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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Applying extracellular vesicles based therapeutics in clinical trials - an ISEV position paper
Thomas Lener,Mario Gimona,Ludwig Aigner,Verena Börger,Edit I. Buzás,Giovanni Camussi,Nathalie Chaput,Devasis Chatterjee,Felipe A. Court,Hernando A. del Portillo,Lorraine O'Driscoll,Stefano Fais,Juan M. Falcón-Pérez,Ursula Felderhoff-Mueser,Lorenzo Fraile,Yong Song Gho,André Görgens,Ramesh C. Gupta,An Hendrix,Dirk M. Hermann,Andrew F. Hill,Fred H. Hochberg,Peter A. Horn,Dominique P.V. de Kleijn,Lambros Kordelas,Boris W. Kramer,Eva-Maria Krämer-Albers,Sandra Laner-Plamberger,Saara Laitinen,Tommaso Leonardi,Magdalena J. Lorenowicz,Sai Kiang Lim,Jan Lötvall,Casey A. Maguire,Antonio Marcilla,Irina Nazarenko,Takahiro Ochiya,Tushar Patel,Shona Pedersen,Gabriella Pocsfalvi,Stefano Pluchino,Peter J. Quesenberry,Ilona G. Reischl,Francisco J. Rivera,Ralf Sanzenbacher,Katharina Schallmoser,Ineke Slaper-Cortenbach,Dirk Strunk,Torsten Tonn,Pieter Vader,Pieter Vader,Bas W.M. van Balkom,Marca H. M. Wauben,Samir El Andaloussi,Samir El Andaloussi,Clotilde Théry,Eva Rohde,Bernd Giebel +57 more
TL;DR: In this paper, the authors summarize recent developments and the current knowledge of extracellular vesicles (EVs) and discuss safety and regulatory requirements that must be considered for pharmaceutical manufacturing and clinical application.
Journal ArticleDOI
ExoCarta: A Web-Based Compendium of Exosomal Cargo
Shivakumar Keerthikumar,David Chisanga,Dinuka Ariyaratne,Haidar Al Saffar,Sushma Anand,Kening Zhao,Monisha Samuel,Mohashin Pathan,Markandeya Jois,Naveen Chilamkurti,Lahiru Gangoda,Suresh Mathivanan +11 more
TL;DR: ExoCarta is described, a manually curated Web-based compendium of exosomal proteins, RNAs and lipids, which features dynamic protein-protein interaction networks and biological pathways of exOSomal proteins.
Journal ArticleDOI
Development of exosome-encapsulated paclitaxel to overcome MDR in cancer cells
Myung Soo Kim,Matthew J. Haney,Yuling Zhao,Vivek Mahajan,I.M. Deygen,Natalia L. Klyachko,Natalia L. Klyachko,Eli Inskoe,Aleksandr Piroyan,Marina Sokolsky,Onyi Okolie,Shawn Hingtgen,Alexander V. Kabanov,Alexander V. Kabanov,Elena V. Batrakova +14 more
TL;DR: A exosome-based system to carry formulation of PTX was developed and showed efficacy in the treatment of multi-drug resistant cancer cells and this novel system may be further developed to carry other chemotherapeutic agents in the future.
Journal ArticleDOI
Quantitative and stoichiometric analysis of the microRNA content of exosomes.
John R. Chevillet,Qing Kang,Ingrid K. Ruf,Hilary A. Briggs,Lucia Vojtech,Sean M. Hughes,Heather H. Cheng,Jason D. Arroyo,Emily K. Meredith,Emily N. Gallichotte,Era L. Pogosova-Agadjanyan,Colm Morrissey,Derek L. Stirewalt,Florian Hladik,Evan Y. Yu,Celestia S. Higano,Muneesh Tewari +16 more
TL;DR: It is shown here that most exosomes derived from standard preparations do not harbor many copies of miRNA molecules, and are, therefore, individually unlikely to be functional as vehicles for miRNA-based communication.
