Austria-Sweden-Netherlands-UK partnership for understanding extracellular vesicle heterogeneity in RNA and protein transfer

Lead Research Organisation: University of Oxford
Department Name: Physiology Anatomy and Genetics

Abstract

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Title Improved extracellular vesicle isolation method from bioreactors 
Description A method was developed to isolate extracellular vesicles produced using hollow fibre bioreactors. Bioreactor-produced extracellular vesicles have several properties that make their isolation with standard methods difficult (e.g. size exclusion chromatography, ultracentrifugation, etc.) due to the high viscosity and high amount of extracellular matrix components, host cell DNA and host cell protein. The method, exploiting the CaptoCore resin matrix, was optimised to yield purified extracellular vesicles with a significantly higher yield and purity as compared to the methods available before. 
Type Of Material Technology assay or reagent 
Year Produced 2020 
Provided To Others? Yes  
Impact The method has been presented at conferences, publication pending. This method allows for the purification of extracellular vesicles from dense bioreactors at high amounts, thus helping to obtain reproducibly extracellular vesicles for use in larger in vivo experiments to study extracellular vesicles as biological drug delivery vehicles. 
 
Title Reporter cell lines for RNA delivery via extracellular vesicles 
Description Reporter cell lines have been developed to study miRNA/siRNA trafficking via EVs between cells, as well as to model extracellular vesicle / cell interactions via the transport of guideRNA in a CRISPR Cas9 model. 
Type Of Material Cell line 
Year Produced 2018 
Provided To Others? No  
Impact Using these reporter models the functional transfer (as opposed to only uptake) of extracellular vesicles can be screened. This allows for the better evaluation of extracellular uptake mechanisms, targeted delivery and it helps to accelerate extracellular vesicle bioengineering. 
 
Description Assistant Prof Pieter Vader 
Organisation University Medical Center Utrecht (UMC)
Country Netherlands 
Sector Academic/University 
PI Contribution The Wood lab has contributed by training in extracellular vesicle isolation and subpopulation/heterogeneity analysis, training of post-doctoral researchers and visiting students, and been part of joint grant applications (BBSRC, John Fell Foundation, EU Horizon 2020). We have collaborated on EV engineering for siRNA delivery. The Wood lab has jointly co-supervised the work of a postdoctoral researcher and a DPhil student.
Collaborator Contribution The partner has contributed by bead-based analysis of extracellular vesicle composition in cell culture and biofluids, the design of a type of engineered extracellular vesicles for CRISPR applications, and providing know-how on the use of native extracellular vesicles for heart regeneration. We have collaborated on EV engineering for siRNA delivery. We've been part of joint grant applications (BBSRC, John Fell Foundation, EU Horizon 2020). Vader has jointly co-supervised the work of a postdoctoral researcher and a DPhil student.
Impact We have co-authored a number of publications: PMID: 29760691, 28392161, 28334866, 27943216, 26931825.
Start Year 2016
 
Description Associate Prof Samir EL Andaloussi, Karolinska Institutet, Sweden 
Organisation Karolinska Institute
Country Sweden 
Sector Academic/University 
PI Contribution The Wood lab has contributed to the work of EL Andaoussi lab by providing access for the analysis of extracellular subpopulations/heterogeneity, bioengineering strategies for RNA loading and delivery by extracellular vesicles, proteomics analysis of extracellular vesicle composition, in vivo immunosuppressive activity of bio-engineered extracellular vesicles, biodistribution and the study of extracellular RNA biology. Moreover, the Wood lab has taken part in EL Andaloussi lab PhD students' co-supervision and been supporting their grant applications and IP.
Collaborator Contribution EL Andaoussi lab by providing access for the analysis of extracellular vesicle transcriptome, providing access to the flow-cytometry based characterisation of extracellular vesicles and their populations, biodistribution and shared know-how on aspects on the design of immunosuppressive engineered extracellular vesicles. Moreover, the EL Andaloussi lab has taken part in PhD students' co-supervision and been supporting our grant applications and IP.
Impact This collaboration has led to a number of publications: PMID: 30411858, 30018314, 28912498, 28392161, 28334866, 27943216, 27466195, 27174238, 26931825, , 26690353, 26153722, 26022510, 25899407, 25659648, and a patent application (WO2017203260) EXOSOMES COMPRISING THERAPEUTIC POLYPEPTIDES. We have collaborated on a number of grant applications (BBSRC, John Fell Foundation, SSMF, MRC, CRUK).
Start Year 2015
 
Description Prof Nicole Meisner-Kober, University of Salzburg 
Organisation Novartis
Country Global 
Sector Private 
PI Contribution Nicole Meisner-Kober is the Professor of Chemical Biology and Biological Therapeutics at the University of Salzburg, Austria, and formerly worked as a senior investigator at Novartis Institutes of Biomedical Research, Basel, Switzerland. Our contribution has been providing the collaborator with extracellular vesicle isolation and characterisation methodologies (including -omics techniques), expertise in extracellular vesicle bioengineering, and extracellular RNA secretion biology. We have been part of joint grant applications for biofunctional analysis of extracellular vesicle uptake.
Collaborator Contribution Prof Meisner-Kober has contributed by enabling the analysis of cellular uptake routes of extracellular vesicles by measuring and tracking the real-time uptake dynamics of extracellular vesicles. Furthermore, she has helped to analyse the cargo copy numbers of extracellular vesicle loaded proteins and provided insights and expertise in the analysis of extracellular vesicle RNA secretion biology. We have been part of joint grant applications for biofunctional analysis of extracellular vesicle uptake.
Impact The Wood lab and Meisner-Kober lab have co-published two peer-reviewed research papers PMID: 27114500, 25659648. Furthermore, there have been joint grant applications (BBSRC, John Fell Fund), and joint supervision of an early-career postdoctoral researcher. Further joint applications and publications are planned. Part of the work has fed into know-how development of a data package that supported spinning out an Oxford University spin-out company Evox Therapeutics.
Start Year 2015
 
Description Prof Nicole Meisner-Kober, University of Salzburg 
Organisation University of Salzburg
Country Austria 
Sector Academic/University 
PI Contribution Nicole Meisner-Kober is the Professor of Chemical Biology and Biological Therapeutics at the University of Salzburg, Austria, and formerly worked as a senior investigator at Novartis Institutes of Biomedical Research, Basel, Switzerland. Our contribution has been providing the collaborator with extracellular vesicle isolation and characterisation methodologies (including -omics techniques), expertise in extracellular vesicle bioengineering, and extracellular RNA secretion biology. We have been part of joint grant applications for biofunctional analysis of extracellular vesicle uptake.
Collaborator Contribution Prof Meisner-Kober has contributed by enabling the analysis of cellular uptake routes of extracellular vesicles by measuring and tracking the real-time uptake dynamics of extracellular vesicles. Furthermore, she has helped to analyse the cargo copy numbers of extracellular vesicle loaded proteins and provided insights and expertise in the analysis of extracellular vesicle RNA secretion biology. We have been part of joint grant applications for biofunctional analysis of extracellular vesicle uptake.
Impact The Wood lab and Meisner-Kober lab have co-published two peer-reviewed research papers PMID: 27114500, 25659648. Furthermore, there have been joint grant applications (BBSRC, John Fell Fund), and joint supervision of an early-career postdoctoral researcher. Further joint applications and publications are planned. Part of the work has fed into know-how development of a data package that supported spinning out an Oxford University spin-out company Evox Therapeutics.
Start Year 2015