Journal ArticleDOI
Comparative proteomics evaluation of plasma exosome isolation techniques and assessment of the stability of exosomes in normal human blood plasma
Hina Kalra,Christopher G. Adda,Michael Liem,Ching-Seng Ang,Adam Mechler,Richard J. Simpson,Mark D. Hulett,Suresh Mathivanan +7 more
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TLDR
The OptiPrepTM density gradient method was superior in isolating pure exosomal populations, devoid of highly abundant plasma proteins, and in the context of cellular uptake, the isolated exosomes were able to fuse with target cells revealing that they were indeed biologically active.Abstract:
Exosomes are nanovesicles released by a variety of cells and are detected in body fluids including blood. Recent studies have highlighted the critical application of exosomes as personalized targeted drug delivery vehicles and as reservoirs of disease biomarkers. While these research applications have created significant interest and can be translated into practice, the stability of exosomes needs to be assessed and exosome isolation protocols from blood plasma need to be optimized. To optimize methods to isolate exosomes from blood plasma, we performed a comparative evaluation of three exosome isolation techniques (differential centrifugation coupled with ultracentrifugation, epithelial cell adhesion molecule immunoaffinity pull-down, and OptiPrep(TM) density gradient separation) using normal human plasma. Based on MS, Western blotting and microscopy results, we found that the OptiPrep(TM) density gradient method was superior in isolating pure exosomal populations, devoid of highly abundant plasma proteins. In addition, we assessed the stability of exosomes in plasma over 90 days under various storage conditions. Western blotting analysis using the exosomal marker, TSG101, revealed that exosomes are stable for 90 days. Interestingly, in the context of cellular uptake, the isolated exosomes were able to fuse with target cells revealing that they were indeed biologically active.read more
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Journal ArticleDOI
Engineered Exosomes as Vehicles for Biologically Active Proteins
TL;DR: The use of the evolutionarily conserved late-domain (L-domain) pathway is reported as a mechanism for loading exogenous proteins into exosomes and it is demonstrated that labeling of a target protein, Cre recombinase, with a WW tag leads to recognition by the L-domain-containing protein Ndfip1, resulting in ubiquitination and loading intoExosomes.
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Analysis of exosome purification methods using a model liposome system and tunable-resistive pulse sensing.
TL;DR: It is concluded that the size distribution profile and characteristics of vesicles are stably maintained during processing and purification, suggesting that reports detailing how exosomes derived from tumour cells differ in size to those from normal cells are reporting a real phenomenon.
Journal ArticleDOI
A System of Cytokines Encapsulated in ExtraCellular Vesicles
Wendy Fitzgerald,Michael L. Freeman,Michael M. Lederman,Elena Vasilieva,Roberto Romero,Leonid Margolis +5 more
TL;DR: It is demonstrated that these two systems of cell–cell communication are not strictly separated, as many cytokines in vitro, ex vivo, and in vivo are released in EV-encapsulated forms and are capable of eliciting biological effects upon contact with sensitive cells.
Journal ArticleDOI
Proteogenomic analysis reveals exosomes are more oncogenic than ectosomes
Shivakumar Keerthikumar,Lahiru Gangoda,Michael Liem,Pamali Fonseka,Ishara Atukorala,Cemil Ozcitti,Adam Mechler,Christopher G. Adda,Ching-Seng Ang,Suresh Mathivanan +9 more
TL;DR: It is ascertained that cancer cells facilitate oncogenesis by the secretion of mutant and oncoproteins into the tumor microenvironment via exosomes and ectosomes.
Journal ArticleDOI
Extracellular vesicles including exosomes are mediators of signal transduction: are they protective or pathogenic?
TL;DR: The involvement of EVs as mediators of signal transduction in neurodegenerative diseases and cancer is discussed and the role of EVs in mediating Wnt and PI3K signaling pathways is also discussed.
References
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Glioblastoma microvesicles transport RNA and proteins that promote tumour growth and provide diagnostic biomarkers
Johan Skog,T. Wurdinger,van Rijn S,Dimphna H. Meijer,Gainche L,Miguel Sena-Esteves,William T. Curry,Bob S. Carter,Anna M. Krichevsky,Xandra O. Breakefield +9 more
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The Human Plasma Proteome History, Character, and Diagnostic Prospects
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Journal ArticleDOI
Delivery of siRNA to the mouse brain by systemic injection of targeted exosomes
TL;DR: It is shown that exosomes—endogenous nano-vesicles that transport RNAs and proteins—can deliver short interfering (si)RNA to the brain in mice, and the therapeutic potential of exosome-mediated siRNA delivery was demonstrated by the strong mRNA and protein knockdown of BACE1, a therapeutic target in Alzheimer's disease, in wild-type mice.