Vésicules extracellulaires, réponses immunes et cancer

Publications de l’équipe

Année de publication : 2020

Rienk Nieuwland, Juan M Falcón-Pérez, Clotilde Théry, Kenneth W Witwer (2020 Dec 21)

Rigor and standardization of extracellular vesicle research: Paving the road towards robustness.

Journal of extracellular vesicles : e12037 : DOI : 10.1002/jev2.12037 En savoir plus
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Felix Royo, Clotilde Théry, Juan M Falcón-Pérez, Rienk Nieuwland, Kenneth W Witwer (2020 Aug 29)

Methods for Separation and Characterization of Extracellular Vesicles: Results of a Worldwide Survey Performed by the ISEV Rigor and Standardization Subcommittee.

Cells : DOI : E1955 En savoir plus
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Research on extracellular vesicles (EVs) is growing exponentially due to an increasing appreciation of EVs as disease biomarkers and therapeutics, an expanding number of EV-containing materials under study, and application of new preparation, detection, and cargo analysis methods. Diversity of both sources and methodologies imposes challenges on the comparison of measurement results between studies and laboratories. While reference guidelines and minimal requirements for EV research have achieved the important objective of assembling community consensus, it is also essential to understand which methodologies and quality controls are currently being applied, and how usage trends are evolving. As an initial response to this need, the International Society for Extracellular Vesicles (ISEV) performed a worldwide survey in 2015 on « Techniques used for the isolation and characterization of extracellular vesicles » and published the results from this survey in 2016. In 2019, a new survey was performed to assess the changing state of the field. The questionnaire received more than 600 full or partial responses, and the present manuscript summarizes the results of this second worldwide survey. The results emphasize that separation methods such as ultracentrifugation and density gradients are still the most commonly used methods, the use of size exclusion chromatography has increased, and techniques based on tangential flow and microfluidics are now being used by more than 10% of respondents. The survey also reveals that most EV researchers still do not perform sample quality controls before or after isolation of EVs. Finally, the majority of EV researchers emphasize that separation and characterization of EVs should receive more attention.

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Guillaume van Niel, Clotilde Théry (2020 Aug 19)

Extracellular vesicles: eat glutamine and spit acidic bubbles.

The EMBO journal : e105119 : DOI : 10.15252/embj.2020105119 En savoir plus
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Extracellular vesicles mediate transfer of diverse molecular content to target cells in order to induce phenotypic changes, which has put them under the spotlight as likely major players in cell-to-cell communication. However, extracellular vesicle heterogeneity in terms of intracellular origin has only recently been recognized as a potential determinant of their activity. Recent work by Fan et al (2020) illustrates how lack of external resources that affect cellular homeostasis and signaling can also modulate EV biogenesis, by inducing the production of a novel subpopulation of exosomes enriched in Rab11a with context-dependent roles in Drosophila gland physiology and cancer cell aggressiveness.

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Federico Cocozza, Eleonora Grisard, Lorena Martin-Jaular, Mathilde Mathieu, Clotilde Théry (2020 Jul 11)

SnapShot: Extracellular Vesicles.

Cell : 262-262.e1 : DOI : S0092-8674(20)30560-2 En savoir plus
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Cells release a variety of extracellular vesicles (EVs; including exosomes, microvesicles, and many others) into their environment. EVs can bud in endosomes or directly at the plasma membrane, carrying a selection of components from the cell and displaying various functional properties. Different techniques can be used to separate EV subtypes and EVs from co-isolated components, resulting in preparations of different abundance and purity.

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Verena Börger, Daniel J Weiss, Johnathon D Anderson, Francesc E Borràs, Benedetta Bussolati, David R F Carter, Massimo Dominici, Juan M Falcón-Pérez, Mario Gimona, Andrew F Hill, Andrew M Hoffman, Dominique de Kleijn, Bruce L Levine, Rebecca Lim, Jan Lötvall, S Alex Mitsialis, Marta Monguió-Tortajada, Maurizio Muraca, Rienk Nieuwland, Anna Nowocin, Lorraine O'Driscoll, Luis A Ortiz, Donald G Phinney, Ilona Reischl, Eva Rohde, Ralf Sanzenbacher, Clotilde Théry, Wei Seong Toh, Kenneth W Witwer, Sai Kiang Lim, Bernd Giebel (2020 May 20)

International Society for Extracellular Vesicles and International Society for Cell and Gene Therapy statement on extracellular vesicles from mesenchymal stromal cells and other cells: considerations for potential therapeutic agents to suppress coronavirus disease-19.

