scholarly journals Navigating the Landscape of Tumor Extracellular Vesicle Heterogeneity

2019 ◽  
Vol 20 (6) ◽  
pp. 1349 ◽  
Author(s):  
Sabrina Roy ◽  
Hsing-Ying Lin ◽  
Chung-Yu Chou ◽  
Chen-Han Huang ◽  
Julia Small ◽  
...  
Keyword(s):  
Extracellular Vesicles ◽  
Intercellular Communication ◽  
Extracellular Vesicle ◽  
Rapid Expansion ◽  
Parent Cell ◽  
Membrane Composition ◽  
Pathological Conditions ◽  
Respective Parent ◽  
Experimental Level

The last decade has seen a rapid expansion of interest in extracellular vesicles (EVs) released by cells and proposed to mediate intercellular communication in physiological and pathological conditions. Considering that the genetic content of EVs reflects that of their respective parent cell, many researchers have proposed EVs as a source of biomarkers in various diseases. So far, the question of heterogeneity in given EV samples is rarely addressed at the experimental level. Because of their relatively small size, EVs are difficult to reliably isolate and detect within a given sample. Consequently, standardized protocols that have been optimized for accurate characterization of EVs are lacking despite recent advancements in the field. Continuous improvements in pre-analytical parameters permit more efficient assessment of EVs, however, methods to more objectively distinguish EVs from background, and to interpret multiple single-EV parameters are lacking. Here, we review EV heterogeneity according to their origin, mode of release, membrane composition, organelle and biochemical content, and other factors. In doing so, we also provide an overview of currently available and potentially applicable methods for single EV analysis. Finally, we examine the latest findings from experiments that have analyzed the issue at the single EV level and discuss potential implications.

scholarly journals Myocardial infarction affects Cx43 content of extracellular vesicles secreted by cardiomyocytes

2020 ◽  
Vol 3 (12) ◽  
pp. e202000821
Author(s):  
Tania Martins-Marques ◽  
Teresa Ribeiro-Rodrigues ◽  
Saskia C de Jager ◽  
Monica Zuzarte ◽  
Cátia Ferreira ◽  
...  
Keyword(s):  
Extracellular Vesicles ◽  
Intercellular Communication ◽  
Connexin 43 ◽  
Extracellular Vesicle ◽  
Therapeutic Strategies ◽  
Target Cells ◽  
Comprehensive Strategy ◽  
Junction Protein ◽  
Gap Junction Protein

Ischemic heart disease has been associated with an impairment on intercellular communication mediated by both gap junctions and extracellular vesicles. We have previously shown that connexin 43 (Cx43), the main ventricular gap junction protein, assembles into channels at the extracellular vesicle surface, mediating the release of vesicle content into target cells. Here, using a comprehensive strategy that included cell-based approaches, animal models and human patients, we demonstrate that myocardial ischemia impairs the secretion of Cx43 into circulating, intracardiac and cardiomyocyte-derived vesicles. In addition, we show that ubiquitin signals Cx43 release in basal conditions but appears to be dispensable during ischemia, suggesting an interplay between ischemia-induced Cx43 degradation and secretion. Overall, this study constitutes a step forward for the characterization of the signals and molecular players underlying vesicle protein sorting, with strong implications on long-range intercellular communication, paving the way towards the development of innovative diagnostic and therapeutic strategies for cardiovascular disorders.

Proteomic Profile of Urinary Extracellular Vesicles Identifies AGP1 as a Potential Biomarker of Primary Aldosteronism

Endocrinology ◽  
2021 ◽  
Vol 162 (4) ◽  
Author(s):  
Eric R Barros ◽  
Juan Pablo Rigalli ◽  
Alejandra Tapia-Castillo ◽  
Andrea Vecchiola ◽  
Morag J Young ◽  
...  
Keyword(s):  
Extracellular Vesicles ◽  
Primary Aldosteronism ◽  
Proteomic Analysis ◽  
Characteristic Curve ◽  
Area Under The Curve ◽  
Extracellular Vesicle ◽  
Parent Cell ◽  
Spot Urine ◽  
Potential Biomarker ◽  
Aldosterone To Renin Ratio

