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Exosomes and Extracellular Vesicles in Health and Diseases 2.0

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A special issue of International Journal of Molecular Sciences (ISSN 1422-0067). This special issue belongs to the section "Molecular Pathology, Diagnostics, and Therapeutics".

Deadline for manuscript submissions: closed (30 January 2024) | Viewed by 20974

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Dr. Francesca Beretti

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Department of Biomedical, Metabolic and Neural Sciences, University of Modena and Reggio Emilia, 41125 Modena, Italy
Interests: exosomes; extracellular vesicles; microRNA; amnion fluid
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Dr. Manuela Zavatti

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Department of Biomedical, Metabolic and Neural Sciences, University of Modena and Reggio Emilia, 41125 Modena, Italy
Interests: molecular mechanism in cancer; PI3K/Akt signaling; adaptive resistance to Akt inhibitors in prostate cancer; immunomodulatory properties of stem cells extracellular vesicle; neurodegenerative disorders
Special Issues, Collections and Topics in MDPI journals

Special Issue Information

Dear Colleagues,

The rising interest in exosomes and extracellular vesicles and their applications is well-documented by the increasing level of research in these fields.

These vesicles are naturally produced by any type of cell and play multiple roles in cell-to-cell communication in both healthy and diseased states. Because they carry different donor-derived cargos, including DNA, RNA, proteins and lipids, they are pivotal for inducing network signals in recipient cells.

In particular, stem-cell-derived extracellular vesicles play roles in regenerative medicine and have the advantage of being a cell-free therapy; therefore, these vesicles are safer than those associated with the transplantation of live cells.

Furthermore, in recent years, exosomes and extracellular vesicles have emerged as promising biomarkers in the diagnosis and prognosis of different diseases, but also as macromolecule delivery carriers in the therapy of a wide spectrum of pathologies.

This Special Issue, entitled “Exosomes and Extracellular Vesicles in Health and Diseases”, welcomes original research articles that contribute to broadening our knowledge of extracellular vesicles and exosomes in different fields of application, starting from the healthy state in order to comprehend mechanisms of individual production, and moving to pathological conditions, such as tumors, neurodegenerative diseases, cardiovascular diseases, chronic and acute inflammation state, etc.

Dr. Francesca Beretti
Dr. Manuela Zavatti
Guest Editors

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Keywords

extracellular vesicles
exosome
biomarkers
cell-to-cell communication
signal transduction
diseases
therapy
regenerative medicine
cargo

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Related Special Issues

Exosomes and Extracellular Vesicles in Health and Diseases in International Journal of Molecular Sciences (9 articles)
Exosomes and Extracellular Vesicles in Health and Diseases 3.0 in International Journal of Molecular Sciences (2 articles)

Published Papers (11 papers)

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Editorial

Jump to: Research, Review, Other

5 pages, 183 KiB

Open AccessEditorial

Special Issue “Exosomes and Extracellular Vesicles in Health and Diseases 2.0”

by Francesca Beretti and Manuela Zavatti

Int. J. Mol. Sci. 2024, 25(22), 12447; https://doi.org/10.3390/ijms252212447 - 20 Nov 2024

Viewed by 216

Abstract

All cells secrete various types of membrane vesicles, known as extracellular vesicles (EVs), and this process is conserved throughout evolution [...] Full article

(This article belongs to the Special Issue Exosomes and Extracellular Vesicles in Health and Diseases 2.0)

Research

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19 pages, 5759 KiB

Open AccessArticle

Autophagy Promotes Enrichment of Raft Components within Extracellular Vesicles Secreted by Human 2FTGH Cells

by Valeria Manganelli, Luciana Dini, Stefano Tacconi, Simone Dinarelli, Antonella Capozzi, Gloria Riitano, Serena Recalchi, Tuba Rana Caglar, Federica Fratini, Roberta Misasi, Maurizio Sorice and Tina Garofalo

Int. J. Mol. Sci. 2024, 25(11), 6175; https://doi.org/10.3390/ijms25116175 - 4 Jun 2024

