Extracellular Vesicles in the Blood of Dogs with Cancer—A Preliminary Study

Extracellular vesicles (EVs) are a heterogeneous population of submicron-sized structures released during the activation, proliferation, or apoptosis of various types of cells. Due to their size, their role in cell-to-cell communication in cancer is currently being discussed. In blood, the most abundant population of EVs is platelet-derived EVs (PEVs). The aim of this study was to estimate the absolute number and the origin of EVs in the blood of healthy dogs and of dogs with various types of cancer. The EV absolute number and cellular origin were examined by flow cytometry technique. EVs were classified on the basis of surface annexin V expression (phosphatidylserine PS+) and co-expression of specific cellular markers (CD61, CD45, CD3, CD21). The number of PEVs was significantly higher in dogs with cancer (median: 409/µL, range: 42–2748/µL vs. median: 170/µL, range: 101–449/µL in controls). The numbers of EVs derived from leukocytes (control median: 86/µL, range: 40–240/µL; cancer median: 443/µL, range: 44–3 352/µL) and T cells (control median: 5/µL, range: 2–66/µL; cancer median: 108/µL, range: 3–1735/µL) were higher in dogs with neoplasia compared to healthy controls. The estimation of PEV and leukocyte-derived EV counts may provide a useful biological marker in dogs with cancer.

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Extracellular Vesicles from the Protozoa Acanthamoeba castellanii: Their Role in Pathogenesis, Environmental Adaptation and Potential Applications

Bioengineering ◽

10.3390/bioengineering6010013 ◽

2019 ◽

Vol 6 (1) ◽

pp. 13 ◽

Cited By ~ 4

Author(s):

Diego Gonçalves ◽

Marina Ferreira ◽

Allan Guimarães

Keyword(s):

Extracellular Vesicles ◽

Cell Communication ◽

Acanthamoeba Castellanii ◽

Living Organism ◽

Biofuel Production ◽

Cellular Origin ◽

Wide Range ◽

Free Living Amoeba ◽

Potential Applications ◽

Multicellular Organisms

Extracellular vesicles (EVs) are membranous compartments of distinct cellular origin and biogenesis, displaying different sizes and include exosomes, microvesicles, and apoptotic bodies. The EVs have been described in almost every living organism, from simple unicellular to higher evolutionary scale multicellular organisms, such as mammals. Several functions have been attributed to these structures, including roles in energy acquisition, cell-to-cell communication, gene expression modulation and pathogenesis. In this review, we described several aspects of the recently characterized EVs of the protozoa Acanthamoeba castellanii, a free-living amoeba (FLA) of emerging epidemiological importance, and compare their features to other parasites’ EVs. These A. castellanii EVs are comprised of small microvesicles and exosomes and carry a wide range of molecules involved in many biological processes like cell signaling, carbohydrate metabolism and proteolytic activity, such as kinases, glucanases, and proteases, respectively. Several biomedical applications of these EVs have been proposed lately, including their use in vaccination, biofuel production, and the pharmaceutical industry, such as platforms for drug delivery.

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Extracellular vesicles as biomarkers for diagnostic and prognostic application in cardiovascular disease

Trillium Exctracellular Vesicles ◽

10.47184/tev.2020.01.05 ◽

2020 ◽

Vol 2 (1) ◽

pp. 36-41

Author(s):

Christoph Lipps

Keyword(s):

Cardiovascular Disease ◽

Extracellular Vesicles ◽

Diagnostic And Prognostic Application ◽

Cell Communication ◽

Current Evidence ◽

Cellular Origin ◽

New Class ◽

Diagnosis And Prognosis ◽

Prognostic Application ◽

Pulmonary Arterial

Cardiovascular disease (CVD) remains the leading cause of morbidity and mortality worldwide. Early diagnosis and the development of a prognosis is important for management or secondary prevention of the disease. In the past few decades, various biomarkers have been identified for improved risk assessment, more accurate diagnosis and prognosis, and a better understanding of the underlying pathophysiology in CVD. Extracellular vesicles (EVs) are thought to be important to cell-to-cell communication in the heart, and EV counts, cellular origin, and EV content have been related to CVD. This review examines current evidence for the potential application of EVs as a new class of biomarkers in CVD. Keywords: extracellular vesicles, biomarker, liquid biopsy, cardiovascular disease, myocardial infarction, heart failure, pulmonary arterial hypertension

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Detection of disease-associated microRNAs — application for autism spectrum disorders