Abstract Context Primary aldosteronism (PA) represents 6% to 10% of all essential hypertension patients and is diagnosed using the aldosterone-to-renin ratio (ARR) and confirmatory studies. The complexity of PA diagnosis encourages the identification of novel PA biomarkers. Urinary extracellular vesicles (uEVs) are a potential source of biomarkers, considering that their cargo reflects the content of the parent cell. Objective We aimed to evaluate the proteome of uEVs from PA patients and identify potential biomarker candidates for PA. Methods Second morning spot urine was collected from healthy controls (n = 8) and PA patients (n = 7). The uEVs were isolated by ultracentrifugation and characterized. Proteomic analysis on uEVs was performed using LC-MS Orbitrap. Results Isolated uEVs carried extracellular vesicle markers, showed a round shape and sizes between 50 and 150 nm. The concentration of uEVs showed a direct correlation with urinary creatinine (r = 0.6357; P = 0.0128). The uEV size mean (167 ± 6 vs 183 ± 4nm) and mode (137 ± 7 vs 171 ± 11nm) was significantly smaller in PA patients than in control subjects, but similar in concentration. Proteomic analysis of uEVs from PA patients identified an upregulation of alpha-1-acid glycoprotein 1 (AGP1) in PA uEVs, which was confirmed using immunoblot. A receiver operating characteristic curve analysis showed an area under the curve of 0.92 (0.82 to 1; P = 0.0055). Conclusion Proteomic and further immunoblot analyses of uEVs highlights AGP1 as potential biomarker for PA.

scholarly journals Emerging role of extracellular vesicles in the regulation of skeletal muscle adaptation

2019 ◽  
Vol 127 (2) ◽  
pp. 645-653 ◽  
Author(s):  
Ivan J. Vechetti
Keyword(s):  
Skeletal Muscle ◽  
Extracellular Vesicles ◽  
Human Body ◽  
Intercellular Communication ◽  
Skeletal Muscle Mass ◽  
Genetic Material ◽  
Muscle Adaptation ◽  
Pathological Conditions ◽  
Isolation Methods

Extracellular vesicles (EVs) were initially characterized as “garbage bags” with the purpose of removing unwanted material from cells. It is now becoming clear that EVs mediate intercellular communication between distant cells through a transfer of genetic material, a process important to the systemic adaptation in physiological and pathological conditions. Although speculative, it has been suggested that the majority of EVs that make it into the bloodstream would be coming from skeletal muscle, since it is one of the largest organs in the human body. Although it is well established that skeletal muscle secretes peptides (currently known as myokines) into the bloodstream, the notion that skeletal muscle releases EVs is in its infancy. Besides intercellular communication and systemic adaptation, EV release could represent the mechanism by which muscle adapts to certain stimuli. This review summarizes the current understanding of EV biology and biogenesis and current isolation methods and briefly discusses the possible role EVs have in regulating skeletal muscle mass.

scholarly journals Emerging roles of extracellular vesicles in the intercellular communication for exercise-induced adaptations

2020 ◽  
Vol 319 (2) ◽  
pp. E320-E329
Author(s):  
Joshua Denham ◽  
Sarah J. Spencer
Keyword(s):  
Extracellular Vesicles ◽  
Intercellular Communication ◽  
Biological Material ◽  
Molecular Mechanisms ◽  
Health Benefits ◽  
Extracellular Vesicle ◽  
Therapeutic Interventions ◽  
Current Evidence ◽  
Future Research ◽  
Age Related

Complex organisms rely heavily on intercellular communication. The rapidly expanding field of extracellular vesicle biology has made it clear that the necessary intercellular communication occurs partly through their paracrine and endocrine actions. Extracellular vesicles are nanoscale lipid membranes (30–2,000 nm in diameter) that shuttle functional biological material between cells. They are released from numerous tissues and are isolated from nearly all biofluids and cell cultures. Although their biogenesis, cell targeting, and functional roles are incompletely understood, they appear to have crucial roles in physiological and disease processes. Their enormous potential to serve as sensitive biomarkers of disease and also new therapeutic interventions for diseases have gained them considerable attention in recent years. Regular physical exercise training confers systemic health benefits and consequently prevents many age-related degenerative diseases. Many of the molecular mechanisms responsible for the salubrious effects of exercise are known, yet a common underlying mechanism potentially responsible for the holistic health benefits of exercise has only recently been explored (i.e., via extracellular vesicle transport of biological material). Here, we provide an overview of extracellular vesicle biology before outlining the current evidence on the capacity for a single bout and chronic exercise to elicit changes in extracellular vesicle content and modulate their molecular cargo (e.g., small RNAs). We highlight areas for future research and emphasize their potential utility as biomarkers and therapeutic strategies of disease and its prevention.

