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A special issue of Membranes (ISSN 2077-0375). This special issue belongs to the section "Biological Membrane Composition and Structures".

Deadline for manuscript submissions: closed (31 May 2022) | Viewed by 40272

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Special Issue Editors

Dr. Ganesh Vilas Shelke

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Chief Guest Editor

1. National Institute of Health-NICHD, USA
2. University of Gothenburg, Gothenburg, Sweden
Interests: extracellular Vesicles; exosomes; cell-biology; membrane trafficking; melanoma; immunology; antigen presentation; immunotherapy

Dr. Stefania Raimondo

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Guest Editor

Department of BioMedicine, Neuroscience and Advanced Diagnostics (Bi.N.D), Section of Biology and Genetics, University of Palermo, 90133 Palermo, Italy
Interests: extracellular vesicles; plants-derived compounds; inflammation and cancer

Dr. Jaewook Lee

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Guest Editor

Department of Life Sciences, Pohang University of Science and Technology (POSTECH), Pohang, Korea
Interests: extracellular vesicles

Special Issue Information

Dear Colleagues,

Cells require constant feedback from their surroundings and respond to cues in order to maintain a homeostatic equilibrium. These secretory signals can be soluble factors (proteins, nucleic acids, and small molecules) that are either free or membrane-associated moieties. In past two decades, cell-derived membranes like extracellular vesicles (e.g Exosomes, Micro-vesicles) have been a topic of interest for many researchers to understand the origin and functions of these secretory cues. Naturally, cell-derived membranes are heterogeneous and are derived from multiple cell types and species like humans, plants, and bacteria. In light of the current knowledge and engineering-technologies, a broad repertoire of cell-derived membranes is being used to obtain artificial membrane vesicles with diverse repertoire/cargo.

The aim of this Special Issue is to cover novel research trends in vesicles derived from cell membranes from both eukaryotic and prokaryotic cells. We request submissions that provide insights into the fundamentals of cell membranes vesicles generation, characterization, functional properties, and application. These cell membranes vesicles can either be naturally produced or artificially engineered. The contributions can be original research papers, reviews, meta-analysis, in-silico analysis, and methodological developments. Areas to be covered may include, but are not limited to:

Artificially derived cell membranes (artificial generated MVs, Hybrid EVs) to create membranes.
Strategies that dissect the heterogeneity of cell membranes vesicles
• Naturally occurring cell-derived membranes vesicles.
• Strategies to manipulate cells to create therapeutic vesicles.
Analysis of cargos derived from cell membranes vesicles using advanced proteomics, lipidomics, metabolomics, and next-generation sequencing tools.

Dr. Ganesh Vilas Shelke
Dr. Stefania Raimondo
Dr. Jaewook Lee
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. Membranes is an international peer-reviewed open access monthly journal published by MDPI.

Please visit the Instructions for Authors page before submitting a manuscript. The Article Processing Charge (APC) for publication in this open access journal is 2700 CHF (Swiss Francs). 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

Extracellular Vesicles Heterogeneity；
Extracellular vesicles Subpopulation；
Exosomes；
Macro-vesicles；
Outer Membrane Vesicles (OMVs)；
Artificial Cell-derived Nano-Vesicles；
Hybrid Vesicles；
EV-like particles；
Engineered Extracellular-vesicles

Published Papers (10 papers)

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Research

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14 pages, 2874 KiB

Open AccessArticle

Hypoxia-Preconditioned Extracellular Vesicles from Mesenchymal Stem Cells Improve Cartilage Repair in Osteoarthritis

by Bocheng Zhang, Xiaoyuan Tian, Zhenan Qu, Jun Hao and Weiguo Zhang

Membranes 2022, 12(2), 225; https://doi.org/10.3390/membranes12020225 - 16 Feb 2022

