Journal of Extracellular Vesicles最新文献

{"title":"Identification of MARVELous Protein Markers for Phytophthora infestans Extracellular Vesicles","authors":"Susan Breen,&nbsp;Hazel McLellan,&nbsp;Wei Wang,&nbsp;Shumei Wang,&nbsp;Lydia Welsh,&nbsp;Jasmine Pham,&nbsp;Stephen C. Whisson,&nbsp;Petra C. Boevink,&nbsp;Paul R. J. Birch","doi":"10.1002/jev2.70101","DOIUrl":"10.1002/jev2.70101","url":null,"abstract":"<p>Extracellular vesicles (EVs) are released from cells by unconventional secretion, but little is known about the biogenesis routes, composition or cargoes of EVs from fungal or oomycete plant pathogens. We investigated the proteome of EV-associated proteins secreted by the oomycete <i>Phytophthora infestans</i>, cause of potato late blight disease. We found that vesicle-associated proteins, transmembrane proteins and RxLR effectors, which are delivered into host cells to suppress immunity, were enriched in the EV proteome. By contrast, the EV-independent secreted proteome was enriched in cell wall modifying enzymes and apoplastic effectors which act outside plant cells. Two proteins, each containing two tetraspanning MARVEL domains, PiMDP1 and PiMDP2, were associated with <i>P. infestans</i> EVs. PiMDP1 and PiMDP2 were co-buoyant with RxLR effectors in sucrose density fractions containing EVs and co-localised frequently with each other and with RxLRs at vesicles within pathogen hyphae grown <i>in vitro</i> and during infection. Interestingly, PiMDP2, which is up-regulated during the early biotrophic phase of infection, accumulates at the haustorial interface, a major site of effector secretion during infection. We argue that PiMDP1 and PiMDP2 are molecular markers that will facilitate studies of the biogenesis and secretion of infection-associated <i>P. infestans</i> EVs.</p>","PeriodicalId":15811,"journal":{"name":"Journal of Extracellular Vesicles","volume":"14 9","pages":""},"PeriodicalIF":14.5,"publicationDate":"2025-09-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://isevjournals.onlinelibrary.wiley.com/doi/epdf/10.1002/jev2.70101","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145129327","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"TurboID-Mediated Profiling of Glioblastoma-Derived Extracellular Vesicle Cargo Proteins","authors":"Marissa N. Russo,&nbsp;Emily S. Norton,&nbsp;Maria José Ulloa-Navas,&nbsp;Natanael Zarco,&nbsp;Weiwei Wang,&nbsp;TuKiet T Lam,&nbsp;Alfredo Quiñones-Hinojosa,&nbsp;Veronique V. Belzil,&nbsp;Hugo Guerrero-Cázares","doi":"10.1002/jev2.70158","DOIUrl":"10.1002/jev2.70158","url":null,"abstract":"<p>Glioblastoma (GBM), the most aggressive primary brain tumour in adults, presents significant challenges due to its universal recurrence and limited survival rates. A key driver of GBM progression is the subpopulation of brain tumour-initiating cells (BTICs), which contribute to therapy resistance and interact with the tumour microenvironment, particularly the cellular components in the subventricular zone (SVZ). Extracellular vesicles (EVs) are critical mediators of intercellular communication, carrying bioactive molecules, such as proteins and RNAs, that can modulate the behaviour of recipient cells. This study investigates the role of EVs in GBM's communication with non-cancer cells. We utilised the proximity-labelling system TurboID to achieve global and unbiased labelling of proteins within GBM-derived EVs. By inducing TurboID expression in primary-cultured human BTICs from GBM patients, we achieved efficient biotinylation of EV proteins without compromising vesicle integrity, and performed proteomic analysis of biotinylated proteins, which revealed a diverse cargo within BTIC-EVs. Our method marks the first implementation of TurboID for unbiased global labelling of EV protein cargo in primary cells. This approach facilitates the investigation of EV-mediated communication and potential therapeutic targets, contributing to the understanding of GBM's complex interactions with the brain's microenvironment and identification of biomarkers for improved diagnosis and treatment response.