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Extracellular Vesicles as a New Source of Liquid Biopsy 2.0
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Extracellular Vesicles as a New Source of Liquid Biopsy 2.0

A special issue of International Journal of Molecular Sciences (ISSN 1422-0067). This special issue belongs to the section "Molecular Endocrinology and Metabolism".

Deadline for manuscript submissions: closed (31 January 2022) | Viewed by 24156

Special Issue Editors

Dr. Paola Lanuti

E-Mail Website
Guest Editor
Department of Medicine and Aging Science, University “G. d’Annunzio” of Chieti-Pescara, Via dei Vestini, 31, 66100 Chieti, Italy
Interests: extracellular vesicles; flow cytometry; biomarkers
Special Issues, Collections and Topics in MDPI journals
Dr. Davide Brocco

E-Mail Website
Guest Editor
Department of Pharmacy, University "G. D'Annunzio" Chieti-Pescara, Chieti, Italy
Interests: Translational research in medical oncology

Special Issue Information

Dear Colleagues,

This Special Issue is the continuation of our previous special issue "Extracellular Vesicles as a New Source of Liquid Biopsy".

Within the past decade, extracellular vesicles have progressively emerged as key regulators of organized communities of cells within multicellular organisms in health and disease. Extracellular vesicles have been described as shuttle vectors or signal transducers that can deliver specific biological information even at long distances. Due to their substantial stability, extracellular vesicles circulate through bodily fluids and have been identified in blood, urine, cerebrospinal fluid, saliva, milk, and tears. It is known that extracellular vesicles are a heterogeneous group of lipidic structures ranging from 30 to 50,000 nm in size, generically subclassified as small, if within 100 nm, and medium/large, if above 100–200 nm.

The extracellular vesicle cargo, consisting of genomic DNA, RNAs, miRNAs, proteins, and lipids dynamically reflecting the characteristics of the cells of origin, makes them an attractive source of biological information. For all of these reasons, extracellular vesicles have high potential as dynamic biomarkers of diseases and/or response to therapies. The deep characterization of extracellular vesicle general characteristics and cargoes related to their roles in tumorigenesis, immunity, and vascular biology may present interesting perspectives in the assessment of extracellular vesicles as a reliable liquid biopsy source.

Dr. Paola Lanuti
Dr. Davide Brocco
Guest Editors

Manuscript Submission Information

Manuscripts should be submitted online at www.mdpi.com by registering and logging in to this website. Once you are registered, click here to go to the submission form. Manuscripts can be submitted until the deadline. All submissions that pass pre-check are peer-reviewed. Accepted papers will be published continuously in the journal (as soon as accepted) and will be listed together on the special issue website. Research articles, review articles as well as short communications are invited. For planned papers, a title and short abstract (about 100 words) can be sent to the Editorial Office for announcement on this website.

Submitted manuscripts should not have been published previously, nor be under consideration for publication elsewhere (except conference proceedings papers). All manuscripts are thoroughly refereed through a single-blind peer-review process. A guide for authors and other relevant information for submission of manuscripts is available on the Instructions for Authors page. International Journal of Molecular Sciences is an international peer-reviewed open access semimonthly journal published by MDPI.

Please visit the Instructions for Authors page before submitting a manuscript. There is an Article Processing Charge (APC) for publication in this open access journal. For details about the APC please see here. Submitted papers should be well formatted and use good English. Authors may use MDPI's English editing service prior to publication or during author revisions.

Keywords

  • Exosomes
  • Microvesicles
  • Apoptotic bodies
  • Large oncosomes
  • Extracellular vesicles
  • Biomarkers
  • Liquid biopsy
  • Intercellular communication
  • Cancer

Related Special Issue

Published Papers (7 papers)