Viewed by 1170

Abstract

Autophagy plays a key role in removing protein aggregates and damaged organelles. In addition to its conventional degradative functions, autophagy machinery contributes to the release of cytosolic proteins through an unconventional secretion pathway. In this research, we analyzed autophagy-induced extracellular vesicles (EVs) in HT1080-derived human fibrosarcoma 2FTGH cells using transmission electron microscopy and atomic force microscopy (AFM). We preliminary observed that autophagy induces the formation of a subset of large heterogeneous intracellular vesicular structures. Moreover, AFM showed that autophagy triggering led to a more visible smooth cell surface with a reduced amount of plasma membrane protrusions. Next, we characterized EVs secreted by cells following autophagy induction, demonstrating that cells release both plasma membrane-derived microvesicles and exosomes. A self-forming iodixanol gradient was performed for cell subfractionation. Western blot analysis showed that endogenous LC3-II co-fractionated with CD63 and CD81. Then, we analyzed whether raft components are enriched within EV cargoes following autophagy triggering. We observed that the raft marker GD3 and ER marker ERLIN1 co-fractionated with LC3-II; dual staining by immunogold electron microscopy and coimmunoprecipitation revealed GD3-LC3-II association, indicating that autophagy promotes enrichment of raft components within EVs. Introducing a new brick in the crosstalk between autophagy and the endolysosomal system may have important implications for the knowledge of pathogenic mechanisms, suggesting alternative raft target therapies in diseases in which the generation of EV is active. Full article

(This article belongs to the Special Issue Exosomes and Extracellular Vesicles in Health and Diseases 2.0)

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19 pages, 4486 KiB

Open AccessArticle

Extracellular Vesicles Generated by Mesenchymal Stem Cells in Stirred Suspension Bioreactors Promote Angiogenesis in Human-Brain-Derived Endothelial Cells

by Jolene Phelps, David A. Hart, Alim P. Mitha, Neil A. Duncan and Arindom Sen

Int. J. Mol. Sci. 2024, 25(10), 5219; https://doi.org/10.3390/ijms25105219 - 10 May 2024

Viewed by 1229

Abstract

Interrupted blood flow in the brain due to ischemic injuries such as ischemic stroke or traumatic brain injury results in irreversible brain damage, leading to cognitive impairment associated with inflammation, disruption of the blood–brain barrier (BBB), and cell death. Since the BBB only allows entry to a small class of drugs, many drugs used to treat ischemia in other tissues have failed in brain-related disorders. The administration of mesenchymal stem cell (MSC)-derived extracellular vesicles (EVs) has shown promise in improving the functional recovery of the brain following cerebral ischemia by inducing blood vessel formation. To facilitate such a treatment approach, it is necessary to develop bioprocesses that can produce therapeutically relevant MSC-EVs in a reproducible and scalable manner. This study evaluated the feasibility of using stirred suspension bioreactors (SSBs) to scale-up the serum-free production of pro-angiogenic MSC-EVs under clinically relevant physioxic conditions. It was found that MSCs grown in SSBs generated EVs that stimulated angiogenesis in cerebral microvascular endothelial cells, supporting the use of SSBs to produce MSC-EVs for application in cerebral ischemia. These properties were impaired at higher cell confluency, outlining the importance of considering the time of harvest when developing bioprocesses to manufacture EV populations. Full article

(This article belongs to the Special Issue Exosomes and Extracellular Vesicles in Health and Diseases 2.0)

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Graphical abstract

21 pages, 5382 KiB

Open AccessArticle

Proteomic Analysis of Salivary Extracellular Vesicles from COVID-19 Patients Reveals a Specific Anti-COVID-19 Response Protein Signature

by Laura Weber, Alfredo Torres, Ornella Realini, María José Bendek, María Luisa Mizgier, Claudia Brizuela, David Herrera, Fermín E. González and Alejandra Chaparro

Int. J. Mol. Sci. 2024, 25(7), 3704; https://doi.org/10.3390/ijms25073704 - 26 Mar 2024

Cited by 1 | Viewed by 1708

Abstract

Despite the understanding of the coronavirus disease-19 (COVID-19), the role of salivary extracellular vesicles (sEVs) in COVID-19 remains unclear. Exploring the proteomic cargo of sEVs could prove valuable for diagnostic and prognostic purposes in assessing COVID-19. The proteomic cargo of sEVs from COVID-19(+) subjects and their healthy close contacts (HCC) was explored. sEVs were isolated by ultracentrifugation from unstimulated saliva samples, and subsequently characterized through nanoparticle tracking, transmission electron microscopy, and Western blot analyses. The proteomic cargo of sEVs was processed by LC-MS/MS. sEVs were morphologically compatible with EVs, with the presence of Syntenin-1 and CD81 EV markers. The sEV pellet showed 1417 proteins: 1288 in COVID-19(+) cases and 1382 in HCC. In total, 124 proteins were differentially expressed in sEVs from COVID-19(+) subjects. “Coronavirus-disease response”, “complement and coagulation cascades”, and “PMN extracellular trap formation” were the most enriched KEGG pathways in COVID-19(+) cases. The most represented biological processes were “Hemoglobin and haptoglobin binding” and “oxygen carrier activity”, and the best-denoted molecular functions were “regulated exocytosis and secretion” and “leucocyte and PMN mediated immunity”. sEV proteomic cargo in COVID-19(+) suggests activity related to immune response processes, oxygen transport, and antioxidant mechanisms. In contrast, in HCC, sEV signature profiles are mainly associated with epithelial homeostasis. Full article