Cytotherapy : 482-485 : DOI : S1465-3249(20)30662-9 En savoir plus
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The International Society for Cellular and Gene Therapies (ISCT) and the International Society for Extracellular Vesicles (ISEV) recognize the potential of extracellular vesicles (EVs, including exosomes) from mesenchymal stromal cells (MSCs) and possibly other cell sources as treatments for COVID-19. Research and trials in this area are encouraged. However, ISEV and ISCT do not currently endorse the use of EVs or exosomes for any purpose in COVID-19, including but not limited to reducing cytokine storm, exerting regenerative effects or delivering drugs, pending the generation of appropriate manufacturing and quality control provisions, pre-clinical safety and efficacy data, rational clinical trial design and proper regulatory oversight.

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Paula Soledad Pérez, María Albertina Romaniuk, Gabriel A Duette, Zezhou Zhao, Yiyao Huang, Lorena Martin-Jaular, Kenneth W Witwer, Clotilde Théry, Matías Ostrowski (2020 Jan 31)

Extracellular vesicles and chronic inflammation during HIV infection.

Journal of extracellular vesicles : 1687275 : DOI : 10.1080/20013078.2019.1687275 En savoir plus
Résumé

Inflammation is a hallmark of HIV infection. Among the multiple stimuli that can induce inflammation in untreated infection, ongoing viral replication is a primary driver. After initiation of effective combined antiretroviral therapy (cART), HIV replication is drastically reduced or halted. However, even virologically controlled patients may continue to have abnormal levels of inflammation. A number of factors have been proposed to cause inflammation in HIV infection: among others, residual (low-level) HIV replication, production of HIV protein or RNA in the absence of replication, microbial translocation from the gut to the circulation, co-infections, and loss of immunoregulatory responses. Importantly, chronic inflammation in HIV-infected individuals increases the risk for a number of non-infectious co-morbidities, including cancer and cardiovascular disease. Thus, achieving a better understanding of the underlying mechanisms of HIV-associated inflammation in the presence of cART is of utmost importance. Extracellular vesicles have emerged as novel actors in intercellular communication, involved in a myriad of physiological and pathological processes, including inflammation. In this review, we will discuss the role of extracellular vesicles in the pathogenesis of HIV infection, with particular emphasis on their role as inducers of chronic inflammation.

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Année de publication : 2019

Clotilde Théry, Yong Song Gho, Peter Quesenberry (2019 Sep 26)

Journal of extracellular vesicles: the seven year itch!

Journal of extracellular vesicles : 1654729 : DOI : 10.1080/20013078.2019.1654729 En savoir plus
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Kenneth W Witwer, Clotilde Théry (2019 Sep 7)

Extracellular vesicles or exosomes? On primacy, precision, and popularity influencing a choice of nomenclature.

Journal of extracellular vesicles : 1648167 : DOI : 10.1080/20013078.2019.1648167 En savoir plus
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Zhaohao Liao, Lorena Martin Jaular, Estelle Soueidi, Mabel Jouve, Dillon C Muth, Tine H Schøyen, Tessa Seale, Norman J Haughey, Matias Ostrowski, Clotilde Théry, Kenneth W Witwer (2019 Jul 16)

Acetylcholinesterase is not a generic marker of extracellular vesicles.

Journal of extracellular vesicles : 1628592 : DOI : 10.1080/20013078.2019.1628592 En savoir plus
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Acetylcholinesterase (AChE) activity is found in abundance in reticulocytes and neurons and was developed as a marker of reticulocyte EVs in the 1970s. Easily, quickly, and cheaply assayed, AChE activity has more recently been proposed as a generic marker for small extracellular vesicles (sEV) or exosomes, and as a negative marker of HIV-1 virions. To evaluate these proposed uses of AChE activity, we examined data from different EV and virus isolation methods using T-lymphocytic (H9, PM1 and Jurkat) and promonocytic (U937) cell lines grown in culture conditions that differed by serum content. When EVs were isolated by differential ultracentrifugation, no correlation between AChE activity and particle count was observed. AChE activity was detected in non-conditioned medium when serum was added, and most of this activity resided in soluble fractions and could not be pelleted by centrifugation. The serum-derived pelletable AChE protein was not completely eliminated from culture medium by overnight ultracentrifugation; however, a serum « extra-depletion » protocol, in which a portion of the supernatant was left undisturbed during harvesting, achieved near-complete depletion. In conditioned medium also, only small percentages of AChE activity could be pelleted together with particles. Furthermore, no consistent enrichment of AChE activity in sEV fractions was observed. Little if any AChE activity is produced by the cells we examined, and this activity was mainly present in non-vesicular structures, as shown by electron microscopy. Size-exclusion chromatography and iodixanol gradient separation showed that AChE activity overlaps only minimally with EV-enriched fractions. AChE activity likely betrays exposure to blood products and not EV abundance, echoing the MISEV 2014 and 2018 guidelines and other publications. Additional experiments may be merited to validate these results for other cell types and biological fluids other than blood.

