Nikki Heath1, Xabier Osteikoetxea1, Taiana Mia de Oliveria2, Elisa Lázaro-Ibáñez3, Olga Shatnyeva3, Christina Schindler4, Natalie Tigue4, Lorenz M Mayr5, Niek Dekker3, Ross Overman1, Rick Davies1. 1. Discovery Biology, Discovery Sciences, BioPharmaceuticals, R&D, AstraZeneca, Alderley Park, UK. 2. Structure, Biophysics & FBLG, Discovery Sciences, Biopharmaceuticals R&D, Cambridge, UK. 3. Discovery Biology, Discovery Sciences, BioPharmaceuticals R&D, AstraZeneca, Gothenburg, Sweden. 4. Antibody Discovery & Protein Engineering, BioPharmaceuticals R&D, AstraZeneca, Granta Park, Cambridge, UK. 5. GE Healthcare Life Sciences, The Grove Centre, White Lion Road, Amersham, UK.
Abstract
Aim: Extracellular vesicles (EVs) are desirable delivery vehicles for therapeutic cargoes. We aimed to load EVs with Cre recombinase protein and determine whether functional delivery to cells could be improved by using endosomal escape enhancing compounds. Materials & methods: Overexpressed CreFRB protein was actively loaded into EVs by rapalog-induced dimerization to CD81FKBP, or passively loaded by overexpression in the absence of rapalog. Functional delivery of CreFRB was analysed using a HEK293 Cre reporter cell line in the absence and presence of endosomal escape enhancing compounds. Results: The EVs loaded with CreFRB by both active and passive mechanisms were able to deliver functional CreFRB to recipient cells only in the presence of endosomal escape enhancing compounds chloroquine and UNC10217938A. Conclusion: The use of endosomal escape enhancing compounds in conjunction with EVs loaded with therapeutic cargoes may improve efficacy of future EV based therapeutics.
Aim: Extracellular vesicles (EVs) are desirable delivery vehicles for therapeutic cargoes. We aimed to load EVs with Cre recombinase protein and determine whether functional delivery to cells could be improved by using endosomal escape enhancing compounds. Materials & methods: Overexpressed CreFRB protein was actively loaded into EVs by rapalog-induced dimerization to CD81FKBP, or passively loaded by overexpression in the absence of rapalog. Functional delivery of CreFRB was analysed using a HEK293 Cre reporter cell line in the absence and presence of endosomal escape enhancing compounds. Results: The EVs loaded with CreFRB by both active and passive mechanisms were able to deliver functional CreFRB to recipient cells only in the presence of endosomal escape enhancing compounds chloroquine and UNC10217938A. Conclusion: The use of endosomal escape enhancing compounds in conjunction with EVs loaded with therapeutic cargoes may improve efficacy of future EV based therapeutics.
Authors: Xabier Osteikoetxea; Andreia Silva; Elisa Lázaro-Ibáñez; Nikki Salmond; Olga Shatnyeva; Josia Stein; Jan Schick; Stephen Wren; Julia Lindgren; Mike Firth; Alexandra Madsen; Lorenz M Mayr; Ross Overman; Rick Davies; Niek Dekker Journal: J Extracell Vesicles Date: 2022-05
Authors: Bethan L Thomas; Suzanne E Eldridge; Babak Nosrati; Mario Alvarez; Anne-Sophie Thorup; Giovanna Nalesso; Sara Caxaria; Aida Barawi; James G Nicholson; Mauro Perretti; Carles Gaston-Massuet; Costantino Pitzalis; Alison Maloney; Adrian Moore; Ray Jupp; Francesco Dell'Accio Journal: J Extracell Vesicles Date: 2021-05-19
Authors: Russell E McConnell; Madeleine Youniss; Bhargavee Gnanasambandam; Palak Shah; Wei Zhang; Jonathan D Finn Journal: J Extracell Vesicles Date: 2022-10