(This article belongs to the Special Issue Exosomes and Extracellular Vesicles in Health and Diseases 2.0)

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16 pages, 9828 KiB

Open AccessArticle

Enhanced Therapeutic Potential of Hybrid Exosomes Loaded with Paclitaxel for Cancer Therapy

by Xuan Wang, Dongdong Li, Gaotian Li, Jinda Chen, Yi Yang, Lijun Bian, Jingying Zhou, Yongge Wu and Yan Chen

Int. J. Mol. Sci. 2024, 25(7), 3645; https://doi.org/10.3390/ijms25073645 - 25 Mar 2024

Cited by 4 | Viewed by 2359

Abstract

The advancement of exosome studies has positioned engineered exosomes as crucial biomaterials for the development of advanced drug delivery systems. This study focuses on developing a hybrid exosome system by fusing mesenchymal stem cells (MSCs) exosomes with folate-targeted liposomes. The aim was to improve the drug loading capacity and target modification of exosome nanocarriers for delivering the first-line chemotherapy drug paclitaxel (PTX) and its effectiveness was assessed through cellular uptake studies to evaluate its ability to deliver drugs to tumor cells in vitro. Additionally, in vivo experiments were conducted using a CT26 tumor-bearing mouse model to assess the therapeutic efficacy of hybrid exosomes loaded with PTX (ELP). Cellular uptake studies demonstrated that ELP exhibited superior drug delivery capabilities to tumor cells in vitro. Moreover, in vivo experiments revealed that ELP significantly suppressed tumor growth in the CT26 tumor-bearing mouse model. Notably, for the first time, we examined the tumor microenvironment following intratumoral administration of ELP. We observed that ELP treatment activated CD4⁺ and CD8⁺ T cells, reduced the expression of M2 type tumor-associated macrophages (TAMs), polarized TAMs towards the M1 type, and decreased regulatory T cells (Tregs). Our research highlights the considerable therapeutic efficacy of ELP and its promising potential for future application in cancer therapy. The development of hybrid exosomes presents an innovative approach to enhance drug delivery and modulate the tumor microenvironment, offering exciting prospects for effective cancer treatment strategies. Full article

(This article belongs to the Special Issue Exosomes and Extracellular Vesicles in Health and Diseases 2.0)

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16 pages, 2249 KiB

Open AccessArticle

Long Interspersed Nuclear Element-1 Analytes in Extracellular Vesicles as Tools for Molecular Diagnostics of Non-Small Cell Lung Cancer

by Emma C. Bowers, Alexandre M. Cavalcante, Kimberly Nguyen, Can Li, Yingshan Wang, Randa El-Zein, Shu-Hsia Chen, Min P. Kim, Brian S. McKay and Kenneth S. Ramos

Int. J. Mol. Sci. 2024, 25(2), 1169; https://doi.org/10.3390/ijms25021169 - 18 Jan 2024

Cited by 1 | Viewed by 1507

Abstract

Aberrant expression of the oncogenic retrotransposon LINE-1 is a hallmark of various cancer types, including non-small cell lung cancers (NSCLCs). Here, we present proof-of-principle evidence that LINE-1 analytes in extracellular vesicles (EVs) serve as tools for molecular diagnostics of NSCLC, with LINE-1 status in tumor cells and tissues mirroring the LINE-1 mRNA and ORF1p cargos of EVs from lung cancer cell culture conditioned media or human plasma. The levels of LINE-1 analytes in plasma EVs from ostensibly healthy individuals were higher in females than males. While the profiles of LINE-1 mRNA and ORF1p in African Americans compared to Hispanics were not significantly different, African Americans showed slightly higher ORF1p content, and 2–3 times greater ranges of LINE-1 values compared to Hispanics. Whole plasma ORF1p levels correlated with EV ORF1p levels, indicating that most of the circulating LINE-1 protein is contained within EVs. EV LINE-1 mRNA levels were elevated in patients with advanced cancer stages and in select patients with squamous cell carcinoma and metastatic tumors compared to adenocarcinomas. The observed EV LINE-1 mRNA profiles paralleled the patterns of ORF1p expression in NSCLC tissue sections suggesting that LINE-1 analytes in plasma EVs may serve to monitor the activity of LINE-1 retroelements in lung cancer. Full article