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Research

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16 pages, 4830 KiB  
Article
Circulating Extracellular Vesicles: Their Role in Patients with Abdominal Aortic Aneurysm (AAA) Undergoing EndoVascular Aortic Repair (EVAR)
by Francesco Lorenzo Serafini, Andrea Delli Pizzi, Pasquale Simeone, Alberto Giammarino, Cristian Mannetta, Michela Villani, Jacopo Izzi, Davide Buca, Giulia Catitti, Piero Chiacchiaretta, Stefano Trebeschi, Sebastiano Miscia, Massimo Caulo and Paola Lanuti
Int. J. Mol. Sci. 2022, 23(24), 16015; https://0-doi-org.brum.beds.ac.uk/10.3390/ijms232416015 - 16 Dec 2022
Cited by 2 | Viewed by 1270
Abstract
Abdominal aortic aneurysm (AAA) is a frequent aortic disease. If the diameter of the aorta is larger than 5 cm, an open surgical repair (OSR) or an endovascular aortic repair (EVAR) are recommended. To prevent possible complications (i.e., endoleaks), EVAR-treated patients need to [...] Read more.
Abdominal aortic aneurysm (AAA) is a frequent aortic disease. If the diameter of the aorta is larger than 5 cm, an open surgical repair (OSR) or an endovascular aortic repair (EVAR) are recommended. To prevent possible complications (i.e., endoleaks), EVAR-treated patients need to be monitored for 5 years following the intervention, using computed tomography angiography (CTA). However, this radiological method involves high radiation exposure in terms of CTA/year. In such a context, the study of peripheral-blood-circulating extracellular vesicles (pbcEVs) has great potential to identify biomarkers for EVAR complications. We analyzed several phenotypes of pbcEVs using polychromatic flow cytometry in 22 patients with AAA eligible for EVAR. From each enrolled patient, peripheral blood samples were collected at AAA diagnosis, and after 1, 6, and 12 months following EVAR implantation, i.e. during the diagnostic follow-up protocol. Patients developing an endoleak displayed a significant decrease in activated-platelet-derived EVs between the baseline condition and 6 months after EVAR intervention. Furthermore, we also observed, that 1 month after EVAR implantation, patients developing an endoleak showed higher concentrations of activated-endothelial-derived EVs than patients who did not develop one, suggesting their great potential as a noninvasive and specific biomarker for early identification of EVAR complications. Full article
(This article belongs to the Special Issue Extracellular Vesicles as a New Source of Liquid Biopsy 2.0)
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12 pages, 1694 KiB  
Article
Human Serum Extracellular Vesicle Proteomic Profile Depends on the Enrichment Method Employed
by Mikel Azkargorta, Ibon Iloro, Iraide Escobes, Diana Cabrera, Juan M. Falcon-Perez, Felix Elortza and Felix Royo
Int. J. Mol. Sci. 2021, 22(20), 11144; https://0-doi-org.brum.beds.ac.uk/10.3390/ijms222011144 - 15 Oct 2021
Cited by 5 | Viewed by 1681
Abstract
The proteomic profiling of serum samples supposes a challenge due to the large abundance of a few blood proteins in comparison with other circulating proteins coming from different tissues and cells. Although the sensitivity of protein detection has increased enormously in the last [...] Read more.
The proteomic profiling of serum samples supposes a challenge due to the large abundance of a few blood proteins in comparison with other circulating proteins coming from different tissues and cells. Although the sensitivity of protein detection has increased enormously in the last years, specific strategies are still required to enrich less abundant proteins and get rid of abundant proteins such as albumin, lipoproteins, and immunoglobulins. One of the alternatives that has become more promising is to characterize circulating extracellular vesicles from serum samples that have great interest in biomedicine. In the present work, we enriched the extracellular vesicles fraction from human serum by applying different techniques, including ultracentrifugation, size-exclusion chromatography, and two commercial precipitation methods based on different mechanisms of action. To improve the performance and efficacy of the techniques to promote purity of the preparations, we have employed a small volume of serum samples (<100 mL). The comparative proteomic profiling of the enriched preparations shows that ultracentrifugation procedure yielded a larger and completely different set of proteins than other techniques, including mitochondrial and ribosome related proteins. The results showed that size exclusion chromatography carries over lipoprotein associated proteins, while a polymer-based precipitation kit has more affinity for proteins associated with granules of platelets. The precipitation kit that targets glycosylation molecules enriches differentially protein harboring glycosylation sites, including immunoglobulins and proteins of the membrane attack complex. Full article