Reviews in the Neurosciences ◽

10.1515/revneuro-2020-0015 ◽

2020 ◽

Vol 31 (7) ◽

pp. 757-769

Author(s):

Barbora Konečná ◽

Jana Radošinská ◽

Petra Keményová ◽

Gabriela Repiská

Keyword(s):

Autism Spectrum Disorders ◽

Extracellular Vesicles ◽

High Throughput Sequencing ◽

Cell Communication ◽

Biological Marker ◽

Storage Conditions ◽

Autism Spectrum ◽

Sample Collection ◽

Spectrum Disorders ◽

Processing And Storage

AbstractAutism spectrum disorders (ASD) diagnostic procedure still lacks a uniform biological marker. This review gathers the information on microRNAs (miRNAs) specifically as a possible source of biomarkers of ASD. Extracellular vesicles, and their subset of exosomes, are believed to be a tool of cell-to-cell communication, and they are increasingly considered to be carriers of such a marker. The interest in studying miRNAs in extracellular vesicles grows in all fields of study and therefore should not be omitted in the field of neurodevelopmental disorders. The summary of miRNAs associated with brain cells and ASD either studied directly in the tissue or biofluids are gathered in this review. The heterogeneity in findings from different studies points out the fact that unified methods should be established, beginning with the determination of the accurate patient and control groups, through to sample collection, processing, and storage conditions. This review, based on the available literature, proposes the standardized approach to obtain the results that would not be affected by technical factors. Nowadays, the method of high-throughput sequencing seems to be the most optimal to analyze miRNAs. This should be followed by the uniformed bioinformatics procedure to avoid misvalidation. At the end, the proper validation of the obtained results is needed. With such an approach as is described in this review, it would be possible to obtain a reliable biomarker that would characterize the presence of ASD.

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Separation of Extracellular Vesicles by DNA-Directed Immunocapturing Followed by Enzymatic Release

10.26434/chemrxiv.12234683 ◽

2020 ◽

Author(s):

Dario Brambilla ◽

Laura Sola ◽

Elisa Chiodi ◽

Natasa Zarovni ◽

Diogo Fortunato ◽

...

Keyword(s):

Cell Culture ◽

Extracellular Vesicles ◽

Culture Media ◽

Biological Fluids ◽

Imaging Techniques ◽

Cell Communication ◽

Disease Diagnosis ◽

Cell Culture Supernatant ◽

Transmission Electron ◽

Downstream Analysis

Extracellular vesicles (EVs) have attracted great interest among researchers due to their role in cell-cell communication, disease diagnosis, and drug delivery. In spite of their potential in the medical field, there is no consensus on the best method for separating microvesicles from cell culture supernatant and complex biological fluids. Obtaining a good recovery yield and preserving physical characteristics is critical for the diagnostic and therapeutic use of EVs. The separation is made complex by the fact that blood and cell culture media, contain a large number of nanoparticles in the same size range. Methods that exploit immunoaffinity capture provide high purity samples and overcome the issues of currently used separation methods. However, the release of captured nanovesicles requires harsh conditions that hinder their use in certain types of downstream analysis. Herein, a novel capture and release approach for small extracellular vesicles (sEVs), based on DNAdirected immobilization of antiCD63 antibody is presented. The flexible DNAlinker increases the capture efficiency and allows releasing of EVs by exploiting the endonucleasic activity of DNAse I. This separation protocol works under mild conditions, enabling the release of intact vesicles that can be successfully analyzed by imaging techniques. In this article sEVs recovered from plasma were characterized by established techniques for EVs analysis including nanoparticle tracking and transmission electron microscopy.<br>

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Multiple Roles of Exosomal Long Noncoding RNAs in Cancers

BioMed Research International ◽

10.1155/2019/1460572 ◽

2019 ◽

Vol 2019 ◽

pp. 1-12 ◽

Cited By ~ 5

Author(s):

Wenyuan Zhao ◽

Yuanqi Liu ◽

Chunfang Zhang ◽

Chaojun Duan

Keyword(s):

Extracellular Vesicles ◽

Cancer Diagnosis ◽

Tumor Angiogenesis ◽

Cell Communication ◽

Noncoding Rnas ◽

Multiple Roles ◽

Long Noncoding Rnas ◽

Cancer Therapies ◽

Transcriptional Noise ◽

Intercellular Exchange

Long noncoding RNAs (lncRNAs) are not transcriptional noise, as previously understood, but are currently considered to be multifunctional. Exosomes are derived from the internal multivesicular compartment and are extracellular vesicles (EVs) with diameters of 30–100 nm. Exosomes play significant roles in the intercellular exchange of information and material. Exosomal lncRNAs may be promising biomarkers for cancer diagnosis and potential targets for cancer therapies, since they are increasingly understood to be involved in tumorigenesis, tumor angiogenesis, and chemoresistance. This review mainly focuses on the roles of emerging exosomal lncRNAs in cancer. In addition, the biogenesis of exosomes, the functions of lncRNAs, and the mechanisms of lncRNAs in exosome-mediated cell-cell communication are also summarized.