Cited by 29 | Viewed by 3344

Abstract

In the past decade, mesenchymal stem cells (MSCs) have been widely used for the treatment of osteoarthritis (OA), and extracellular vesicles (EVs) may play a major role in the efficacy of this treatment. Hypoxia can change the cargo and biological functions of MSC-derived EVs (MSC-EVs). The aim of the present study was to determine whether the effects of hypoxia-preconditioned MSC-EVs on OA cartilage repair are superior to normoxia-preconditioned MSC-EVs. By using in vitro and in vivo OA models, we verified that hypoxia-preconditioned MSC-EVs improved chondrocyte proliferation and migration and suppressed chondrocyte apoptosis to a greater extent than normoxia-preconditioned MSC-EVs. Furthermore, we found that hypoxia altered the microRNA expression in MSC-EVs and identified four differentially expressed microRNAs: hsa-miR-181c-5p, hsa-miR-18a-3p, hsa-miR-376a-5p, and hsa-miR-337-5p. Bioinformatics analysis revealed that hypoxic pretreatment may promote cartilage repair by stimulating chondrocyte proliferation and migration and suppressing chondrocyte apoptosis through the miRNA-18-3P/JAK/STAT or miRNA-181c-5p/MAPK signaling pathway. Therefore, hypoxia-preconditioned EVs may be a novel treatment for OA. Full article

(This article belongs to the Special Issue Cell Membrane Vesicles)

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11 pages, 1280 KiB

Open AccessArticle

Putative Internal Control Genes in Bovine Milk Small Extracellular Vesicles Suitable for Normalization in Quantitative Real Time-Polymerase Chain Reaction

by Md. Matiur Rahman, Shigeo Takashima, Yuji O. Kamatari, Yassien Badr, Kaori Shimizu, Ayaka Okada and Yasuo Inoshima

Membranes 2021, 11(12), 933; https://doi.org/10.3390/membranes11120933 - 26 Nov 2021

Cited by 4 | Viewed by 1722

Abstract

Bovine milk small extracellular vesicles (sEVs) contain many biologically important molecules, including mRNAs. Quantitative real-time polymerase chain reaction (qRT-PCR) is a widely used method for quantifying mRNA in tissues and cells. However, the use, selection, and stability of suitable putative internal control genes in bovine milk sEVs for normalization in qRT-PCR have not yet been identified. Thus, the aim of the present study was to determine suitable putative internal control genes in milk sEVs for the normalization of qRT-PCR data. Milk sEVs were isolated from six healthy Holstein-Friesian cattle, followed by RNA extraction and cDNA synthesis. In total, 17 mRNAs were selected for investigation and quantification using qRT-PCR; they were further evaluated using geNorm, NormFinder, BestKeeper, and ∆CT algorithms to identify those that were highly stable putative internal control genes in milk sEVs. The final ranking of suitable putative internal control genes was determined using RefFinder. The mRNAs from TUB, ACTB, DGKZ, ETFDH, YWHAZ, STATH, DCAF11, and EGFLAM were detected in milk sEVs from six cattle by qRT-PCR. RefFinder demonstrated that TUB, ETFDH, and ACTB were highly stable in milk sEVs, and thus suitable for normalization of qRT-PCR data. The present study suggests that the use of these genes as putative internal control genes may further enhance the robustness of qRT-PCR in bovine milk sEVs. Since these putative internal control genes apply to healthy bovines, it would be helpful to include that the genes were stable in sEVs under “normal or healthy conditions”. Full article

(This article belongs to the Special Issue Cell Membrane Vesicles)

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11 pages, 754 KiB

Open AccessArticle

In Vivo Secretion of β-Lactamase-Carrying Outer Membrane Vesicles as a Mechanism of β-Lactam Therapy Failure

by Martina Bielaszewska, Ondřej Daniel, Otakar Nyč and Alexander Mellmann

Membranes 2021, 11(11), 806; https://doi.org/10.3390/membranes11110806 - 23 Oct 2021

