Engineering of extracellular vesicles for efficient intracellular delivery of multimodal therapeutics including genome editors
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.
Authors
- Oscar P. B. Wiklander (ORCID: https://orcid.org/0000-0003-1176-8114)
- Xiuming Liang (ORCID: https://orcid.org/0000-0003-0202-1211)
- André Görgens (ORCID: https://orcid.org/0000-0001-9198-0857)
- Junhua Xie (ORCID: https://orcid.org/0000-0003-0684-8515)
- Rim Jawad Wiklander (ORCID: https://orcid.org/0000-0002-4216-0913)
- Lien Van Hoecke (ORCID: https://orcid.org/0000-0001-8602-4211)
- Elin K. Esbjörner (ORCID: https://orcid.org/0000-0002-1253-6342)
- Daniel W. Hagey (ORCID: https://orcid.org/0000-0001-9246-6235)
- Radka Slovak (ORCID: https://orcid.org/0000-0002-9963-5975)
- Marziyeh Ghaeidamini
- Elien Van Wonterghem (ORCID: https://orcid.org/0009-0008-0027-485X)
- Julia Rädler
- Houze Zhou
- Doste R. Mamand
- Guannan Zhou (ORCID: https://orcid.org/0000-0001-9353-1908)
- Joel Z. Nordin (ORCID: https://orcid.org/0000-0002-3653-7710)
- Samantha Roudi (ORCID: https://orcid.org/0000-0003-3923-3410)
- Antje M. Zickler (ORCID: https://orcid.org/0000-0001-5001-6586)
- Carla Martín Pérez (ORCID: https://orcid.org/0000-0002-9852-1976)
- Antonin de Fougerolles (ORCID: https://orcid.org/0000-0002-7164-7261)
- Roosmarijn E. Vandenbroucke (ORCID: https://orcid.org/0000-0002-8327-620X)
- Thomas C. Roberts (ORCID: https://orcid.org/0000-0002-3313-7631)
- Jeremy Bost (ORCID: https://orcid.org/0000-0002-3804-0521)
- Aileen Geobee Uy
- H. Yesid Estupiñán (ORCID: https://orcid.org/0000-0002-9388-5957)
- Samir EL Andaloussi (ORCID: https://orcid.org/0000-0003-4468-9113)
- Vincent Hou
- Wenyi Zheng (ORCID: https://orcid.org/0000-0003-2416-5822)
- Zheyu Niu (ORCID: https://orcid.org/0000-0002-3866-5716)
- Dhanu Gupta (ORCID: https://orcid.org/0000-0003-0163-2678)
- Pieter Vader
- Justin Hean (ORCID: https://orcid.org/0000-0002-7291-080X)
- Olivier G. de Jong (ORCID: https://orcid.org/0000-0002-5952-9617)
- Dave Carter (ORCID: https://orcid.org/0000-0003-3503-7615)
- Yuanyuan Zong
- Matthew J. A. Wood (ORCID: https://orcid.org/0000-0002-5436-6011)
- Imre Mäger (ORCID: https://orcid.org/0000-0003-2242-7227)
- Rui He (ORCID: https://orcid.org/0000-0003-3950-118X)
Institutions
- Universidad Industrial de Santander (CO)
- Uppsala University (SE)
- Karolinska University Hospital (SE)
- Utrecht University (NL)
- Karolinska Institutet (SE)
- University Medical Center Utrecht (NL)
- Ghent University (BE)
- University of Oxford (GB)
- PsiOxus Therapeutics (United Kingdom) (GB)
- Shandong Provincial Hospital (CN)
- VIB-UGent Center for Inflammation Research (BE)
- Obstetrics and Gynecology Hospital of Fudan University (CN)
- Oxford BioMedica (United Kingdom) (GB)
- Shandong First Medical University (CN)
- University of Tartu (EE)
- University of Duisburg-Essen (DE)
- Chalmers University of Technology (SE)
Publication Details
- Journal
- Nature Communications
- Published
- 2025-04-29
- DOI
- https://doi.org/10.1038/s41467-025-59377-y
- Citations
- 49
- Primary Topic
- Extracellular vesicles in disease
- Type
- article
- Field-Weighted Citation Impact
- 39.49