</p>","PeriodicalId":15811,"journal":{"name":"Journal of Extracellular Vesicles","volume":"14 9","pages":""},"PeriodicalIF":14.5,"publicationDate":"2025-09-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://isevjournals.onlinelibrary.wiley.com/doi/epdf/10.1002/jev2.70158","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145124665","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Selective EV Protein Sorting and Pathway Perturbation in AML Upon Synergistic FLT3 and Hedgehog Pathway Inhibition","authors":"Constantin Blöchl,&nbsp;Gabriele Blümel,&nbsp;Martin Wolf,&nbsp;Christof Regl,&nbsp;Heide-Marie Binder,&nbsp;Suzana Tesanovic,&nbsp;Daniel Lankes,&nbsp;Nicole Maeding,&nbsp;Peter W. Krenn,&nbsp;Dirk Strunk,&nbsp;Fritz Aberger,&nbsp;Christian G. Huber","doi":"10.1002/jev2.70163","DOIUrl":"10.1002/jev2.70163","url":null,"abstract":"<p>Acute myeloid leukaemia (AML) is a haematologic malignancy with high relapse incidence and mortality. Approximately one-third of AML patients carry an fms-like tyrosine kinase 3 (FLT3) mutation, often associated with GLI expression and Hedgehog signalling. AML cells shape their microenvironment into a leukaemia-permissive space by releasing extracellular vesicles (EVs). EVs can transfer chemoresistance and thereby play an important role in refractory and relapsing diseases. Here, we discovered a synergistic effect of combined treatment with the FLT3 inhibitor Crenolanib and the Hedgehog pathway inhibitor HPI-1 in the AML cell lines MOLM-14 and MV4-11. In-depth comparative proteomics revealed alterations in the cellular and the EV proteome upon single or combined inhibition of FLT3 and GLI, highlighting affected pathways. By comparing cellular and EV proteomes, we found that transport of ribosomal proteins, such as RPS26 and RPL27A, and ErbB pathway members such as GAB1, GRB2 and SHC1 to EVs, is selectively avoided upon treatment with Crenolanib. These findings were corroborated by comparative proteomics of EVs derived from AML patients and healthy donors. Ribosomal and ErbB signalling pathway proteins may play an important role in microenvironmental modulation by EVs, and Crenolanib treatment potentially acts by interfering with leukaemia niche formation.</p>","PeriodicalId":15811,"journal":{"name":"Journal of Extracellular Vesicles","volume":"14 9","pages":""},"PeriodicalIF":14.5,"publicationDate":"2025-09-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://isevjournals.onlinelibrary.wiley.com/doi/epdf/10.1002/jev2.70163","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145124746","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Sphingolipids in Extracellular Vesicles Released From the Skeletal Muscle Plasma Membrane Control Muscle Stem Cell Fate During Muscle Regeneration","authors":"Rhyma Hakkar,&nbsp;Caroline E. Brun,&nbsp;Pascal Leblanc,&nbsp;Emmanuelle Meugnier,&nbsp;Emmanuelle Berger-Danty,&nbsp;Olivier Blanc-Brude,&nbsp;Stefano Tacconi,&nbsp;Audrey Jalabert,&nbsp;Laura Reininger,&nbsp;Sandra Pesenti,&nbsp;Catherine Calzada,&nbsp;Vincent Gache,&nbsp;Sanjay B. Vasan,&nbsp;Julien Pichon,&nbsp;Thibaut Larcher,&nbsp;Elizabeth Errazuriz-Cerda,&nbsp;Christelle Cassin,&nbsp;Bong Hwan Sung,&nbsp;Alissa Weaver,&nbsp;Antonella Bongiovanni,&nbsp;Karl Rouger,&nbsp;Jean-Paul Pais de Barros,&nbsp;karim Bouzakri,&nbsp;Sophie Rome","doi":"10.1002/jev2.70164","DOIUrl":"10.1002/jev2.70164","url":null,"abstract":"<p>Extracellular vesicles (EVs) represent a cytokine-independent pathway though which skeletal muscle (SkM) cells influence the fate of neighbouring cells, thereby regulating SkM metabolic homeostasis and regeneration. Although SkM-EVs are increasingly being explored as a therapeutic strategy to enhance muscle regeneration or to induce the myogenic differentiation of induced pluripotent stem cells (iPSCs), the mechanisms governing their release from muscle cells remain poorly described. Moreover, because muscle regeneration involves a tightly regulated inflammatory response it also important to determine how inflammation alters SkM-EV cargo and function in order to design more effective EV-based therapies. To address this knowledge gap, we isolated and characterized the large and small EVs (lEVs, sEVs) released from SkM cells under basal conditions and in response to TNF-α, a well-established inflammatory mediator elevated in both acute muscle injury and chronic inflammatory conditions such as type 2 diabetes. We then evaluated the regenerative roles of these EV subtypes in vivo using a mouse model of cardiotoxin-induced muscle injury, with a specific focus on