Cited by 6 | Viewed by 3481

Abstract

Outer membrane vesicles carrying β-lactamase (βLOMVs) protect bacteria against β-lactam antibiotics under experimental conditions, but their protective role during a patient’s treatment leading to the therapy failure is unknown. We investigated the role of βLOMVs in amoxicillin therapy failure in a patient with group A Streptococcus pyogenes (GAS) pharyngotonsillitis. The patient’s throat culture was examined by standard microbiological procedures. Bacterial vesicles were analyzed for β-lactamase by immunoblot and the nitrocefin assay, and in vivo secretion of βLOMVs was detected by electron microscopy. These analyses demonstrated that the patient’s throat culture grew, besides amoxicillin-susceptible GAS, an amoxicillin-resistant nontypeable Haemophilus influenzae (NTHi), which secreted βLOMVs. Secretion and β-lactamase activity of NTHi βLOMVs were induced by amoxicillin concentrations reached in the tonsils during therapy. The presence of NTHi βLOMVs significantly increased the minimal inhibitory concentration of amoxicillin for GAS and thereby protected GAS against bactericidal concentrations of amoxicillin. NTHi βLOMVs were identified in the patient’s pharyngotonsillar swabs and saliva, demonstrating their secretion in vivo at the site of infection. We conclude that the pathogen protection via βLOMVs secreted by the flora colonizing the infection site represents a yet underestimated mechanism of β-lactam therapy failure that warrants attention in clinical studies. Full article

(This article belongs to the Special Issue Cell Membrane Vesicles)

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18 pages, 1734 KiB

Open AccessArticle

Skeletal Muscle Cell Growth Alters the Lipid Composition of Extracellular Vesicles

by Taylor R. Valentino, Blake D. Rule, C. Brooks Mobley, Mariana Nikolova-Karakashian and Ivan J. Vechetti

Membranes 2021, 11(8), 619; https://doi.org/10.3390/membranes11080619 - 12 Aug 2021

Cited by 7 | Viewed by 3049

Abstract

We sought to characterize the lipid profile of skeletal muscle cell-derived Extracellular Vesicles (EVs) to determine if a hypertrophic stimulus would affect the lipid composition of C2C12 myotube-derived EVs. Analyses included C2C12 murine myoblasts differentiated into myotubes and treated with Insulin-Like Growth Factor 1 (IGF-1) for 24 h to induce hypertrophic growth. EVs were isolated from cell culture media, quantified using Nanoparticle Tracking Analysis (NTA) and analyzed using Transmission Electron Microscopy (TEM). EVs were homogenized and lipids extracted for quantification by Mass Spectrometry followed by downstream lipid class enrichment and lipid chain analysis. IGF-1 treatment elicited an increase in CD63 and CD81 levels (39% and 21%) compared to the controls (16%), respectively. Analysis revealed that skeletal muscle-derived EVs are enriched in bioactive lipids that are likely selectively incorporated into EVs during hypertrophic growth. IGF-1 treatment of myotubes had a significant impact on the levels of diacylglycerol (DG) and ceramide (Cer) in secreted EVs. Specifically, the proportion of unsaturated DG was two- to three-fold higher in EVs derived from IGF-treated cells, as compared to those from control cells. The levels of saturated DG were unaffected. Selective increases were similarly seen in C16- and C24-Cer but not in other species. Levels of free sphingoid bases tended to decrease, while those of sphingosine-1-phosphate was unaffected. Our results suggest that the lipid composition and biogenesis of skeletal muscle-derived EVs, are specific and highly selective during hypertrophic growth. Full article

(This article belongs to the Special Issue Cell Membrane Vesicles)

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11 pages, 2705 KiB

Open AccessArticle

Mesenchymal Stem Cells Do Not Lose Direct Labels Including Iron Oxide Nanoparticles and DFO-⁸⁹Zr Chelates through Secretion of Extracellular Vesicles

by Yue Gao, Anna Jablonska, Chengyan Chu, Piotr Walczak and Miroslaw Janowski

Membranes 2021, 11(7), 484; https://doi.org/10.3390/membranes11070484 - 29 Jun 2021

Cited by 2 | Viewed by 2126

Abstract

Rapidly ageing populations are beset by tissue wear and damage. Stem cell-based regenerative medicine is considered a solution. Years of research point to two important aspects: (1) the use of cellular imaging to achieve sufficient precision of therapeutic intervention, and the fact that (2) many therapeutic actions are executed through extracellular vesicles (EV), released by stem cells. Therefore, there is an urgent need to interrogate cellular labels in the context of EV release. We studied clinically applicable cellular labels: superparamagnetic iron oxide nanoparticles (SPION), and radionuclide detectable by two main imaging modalities: MRI and PET. We have demonstrated effective stem cell labeling using both labels. Then, we obtained EVs from cell cultures and tested for the presence of cellular labels. We did not find either magnetic or radioactive labels in EVs. Therefore, we report that stem cells do not lose labels in released EVs, which indicates the reliability of stem cell magnetic and radioactive labeling, and that there is no interference of labels with EV content. In conclusion, we observed that direct cellular labeling seems to be an attractive approach to monitoring stem cell delivery, and that, importantly, labels neither locate in EVs nor affect their basic properties. Full article