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Emeline Bonsergent, Gregory Lavieu (2019 Jun 9)

Content release of extracellular vesicles in a cell-free extract.

FEBS letters : DOI : 10.1002/1873-3468.13472 En savoir plus
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Extracellular Vesicles (EVs) transfer molecules from donor to acceptor cells. The EV-content delivery process within the acceptor cell is poorly characterized. We developed a new cell-free assay to assess EV-content release in vitro. We found that EV-cytosolic cargoes are released from EVs when isolated vesicles are incubated with purified plasma membrane sheets at acidic pH, a characteristic of the endo/lysosomal environment. This process is protein dependent. Our results suggest that EV-content delivery occurs within the endo/lysosomes of acceptor cells and is triggered by acidification. This process resembles virus content delivery and may require membrane fusion. The assay presented here will facilitate investigations into the core machinery and mechanisms underlying EV content delivery. This article is protected by copyright. All rights reserved.

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Daniel E Murphy, Olivier G de Jong, Maarten Brouwer, Matthew J Wood, Grégory Lavieu, Raymond M Schiffelers, Pieter Vader (2019 Mar 16)

Extracellular vesicle-based therapeutics: natural versus engineered targeting and trafficking.

Experimental & molecular medicine : 32 : DOI : 10.1038/s12276-019-0223-5 En savoir plus
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Extracellular vesicles (EVs) are increasingly being recognized as mediators of intercellular signaling via the delivery of effector molecules. Interestingly, certain types of EVs are also capable of inducing therapeutic responses. For these reasons, the therapeutic potential of EVs is a topic of intense research, both in the context of drug delivery and regenerative medicine. However, to fully utilize EVs for therapeutic purposes, an improved understanding of the mechanisms by which they function would be highly advantageous. Here, the current state of knowledge regarding the cellular uptake and trafficking of EVs is reviewed, along with a consideration of how these pathways potentially influence the functions of therapeutic EVs. Furthermore, the natural cell-targeting abilities, biodistribution profiles, and pharmacokinetics of exogenously administered EVs, along with the components responsible for these features are discussed. An overview of the potential clinical applications and preclinical examples of their successful use is also provided. Finally, examples of EV modifications that have successfully been employed to improve their therapeutic characteristics receive a particular focus. We suggest that, in addition to investigation of EV cell targeting and routes of uptake, future research into the routes of intracellular trafficking in recipient cells is required to optimally utilize EVs for therapeutic purposes.

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Mathilde Mathieu, Lorena Martin-Jaular, Grégory Lavieu, Clotilde Théry (2019 Jan 4)

Specificities of secretion and uptake of exosomes and other extracellular vesicles for cell-to-cell communication.

Nature cell biology : 9-17 : DOI : 10.1038/s41556-018-0250-9 En savoir plus
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The ability of exosomes to transfer cargo from donor to acceptor cells, thereby triggering phenotypic changes in the latter, has generated substantial interest in the scientific community. However, the extent to which exosomes differ from other extracellular vesicles in terms of their biogenesis and functions remains ill-defined. Here, we discuss the current knowledge on the specificities of exosomes and other types of extracellular vesicles, and their roles as important agents of cell-to-cell communication.

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Année de publication : 2018

Clotilde Théry, Kenneth W Witwer, Elena Aikawa, Maria Jose Alcaraz, Gregory Lavieu, ... Lorena Martin-Jaular, … Mathilde Mathieu, … Mercedes Tkach,… , Ewa K Zuba-Surma (2018 Nov 23)

Minimal information for studies of extracellular vesicles 2018 (MISEV2018): a position statement of the International Society for Extracellular Vesicles and update of the MISEV2014 guidelines.

