TY - JOUR
T1 - TOP-EVs
T2 - Technology of Protein delivery through Extracellular Vesicles is a versatile platform for intracellular protein delivery
AU - Ilahibaks, Nazma F.
AU - Ardisasmita, Arif I.
AU - Xie, Songpu
AU - Gunnarsson, Anders
AU - Brealey, Joseph
AU - Vader, Pieter
AU - de Jong, Olivier G.
AU - de Jager, Saskia
AU - Dekker, Niek
AU - Peacock, Ben
AU - Schiffelers, Raymond M.
AU - Sluijter, Joost P.G.
AU - Lei, Zhiyong
N1 - Funding Information:
Funding for the present work was provided by the Project EVICARE [No. 725229] of the European Research Council (ERC) to J.P.G.S, co-funded by the Project SMARTCARE-II of the BioMedicalMaterials institute to J.P.G.S, the ZonMw-TAS program [No. 116002016 ] to J.P.G.S./Z.L, PPS grant [No. 2018B014 ] to J.P.G.S./Z.L/P.V./S.J., the Dutch Ministry of Economic Affairs, Agriculture and Innovation and the Netherlands CardioVascular Research Initiative (CVON) : the Dutch Heart Foundation to J.P.G.S, Dutch Federations of University Medical Centers , the Netherlands Organization for Health Research and Development , and the Royal Netherlands Academy of Sciences .
Publisher Copyright:
© 2023 The Authors
PY - 2023/3
Y1 - 2023/3
N2 - Extracellular vesicles (EVs) have emerged as biocompatible drug delivery vehicles due to their native ability to deliver bioactive cargo to recipient cells. However, the application of EVs as a therapeutic delivery vehicle is hampered by effective methods for endogenously loading target proteins inside EVs and unloading proteins after delivery to recipient cells. Most EV-based engineered loading methods have a limited delivery efficiency owing to their inefficient endosomal escape or cargo release from the intraluminal attachment from the EV membrane. Here, we describe the 'Technology Of Protein delivery through Extracellular Vesicles’ (TOP-EVs) as a tool for efficient intracellular delivery of target proteins mediated via EVs. The vesicular stomatitis virus glycoprotein and the rapamycin-heterodimerization of the FKBP12/T82L mutant FRB proteins were both important for the effective protein delivery through TOP-EVs. We showed that TOP-EVs could efficiently deliver Cre recombinase and CRISPR/Cas9 ribonucleoprotein complex in vitro. Moreover, our results demonstrated that the capacity of TOP-EVs to deliver intracellular proteins in recipient cells was not an artifact of plasmid contamination or direct plasmid loading into EVs. Finally, we showed that TOP-EVs could successfully mediate intracellular protein delivery in the liver in vivo. Taken together, TOP-EVs are a versatile platform for efficient intracellular protein delivery in vitro and in vivo, which can be applied to advance the development of protein-based therapeutics.
AB - Extracellular vesicles (EVs) have emerged as biocompatible drug delivery vehicles due to their native ability to deliver bioactive cargo to recipient cells. However, the application of EVs as a therapeutic delivery vehicle is hampered by effective methods for endogenously loading target proteins inside EVs and unloading proteins after delivery to recipient cells. Most EV-based engineered loading methods have a limited delivery efficiency owing to their inefficient endosomal escape or cargo release from the intraluminal attachment from the EV membrane. Here, we describe the 'Technology Of Protein delivery through Extracellular Vesicles’ (TOP-EVs) as a tool for efficient intracellular delivery of target proteins mediated via EVs. The vesicular stomatitis virus glycoprotein and the rapamycin-heterodimerization of the FKBP12/T82L mutant FRB proteins were both important for the effective protein delivery through TOP-EVs. We showed that TOP-EVs could efficiently deliver Cre recombinase and CRISPR/Cas9 ribonucleoprotein complex in vitro. Moreover, our results demonstrated that the capacity of TOP-EVs to deliver intracellular proteins in recipient cells was not an artifact of plasmid contamination or direct plasmid loading into EVs. Finally, we showed that TOP-EVs could successfully mediate intracellular protein delivery in the liver in vivo. Taken together, TOP-EVs are a versatile platform for efficient intracellular protein delivery in vitro and in vivo, which can be applied to advance the development of protein-based therapeutics.
KW - Active protein loading
KW - Extracellular vesicles
KW - Functional intracellular protein delivery
KW - Genome editing
KW - TOP-EV
UR - http://www.scopus.com/inward/record.url?scp=85148081463&partnerID=8YFLogxK
U2 - 10.1016/j.jconrel.2023.02.003
DO - 10.1016/j.jconrel.2023.02.003
M3 - Article
C2 - 36746337
AN - SCOPUS:85148081463
SN - 0168-3659
VL - 355
SP - 579
EP - 592
JO - Journal of Controlled Release
JF - Journal of Controlled Release
ER -