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New Insights into Extracellular Vesicles in Health and Disease
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New Insights into Extracellular Vesicles in Health and Disease

A special issue of Life (ISSN 2075-1729). This special issue belongs to the section "Pharmaceutical Science".

Deadline for manuscript submissions: closed (31 October 2023) | Viewed by 21201

Special Issue Editor

Dr. Juan Pablo Rigalli

E-Mail Website
Guest Editor
Department of Clinical Pharmacology and Pharmacoepidemiology, Heidelberg University Hospital, Heidelberg, Germany
Interests: extracellular vesicles; drug transporters; drug–drug interactions; purinergic signaling; nuclear receptors

Special Issue Information

Dear Colleagues,

Extracellular vesicles (EVs) are nanoparticles released by most types of cells. In the past two decades, EVs were demonstrated to mediate the transfer of information between cells. In this way, EVs participate in different physiological processes such as antigen presentation, tissue regeneration, gut–brain communication and the regulation of the electrolyte balance, among others. In addition, participation of EVs in pathophysiological processes such as cancer, atherosclerosis, inflammation, diabetes, polycystic kidney disease and microbial and viral infections, among other disorders, has also been established. In the past few years, strong evidence supporting the role of EVs in the pathophysiological mechanisms underlying the infection of SARS-CoV-2 has also been obtained. All these recently elucidated molecular and cellular mechanisms confer EVs as a significant therapeutic potential. Moreover, due to their lower immunogenicity and increased tissue-specificity with respect to other nanoparticles, the use of EVs as carriers for therapeutic agents is being investigated. Last, but not least, EVs can be easily isolated from biofluids such as the blood and the urine. This, together with the fact that the constitution of EVs partially resembles the proteome and the transcriptome of the cells of origin, makes them a promising source of biomarkers. This Special Issue will address the participation of EVs in physiological and pathophysiological processes, their application as a source of biomarkers and the pharmacological and pharmaceutical relevance of EVs, both as druggable targets and drug carriers.

Dr. Juan Pablo Rigalli
Guest Editor

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Keywords

  • extracellular vesicles
  • exosomes
  • microvesicles
  • nanovesicles
  • cell–cell communication
  • EVs

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Published Papers (10 papers)

