, Li L, Marriott G, Lu B
Formato:	article
Lenguaje:	EN
Publicado:	Dove Medical Press 2017
Materias:	Exosome VSVG Protein delivery nanotechnology Medicine (General) R5-920
Acceso en línea:	https://doaj.org/article/e62447167c754ee7a16aa00838c5e6cb
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spelling	oai:doaj.org-article:e62447167c754ee7a16aa00838c5e6cb2021-12-02T00:39:20ZPseudotyping exosomes for enhanced protein delivery in mammalian cells1178-2013https://doaj.org/article/e62447167c754ee7a16aa00838c5e6cb2017-04-01T00:00:00Zhttps://www.dovepress.com/pseudotyping-exosomes-for-enhanced-protein-delivery-in-mammalian-cells-peer-reviewed-article-IJNhttps://doaj.org/toc/1178-2013Conary Meyer,1,* Joseph Losacco,1,* Zachary Stickney,1 Lingxuan Li,2 Gerard Marriott,3 Biao Lu1 1Department of Bioengineering, Santa Clara University, Santa Clara, 2Crown College, University of California at Santa Cruz, Santa Cruz, 3Department of Bioengineering, University of California at Berkeley, Berkeley, CA, USA *These authors contributed equally to this work Abstract: Exosomes are cell-derived nanovesicles that hold promise as living vehicles for intracellular delivery of therapeutics to mammalian cells. This potential, however, is undermined by the lack of effective methods to load exosomes with therapeutic proteins and to facilitate their uptake by target cells. Here, we demonstrate how a vesicular stomatitis virus glycoprotein (VSVG) can both load protein cargo onto exosomes and increase their delivery ability via a pseudotyping mechanism. By fusing a set of fluorescent and luminescent reporters with VSVG, we show the successful targeting and incorporation of VSVG fusions into exosomes by gene transfection and fluorescence tracking. We subsequently validate our system by live cell imaging of VSVG and its participation in endosomes/exosomes that are ultimately released from transfected HEK293 cells. We show that VSVG pseudotyping of exosomes does not affect the size or distributions of the exosomes, and both the full-length VSVG and the VSVG without the ectodomain are shown to integrate into the exosomal membrane, suggesting that the ectodomain is not required for protein loading. Finally, exosomes pseudotyped with full-length VSVG are internalized by multiple-recipient cell types to a greater degree compared to exosomes loaded with VSVG without the ectodomain, confirming a role of the ectodomain in cell tropism. In summary, our work introduces a new genetically encoded pseudotyping platform to load and enhance the intracellular delivery of therapeutic proteins via exosome-based vehicles to target cells. Keywords: exosome, VSVG, protein delivery, nanotechnologyMeyer CLosacco JStickney ZLi LMarriott GLu BDove Medical PressarticleExosomeVSVGProtein deliverynanotechnologyMedicine (General)R5-920ENInternational Journal of Nanomedicine, Vol Volume 12, Pp 3153-3170 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Exosome VSVG Protein delivery nanotechnology Medicine (General) R5-920
spellingShingle	Exosome VSVG Protein delivery nanotechnology Medicine (General) R5-920 Meyer C Losacco J Stickney Z Li L Marriott G Lu B Pseudotyping exosomes for enhanced protein delivery in mammalian cells
description	Conary Meyer,1,* Joseph Losacco,1,* Zachary Stickney,1 Lingxuan Li,2 Gerard Marriott,3 Biao Lu1 1Department of Bioengineering, Santa Clara University, Santa Clara, 2Crown College, University of California at Santa Cruz, Santa Cruz, 3Department of Bioengineering, University of California at Berkeley, Berkeley, CA, USA *These authors contributed equally to this work Abstract: Exosomes are cell-derived nanovesicles that hold promise as living vehicles for intracellular delivery of therapeutics to mammalian cells. This potential, however, is undermined by the lack of effective methods to load exosomes with therapeutic proteins and to facilitate their uptake by target cells. Here, we demonstrate how a vesicular stomatitis virus glycoprotein (VSVG) can both load protein cargo onto exosomes and increase their delivery ability via a pseudotyping mechanism. By fusing a set of fluorescent and luminescent reporters with VSVG, we show the successful targeting and incorporation of VSVG fusions into exosomes by gene transfection and fluorescence tracking. We subsequently validate our system by live cell imaging of VSVG and its participation in endosomes/exosomes that are ultimately released from transfected HEK293 cells. We show that VSVG pseudotyping of exosomes does not affect the size or distributions of the exosomes, and both the full-length VSVG and the VSVG without the ectodomain are shown to integrate into the exosomal membrane, suggesting that the ectodomain is not required for protein loading. Finally, exosomes pseudotyped with full-length VSVG are internalized by multiple-recipient cell types to a greater degree compared to exosomes loaded with VSVG without the ectodomain, confirming a role of the ectodomain in cell tropism. In summary, our work introduces a new genetically encoded pseudotyping platform to load and enhance the intracellular delivery of therapeutic proteins via exosome-based vehicles to target cells. Keywords: exosome, VSVG, protein delivery, nanotechnology
format	article
author	Meyer C Losacco J Stickney Z Li L Marriott G Lu B
author_facet	Meyer C Losacco J Stickney Z Li L Marriott G Lu B
author_sort	Meyer C
title	Pseudotyping exosomes for enhanced protein delivery in mammalian cells
title_short	Pseudotyping exosomes for enhanced protein delivery in mammalian cells
title_full	Pseudotyping exosomes for enhanced protein delivery in mammalian cells
title_fullStr	Pseudotyping exosomes for enhanced protein delivery in mammalian cells
title_full_unstemmed	Pseudotyping exosomes for enhanced protein delivery in mammalian cells
title_sort	pseudotyping exosomes for enhanced protein delivery in mammalian cells
publisher	Dove Medical Press
publishDate	2017
url	https://doaj.org/article/e62447167c754ee7a16aa00838c5e6cb
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_version_	1718403589905317888

Pseudotyping exosomes for enhanced protein delivery in mammalian cells

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