Identifying extracellular vesicle populations from single cells

Nikoloff, Jonas; Saucedo-Espinosa, Mario A.; Kling, André; Dittrich, Petra S.

doi:10.1073/pnas.2106630118

Cited by 47 publications

(44 citation statements)

References 61 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…With the majority of EV analyses carried out on bulk populations, the nuance of EVs from each cell is lost in the milieu of millions of cells and with recent evidence of communication via EV secretion at the immune synapse, understanding the exchange of EVs between individual cells is of great importance (Choudhuri et al., 2014; Hoen et al., 2009; Saliba et al., 2019; Tetta et al., 2013). Microfluidic devices have been effectively employed to isolate single cells and subsequently their EVs, but are comparatively complex, only utilise low‐resolution imaging, are not applicable to all cell types and cannot directly observe the events of secretion (Ji et al., 2019; Nikoloff et al., 2021; Son et al., 2016).…”

Section: Discussionmentioning

confidence: 99%

Heterogeneity in extracellular vesicle secretion by single human macrophages revealed by super‐resolution microscopy

Dechantsreiter

Ambrose

Worboys

et al. 2022

J of Extracellular Vesicle

View full text Add to dashboard Cite

The diverse origins, nanometre-scale and invasive isolation procedures associated with extracellular vesicles (EVs) mean they are usually studied in bulk and disconnected from their parental cell. Here, we used super-resolution microscopy to directly compare EVs secreted by individual human monocyte-derived macrophages (MDMs). MDMs were differentiated to be M0-, M1-or M2-like, with all three secreting EVs at similar densities following activation. However, M0-like cells secreted larger EVs than M1-and M2-like macrophages. Proteomic analysis revealed variations in the contents of differently sized EVs as well as between EVs secreted by different MDM phenotypes. Super resolution microscopy of single-cell secretions identified that the class II MHC protein, HLA-DR, was expressed on ∼40% of EVs secreted from M1-like MDMs, which was double the frequency observed for M0-like and M2like EVs. Strikingly, human macrophages, isolated from the resected lungs of cancer patients, secreted EVs that expressed HLA-DR at double the frequency and with greater intensity than M1-like EVs. Quantitative analysis of single-cell EV profiles from all four macrophage phenotypes revealed distinct secretion types, five of which were consistent across multiple sample cohorts. A sub-population of M1-like MDMs secreted EVs similar to lung macrophages, suggesting an expansion or recruitment of cells with a specific EV secretion profile within the lungs of cancer patients. Thus, quantitative analysis of EV heterogeneity can be used for single cell profiling and to reveal novel macrophage biology.

show abstract

Section: Discussionmentioning

confidence: 99%

Heterogeneity in extracellular vesicle secretion by single human macrophages revealed by super‐resolution microscopy

Dechantsreiter

Ambrose

Worboys

et al. 2022

J of Extracellular Vesicle

View full text Add to dashboard Cite

show abstract

“…In addition, recent evidence has demonstrated the ability of EVs to establish surface protein-protein interactions. It appears, therefore, that at least a fraction of EVs tends to show functionally integrated complexes ( Leidal et al, 2020 ; Levy et al, 2021 ; Nikoloff et al, 2021 ; Razzauti and Laurent, 2021 ). At present, populations of single EV type can be isolated by various techniques based on distinct approaches, including monoclonal antibodies ( Levy et al, 2021 ; Lim et al, 2021 ).…”

Section: Evs: Origin Navigation and Fusion With Target Cellsmentioning

confidence: 99%

“…The state of knowledge and techniques are already advanced. Additional developments are expected for the future including their markers and signatures, useful for the identification of subtype-specific EVs and the unconventional secretion of their proteins ( Garcia-Martin et al, 2021 ; Levy et al, 2021 ; Nikoloff et al, 2021 ; Razzauti and Laurent, 2021 ).…”

Section: Evs: Origin Navigation and Fusion With Target Cellsmentioning

confidence: 99%

Unconventional Protein Secretion Dependent on Two Extracellular Vesicles: Exosomes and Ectosomes

Meldolesi

2022

Front. Cell Dev. Biol.