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Research

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13 pages, 6706 KiB  
Article
Resistin Induces Migration and Invasion in PC3 Prostate Cancer Cells: Role of Extracellular Vesicles
by Mario Israel Oregel-Cortez, Héctor Frayde-Gómez, Georgina Quintana-González, Victor García-González, Jose Gustavo Vazquez-Jimenez and Octavio Galindo-Hernández
Life 2023, 13(12), 2321; https://doi.org/10.3390/life13122321 - 10 Dec 2023
Cited by 4 | Viewed by 2333
Abstract
Resistin is an adipokine with metabolic and inflammatory functions. Epidemiological and translational studies report that an increase in plasma levels and tissue expression of resistin increases the aggressiveness of prostate tumor cells. Extracellular vesicles (EVs) are secreted constitutively and induced by cytokines, growth [...] Read more.
Resistin is an adipokine with metabolic and inflammatory functions. Epidemiological and translational studies report that an increase in plasma levels and tissue expression of resistin increases the aggressiveness of prostate tumor cells. Extracellular vesicles (EVs) are secreted constitutively and induced by cytokines, growth factors, and calcium and are found in multiple biological fluids such as saliva, serum, semen, and urine. In particular, EVs have been shown to promote tumor progression through the induction of proliferation, growth, angiogenesis, resistance to chemotherapy, and metastasis. However, the role of resistin in the migration, invasion, and secretion of EVs in invasive prostate tumor cells remains to be studied. In the present study, we demonstrate that resistin induces increased migration and invasion in PC3 cells. In addition, these phenomena are accompanied by increased p-FAK levels and increased secretion of MMP-2 and MMP-9 in resistin-treated PC3 cells. Interestingly, EVs isolated from supernatants of PC3 cells treated with resistin induce an increase in migration and invasion accompanied by high MMP-2 and MMP-9 secretion in an autocrine stimulation model. In summary, our data for the first time demonstrate that resistin induces migration and invasion, partly through the secretion of EVs with pro-invasive characteristics in PC3 cells. Full article
(This article belongs to the Special Issue New Insights into Extracellular Vesicles in Health and Disease)
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21 pages, 1882 KiB  
Article
A Comparative Analysis of the Protein Cargo of Extracellular Vesicles from Helminth Parasites
by María Eugenia Ancarola, Lucas L. Maldonado, Lucía C. A. García, Gisela R. Franchini, Gustavo Mourglia-Ettlin, Laura Kamenetzky and Marcela A. Cucher
Life 2023, 13(12), 2286; https://doi.org/10.3390/life13122286 - 30 Nov 2023
Cited by 1 | Viewed by 1902
Abstract
Helminth parasites cause debilitating—sometimes fatal—diseases in humans and animals. Despite their impact on global health, mechanisms underlying host–parasite interactions are still poorly understood. One such mechanism involves the exchange of extracellular vesicles (EVs), which are membrane-enclosed subcellular nanoparticles. To date, EV secretion has [...] Read more.
Helminth parasites cause debilitating—sometimes fatal—diseases in humans and animals. Despite their impact on global health, mechanisms underlying host–parasite interactions are still poorly understood. One such mechanism involves the exchange of extracellular vesicles (EVs), which are membrane-enclosed subcellular nanoparticles. To date, EV secretion has been studied in helminth parasites, including EV protein content. However, information is highly heterogeneous, since it was generated in multiple species, using varied protocols for EV isolation and data analysis. Here, we compared the protein cargo of helminth EVs to identify common markers for each taxon. For this, we integrated published proteomic data and performed a comparative analysis through an orthology approach. Overall, only three proteins were common in the EVs of the seven analyzed species. Additionally, varied repertoires of proteins with moonlighting activity, vaccine antigens, canonical and non-canonical proteins related to EV biogenesis, taxon-specific proteins of unknown function and RNA-binding proteins were observed in platyhelminth and nematode EVs. Despite the lack of consensus on EV isolation protocols and protein annotation, several proteins were shown to be consistently detected in EV preparations from organisms at different taxa levels, providing a starting point for a selective biochemical characterization. Full article