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CSF extracellular vesicles and risk of disease activity after a first demyelinating event

Multiple Sclerosis Journal ◽

10.1177/1352458520987542 ◽

2021 ◽

pp. 135245852098754

Author(s):

Gloria Dalla Costa ◽

Tommaso Croese ◽

Marco Pisa ◽

Annamaria Finardi ◽

Lorena Fabbella ◽

...

Keyword(s):

Disease Activity ◽

Extracellular Vesicles ◽

Prognostic Markers ◽

Cell Communication ◽

Extracellular Vesicle ◽

Intervention Strategies ◽

Targeted Intervention ◽

Improve Risk Stratification ◽

Risk Of Disease ◽

In Cis

Background: Extracellular vesicles (EVs), a recently described mechanism of cell communication, are released from activated microglial cells and macrophages and are a candidate biomarker in diseases characterized by chronic inflammatory process such as multiple sclerosis (MS). Methods: We explored cerebrospinal fluid extracellular vesicle (CSF EV) of myeloid origin (MEVs), cytokine and chemokine levels in patients with clinically isolated syndrome (CIS). Results: We found that CSF MEVs were significantly higher in CIS patients than in controls and were inversely correlated to CSF CCL2 levels. MEVs level were significantly associated with an shorter time to evidence of disease activity (hazard ratio: 1.01, 95% confidence interval: 1.00–1.02, p < 0.01) independently from other known prognostic markers. Conclusion: After a first demyelinating event, CSF EVs may improve risk stratification of these patients and allow more targeted intervention strategies.

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Glycosylation of Cancer Extracellular Vesicles: Capture Strategies, Functional Roles and Potential Clinical Applications

Cells ◽

10.3390/cells10010109 ◽

2021 ◽

Vol 10 (1) ◽

pp. 109

Author(s):

Álvaro M. Martins ◽

Cátia C. Ramos ◽

Daniela Freitas ◽

Celso A. Reis

Keyword(s):

Extracellular Vesicles ◽

Biomarker Discovery ◽

De Novo ◽

Recipient Cell ◽

Cell Communication ◽

Cancer Biomarkers ◽

Functional Roles ◽

Molecular Information ◽

Glycosylation Pathway ◽

Organ Tropism

Glycans are major constituents of extracellular vesicles (EVs). Alterations in the glycosylation pathway are a common feature of cancer cells, which gives rise to de novo or increased synthesis of particular glycans. Therefore, glycans and glycoproteins have been widely used in the clinic as both stratification and prognosis cancer biomarkers. Interestingly, several of the known tumor-associated glycans have already been identified in cancer EVs, highlighting EV glycosylation as a potential source of circulating cancer biomarkers. These particles are crucial vehicles of cell–cell communication, being able to transfer molecular information and to modulate the recipient cell behavior. The presence of particular glycoconjugates has been described to be important for EV protein sorting, uptake and organ-tropism. Furthermore, specific EV glycans or glycoproteins have been described to be able to distinguish tumor EVs from benign EVs. In this review, the application of EV glycosylation in the development of novel EV detection and capture methodologies is discussed. In addition, we highlight the potential of EV glycosylation in the clinical setting for both cancer biomarker discovery and EV therapeutic delivery strategies.

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The Cellular and Biological Impact of Extracellular Vesicles in Pancreatic Cancer

Cancers ◽

10.3390/cancers13123040 ◽

2021 ◽

Vol 13 (12) ◽

pp. 3040

Author(s):

Zainab Hussain ◽

Jeremy Nigri ◽

Richard Tomasini

Keyword(s):