scholarly journals Antibody-Free Labeling of Malaria-Derived Extracellular Vesicles Using Flow Cytometry

Biomedicines ◽  
2020 ◽  
Vol 8 (5) ◽  
pp. 98
Author(s):  
Elya Dekel ◽  
Paula Abou Karam ◽  
Yael Ohana-Daniel ◽  
Mirit Biton ◽  
Neta Regev-Rudzki ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Extracellular Vesicles ◽  
Intercellular Communication ◽  
Malaria Parasite ◽  
Flow Cytometer ◽  
Cellular Processes ◽  
Parasite Plasmodium ◽  
Membrane Bound ◽  
The Way

Extracellular vesicles (EVs) are cell-derived membrane-bound structures that are believed to play a major role in intercellular communication by allowing cells to exchange proteins and genetic cargo between them. In particular, pathogens, such as the malaria parasite Plasmodium (P.) falciparum, utilize EVs to promote their growth and to alter their host’s response. Thus, better characterization of these secreted organelles will enhance our understanding of the cellular processes that govern EVs’ biology and pathological functions. Here we present a method that utilizes a high-end flow cytometer system to characterize small EVs, i.e., with a diameter less than 200 nm. Using this method, we could evaluate different parasite-derived EV populations according to their distinct cargo by using antibody-free labeling. It further allows to closely monitor a sub-population of vesicles carrying parasitic DNA cargo. This ability paves the way to conducting a more ‘educated’ analysis of the various EV cargo components.

scholarly journals 3249 Defining the Extracellular Vesicle Content of Interstitial Fluid for Blood-Free Diagnostics; Extraction Methods and Initial Characterization

2019 ◽  
Vol 3 (s1) ◽  
pp. 7-7
Author(s):  
Justin Baca Robert Taylor ◽  
Srinivasa Rao Gadam ◽  
Lauren Perez
Keyword(s):  
Electron Microscopy ◽  
Minimally Invasive ◽  
Intercellular Communication ◽  
Interstitial Fluid ◽  
Human Subjects ◽  
Extracellular Vesicle ◽  
Vis Spectroscopy ◽  
Initial Characterization

OBJECTIVES/SPECIFIC AIMS: Recent advances in microneedle technology have enabled practical, in vivo dermal interstitial fluid (ISF) sampling. These minimally-invasive techniques allow for collection of ISF without damage to adjacent tissues and do not rely on blister formation. Initial reports of extracellular vesicle (EV) isolation from dermal ISF and paired blood samples suggest that EVs may be more abundant in ISF. Analysis of ISF-derived EVs may allow for more detailed study of intercellular communication at the tissue level, particularly in acute inflammatory conditions. The objective of this study is to describe the isolation and initial characterization of interstitial fluid-derived exosomes. METHODS/STUDY POPULATION: We apply electron microscopy, nanoparticle tracking analysis (NTA), immuochemical, and sequencing methods to describe and distinguish the EV content of interstitial fluid. We include apparently healthy adult human subjects with no active skin disease. We also study immunocompetent, CD-hairless rats to demonstrate the generalizability of the methods. RESULTS/ANTICIPATED RESULTS: We successfully isolated EVs from human and rat interstitial fluid using commercially available precipitation methods. The EVs were initially characterized using UV/Vis spectroscopy, electron microscopy, and NTA. While the study is ongoing, initial results suggest that the concentration and size distribution of EVs differs significantly between blood fractions and ISF. Further immunochemical and sequencing characterization is ongoing. DISCUSSION/SIGNIFICANCE OF IMPACT: We present here the initial characterization of EVs isolated from dermal interstitial fluid. This appears to be the first report of EV characterization using ISF collection methods that do not perturb adjacent tissues (such as with blister or microdialysis methods). The present study lays a foundation for further examination of ISF-derived EVs in acute inflammatory disease such as cellulitis or infectious neuritis. This may enable minimally invasive diagnostics and new research tools to understand intercellular communication in living organisms with increased spatial and temporal resolution.