their bioactive sphingolipid content. Using transmission, scanning or cryo-electron microscopy, lipidomic profiling and an adenoviral construct to express labelled CD63 in myotubes, we demonstrated that SkM cells release both sEVs and lEVs primarily from the plasma membrane. Notably, sEVs were generated from specialized membrane folds enriched in the EV markers ALIX (ALG-2 interacting protein X) and TSG101, as well as lipid raft-associated lipids. During regeneration, sEVs promoted M1 macrophage polarization and migration and muscle stem cell (MuSC) differentiation, thereby accelerating muscle repair. In contrast, lEVs inhibited and promoted MuSC proliferation and impaired the transition from the pro-inflammatory to the anti-inflammatory response, an essential step for promoting MuSC differentiation. Treatment of isolated muscle fibres with SkM-EVs revealed that the distinct effects of sEVs and lEVs on MuSC behaviour and macrophage phenotype could be largely explained by differences in their lipid composition, particularly the ratio of sphingosine-1-phosphate (S1P) subspecies. However, TNF-α exposure altered these ratios in sEVs and impaired their regenerative functions on MuSC and their effect on macrophage migration and polarization. These results demonstrate for the first time the importance of the sphingolipid content of EVs released by skeletal muscle in their regenerative function within muscle tissue, largely explained by their role as carriers of different subspecies of sphingosine-1-phosphate. This suggests that modulating the sphingolipid composition of EVs could be a viable strategy to enhance the regenerative potential of muscle tissue in addition to therapeutic interventions.</p>","PeriodicalId":15811,"journal":{"name":"Journal of Extracellular Vesicles","volume":"14 9","pages":""},"PeriodicalIF":14.5,"publicationDate":"2025-09-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://isevjournals.onlinelibrary.wiley.com/doi/epdf/10.1002/jev2.70164","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145110973","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Epidermal Stem Cell-Derived Extracellular Vesicles Induce Fibroblasts Mesenchymal-Epidermal Transition to Alleviate Hypertrophic Scar Formation via miR-200s Inhibition of ZEB1 and 2","authors":"Miao Zhen,&nbsp;Juntao Xie,&nbsp;Rui Yang,&nbsp;Lijuan Liu,&nbsp;Hengdeng Liu,&nbsp;Xuefeng He,&nbsp;Suyue Gao,&nbsp;Junyou Zhu,&nbsp;Jingting Li,&nbsp;Bin Shu,&nbsp;Peng Wang","doi":"10.1002/jev2.70160","DOIUrl":"10.1002/jev2.70160","url":null,"abstract":"<p>Hypertrophic scar (HS) is a prevalent yet unresolved wound healing complication characterized by persistent hyperactive and proliferative fibroblasts, leading to excessive extracellular matrix (ECM) synthesis and collagen contraction. Our previous studies have identified epidermal stem cells (ESCs) as critical for wound healing and HS remodelling, with its extracellular vesicles (EVs) playing a vital role. However, the specific mechanisms remain unclear. In this study, we first discovered that ESC-EVs could effectively induce the mesenchymal-epidermal transition (MET) of HS fibroblasts (HSFs) and inhibit their biological activity. Furthermore, by next-generation sequencing and multiplexed CRISPR/Cas9 system, we elucidated that this therapeutic effect is mediated by the miR-200 family (miR-200s) encapsulated in ESC-EVs, which targeted and inhibited ZEB1 and ZEB2 in HSFs. This vital role and mechanism have been thoroughly validated in both in vitro cell experiments and in vivo rat tail HS (RHS) models. These findings not only shed light on a previously unidentified mechanism of ESC-EVs for HS, but also provide potential novel targets and strategies for its precise treatment.</p>","PeriodicalId":15811,"journal":{"name":"Journal of Extracellular Vesicles","volume":"14 9","pages":""},"PeriodicalIF":14.5,"publicationDate":"2025-09-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12438467/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145069708","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Outer Membrane Vesicles as a Versatile Platform for Vaccine Development: Engineering Strategies, Applications and Challenges","authors":"Asja Garling,&nbsp;Frédéric Auvray,&nbsp;Mathieu Epardaud,&nbsp;Éric Oswald,&nbsp;Priscilla Branchu","doi":"10.1002/jev2.70150","DOIUrl":"10.1002/jev2.70150","url":null,"abstract":"<p>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.