(This article belongs to the Special Issue Extracellular Vesicles as a New Source of Liquid Biopsy 2.0)
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19 pages, 1986 KiB  
Article
Burn Injury Induces Proinflammatory Plasma Extracellular Vesicles That Associate with Length of Hospital Stay in Women: CRP and SAA1 as Potential Prognostic Indicators
by Robert Maile, Micah L. Willis, Laura E. Herring, Alex Prevatte, Cressida Mahung, Bruce Cairns, Shannon Wallet and Leon G. Coleman, Jr.
Int. J. Mol. Sci. 2021, 22(18), 10083; https://0-doi-org.brum.beds.ac.uk/10.3390/ijms221810083 - 18 Sep 2021
Cited by 9 | Viewed by 2685
Abstract
Severe burn injury is a devastating form of trauma that results in persistent immune dysfunction with associated morbidity and mortality. The underlying drivers of this immune dysfunction remain elusive, and there are no prognostic markers to identify at-risk patients. Extracellular vesicles (EVs) are [...] Read more.
Severe burn injury is a devastating form of trauma that results in persistent immune dysfunction with associated morbidity and mortality. The underlying drivers of this immune dysfunction remain elusive, and there are no prognostic markers to identify at-risk patients. Extracellular vesicles (EVs) are emerging as drivers of immune dysfunction as well as biomarkers. We investigated if EVs after burn injury promote macrophage activation and assessed if EV contents can predict length of hospital stay. EVs isolated early from mice that received a 20% total body surface area (TBSA) burn promoted proinflammatory responses in cultured splenic macrophages. Unbiased LC-MS/MS proteomic analysis of early EVs (<72 h post-injury) from mice and humans showed some similarities including enrichment of acute phase response proteins such as CRP and SAA1. Semi-unbiased assessment of early human burn patient EVs found alterations consistent with increased proinflammatory signaling and loss of inhibition of CRP expression. In a sample of 50 patients with large burn injury, EV SAA1 and CRP were correlated with TBSA injury in both sexes and were correlated with length of hospital stay in women. These findings suggest that EVs are drivers of immune responses after burn injury and their content may predict hospital course. Full article
(This article belongs to the Special Issue Extracellular Vesicles as a New Source of Liquid Biopsy 2.0)
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18 pages, 2297 KiB  
Article
Flow Cytometry Analysis of Circulating Extracellular Vesicle Subtypes from Fresh Peripheral Blood Samples
by Marco Marchisio, Pasquale Simeone, Giuseppina Bologna, Eva Ercolino, Laura Pierdomenico, Damiana Pieragostino, Alessia Ventrella, Francesca Antonini, Genny Del Zotto, Daniele Vergara, Christian Celia, Luisa Di Marzio, Piero Del Boccio, Antonella Fontana, Domenico Bosco, Sebastiano Miscia and Paola Lanuti
Int. J. Mol. Sci. 2021, 22(1), 48; https://0-doi-org.brum.beds.ac.uk/10.3390/ijms22010048 - 23 Dec 2020
Cited by 47 | Viewed by 7382
Abstract
Extracellular vesicles (EVs) are released by shedding during different physiological processes and are increasingly thought to be new potential biomarkers. However, the impact of pre-analytical processing phases on the final measurement is not predictable and for this reason, the translation of basic research [...] Read more.
Extracellular vesicles (EVs) are released by shedding during different physiological processes and are increasingly thought to be new potential biomarkers. However, the impact of pre-analytical processing phases on the final measurement is not predictable and for this reason, the translation of basic research into clinical practice has been precluded. Here we have optimized a simple procedure in combination with polychromatic flow cytometry (PFC), to identify, classify, enumerate, and separate circulating EVs from different cell origins. This protocol takes advantage of a lipophilic cationic dye (LCD) able to probe EVs. Moreover, the application of the newly optimized PFC protocol here described allowed the obtainment of repeatable EVs counts. The translation of this PFC protocol to fluorescence-activated cell sorting allowed us to separate EVs from fresh peripheral blood samples. Sorted EVs preparations resulted particularly suitable for proteomic analyses, which we applied to study their protein cargo. Here we show that LCD staining allowed PFC detection and sorting of EVs from fresh body fluids, avoiding pre-analytical steps of enrichment that could impact final results. Therefore, LCD staining is an essential step towards the assessment of EVs clinical significance. Full article
(This article belongs to the Special Issue Extracellular Vesicles as a New Source of Liquid Biopsy 2.0)
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Review