scholarly journals Extracellular Vesicle Quantification and Characterization: Common Methods and Emerging Approaches

2019 ◽  
Vol 6 (1) ◽  
pp. 7 ◽  
Author(s):  
Thomas Hartjes ◽  
Serhii Mytnyk ◽  
Guido Jenster ◽  
Volkert van Steijn ◽  
Martin van Royen
Keyword(s):  
Physical Properties ◽  
Extracellular Vesicles ◽  
Intercellular Communication ◽  
Therapeutic Potential ◽  
Membrane Vesicles ◽  
Extracellular Vesicle ◽  
Medical Applications ◽  
Accurate Assessment ◽  
Further Development

Extracellular vesicles (EVs) are a family of small membrane vesicles that carry information about cells by which they are secreted. Growing interest in the role of EVs in intercellular communication, but also in using their diagnostic, prognostic and therapeutic potential in (bio) medical applications, demands for accurate assessment of their biochemical and physical properties. In this review, we provide an overview of available technologies for EV analysis by describing their working principles, assessing their utility in EV research and summarising their potential and limitations. To emphasise the innovations in EV analysis, we also highlight the unique possibilities of emerging technologies with high potential for further development.

scholarly journals Tetraspanins, More than Markers of Extracellular Vesicles in Reproduction

2020 ◽  
Vol 21 (20) ◽  
pp. 7568
Author(s):  
Jana Jankovičová ◽  
Petra Sečová ◽  
Katarína Michalková ◽  
Jana Antalíková
Keyword(s):  
Extracellular Vesicles ◽  
Reproductive System ◽  
Extracellular Vesicle ◽  
Cell Uptake ◽  
Cell Contact ◽  
Cellular Processes ◽  
Pathological Conditions ◽  
Vesicle Biogenesis ◽  
Future Direction ◽  
Insight Into

The participation of extracellular vesicles in many cellular processes, including reproduction, is unquestionable. Although currently, the tetraspanin proteins found in extracellular vesicles are mostly applied as markers, increasing evidence points to their role in extracellular vesicle biogenesis, cargo selection, cell targeting, and cell uptake under both physiological and pathological conditions. In this review, we bring other insight into the involvement of tetraspanin proteins in extracellular vesicle physiology in mammalian reproduction. We provide knowledge regarding the involvement of extracellular vesicle tetraspanins in these processes in somatic cells. Furthermore, we discuss the future direction towards an understanding of their functions in the tissues and fluids of the mammalian reproductive system in gamete maturation, fertilization, and embryo development; their involvement in mutual cell contact and communication in their complexity.

scholarly journals From Tumor Metastasis towards Cerebral Ischemia—Extracellular Vesicles as a General Concept of Intercellular Communication Processes

2019 ◽  
Vol 20 (23) ◽  
pp. 5995 ◽  
Author(s):  
Xuan Zheng ◽  
Mathias Bähr ◽  
Thorsten R. Doeppner
Keyword(s):  
Cerebral Ischemia ◽  
Extracellular Vesicles ◽  
Intercellular Communication ◽  
Short Review ◽  
General Concept ◽  
Tumor Formation ◽  
Pathological Conditions ◽  
Communication Processes ◽  
Reported Data

Extracellular vesicles (EVs) have been tremendous carriers in both experimental and translational science. These vesicles—formerly regarded as artifacts of in vitro research—have a heterogeneous population of vesicles derived from virtually all eukaryotic cells. EVs consist of a bilayer lipid structure with a diameter of about 30 to 1000 nm and have a characteristic protein and non-coding RNA content that make up different forms of EVs such as exosomes, microvesicles, and others. Despite recent progress in the EV field, which is known to serve as potential biomarkers and therapeutic tools under various pathological conditions, fundamental questions are yet to be answered. This short review focuses on recently reported data regarding EVs under pathological conditions with a particular emphasis on the role of EVs under such different conditions like tumor formation and cerebral ischemia. The review strives to point out general concepts of EV intercellular communication processes that might be vital to both diagnostic and therapeutic strategies in the long run.

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