(This article belongs to the Special Issue Exosomes and Extracellular Vesicles in Health and Diseases 2.0)

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23 pages, 4202 KiB

Open AccessArticle

Circulating Extracellular Vesicles in Subarachnoid Hemorrhage Patients: Characterization and Cellular Effects

by Elena Grossini, Teresa Esposito, Michela Viretto, Sakthipriyan Venkatesan, Ilaria Licari, Daniela Surico, Francesco Della Corte, Luigi Castello, Stefania Bruno, Marco Quaglia, Cristoforo Comi, Vincenzo Cantaluppi and Rosanna Vaschetto

Int. J. Mol. Sci. 2023, 24(19), 14913; https://doi.org/10.3390/ijms241914913 - 5 Oct 2023

Cited by 2 | Viewed by 1557

Abstract

Circulating extracellular vesicles (EVs) may play a pathophysiological role in the onset of complications of subarachnoid hemorrhage (SAH), potentially contributing to the development of vasospasm (VP). In this study, we aimed to characterize circulating EVs in SAH patients and examine their effects on endothelial and smooth muscle cells (SMCs). In a total of 18 SAH patients, 10 with VP (VP), 8 without VP (NVP), and 5 healthy controls (HC), clinical variables were recorded at different time points. EVs isolated from plasma samples were characterized and used to stimulate human vascular endothelial cells (HUVECs) and SMCs. We found that EVs from SAH patients expressed markers of T-lymphocytes and platelets and had a larger size and a higher concentration compared to those from HC. Moreover, EVs from VP patients reduced cell viability and mitochondrial membrane potential in HUVECs and increased oxidants and nitric oxide (NO) release. Furthermore, EVs from SAH patients increased intracellular calcium levels in SMCs. Altogether, our findings reveal an altered pattern of circulating EVs in SAH patients, suggesting their pathogenic role in promoting endothelial damage and enhancing smooth muscle reactivity. These results have significant implications for the use of EVs as potential diagnostic/prognostic markers and therapeutic tools in SAH management. Full article

(This article belongs to the Special Issue Exosomes and Extracellular Vesicles in Health and Diseases 2.0)

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Review

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20 pages, 3234 KiB

Open AccessReview

Analysis and Characterization of the Extracellular Vesicles Released in Non-Cancer Diseases Using Matrix-Assisted Laser Desorption Ionization/Mass Spectrometry

by Antonella Maria Aresta, Nicoletta De Vietro and Carlo Zambonin

Int. J. Mol. Sci. 2024, 25(8), 4490; https://doi.org/10.3390/ijms25084490 - 19 Apr 2024

Cited by 1 | Viewed by 1560

Abstract

The extracellular vesicles (EVs) released by cells play a crucial role in intercellular communications and interactions. The direct shedding of EVs from the plasma membrane represents a fundamental pathway for the transfer of properties and information between cells. These vesicles are classified based on their origin, biogenesis, size, content, surface markers, and functional features, encompassing a variety of bioactive molecules that reflect the physiological state and cell type of origin. Such molecules include lipids, nucleic acids, and proteins. Research efforts aimed at comprehending EVs, including the development of strategies for their isolation, purification, and characterization, have led to the discovery of new biomarkers. These biomarkers are proving invaluable for diagnosing diseases, monitoring disease progression, understanding treatment responses, especially in oncology, and addressing metabolic, neurological, infectious disorders, as well as advancing vaccine development. Matrix-Assisted Laser Desorption Ionization (MALDI)/Mass Spectrometry (MS) stands out as a leading tool for the analysis and characterization of EVs and their cargo. This technique offers inherent advantages such as a high throughput, minimal sample consumption, rapid and cost-effective analysis, and user-friendly operation. This review is mainly focused on the primary applications of MALDI–time-of-flight (TOF)/MS in the analysis and characterization of extracellular vesicles associated with non-cancerous diseases and pathogens that infect humans, animals, and plants. Full article

(This article belongs to the Special Issue Exosomes and Extracellular Vesicles in Health and Diseases 2.0)

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35 pages, 3878 KiB

Open AccessReview

Therapeutic Applications of Stem Cell-Derived Exosomes

by Omar Abdulhakeem Ahmed Yusuf Abdulmalek, Khaled Hameed Husain, Haya Khaled Ali Abdulla AlKhalifa, Mariam Masood Abdulkarim Bahrooz Alturani, Alexandra E. Butler and Abu Saleh Md Moin