(This article belongs to the Special Issue New Insights into Extracellular Vesicles in Health and Disease)
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14 pages, 4509 KiB  
Article
Extracellular Vesicles of Porphyromonas gingivalis Disrupt Trophoblast Cell Interaction with Vascular and Immune Cells in an In Vitro Model of Early Placentation
by Brenda Lara, Matías Sassot, Guillermina Calo, Daniel Paparini, Laura Gliosca, Gabriela Chaufan, Iñaki Loureiro, Daiana Vota, Rosanna Ramhorst, Claudia Pérez Leirós and Vanesa Hauk
Life 2023, 13(10), 1971; https://doi.org/10.3390/life13101971 - 27 Sep 2023
Cited by 2 | Viewed by 1644
Abstract
Extracellular vesicles released by the primary pathogen of periodontal disease Porphyromonas gingivalis (Pg), referred to as outer membrane vesicles (OMVs), have been associated with the pathogenesis of systemic diseases like cardiovascular disease, rheumatoid arthritis, and Alzheimer’s disease. A pathogenic role for [...] Read more.
Extracellular vesicles released by the primary pathogen of periodontal disease Porphyromonas gingivalis (Pg), referred to as outer membrane vesicles (OMVs), have been associated with the pathogenesis of systemic diseases like cardiovascular disease, rheumatoid arthritis, and Alzheimer’s disease. A pathogenic role for Pg by disrupting placental homeostasis was proposed in the association between periodontal disease and adverse pregnancy outcomes. On the basis that trophoblast-derived factors modulate endothelial and immune cell profiles in normal pregnancy and the scarce presence of Pg in placenta, we hypothesized that OMVs from Pg affect trophoblast cell phenotype, impairing trophoblast–endothelium and trophoblast–neutrophil interactions. By means of in vitro designs with first-trimester human trophoblast cells, endothelial cells, and freshly isolated neutrophils, we showed that Pg OMVs are internalized by trophoblast cells and modulate the activity and expression of functional markers. Trophoblast cells primed with Pg OMVs enhanced neutrophil chemoattraction and lost their anti-inflammatory effect. In addition, reduced migration with enhanced adhesion of monocytes was found in endothelial cells upon incubation with the media from trophoblast cells pretreated with Pg OMVs. Taken together, the results support a pathogenic role of Pg OMVs at early stages of pregnancy and placentation through disruption of trophoblast contribution to vascular transformation and immune homeostasis maintenance. Full article
(This article belongs to the Special Issue New Insights into Extracellular Vesicles in Health and Disease)
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15 pages, 3087 KiB  
Article
Total and Extracellular Vesicle cAMP Contents in Urine Are Associated with Autosomal Dominant Polycystic Kidney Disease (ADPKD) Progression
by María Lucía Rosenberg, Agustín Yaneff, Gonzalo Manuel Ferradás, Margarita Paz Villafañe Tapia, Carlos Alberto Davio, Nora Paula Goette, Sandra Gabriela Vlachovsky, Roxana Noemí Peroni, Elisabet Mónica Oddo and Pablo Javier Azurmendi
Life 2023, 13(9), 1817; https://doi.org/10.3390/life13091817 - 28 Aug 2023
Cited by 2 | Viewed by 2189
Abstract
ADPKD is the most common genetic renal disease, characterized by the presence of multiple cysts which, through slow and gradual growth, lead to glomerular filtration rate (GFR) decline and end-stage renal disease. Cystic growth is associated with increased intracellular levels of 3′,5′-cyclic adenosine [...] Read more.