View full text Add to dashboard Cite

In addition to conventional protein secretion, dependent on the specific cleavage of signal sequences, proteins are secreted by other processes, all together called unconventional. Among the mechanisms operative in unconventional secretion, some are based on two families of extracellular vesicle (EVs), expressed by all types of cells: the exosomes (before secretion called ILVs) and ectosomes (average diameters ∼70 and ∼250 nm). The two types of EVs have been largely characterized by extensive studies. ILVs are assembled within endocytic vacuoles by inward budding of small membrane microdomains associated to cytosolic cargos including unconventional secretory proteins. The vacuoles containing ILVs are called multivesicular bodies (MVBs). Upon their possible molecular exchange with autophagosomes, MVBs undergo two alternative forms of fusion: 1. with lysosomes, followed by large digestion of their cargo molecules; and 2. with plasma membrane (called exocytosis), followed by extracellular diffusion of exosomes. The vesicles of the other type, the ectosomes, are differently assembled. Distinct plasma membrane rafts undergo rapid outward budding accompanied by accumulation of cytosolic/secretory cargo molecules, up to their sewing and pinching off. Both types of EV, released to the extracellular fluid in their complete forms including both membrane and cargo, start navigation for various times and distances, until their fusion with target cells. Release/navigation/fusion of EVs establish continuous tridimensional networks exchanging molecules, signals and information among cells. The proteins unconventionally secreted via EVs are a few hundreds. Some of them are functionally relevant (examples FADD, TNF, TACE), governing physiological processes and important diseases. Such proteins, at present intensely investigated, predict future discoveries and innovative developments, relevant for basic research and clinical practice.

show abstract

“…These nano-sized vesicles contain abundant signaling molecules of proteins, lipids, or nucleic acids, serving as important mediators of intercellular communication (2). The size, concentration, and composition of EVs have been reported to reliably reflect the function and pathological status of their cells of origin (3). For instance, activated leukocytes secrete more EVs that are enriched in cytokines and chemokines (4).…”

Section: Introductionmentioning

confidence: 99%

Burn Injury-Induced Extracellular Vesicle Production and Characteristics

Yang

Chatterjee

Zheng

et al. 2022

Shock

View full text Add to dashboard Cite

Extracellular vesicles (EVs) are nano-sized membrane-bound particles containing biologically active cargo molecules. The production and molecular composition of EVs reflect the physiological state of parent cells, and once released into the circulation, they exert pleiotropic functions via transferring cargo contents. Thus, circulating EVs not only serve as biomarkers, but also mediators in disease processes or injury responses. In the present study, we performed a comprehensive analysis of plasma EVs from burn patients and healthy subjects, characterizing their size distribution, concentration, temporal changes, cell origins, and cargo protein contents. Our results indicated that burn injury induced a significant increase in circulating EVs, the response peaked at the time of admission and declined over the course of recovery. Importantly, EV production correlated with injury severity, as indicated by the total body surface area and depth of burn, requirement for critical care/ICU stay, hospitalization length, wound infection, and concurrence of sepsis. Burn patients with inhalation injury showed a higher level of EVs than those without inhalation injury. We also evaluated patient demographics (age and sex) and pre-existing conditions (hypertension, obesity, and smoking) and found no significant correlation between these conditions and overall EV production. At the molecular level, flow cytometric analysis showed that the burn-induced EVs were largely derived from leukocytes and endothelial cells (ECs), which are known to be activated postburn. Additionally, a high level of zona-occludens-1 (ZO-1), a major constituent of tight junctions, was identified in burn EV cargos, indicative of injury in tissues that form barriers via tight junctions. Moreover, when applied to endothelial cell monolayers, burn EVs caused significant barrier dysfunction, characterized by decreased transcellular barrier resistance and disrupted cell–cell junction continuity. Taken together, these data suggest that burn injury promotes the production of EVs containing unique cargo proteins in a time-dependent manner; the response correlates with injury severity and worsened clinical outcomes. Functionally, burn EVs serve as a potent mediator capable of reducing endothelial barrier resistance and impairing junction integrity, a pathophysiological process underlying burn-associated tissue dysfunction. Thus, further in-depth characterization of circulating EVs will contribute to the development of new prognostic tools or therapeutic targets for advanced burn care.

show abstract

Identifying extracellular vesicle populations from single cells

Cited by 47 publications

References 61 publications

Heterogeneity in extracellular vesicle secretion by single human macrophages revealed by super‐resolution microscopy

Heterogeneity in extracellular vesicle secretion by single human macrophages revealed by super‐resolution microscopy

Unconventional Protein Secretion Dependent on Two Extracellular Vesicles: Exosomes and Ectosomes

Burn Injury-Induced Extracellular Vesicle Production and Characteristics

Contact Info

Product

Resources

About