Int. J. Mol. Sci. 2024, 25(6), 3562; https://doi.org/10.3390/ijms25063562 - 21 Mar 2024

Cited by 9 | Viewed by 4376

Abstract

Exosomes are extracellular vesicles of endosomal origin, ranging from 30 to 150 nm in diameter, that mediate intercellular transfer of various biomolecules, such as proteins, lipids, nucleic acids, and metabolites. They modulate the functions of recipient cells and participate in diverse physiological and pathological processes, such as immune responses, cell–cell communication, carcinogenesis, and viral infection. Stem cells (SCs) are pluripotent or multipotent cells that can differentiate into various cell types. SCs can also secrete exosomes, which exhibit remarkable therapeutic potential for various diseases, especially in the field of regenerative medicine. For example, exosomes derived from mesenchymal stem cells (MSCs) contain proteins, lipids, and miRNAs that can ameliorate endocrine disorders, such as diabetes and cancer. Exosomes from SCs (sc-exos) may offer similar advantages as SCs, but with reduced risks and challenges. Sc-exos have lower tumorigenicity, immunogenicity, and infectivity. They can also deliver drugs more efficiently and penetrate deeper into tissues. In this review, we provide an overview of the recent advances in sc-exos and their therapeutic applications in various diseases, such as diabetes and cancer. We also elucidate how the biological effects of sc-exos depend on their molecular composition. We also address the current challenges and future directions of using sc-exos. Full article

(This article belongs to the Special Issue Exosomes and Extracellular Vesicles in Health and Diseases 2.0)

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22 pages, 2085 KiB

Open AccessReview

Exosomes Interactions with the Blood–Brain Barrier: Implications for Cerebral Disorders and Therapeutics

by Zaynab Osaid, Mohamed Haider, Rifat Hamoudi and Rania Harati

Int. J. Mol. Sci. 2023, 24(21), 15635; https://doi.org/10.3390/ijms242115635 - 26 Oct 2023

Cited by 7 | Viewed by 2810

Abstract

The Blood–Brain Barrier (BBB) is a selective structural and functional barrier between the circulatory system and the cerebral environment, playing an essential role in maintaining cerebral homeostasis by limiting the passage of harmful molecules. Exosomes, nanovesicles secreted by virtually all cell types into body fluids, have emerged as a major mediator of intercellular communication. Notably, these vesicles can cross the BBB and regulate its physiological functions. However, the precise molecular mechanisms by which exosomes regulate the BBB remain unclear. Recent research studies focused on the effect of exosomes on the BBB, particularly in the context of their involvement in the onset and progression of various cerebral disorders, including solid and metastatic brain tumors, stroke, neurodegenerative, and neuroinflammatory diseases. This review focuses on discussing and summarizing the current knowledge about the role of exosomes in the physiological and pathological modulation of the BBB. A better understanding of this regulation will improve our understanding of the pathogenesis of cerebral diseases and will enable the design of effective treatment strategies. Full article

(This article belongs to the Special Issue Exosomes and Extracellular Vesicles in Health and Diseases 2.0)

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Other

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7 pages, 960 KiB

Open AccessOpinion

Specific Extracellular Vesicles, Generated and Operating at Synapses, Contribute to Neuronal Effects and Signaling

by Jacopo Meldolesi

Int. J. Mol. Sci. 2024, 25(10), 5103; https://doi.org/10.3390/ijms25105103 - 7 May 2024

Viewed by 1468

Abstract

In all cell types, small EVs, very abundant extracellular vesicles, are generated and accumulated within MVB endocytic cisternae. Upon MVB fusion and exocytosis with the plasma membrane, the EVs are released to the extracellular space. In the central nervous system, the release of neuronal EVs was believed to occur only from the surface of the body and dendrites. About 15 years ago, MVB cisternae and EVs were shown to exist and function at synaptic boutons, the terminals’ pre- and post-synaptic structures essential for canonical neurotransmitter release. Recent studies have revealed that synaptic EVs are peculiar in many respects and heterogeneous with respect to other neuronal EVs. The distribution of synaptic EVs and the effect of their specific molecules are found at critical sites of their distribution. The role of synaptic EVs could consist of the modulation of canonical neurotransmitter release or a distinct, non-canonical form of neurotransmission. Additional roles of synaptic EVs are still not completely known. In the future, additional investigations will clarify the role of synaptic EVs in pathology, concerning, for example, circuits, trans-synaptic transmission, diagnosis and the therapy of diseases. Full article

(This article belongs to the Special Issue Exosomes and Extracellular Vesicles in Health and Diseases 2.0)

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