ADPKD is the most common genetic renal disease, characterized by the presence of multiple cysts which, through slow and gradual growth, lead to glomerular filtration rate (GFR) decline and end-stage renal disease. Cystic growth is associated with increased intracellular levels of 3′,5′-cyclic adenosine monophosphate (cAMP). Extracellular vesicles (EVs) are proposed to participate in “remote sensing” by transporting different cargoes, but their relevance to ADPKD progression is poorly understood. This study aimed to determine whether cAMP is contained in urinary EVs and, if so, how total and/or EV cAMP contents participate in disease progression. Fourteen ADPKD patients, naïve for V2 receptor antagonism treatment, and seven controls were studied. Progression was evaluated by estimating GFR (eGFR) and height-adjusted total kidney volume (htTKV). Fresh morning urine was collected to determine cAMP by the competitive radioligand assay. Urine EVs were isolated using an adapted centrifugation method and characterized by electron microscopy, dynamic light scanning, flow cytometry with FITC CD63 labeling, protein and RNA content, and AQP2 and GAPDH mRNA detection. Total and EV cAMP was measurable in both control and patient urine samples. Total cAMP was significantly correlated with eGFR and its annual change but inversely correlated with htTKV. The cAMP-EVs showed a bimodal pattern with htTKV, increasing to ~1 L/m and falling at larger sizes. Our results demonstrate that urine cAMP correlates with ADPKD progression markers, and that its extracellular delivery by EVs could reflect the architectural disturbances of the organ. Full article
(This article belongs to the Special Issue New Insights into Extracellular Vesicles in Health and Disease)
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15 pages, 2609 KiB  
Article
Diagnosis of Prostate Cancer through the Multi-Ligand Binding of Prostate-Derived Extracellular Vesicles and miRNA Analysis
by Lidia Zabegina, Ilya Zyatchin, Margarita Kniazeva, Andrey Shalaev, Maria Berkut, Vladimir Sharoyko, Vladimir Mikhailovskii, Kirill Kondratov, Sergey Reva, Alexandr Nosov and Anastasia Malek
Life 2023, 13(4), 885; https://doi.org/10.3390/life13040885 - 27 Mar 2023
Cited by 3 | Viewed by 2000
Abstract
Background: The development of new non-invasive markers for prostate cancer (PC) diagnosis, prognosis, and management is an important issue that needs to be addressed to decrease PC mortality. Small extracellular vesicles (SEVs) secreted by prostate gland or prostate cancer cells into the plasma [...] Read more.
Background: The development of new non-invasive markers for prostate cancer (PC) diagnosis, prognosis, and management is an important issue that needs to be addressed to decrease PC mortality. Small extracellular vesicles (SEVs) secreted by prostate gland or prostate cancer cells into the plasma are considered next-generation diagnostic tools because their chemical composition might reflect the PC development. The population of plasma vesicles is extremely heterogeneous. The study aimed to explore a new approach for prostate-derived SEV isolation followed by vesicular miRNA analysis. Methods: We used superparamagnetic particles functionalized by five types of DNA-aptamers binding the surface markers of prostate cells. Specificity of binding was assayed by AuNP-aptasensor. Prostate-derived SEVs were isolated from the plasma of 36 PC patients and 18 healthy donors and used for the assessment of twelve PC-associated miRNAs. The amplification ratio (amp-ratio) value was obtained for all pairs of miRNAs, and the diagnostic significance of these parameters was evaluated. Results: The multi-ligand binding approach doubled the efficiency of prostate-derived SEVs’ isolation and made it possible to purify a sufficient amount of vesicular RNA. The neighbor clusterization, using three pairs of microRNAs (miR-205/miR-375, miR-26b/miR375, and miR-20a/miR-375), allowed us to distinguish PC patients and donors with sensitivity—94%, specificity—76%, and accuracy—87%. Moreover, the amp-ratios of other miRNAs pairs reflected such parameters as plasma PSA level, prostate volume, and Gleason score of PC. Conclusions: Multi-ligand isolation of prostate-derived vesicles followed by vesicular miRNA analysis is a promising method for PC diagnosis and monitoring. Full article
(This article belongs to the Special Issue New Insights into Extracellular Vesicles in Health and Disease)
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Review