Pancreatic Cancer ◽

Tumor Cells ◽

Extracellular Vesicles ◽

Cell Communication ◽

Pancreatic Tumors ◽

Mediated Communication ◽

Biological Impact ◽

Physiological Relevance ◽

Cellular Components ◽

Tumoral Cells

Deciphering the interactions between tumor and stromal cells is a growing field of research to improve pancreatic cancer-associated therapies and patients’ care. Indeed, while accounting for 50 to 90% of the tumor mass, many pieces of evidence reported that beyond their structural role, the non-tumoral cells composing the intra-tumoral microenvironment influence tumor cells’ proliferation, metabolism, cell death and resistance to therapies, among others. Simultaneously, tumor cells can influence non-tumoral neighboring or distant cells in order to shape a tumor-supportive and immunosuppressive environment as well as influencing the formation of metastatic niches. Among intercellular modes of communication, extracellular vesicles can simultaneously transfer the largest variety of signals and were recently reported as key effectors of cell–cell communication in pancreatic cancer, from its development to its evolution as well as its ability to resist available treatments. This review focuses on extracellular vesicles-mediated communication between different cellular components of pancreatic tumors, from the modulation of cellular activities and abilities to their biological and physiological relevance. Taking into consideration the intra-tumoral microenvironment and its extracellular-mediated crosstalk as main drivers of pancreatic cancer development should open up new therapeutic windows.

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Extracellular Vesicles-Based Drug Delivery Systems: A New Challenge and the Exemplum of Malignant Pleural Mesothelioma

International Journal of Molecular Sciences ◽

10.3390/ijms21155432 ◽

2020 ◽

Vol 21 (15) ◽

pp. 5432 ◽

Cited By ~ 3

Author(s):

Stefano Burgio ◽

Leila Noori ◽

Antonella Marino Gammazza ◽

Claudia Campanella ◽

Mariantonia Logozzi ◽

...

Keyword(s):

Drug Delivery ◽

Early Diagnosis ◽

Extracellular Vesicles ◽

Delivery System ◽

Malignant Pleural Mesothelioma ◽

Cell Communication ◽

Cell Types ◽

Delivery Systems ◽

Pleural Mesothelioma ◽

Potential Use

Research for the most selective drug delivery to tumors represents a fascinating key target in science. Alongside the artificial delivery systems identified in the last decades (e.g., liposomes), a family of natural extracellular vesicles (EVs) has gained increasing focus for their potential use in delivering anticancer compounds. EVs are released by all cell types to mediate cell-to-cell communication both at the paracrine and the systemic levels, suggesting a role for them as an ideal nano-delivery system. Malignant pleural mesothelioma (MPM) stands out among currently untreatable tumors, also due to the difficulties in achieving an early diagnosis. Thus, early diagnosis and treatment of MPM are both unmet clinical needs. This review looks at indirect and direct evidence that EVs may represent both a new tool for allowing an early diagnosis of MPM and a potential new delivery system for more efficient therapeutic strategies. Since MPM is a relatively rare malignant tumor and preclinical MPM models developed to date are very few and not reliable, this review will report data obtained in other tumor types, suggesting the potential use of EVs in mesothelioma patients as well.

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Cell-to-Cell Communication by Host-Released Extracellular Vesicles in the Gut: Implications in Health and Disease

International Journal of Molecular Sciences ◽

10.3390/ijms22042213 ◽

2021 ◽

Vol 22 (4) ◽

pp. 2213

Author(s):

Natalia Diaz-Garrido ◽

Cecilia Cordero ◽

Yenifer Olivo-Martinez ◽

Josefa Badia ◽

Laura Baldomà

Keyword(s):

Extracellular Vesicles ◽

Intestinal Mucosa ◽

Current Knowledge ◽

Cell Communication ◽

Bioactive Molecules ◽

Target Cells ◽

Immune Functions ◽

Intestinal Homeostasis ◽

General Terms ◽

Health And Disease

Communication between cells is crucial to preserve body homeostasis and health. Tightly controlled intercellular dialog is particularly relevant in the gut, where cells of the intestinal mucosa are constantly exposed to millions of microbes that have great impact on intestinal homeostasis by controlling barrier and immune functions. Recent knowledge involves extracellular vesicles (EVs) as mediators of such communication by transferring messenger bioactive molecules including proteins, lipids, and miRNAs between cells and tissues. The specific functions of EVs principally depend on the internal cargo, which upon delivery to target cells trigger signal events that modulate cellular functions. The vesicular cargo is greatly influenced by genetic, pathological, and environmental factors. This finding provides the basis for investigating potential clinical applications of EVs as therapeutic targets or diagnostic biomarkers. Here, we review current knowledge on the biogenesis and cargo composition of EVs in general terms. We then focus the attention to EVs released by cells of the intestinal mucosa and their impact on intestinal homeostasis in health and disease. We specifically highlight their role on epithelial barrier integrity, wound healing of epithelial cells, immunity, and microbiota shaping. Microbiota-derived EVs are not reviewed here.

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