Journal of Extracellular Vesicles最新文献

Outer membrane vesicles (OMVs) are nanosized vesicles naturally secreted by Gram-negative bacteria and represent a promising platform for vaccine development. OMVs possess inherent immunostimulatory properties due to the presence of pathogen-associated molecular patterns (PAMPs), providing self-adjuvanting capabilities and the ability to elicit both innate and adaptive immune responses. This review outlines the advantages of OMVs over traditional vaccine strategies, including their safety, modularity, and the potential for genetic engineering to enable targeted antigen delivery. We describe approaches to enhance OMVs yield and immunogenicity, such as modifications to reduce lipopolysaccharide (LPS) toxicity and systems enabling antigen localization—either on the surface or within the lumen—using fusion constructs like ClyA, Lpp-OmpA, AIDA-I, Hbp, and Sec/Tat signal peptides. We further summarize preclinical applications of OMVs-based vaccines targeting bacterial pathogens, viral infections, and cancer. In addition, we address key challenges in large-scale production, purification, and long-term stability, and explore strategies for conjugating or encapsulating heterologous antigens. Overall, OMVs offer a versatile and scalable extracellular vesicle-based platform with strong potential for next-generation vaccines targeting diverse infectious diseases and beyond.

Osteoarthritis (OA), the prevalent debilitating joint disorder, is accelerated by dysregulated intercellular crosstalk, yet the role of fibroblast-like synoviocyte (FLS)-derived extracellular vesicles and particles (EVPs) in disease progression remains to be elucidated. Here, integrative analysis of clinical specimens, animal models, and publicly available datasets revealed significant alterations in exosomal pathways within OA synovium. Proteomic profiling revealed distinct molecular signatures in EVPs derived from inflammatory and senescent FLSs, reflecting the pathophysiological status of their parent cells. We demonstrated that FLSs under inflammatory and senescent states in OA secreted pathogenic EVPs that propagated joint degeneration by disrupting chondrocyte homeostasis, polarizing macrophages towards a pro-inflammatory phenotype, and impairing chondrogenesis of mesenchymal stem cells. To therapeutically target these pathogenic EVPs, we engineered an adeno-associated virus 9 (AAV9) vector fused with a synovium-affinity peptide (HAP-1) to deliver shRNA against Rab27a, a key regulator of EVP secretion. Intra-articular administration of the engineered AAV9 in a murine OA model induced by destabilization of the medical meniscus significantly reduced synovial hyperplasia, cartilage degradation and inflammatory responses, while demonstrating satisfactory systemic biosafety. Our findings establish FLS-derived EVPs as critical mediators of OA pathogenesis and propose a targeted strategy to block their secretion, offering a promising disease-modifying therapeutic avenue for OA.

Tumour progression depends on the bidirectional interactions between cancer and stroma in the heterogeneous tumour microenvironment (TME) partially through extracellular vesicles (EVs). However, the secretary mechanism and biological effect of cancer cell derived EVs on tumour survival under starvation is poorly defined. Here, we identify cancer cells selectively secrete miR-33a with the assistance of aconitase 1 (ACO1), an iron-responsive RNA binding protein, under glucose starvation and lower iron level, which affiliates the binding capability of miR-33a and ACO1. Exosomal miR-33a suppresses putrescine biosynthesis by targeting AGMAT in cancer-associated fibroblasts (CAFs) from tumour core region, where putrescine inhibits the expression of demethylase KDM5C. TIA1 gene, stress granule (SG) marker, is tightly regulated by miR-33a/KDM5C/H3K4me3 axis and exosomal miR-33a diminishes the formation of stromal SGs in CAFs. Collectively, our study reveals tumour selectively secretes miR-33a-5p through EVs to remodel the stromal SG formation and gain survival possibility for cancer cells in tumour core region, highlighting a novel regulatory mechanism of iron and nutrient level on EV secretion and the function of polyamine metabolism in reshaping epigenetic profiles.