(This article belongs to the Special Issue Cell Membrane Vesicles)

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Review

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20 pages, 1750 KiB

Open AccessReview

An Updated View of the Importance of Vesicular Trafficking and Transport and Their Role in Immune-Mediated Diseases: Potential Therapeutic Interventions

by Miguel A. Ortega, Oscar Fraile-Martinez, Cielo Garcia-Montero, Miguel Angel Alvarez-Mon, Ana Maria Gomez-Lahoz, Agustin Albillos, Guillermo Lahera, Javier Quintero, Jorge Monserrat, Luis G. Guijarro and Melchor Alvarez-Mon

Membranes 2022, 12(6), 552; https://doi.org/10.3390/membranes12060552 - 25 May 2022

Cited by 7 | Viewed by 5355

Abstract

Cellular trafficking is the set of processes of distributing different macromolecules by the cell. This process is highly regulated in cells, involving a system of organelles (endomembranous system), among which are a great variety of vesicles that can be secreted from the cell, giving rise to different types of extracellular vesicles (EVs) that can be captured by other cells to modulate their function. The cells of the immune system are especially sensitive to this cellular traffic, producing and releasing different classes of EVs, especially in disease states. There is growing interest in this field due to the therapeutic and translational possibilities it offers. Different ways of taking advantage of the understanding of cell trafficking and EVs are being investigated, and their use as biomarkers or therapeutic targets is being investigated. The objective of this review is to collect the latest results and knowledge in this area with a specific focus on immune-mediated diseases. Although some promising results have been obtained, further knowledge is still needed, at both the basic and translational levels, to understand and modulate cellular traffic and EVs for better clinical management of these patients. Full article

(This article belongs to the Special Issue Cell Membrane Vesicles)

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16 pages, 1922 KiB

Open AccessReview

Plant-RNA in Extracellular Vesicles: The Secret of Cross-Kingdom Communication

by Ornella Urzì, Roberta Gasparro, Nima Rabienezhad Ganji, Riccardo Alessandro and Stefania Raimondo

Membranes 2022, 12(4), 352; https://doi.org/10.3390/membranes12040352 - 23 Mar 2022

Cited by 22 | Viewed by 4841

Abstract

The release of extracellular vesicles (EVs) is a common language, used by living organisms from different kingdoms as a means of communication between them. Extracellular vesicles are lipoproteic particles that contain many biomolecules, such as proteins, nucleic acids, and lipids. The primary role of EVs is to convey information to the recipient cells, affecting their function. Plant-derived extracellular vesicles (PDEVs) can be isolated from several plant species, and the study of their biological properties is becoming an essential starting point to study cross-kingdom communication, especially between plants and mammalians. Furthermore, the presence of microRNAs (miRNAs) in PDEVs represents an interesting aspect for understanding how PDEVs can target the mammalian genes involved in pathological conditions such as cancer, inflammation, and oxidative stress. In particular, this review focuses on the history of PDEVs, from their discovery, to purification from various matrices, and on the functional role of PDEV-RNAs in cross-kingdom interactions. It is worth noting that miRNAs packaged in PDEVs can be key modulators of human gene expression, representing potential therapeutic agents. Full article

(This article belongs to the Special Issue Cell Membrane Vesicles)

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17 pages, 1682 KiB

Open AccessReview

Targeted Delivery of Exosomes Armed with Anti-Cancer Therapeutics

by Hojun Choi, Hwayoung Yim, Cheolhyoung Park, So-Hee Ahn, Yura Ahn, Areum Lee, Heekyoung Yang and Chulhee Choi

Membranes 2022, 12(1), 85; https://doi.org/10.3390/membranes12010085 - 13 Jan 2022

