Exosome Display technology: Applications to the development of new diagnostics and therapeutics

Delcayre, Alain; Estellés, Angeles; Sperinde, Jeffrey; Roulon, Thibaut; Paz, Pedro; Aguilar, Barbara; Villanueva, Janeth C.; Khine, SuSu; Pecq, Jean-Bernard Le

doi:10.1016/j.bcmd.2005.07.003

Cited by 162 publications

(153 citation statements)

References 32 publications

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“…Fusion of target proteins to mouse lactadherin has been shown to increase EV association, due to the binding of this protein to phosphatidylserine, a lipid that is enriched in EV [18]. To maximize the amount of hetIL-15 associated with EV, we generated a DNA construct encoding a fully human, chimeric molecule consisting of the IL-15Rα ectodomain fused at its C-terminus with the C1C2 domains of lactadherin.…”

Section: Resultsmentioning

confidence: 99%

Scalable, cGMP‐compatible purification of extracellular vesicles carrying bioactive human heterodimeric IL‐15/lactadherin complexes

Watson

Yung

Bergamaschi

et al. 2018

J of Extracellular Vesicle

113

100

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The development of extracellular vesicles (EV) for therapeutic applications is contingent upon the establishment of reproducible, scalable, and high-throughput methods for the production and purification of clinical grade EV. Methods including ultracentrifugation (U/C), ultrafiltration, immunoprecipitation, and size-exclusion chromatography (SEC) have been employed to isolate EV, each facing limitations such as efficiency, particle purity, lengthy processing time, and/or sample volume. We developed a cGMP-compatible method for the scalable production, concentration, and isolation of EV through a strategy involving bioreactor culture, tangential flow filtration (TFF), and preparative SEC. We applied this purification method for the isolation of engineered EV carrying multiple complexes of a novel human immunostimulatory cytokine-fusion protein, heterodimeric IL-15 (hetIL-15)/lactadherin. HEK293 cells stably expressing the fusion cytokine were cultured in a hollow-fibre bioreactor. Conditioned medium was collected and EV were isolated comparing three procedures: U/C, SEC, or TFF + SEC. SEC demonstrated comparable particle recovery, size distribution, and hetIL-15 density as U/C purification. Relative to U/C, SEC preparations achieved a 100-fold reduction in ferritin concentration, a major protein-complex contaminant. Comparative proteomics suggested that SEC additionally decreased the abundance of cytoplasmic proteins not associated with EV. Combination of TFF and SEC allowed for bulk processing of large starting volumes, and resulted in bioactive EV, without significant loss in particle yield or changes in size, morphology, and hetIL-15/lactadherin density. Taken together, the combination of bioreactor culture with TFF + SEC comprises a scalable, efficient method for the production of highly purified, bioactive EV carrying hetIL-15/lactadherin, which may be useful in targeted cancer immunotherapy approaches.

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Section: Resultsmentioning

confidence: 99%

Scalable, cGMP‐compatible purification of extracellular vesicles carrying bioactive human heterodimeric IL‐15/lactadherin complexes

Watson

Yung

Bergamaschi

et al. 2018

J of Extracellular Vesicle

113

100

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“…In addition, exosome-expressed tumour-specific antigens show strong antigen immunogenicity, and thus could be used to develop cancer vaccines and produce monoclonal antibodies [76,89]. …”

Section: Therapeutic Applications Of Surface-modified Evsmentioning

confidence: 99%

Extracellular vesicles as a platform for membrane‐associated therapeutic protein delivery

Yang

Hong

Cho

et al. 2018

J of Extracellular Vesicle

181

140

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Membrane proteins are of great research interest, particularly because they are rich in targets for therapeutic application. The suitability of various membrane proteins as targets for therapeutic formulations, such as drugs or antibodies, has been studied in preclinical and clinical studies. For therapeutic application, however, a protein must be expressed and purified in as close to its native conformation as possible. This has proven difficult for membrane proteins, as their native conformation requires the association with an appropriate cellular membrane. One solution to this problem is to use extracellular vesicles as a display platform. Exosomes and microvesicles are membranous extracellular vesicles that are released from most cells. Their membranes may provide a favourable microenvironment for membrane proteins to take on their proper conformation, activity, and membrane distribution; moreover, membrane proteins can cluster into microdomains on the surface of extracellular vesicles following their biogenesis. In this review, we survey the state-of-the-art of extracellular vesicle (exosome and small-sized microvesicle)-based therapeutics, evaluate the current biological understanding of these formulations, and forecast the technical advances that will be needed to continue driving the development of membrane protein therapeutics.

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“…Earlier exosome display technology work by Delcayre et al demonstrated that the C1C2 domains of lactadherin preferentially localize to exosome membranes [41]. Exosomes expressing lactadherin were more effective at immunizing mice against lactadherin than free lactadherin.…”

Section: Optimizing Formulation Of Exosomal Nanocarriersmentioning

confidence: 99%

Post Isolation Modification of Exosomes for Nanomedicine Applications

Hood

2016

Nanomedicine (Lond.)

151

115

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Exosomes are extracellular nanovesicles. They innately possess ideal structural and biocompatible nanocarrier properties. Exosome components can be engineered at the cellular level. Alternatively, when exosome source cells are unavailable for customized exosome production, exosomes derived from a variety of biological origins can be modified post isolation which is the focus of this article. Modification of exosome surface structures allows for exosome imaging and tracking in vivo. Exosome membranes can be loaded with hydrophobic therapeutics to increase drug stability and efficacy. Hydrophilic therapeutics such as RNA can be encapsulated in exosomes to improve cellular delivery. Despite advances in post isolation exosome modification strategies, many challenges to effectively harnessing their therapeutic potential remain. Future topics of exploration include: matching exosome subtypes with nanomedicine applications, optimizing exosomal nanocarrier formulation and investigating how modified exosomes interface with the immune system. Research into these areas will greatly facilitate personalized exosome-based nanomedicine endeavors.

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Exosome Display technology: Applications to the development of new diagnostics and therapeutics

Cited by 162 publications

References 32 publications

Scalable, cGMP‐compatible purification of extracellular vesicles carrying bioactive human heterodimeric IL‐15/lactadherin complexes

Scalable, cGMP‐compatible purification of extracellular vesicles carrying bioactive human heterodimeric IL‐15/lactadherin complexes

Extracellular vesicles as a platform for membrane‐associated therapeutic protein delivery

Post Isolation Modification of Exosomes for Nanomedicine Applications

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