Extracellular vesicles (EVs) are membrane-bound nanoparticles secreted by cells that are involved in multiple forms of intercellular communication and show promising potential for biotechnological applications. Arthropod-derived EV research remains relatively fragmented in contrast to the extensively studied mammalian EV field. In this review, we present a comprehensive synthesis of over 100 studies exploring EV biology across arthropods - including insects, arachnids and crustaceans. Specifically, we summarise the key proteins involved in EV biogenesis and trafficking and provide an overview of the diverse biological roles of EVs in arthropod systems. These include (i) roles in developmental, neurobiological and reproductive processes, as well as in ageing, starvation and protein homeostasis; and (ii) involvement in immunity, vector-pathogen-host dynamics and host-parasite interactions. In addition, we provide an overview of current EV isolation methodologies and their application in arthropod studies, as well as explore the emerging biotechnological potential of arthropod-derived EVs. Finally, we address key challenges in the field, including technical limitations in EV isolation, existing knowledge gaps and opportunities for biotechnological applications. By identifying technical limitations and knowledge gaps, as well as proposing directions for future research, we provide a timely and comprehensive resource to guide the progress of arthropod EV research.

Although the field of plant EVs (PEVs) is experiencing exponential growth, rigorous characterisation complying with MISEV guidelines has not been yet implemented due to the lack of bona fide reference markers. In this work, we have paved the way for the standardisation of PEV markers, providing the most profound proteomic data so far from apoplastic washing fluid-EVs, a sample enriched in genuine extracellular vesicles from plant tissue of two reference plant species: Arabidopsis thaliana (Arath-EVs) and Brassica oleracea (Braol-EVs). Besides, we analysed the protein content of the soluble fraction of the apoplast and calculated the enrichment of the potential markers studied in EVs. Additionally, we have conducted an exhaustive analysis of the proteomic data available so far from genuine EVs from any plant species, evaluating current potential markers, together with those found in our proteomic analyses. Our results provide evidence supporting the potential use of the following families as PEV markers: aquaporins, vacuolar-type ATPase complex subunits, some fasciclin-like arabinogalactan proteins (FLAs), tetraspanins, syntaxins, germin-like proteins and calreticulins. Next, we analysed the presence of orthologues and their degree of conservation throughout plant taxa, as well as in 2 reference species from the animal kingdom: human and mouse. Their degree of conservation was compared with that of current animal EVs: CD63, CD81 and CD9. Among the protein families with potential to be used as PEV markers, 2 were found to be plant-specific: FLAs and germin-like proteins. On the other hand, aquaporins and vacuolar-type ATPase complex subunits showed the greatest degree of conservation across plant and animal kingdoms. Our results provide key insights on several aspects of classical and novel protein identity markers for PEVs to assist in the selection of the best candidates for standardisation: (1) species-specific abundance, (2) specificity for PEVs and (3) conservation and plant specificity.

Extracellular vesicles (EVs) are small anuclear cellular membrane encapsulated fragments of importance for cellular interaction and transfer of information. These small vesicles, diverse in size and functionality, can be obtained from cells, tissues and bodily fluids. A complicated step for obtaining EVs from whole organs is understanding the optimal methodology for organ processing. In this study, we have examined two different techniques: one enzymatic and one novel non-enzymatic automated tissue dissociation (ATD) machine. Animals were perfused, organs extracted, and techniques comparatively applied. We have used these techniques for organ-based dissociation followed by EV isolation from the dissociated tissues (heart, kidney, lung). While both approaches allow isolation of intact EVs there are distinct differences in overall cell and particle yields. Our study highlights tissue specific inter-organ variability and differential impact of dissociation strategies on organ-based EV profiles, as well as cellular characteristics. Our findings indicate that EV yields and characteristics varies between enzymatic and ATD techniques as well as between organs with highest EV yield obtained from kidneys following enzymatic dissociation. Our findings can be rapidly transferred to other setups or developed to enable enumeration and characterization of EVs obtained from whole organs in physiological and pathological settings.