Cited by 17 | Viewed by 5517

Abstract

Among extracellular vesicles, exosomes have gained great attention for their role as therapeutic vehicles for delivering various active pharmaceutical ingredients (APIs). Exosomes “armed” with anti-cancer therapeutics possess great potential for an efficient intracellular delivery of anti-cancer APIs and enhanced targetability to tumor cells. Various technologies are being developed to efficiently incorporate anti-cancer APIs such as genetic materials (miRNA, siRNA, mRNA), chemotherapeutics, and proteins into exosomes and to induce targeted delivery to tumor burden by exosomal surface modification. Exosomes can incorporate the desired therapeutic molecules via direct exogenous methods (e.g., electroporation and sonication) or indirect methods by modifying cells to produce “armed” exosomes. The targeted delivery of “armed” exosomes to tumor burden could be accomplished either by “passive” targeting using the natural tropism of exosomes or by “active” targeting via the surface engineering of exosomal membranes. Although anti-cancer exosome therapeutics demonstrated promising results in preclinical studies, success in clinical trials requires thorough validation in terms of chemistry, manufacturing, and control techniques. While exosomes possess multiple advantages over synthetic nanoparticles, challenges remain in increasing the loading efficiency of anti-cancer agents into exosomes, as well as establishing quantitative and qualitative analytical methods for monitoring the delivery of in vivo administered exosomes and exosome-incorporated anti-cancer agents to the tumor parenchyma. Full article

(This article belongs to the Special Issue Cell Membrane Vesicles)

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15 pages, 1111 KiB

Open AccessReview

Extracellular Vesicles in Cervical Cancer and HPV Infection

by Víctor Acevedo-Sánchez, Ruth M. Rodríguez-Hernández, Sergio R. Aguilar-Ruíz, Honorio Torres-Aguilar and María de los A. Romero-Tlalolini

Membranes 2021, 11(6), 453; https://doi.org/10.3390/membranes11060453 - 20 Jun 2021

Cited by 16 | Viewed by 3482

Abstract

Since their description, extracellular vesicles (EVs) have shown growing relevance in cancer progression. These cell structures contain and transfer molecules such as nucleic acids (including DNA and RNA), proteins, and lipids. Despite the rising information about EVs’ relationship with cancer, there is still scarce evidence about their content and function in cervical cancer. Interestingly, the composition and purposes of some cellular molecules and the expression of oncogenic proteins packaged in EVs seem modified in HPV-infected cells; and, although only the E6 oncogenic protein has been detected in exosomes from HPV-positive cells, both E6/E7 oncogenes mRNA has been identified in EVs; however, their role still needs to be clarified. Given that EVs internalizing into adjacent or distant cells could modify their cellular behavior or promote cancer-associated events like apoptosis, proliferation, migration, or angiogenesis in receptor cells, their comprehensive study will reveal EV-associated mechanisms in cervical cancer. This review summarizes the current knowledge in composition and functions of cervical cancer and HPV Infection-derived EVs. Full article

(This article belongs to the Special Issue Cell Membrane Vesicles)

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9 pages, 1030 KiB

Open AccessReview

Plant-Derived Extracellular Vesicles: Current Findings, Challenges, and Future Applications

by Nader Kameli, Anya Dragojlovic-Kerkache, Paul Savelkoul and Frank R. Stassen

Membranes 2021, 11(6), 411; https://doi.org/10.3390/membranes11060411 - 29 May 2021

Cited by 43 | Viewed by 6171

Abstract

In recent years, plant-derived extracellular vesicles (PDEVs) have gained the interest of many experts in fields such as microbiology and immunology, and research in this field has exponentially increased. These nano-sized particles have provided researchers with a number of interesting findings, making their application in human health and disease very promising. Both in vitro and in vivo experiments have shown that PDEVs can exhibit a multitude of effects, suggesting that these vesicles may have many potential future applications, including therapeutics and nano-delivery of compounds. While the preliminary results are promising, there are still some challenges to face, such as a lack of protocol standardization, as well as knowledge gaps that need to be filled. This review aims to discuss various aspects of PDEV knowledge, including their preliminary findings, challenges, and future uses, giving insight into the complexity of conducting research in this field. Full article

(This article belongs to the Special Issue Cell Membrane Vesicles)

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