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30 pages, 2850 KiB  
Review
Exosomes in Cancer Progression and Therapy Resistance: Molecular Insights and Therapeutic Opportunities
by Madita Wandrey, Jadwiga Jablonska, Roland H. Stauber and Désirée Gül
Life 2023, 13(10), 2033; https://doi.org/10.3390/life13102033 - 9 Oct 2023
Cited by 13 | Viewed by 3178
Abstract
The development of therapy resistance still represents a major hurdle in treating cancers, leading to impaired treatment success and increased patient morbidity. The establishment of minimally invasive liquid biopsies is a promising approach to improving the early diagnosis, as well as therapy monitoring, [...] Read more.
The development of therapy resistance still represents a major hurdle in treating cancers, leading to impaired treatment success and increased patient morbidity. The establishment of minimally invasive liquid biopsies is a promising approach to improving the early diagnosis, as well as therapy monitoring, of solid tumors. Because of their manifold functions in the tumor microenvironment, tumor-associated small extracellular vesicles, referred to as exosomes, have become a subject of intense research. Besides their important roles in cancer progression, metastasis, and the immune response, it has been proposed that exosomes also contribute to the acquisition and transfer of therapy resistance, mainly by delivering functional proteins and RNAs, as well as facilitating the export of active drugs or functioning as extracellular decoys. Extensive research has focused on understanding the molecular mechanisms underlying the occurrence of resistance and translating these into strategies for early detection. With this review, we want to provide an overview of the current knowledge about the (patho-)biology of exosomes, as well as state-of-the-art methods of isolation and analysis. Furthermore, we highlight the role of exosomes in tumorigenesis and cancer treatment, where they can function as therapeutic agents, biomarkers, and/or targets. By focusing on their roles in therapy resistance, we will reveal new paths of exploiting exosomes for cancer diagnosis and treatment. Full article
(This article belongs to the Special Issue New Insights into Extracellular Vesicles in Health and Disease)
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17 pages, 5249 KiB  
Review
A Review Study of the Participation of Late Domains in Sorting and Transport of Viral Factors to Exosomes
by Manuel Adrián Velázquez-Cervantes, Yazmín Rocío Benítez-Zeferino, Arturo Flores-Pliego, Addy Cecilia Helguera-Repetto, David Eduardo Meza-Sánchez, José Luis Maravillas-Montero, Guadalupe León-Reyes, Javier Mancilla-Ramírez, Jorge Francisco Cerna-Cortés, María Isabel Baeza-Ramírez and Moises León-Juaárez
Life 2023, 13(9), 1842; https://doi.org/10.3390/life13091842 - 31 Aug 2023
Cited by 1 | Viewed by 1311
Abstract
Cellular communication depends heavily on the participation of vesicular systems generated by most cells of an organism. Exosomes play central roles in this process. Today, these vesicles have been characterized, and it has been determined that the cargo they transport is not within [...] Read more.
Cellular communication depends heavily on the participation of vesicular systems generated by most cells of an organism. Exosomes play central roles in this process. Today, these vesicles have been characterized, and it has been determined that the cargo they transport is not within a random system. In fact, it depends on various molecular signals and the recruitment of proteins that participate in the biogenesis of exosomes. It has also been shown that multiple viruses can recruit these vesicles to transport viral factors such as genomes or proteins. It has been shown that the late domains present in viral proteins are critical for the exosomal selection and biogenesis systems to recognize these viral proteins and introduce them into the exosomes. In this review, the researchers discuss the evidence related to the characterization of these late domains and their role in exosome recruitment during viral infection. Full article
(This article belongs to the Special Issue New Insights into Extracellular Vesicles in Health and Disease)
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22 pages, 1361 KiB  
Review
Extracellular Vesicles as Surrogates for Drug Metabolism and Clearance: Promise vs. Reality
by Anna Gagliardi, Gzona Bajraktari-Sylejmani, Elisabetta Barocelli, Johanna Weiss and Juan Pablo Rigalli
Life 2023, 13(8), 1745; https://doi.org/10.3390/life13081745 - 14 Aug 2023
Cited by 3 | Viewed by 1969
Abstract
Drug-metabolizing enzymes (DMEs) and transporters play a major role in drug efficacy and safety. They are regulated at multiple levels and by multiple factors. Estimating their expression and activity could contribute to predicting drug pharmacokinetics and their regulation by drugs or pathophysiological situations. [...] Read more.
Drug-metabolizing enzymes (DMEs) and transporters play a major role in drug efficacy and safety. They are regulated at multiple levels and by multiple factors. Estimating their expression and activity could contribute to predicting drug pharmacokinetics and their regulation by drugs or pathophysiological situations. Determining the expression of these proteins in the liver, intestine, and kidney requires the collection of biopsy specimens. Instead, the isolation of extracellular vesicles (EVs), which are nanovesicles released by most cells and present in biological fluids, could deliver this information in a less invasive way. In this article, we review the use of EVs as surrogates for the expression and activity of DMEs, uptake, and efflux transporters. Preliminary evidence has been provided for a correlation between the expression of some enzymes and transporters in EVs and the tissue of origin. In some cases, data obtained in EVs reflect the induction of phase I-DMEs in the tissues. Further studies are required to elucidate to what extent the regulation of other DMEs and transporters in the tissues reflects in the EV cargo. If an association between tissues and their EVs is firmly established, EVs may represent a significant advancement toward precision therapy based on the biotransformation and excretion capacity of each individual. Full article
(This article belongs to the Special Issue New Insights into Extracellular Vesicles in Health and Disease)
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33 pages, 1277 KiB  
Review
Extracellular Vesicles and Cancer Multidrug Resistance: Undesirable Intercellular Messengers?
by María Bucci-Muñoz, Aldana Magalí Gola, Juan Pablo Rigalli, María Paula Ceballos and María Laura Ruiz
Life 2023, 13(8), 1633; https://doi.org/10.3390/life13081633 - 27 Jul 2023
Cited by 3 | Viewed by 2066
Abstract
Cancer multidrug resistance (MDR) is one of the main mechanisms contributing to therapy failure and mortality. Overexpression of drug transporters of the ABC family (ATP-binding cassette) is a major cause of MDR. Extracellular vesicles (EVs) are nanoparticles released by most cells of the [...] Read more.
Cancer multidrug resistance (MDR) is one of the main mechanisms contributing to therapy failure and mortality. Overexpression of drug transporters of the ABC family (ATP-binding cassette) is a major cause of MDR. Extracellular vesicles (EVs) are nanoparticles released by most cells of the organism involved in cell–cell communication. Their cargo mainly comprises, proteins, nucleic acids, and lipids, which are transferred from a donor cell to a target cell and lead to phenotypical changes. In this article, we review the scientific evidence addressing the regulation of ABC transporters by EV-mediated cell–cell communication. MDR transfer from drug-resistant to drug-sensitive cells has been identified in several tumor entities. This was attributed, in some cases, to the direct shuttle of transporter molecules or its coding mRNA between cells. Also, EV-mediated transport of regulatory proteins (e.g., transcription factors) and noncoding RNAs have been indicated to induce MDR. Conversely, the transfer of a drug-sensitive phenotype via EVs has also been reported. Additionally, interactions between non-tumor cells and the tumor cells with an impact on MDR are presented. Finally, we highlight uninvestigated aspects and possible approaches to exploiting this knowledge toward the identification of druggable processes and molecules and, ultimately, the development of novel therapeutic strategies. Full article
(This article belongs to the Special Issue New Insights into Extracellular Vesicles in Health and Disease)
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Other

