Engineering of extracellular vesicles for efficient intracellular delivery of multimodal therapeutics including genome editors
Abstract
Abstract Intracellular delivery of protein and RNA therapeutics represents a major challenge. Here, we develop highly potent engineered extracellular vesicles (EVs) by incorporating bio-inspired attributes required for effective delivery. These comprise an engineered mini-intein protein with self-cleavage activity for active cargo loading and release, and fusogenic VSV-G protein for endosomal escape. Combining these components allows high efficiency recombination and genome editing in vitro following EV-mediated delivery of Cre recombinase and Cas9/sgRNA RNP cargoes, respectively. In vivo, infusion of a single dose Cre loaded EVs into the lateral ventricle in brain of Cre-LoxP R26-LSL-tdTomato reporter mice results in greater than 40% and 30% recombined cells in hippocampus and cortex respectively. In addition, we demonstrate therapeutic potential of this platform by showing inhibition of LPS-induced systemic inflammation via delivery of a super-repressor of NF-ĸB activity. Our data establish these engineered EVs as a platform for effective delivery of multimodal therapeutic cargoes, including for efficient genome editing.
Article Details
Authors (38)
Xiuming Liang
Dhanu Gupta
Junhua Xie
Elien Van Wonterghem
Lien Van Hoecke
Justin Hean
Zheyu Niu
Marziyeh Ghaeidamini
Oscar P. B. Wiklander
Wenyi Zheng
Rim Jawad Wiklander
Rui He
Tianjin Key Laboratory of Structure and Performance for Functional Molecules, College of Chemistry
Doste R. Mamand
Department of Laboratory Medicine
Jeremy Bost
Guannan Zhou
Houze Zhou
Samantha Roudi
H. Yesid Estupiñán
Julia Rädler
Antje M. Zickler
André Görgens
Department of Laboratory Medicine
Vicky W. Q. Hou
Radka Slovak
Daniel W. Hagey
Olivier G. de Jong
Aileen Geobee Uy
Yuanyuan Zong
Imre Mäger
Carla Martín Pérez
Thomas C. Roberts
Dave Carter
Pieter Vader
Elin K. Esbjörner
Antonin de Fougerolles
Matthew J. A. Wood
Institute for Developmental and Regenerative Medicine, Department of Paediatrics, University of Oxford
Roosmarijn E. Vandenbroucke
Joel Z. Nordin
Samir El Andaloussi