</p>","PeriodicalId":15811,"journal":{"name":"Journal of Extracellular Vesicles","volume":"14 9","pages":""},"PeriodicalIF":14.5,"publicationDate":"2025-09-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://isevjournals.onlinelibrary.wiley.com/doi/epdf/10.1002/jev2.70150","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145008002","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Targeted Blockage of Pathological Extracellular Vesicles and Particles From Fibroblast-Like Synoviocytes for Osteoarthritis Relief: Proteomic Analysis and Cellular Effect","authors":"Bin Liu,&nbsp;Yansi Xian,&nbsp;Tao Shen,&nbsp;Yu Ben,&nbsp;Wenshu Wu,&nbsp;Yong Shi,&nbsp;Xueying An,&nbsp;Rui Peng,&nbsp;Wentian Gao,&nbsp;Wang Gong,&nbsp;Xiang Chen,&nbsp;Baosheng Guo,&nbsp;Qing Jiang","doi":"10.1002/jev2.70162","DOIUrl":"10.1002/jev2.70162","url":null,"abstract":"<p>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 <i>Rab27a</i>, 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.</p>","PeriodicalId":15811,"journal":{"name":"Journal of Extracellular Vesicles","volume":"14 9","pages":""},"PeriodicalIF":14.5,"publicationDate":"2025-09-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://isevjournals.onlinelibrary.wiley.com/doi/epdf/10.1002/jev2.70162","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145012980","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Cancer Cell-Secreted miR-33a Reduces Stress Granule Formation by Targeting Polyamine Metabolism in Stroma to Promote Tumourigenesis","authors":"Sheng Hu,&nbsp;Xu Li,&nbsp;Qixin Hu,&nbsp;Chenyu Wang,&nbsp;Ao Hua,&nbsp;Gang Deng,&nbsp;Wenda Huang,&nbsp;Xiaoyu Fu,&nbsp;Haifeng Zhou,&nbsp;Xiaohui Zhang,&nbsp;Meixin Li,&nbsp;Juan Wu,&nbsp;Mingzhou Chen,&nbsp;Xiaolu Zhao,&nbsp;Lianyun Li,&nbsp;Zifu Li,&nbsp;Min Wu,&nbsp;Juanjuan Li,&nbsp;Wei Yan","doi":"10.1002/jev2.70153","DOIUrl":"10.1002/jev2.70153","url":null,"abstract":"<p>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.</p>","PeriodicalId":15811,"journal":{"name":"Journal of Extracellular Vesicles","volume":"14 9","pages":""},"PeriodicalIF":14.5,"publicationDate":"2025-09-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://isevjournals.onlinelibrary.wiley.com/doi/epdf/10.1002/jev2.70153","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144935092","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Extracellular Vesicles in Arthropods: Biogenesis, Functions, Isolation Methods and Applications","authors":"Simon Remans,&nbsp;Stijn Van den Brande,&nbsp;Jozef Vanden Broeck,&nbsp;Dulce Santos","doi":"10.1002/jev2.70156","DOIUrl":"10.1002/jev2.70156","url":null,"abstract":"<p>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.</p>","PeriodicalId":15811,"journal":{"name":"Journal of Extracellular Vesicles","volume":"14 9","pages":""},"PeriodicalIF":14.5,"publicationDate":"2025-09-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://isevjournals.onlinelibrary.wiley.com/doi/epdf/10.1002/jev2.70156","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144935089","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A Compendium of Bona Fide Reference Markers for Genuine Plant Extracellular Vesicles and Their Degree of Phylogenetic Conservation","authors":"Miriam M. Rodríguez de Lope,&nbsp;Ibone R. Sánchez-Pajares,&nbsp;Estela Herranz,&nbsp;Cristina M. López-Vázquez,&nbsp;Ainara González-Moro,&nbsp;Alan Rivera-Tenorio,&nbsp;Carlos González-Sanz,&nbsp;Soledad Sacristán,&nbsp;Eduardo Chicano-Gálvez,&nbsp;Fernando de la Cuesta","doi":"10.1002/jev2.70147","DOIUrl":"10.1002/jev2.70147","url":null,"abstract":"<p>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 <i>bona fide</i> 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: <i>Arabidopsis thaliana</i> (Arath-EVs) and <i>Brassica oleracea</i> (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.</p>","PeriodicalId":15811,"journal":{"name":"Journal of Extracellular Vesicles","volume":"14 9","pages":""},"PeriodicalIF":14.5,"publicationDate":"2025-09-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://isevjournals.onlinelibrary.wiley.com/doi/epdf/10.1002/jev2.70147","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144935093","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}