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10 pages, 1458 KiB  
Brief Report
An Improved Method to Enrich Large Extracellular Vesicles Derived from Giardia intestinalis through Differential Centrifugation
by Abel Sana, Izadora Volpato Rossi, Bruna Sabatke, Letícia Bassani Bonato, Lia Carolina Soares Medeiros and Marcel Ivan Ramirez
Life 2023, 13(9), 1799; https://doi.org/10.3390/life13091799 - 24 Aug 2023
Cited by 2 | Viewed by 1608
Abstract
Giardia intestinalis is a flagellated unicellular protozoan that colonizes the small intestine, causing the diarrheal disease called giardiasis. The production of extracellular vesicles (EVs) by G. intestinalis and the role of these EVs in the parasite’s interaction with the host have been described. [...] Read more.
Giardia intestinalis is a flagellated unicellular protozoan that colonizes the small intestine, causing the diarrheal disease called giardiasis. The production of extracellular vesicles (EVs) by G. intestinalis and the role of these EVs in the parasite’s interaction with the host have been described. According to biogenesis, EVs are grouped mainly into large (microvesicles—derived from the plasma membrane) and small (exosomes—derived from multivesicular bodies). Populations of EVs are heterogeneous, and improved methods to separate and study them are needed to understand their roles in cell physiology and pathologies. This work aimed to enrich the large extracellular vesicles (LEVs) of G. intestinalis in order to better understand the roles of these vesicles in the interaction of the parasite with the host. To achieve the enrichment of the LEVs, we have modified our previously described method and compared it by protein dosage and using Nano tracking analysis. Giardia intestinalis vesiculation was induced by incubation in a TYI-S-33 medium without serum, to which 1 mM of CaCl2 was added at 37 °C for 1 h. Then, the supernatant was centrifuged at 15,000× g for 1 h (15 K 1 h pellet), 15,000× g for 4 h (15 K 4 h pellet) and 100,000× g for 1.5 h (100 K 1h30 pellet). The pellet (containing EVs) was resuspended in 1× PBS and stored at 4 °C for later analysis. The EVs were quantified based on their protein concentrations using the Pierce BCA assay, and by nanoparticle tracking analysis (NTA), which reports the concentration and size distribution of the particles. The NTA showed that direct ultracentrifugation at 100,000× g for 1.5 h and centrifugation at 15,000× g for 4 h concentrated more EVs compared to centrifugation at 15,000× g for 1 h. Additionally, it revealed that centrifugation at 15,000× g 4 h was able to concentrate at the same particle concentration levels as a direct ultracentrifugation at 100,000× g for 1.5 h. As for the enrichment of LEVs, the NTA has shown a higher concentration of LEVs in direct ultracentrifugation at 100,000× g for 1.5 h, and in centrifugation at 15,000× g for 4 h, compared to centrifugation at 15,000× g for 1 h. Our results have shown that the most used method at 15,000× g for 1 h is not enough to obtain a representative population of large EVs, and we suggest that LEVs released by G. intestinalis can be better enriched by direct ultracentrifugation at 100,000× g for 1.5 h, or by centrifugation at 15,000× g for 4 h. Full article
(This article belongs to the Special Issue New Insights into Extracellular Vesicles in Health and Disease)
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