Journal ArticleDOI
EV-TRACK: transparent reporting and centralizing knowledge in extracellular vesicle research
Jan Van Deun,Pieter Mestdagh,Patrizia Agostinis,Özden Akay,Sushma Anand,Jasper Anckaert,Zoraida Andreu Martinez,Tine Baetens,Els Beghein,Laurence Bertier,Geert Berx,Janneke Boere,Stephanie Boukouris,Michel Bremer,Dominik Buschmann,James Brian Byrd,Clara Casert,Lesley Cheng,Anna Cmoch,Delphine Daveloose,Eva De Smedt,Seyma Demirsoy,Victoria Depoorter,Bert Dhondt,Tom A. P. Driedonks,Aleksandra M. Dudek,Abdou ElSharawy,Ilaria Floris,Andrew D Foers,Kathrin Gärtner,Abhishek D. Garg,Edward Geeurickx,Jan Gettemans,Farzaneh Ghazavi,Bernd Giebel,Tom Groot Kormelink,Grace V. Hancock,Hetty Helsmoortel,Andrew F. Hill,Vincent Hyenne,Hina Kalra,David Kim,Joanna Kowal,Joanna Kowal,Sandra Kraemer,Petra Leidinger,Carina Leonelli,Yaxuan Liang,Lien Lippens,Shu Liu,Alessandra Lo Cicero,Alessandra Lo Cicero,Shaun Martin,Suresh Mathivanan,Prabhu Mathiyalagan,Tamás Matusek,Gloria Milani,Marta Monguió-Tortajada,Liselot Mus,Dillon C. Muth,Andrea Németh,Esther N. M. Nolte-‘t Hoen,Lorraine O'Driscoll,Roberta Palmulli,Roberta Palmulli,Michael W. Pfaffl,Bjarke Primdal-Bengtson,Bjarke Primdal-Bengtson,Erminia Romano,Quentin Rousseau,Susmita Sahoo,Natalia G. Sampaio,Monisha Samuel,Benjamin J. Scicluna,Bieke Soen,Anneleen Steels,Johannes V. Swinnen,Maarit Takatalo,Safia Thaminy,Clotilde Théry,Clotilde Théry,Joeri Tulkens,Isabel Van Audenhove,Susanne G. van der Grein,Alan Van Goethem,Martijn J. C. van Herwijnen,Guillaume van Niel,Guillaume van Niel,Nadine Van Roy,Alexander R. van Vliet,Niels Vandamme,Suzanne Vanhauwaert,Glenn Vergauwen,Frederik J. Verweij,Frederik J. Verweij,Annelynn Wallaert,Marca H. M. Wauben,Kenneth W. Witwer,Marijke I. Zonneveld,Olivier De Wever,Jo Vandesompele,An Hendrix +101 more
TL;DR: It is argued that the field of extracellular vesicle (EV) biology needs more transparent reporting to facilitate interpretation and replication of experiments and EV-TRACK, a crowdsourcing knowledgebase that centralizes EV biology and methodology, is described.
References
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Journal ArticleDOI
Exosome-mediated transfer of mRNAs and microRNAs is a novel mechanism of genetic exchange between cells
TL;DR: It is shown that exosomes contain both mRNA and microRNA, which can be delivered to another cell, and can be functional in this new location, and it is proposed that this RNA is called “exosomal shuttle RNA” (esRNA).
Journal ArticleDOI
Exosomes: composition, biogenesis and function
TL;DR: The physical properties that define exosomes as a specific population of secreted vesicles are described, their biological effects, particularly on the immune system, are summarized, and the potential roles that secretedvesicles could have as intercellular messengers are discussed.
Journal ArticleDOI
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
TL;DR: Tumour-derived microvesicles may provide diagnostic information and aid in therapeutic decisions for cancer patients through a blood test by incorporating an mRNA for a reporter protein into them, and it is demonstrated that messages delivered by microvesicle are translated by recipient cells.
Journal ArticleDOI
The Human Plasma Proteome History, Character, and Diagnostic Prospects
TL;DR: This work speculates on the reasons behind this large discrepancy between the expectations arising from proteomics and the realities of clinical diagnostics and suggests approaches by which protein-disease associations may be more effectively translated into diagnostic tools in the future.
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.