Intervertebral disc degeneration (IVDD) is a common age-related disorder associated with inflammation, pain and impaired mobility. In this study, we developed a therapeutic system using silk fibroin (SF) hydrogel loaded with mRNA-engineered extracellular vesicles derived from murine bone marrow mesenchymal stem cells (BMSCs-EVs) to modulate macrophage polarization and alleviate IVDD. BMSCs were isolated from 6-week-old C57BL/6 mice, and an acute IVDD model was established via 18G needle puncture of the coccygeal discs (Co7-Co10). RAW 264.7 murine macrophages were used for in vitro assays, with M1 polarization induced by LPS and IFN-γ. The SF/EVs complex was characterized by SEM, FTIR and rheology, confirming its structural suitability for EV delivery. Functionally, SF hydrogel not only served as a biocompatible carrier but also enabled sustained release of EVs, enhancing their anti-inflammatory effects. In vitro, SF/EVs inhibited M1 polarization and promoted M2 marker expression. In vivo implantation improved disc histology and reduced inflammatory macrophage infiltration. High-throughput RNA sequencing identified S100B as a key functional cargo within EVs. Lentivirus-mediated overexpression and knockdown experiments confirmed that EV-derived S100B suppresses M1 polarization and mitigates IVDD progression. In summary, SF hydrogel loaded with S100B-enriched BMSCs-EVs offers a promising strategy to reshape the inflammatory microenvironment and promote disc regeneration in IVDD.

Giardia lamblia, a eukaryotic intestinal parasite, produces small extracellular vesicles (sEVs) as a conserved evolutionary mechanism. This study investigates the functional role of sEVs in modulating drug response traits among G. lamblia parasites. Here, we showed that sEVs derived from metronidazole (MTZ)–resistant clones modify the expression of enzymes involved in MTZ metabolism and the production of reactive oxygen species (ROS) in recipient wild type parasites. The transfer efficiency and phenotypic impact vary depending on the genetic background of the isolates, highlighting a genotype-specific mechanism. Our findings reveal that sEVs act as mediators of phenotypic adaptation in G. lamblia, enhancing parasite survival under drug-induced stress. This study highlights the significance of sEVs in drug-sensitive dynamics and lays the groundwork for investigating therapeutic interventions that target EV-mediated sensitivity in giardiasis.

Mesenchymal stem cell (MSC) transplantation is considered one of the most promising regenerative strategies for treating degenerative musculoskeletal diseases, yet its underlying therapeutic mechanisms remain incompletely understood. In this study, we demonstrate that transplanted MSCs regulate apoptosis and DNA damage repair (DDR) in senescent nucleus pulposus cells (NPCs) by releasing apoptotic extracellular vesicles (ApoEVs), thereby delaying the process of intervertebral disc degeneration (IVDD). Mechanistically, we found that NPCs in degenerated discs exhibit abnormal subcellular localization of the deubiquitinase ubiquitin specific peptidase 5 (USP5), with excessive cytoplasmic retention leading to aberrant ubiquitination and degradation of the E2F transcription factor 1 (E2F1). Following transplantation into the degenerative disc microenvironment, MSCs undergo extensive apoptosis in the short-term and release ApoEVs enriched in highly acetylated USP5. These vesicles promote nuclear translocation of USP5 in NPCs, which stabilizes E2F1 by preventing its ubiquitin-mediated degradation. This cascade reduces DNA damage and apoptosis in NPCs and enhances their functional activity. Overall, our findings reveal a previously unrecognized mechanism by which apoptotic donor MSCs exert therapeutic effects through intercellular communication, specifically by modulating recipient NPCs apoptosis and DDR pathways. This study underscores the critical role of donor cell apoptosis in the therapeutic efficacy of stem cell transplantation and provides new insights for optimizing regenerative medicine strategies.

Extracellular vesicle (EV)-enriched secretomes are emerging as a new and innovative therapeutic option in the field of regenerative medicine. The clinical use of EV-enriched secretome-based products requires manufacturing processes and quality control (QC) testing that comply with current good manufacturing practice (GMP). The goal of this work was to develop a robust and reproducible large-scale GMP-compliant process for the production of an EV-enriched secretome derived from cardiovascular progenitor cells (CPC), including the vesiculation of CPC, purification and concentration of the product; and sterilising filtration. QC strategies for in-process and release testing of an investigational medicinal product (IMP) were developed to guarantee quantity, safety, purity and identity. The IMP showed biological activity and was non-immunogenic in vitro, and showed no signs of toxicity or tumour development in vivo. The IMP was approved for use in a single-centre Phase I clinical trial by the French National Agency for Medicines and Health (ANSM) for the treatment of heart failure. The IMP is stored between –65°C and –85°C and can be easily diluted by the hospital pharmacy for infusion to the patient. This work represents a major advance for the use of CPC derived EV-enriched secretomes as a biological drug for cardiac clinical applications.

Trial Registration: ClinicalTrials.gov identifier: NCT05774509