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12 pages, 1017 KiB  
Review
Extracellular Vesicles as Players in the Anti-Inflammatory Inter-Cellular Crosstalk Induced by Exercise Training
by Giulia Catitti, Domenico De Bellis, Simone Vespa, Pasquale Simeone, Barbara Canonico and Paola Lanuti
Int. J. Mol. Sci. 2022, 23(22), 14098; https://0-doi-org.brum.beds.ac.uk/10.3390/ijms232214098 - 15 Nov 2022
Cited by 4 | Viewed by 2121
Abstract
Extracellular Vesicles (EVs) are circulating particles surrounded by a plasma membrane carrying a cargo consisting of proteins, lipids, RNAs, and DNA fragments, stemming from the cells from which they originated. EV factors (i.e., miRNAs) play relevant roles in intercellular crosstalk, both locally and [...] Read more.
Extracellular Vesicles (EVs) are circulating particles surrounded by a plasma membrane carrying a cargo consisting of proteins, lipids, RNAs, and DNA fragments, stemming from the cells from which they originated. EV factors (i.e., miRNAs) play relevant roles in intercellular crosstalk, both locally and systemically. As EVs increasingly gained attention as potential carriers for targeted genes, the study of EV effects on the host immune response became more relevant. It has been demonstrated that EVs regulate the host immune response, executing both pro- and anti-inflammatory functions. It is also known that physical exercise triggers anti-inflammatory effects. This review underlines the role of circulating EVs as players in the anti-inflammatory events associated with the regulation of the host’s immune response to physical exercise. Full article
(This article belongs to the Special Issue Extracellular Vesicles as a New Source of Liquid Biopsy 2.0)
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30 pages, 908 KiB  
Review
Pregnancy-Related Extracellular Vesicles Revisited
by Carmen Elena Condrat, Valentin Nicolae Varlas, Florentina Duică, Panagiotis Antoniadis, Cezara Alina Danila, Dragos Cretoiu, Nicolae Suciu, Sanda Maria Crețoiu and Silviu Cristian Voinea
Int. J. Mol. Sci. 2021, 22(8), 3904; https://0-doi-org.brum.beds.ac.uk/10.3390/ijms22083904 - 09 Apr 2021
Cited by 26 | Viewed by 4872
Abstract
Extracellular vesicles (EVs) are small vesicles ranging from 20–200 nm to 10 μm in diameter that are discharged and taken in by many different types of cells. Depending on the nature and quantity of their content—which generally includes proteins, lipids as well as [...] Read more.
Extracellular vesicles (EVs) are small vesicles ranging from 20–200 nm to 10 μm in diameter that are discharged and taken in by many different types of cells. Depending on the nature and quantity of their content—which generally includes proteins, lipids as well as microRNAs (miRNAs), messenger-RNA (mRNA), and DNA—these particles can bring about functional modifications in the receiving cells. During pregnancy, placenta and/or fetal-derived EVs have recently been isolated, eliciting interest in discovering their clinical significance. To date, various studies have associated variations in the circulating levels of maternal and fetal EVs and their contents, with complications including gestational diabetes and preeclampsia, ultimately leading to adverse pregnancy outcomes. Furthermore, EVs have also been identified as messengers and important players in viral infections during pregnancy, as well as in various congenital malformations. Their presence can be detected in the maternal blood from the first trimester and their level increases towards term, thus acting as liquid biopsies that give invaluable insight into the status of the feto-placental unit. However, their exact roles in the metabolic and vascular adaptations associated with physiological and pathological pregnancy is still under investigation. Analyzing peer-reviewed journal articles available in online databases, the purpose of this review is to synthesize current knowledge regarding the utility of quantification of pregnancy related EVs in general and placental EVs in particular as non-invasive evidence of placental dysfunction and adverse pregnancy outcomes, and to develop the current understanding of these particles and their applicability in clinical practice. Full article
(This article belongs to the Special Issue Extracellular Vesicles as a New Source of Liquid Biopsy 2.0)
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14 pages, 289 KiB  
Review
MicroRNAs as Biomarkers for Nephrotic Syndrome
by Kenji Tsuji, Shinji Kitamura and Jun Wada
Int. J. Mol. Sci. 2021, 22(1), 88; https://doi.org/10.3390/ijms22010088 - 23 Dec 2020
Cited by 16 | Viewed by 3050
Abstract
Nephrotic syndrome represents the clinical situation characterized by presence of massive proteinuria and low serum protein caused by a variety of diseases, including minimal change nephrotic syndrome (MCNS), focal segmental glomerulosclerosis (FSGS) and membranous glomerulonephropathy. Differentiating between diagnoses requires invasive renal biopsies in [...] Read more.
Nephrotic syndrome represents the clinical situation characterized by presence of massive proteinuria and low serum protein caused by a variety of diseases, including minimal change nephrotic syndrome (MCNS), focal segmental glomerulosclerosis (FSGS) and membranous glomerulonephropathy. Differentiating between diagnoses requires invasive renal biopsies in general. Even with the biopsy, we encounter difficulties to differentiate MCNS and FSGS in some cases. There is no other better option currently available for the diagnosis other than renal biopsy. MicroRNAs (miRNAs) are no-coding RNAs of approximately 20 nucleotides in length, which regulate target genes in the post-transcriptional processes and have essential roles in many diseases. MiRNAs in serum and urine have been shown as non-invasive biomarkers in multiple diseases, including renal diseases. In this article, we summarize the current knowledge of miRNAs as the promising biomarkers for nephrotic syndrome. Full article
(This article belongs to the Special Issue Extracellular Vesicles as a New Source of Liquid Biopsy 2.0)
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