Journal of extracellular vesicles : 1535750 : DOI : 10.1080/20013078.2018.1535750 En savoir plus
Résumé

The last decade has seen a sharp increase in the number of scientific publications describing physiological and pathological functions of extracellular vesicles (EVs), a collective term covering various subtypes of cell-released, membranous structures, called exosomes, microvesicles, microparticles, ectosomes, oncosomes, apoptotic bodies, and many other names. However, specific issues arise when working with these entities, whose size and amount often make them difficult to obtain as relatively pure preparations, and to characterize properly. The International Society for Extracellular Vesicles (ISEV) proposed Minimal Information for Studies of Extracellular Vesicles (« MISEV ») guidelines for the field in 2014. We now update these « MISEV2014 » guidelines based on evolution of the collective knowledge in the last four years. An important point to consider is that ascribing a specific function to EVs in general, or to subtypes of EVs, requires reporting of specific information beyond mere description of function in a crude, potentially contaminated, and heterogeneous preparation. For example, claims that exosomes are endowed with exquisite and specific activities remain difficult to support experimentally, given our still limited knowledge of their specific molecular machineries of biogenesis and release, as compared with other biophysically similar EVs. The MISEV2018 guidelines include tables and outlines of suggested protocols and steps to follow to document specific EV-associated functional activities. Finally, a checklist is provided with summaries of key points.

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Mercedes Tkach, Joanna Kowal, Clotilde Théry (2018 Jan 5)

Why the need and how to approach the functional diversity of extracellular vesicles.

Philosophical transactions of the Royal Society of London. Series B, Biological sciences : DOI : 20160479 En savoir plus
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In the past decade, cell-to-cell communication mediated by exosomes has attracted growing attention from biomedical scientists and physicians, leading to several recent publications in top-tier journals. Exosomes are generally defined as secreted membrane vesicles, or extracellular vesicles (EVs), corresponding to the intraluminal vesicles of late endosomal compartments, which are secreted upon fusion of multi-vesicular endosomes with the cell’s plasma membrane. Cells, however, were shown to release other types of EVs, for instance, by direct budding off their plasma membrane. Some of these EVs share with exosomes major biophysical and biochemical characteristics, such as size, density and membrane orientation, which impose difficulties in their efficient separation. Despite frequent claims in the literature, whether exosomes really display more important patho/physiological functions, or are endowed with higher potential as diagnostic or therapeutic tools than other EVs, is not yet convincingly demonstrated. In this opinion article, we describe reasons for this lack of precision knowledge in the current stage of the EV field, we review recently described approaches to overcome these caveats, and we propose ways to improve our knowledge on the respective functions of distinct EVs, which will be crucial for future development of well-designed EV-based clinical applications.This article is part of the discussion meeting issue ‘Extracellular vesicles and the tumour microenvironment’.

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Année de publication : 2017

Mercedes Tkach, Joanna Kowal, Andres E Zucchetti, Lotte Enserink, Mabel Jouve, Danielle Lankar, Michael Saitakis, Lorena Martin-Jaular, Clotilde Théry (2017 Sep 20)

Qualitative differences in T-cell activation by dendritic cell-derived extracellular vesicle subtypes.

The EMBO journal : DOI : e201696003 En savoir plus
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Exosomes, nano-sized secreted extracellular vesicles (EVs), are actively studied for their diagnostic and therapeutic potential. In particular, exosomes secreted by dendritic cells (DCs) have been shown to carry MHC-peptide complexes allowing efficient activation of T lymphocytes, thus displaying potential as promoters of adaptive immune responses. DCs also secrete other types of EVs of different size, subcellular origin and protein composition, whose immune capacities have not been yet compared to those of exosomes. Here, we show that large EVs (lEVs) released by human DCs are as efficient as small EVs (sEVs), including exosomes, to induce CD4(+) T-cell activation in vitro When released by immature DCs, however, lEVs and sEVs differ in their capacity to orient T helper (Th) cell responses, the former favouring secretion of Th2 cytokines, whereas the latter promote Th1 cytokine secretion (IFN-γ). Upon DC maturation, however, these functional differences are abolished, and all EVs become able to induce IFN-γ. Our results highlight the need to comprehensively compare the functionalities of EV subtypes in all patho/physiological systems where exosomes are claimed to perform critical roles.

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