Journal of extracellular biology最新文献

{"title":"CD147 Mediates the Metabolic Reprogramming of Cancer Associated Fibroblasts Induced by EVs Released by Differentiating Cancer Stem Cells","authors":"Filomena Colella,&nbsp;Federica Calapà,&nbsp;Giulia Artemi,&nbsp;Erica Pazzaglia,&nbsp;Rita Colonna,&nbsp;Sara Vitale,&nbsp;Giacomo Lazzarino,&nbsp;Federica Vincenzoni,&nbsp;Micol Eleonora Fiori,&nbsp;Ruggero De Maria,&nbsp;Sara Lucchisani,&nbsp;Giannicola Genovese,&nbsp;Luigi Perelli,&nbsp;Barbara Tavazzi,&nbsp;Alessandro Sgambato,&nbsp;Donatella Lucchetti","doi":"10.1002/jex2.70039","DOIUrl":"https://doi.org/10.1002/jex2.70039","url":null,"abstract":"<p>Several reports have demonstrated that CD147, an N-glycosylated protein that is exchanged by cells in soluble form or through small extracellular vesicles (sEVs), can promote cancer progression. However, its activity related to EVs in colorectal cancer (CRC) is still not fully understood. Previously, we showed that sEV secretion during CRC stem cell (CR-CSCs) differentiation is partially controlled by CD147, and that CD147-expressing sEVs (sEVs-CD147) activate a signalling cascade in recipient cells, inducing molecular invasive features in CR-CSCs. In the present study, we demonstrated that sEVs-CD147 increase the expression of myofibroblast and activation markers in cancer-associated fibroblasts (CAF). In sEVs-CD147-activated CAF, aerobic glycolysis was also triggered by the β-catenin signalling pathway and induced lactate release. These effects were associated with NFKβ upregulation and NO secretion that caused increased cytokines production and VEGF release, respectively. Furthermore, co-culture with CAF promoted CR-CSC invasivity in vitro and tumour growth in vivo. Spatial proteomics analysis confirmed in vivo the activation of fibroblasts and the modulation of their metabolic features, within their biological context, after their conditioning with CD147-expressing sEVs. Our findings indicate that sEV-packaged CD147 is involved in CAF activation, thus promoting tumour progression via stroma metabolism modification.</p>","PeriodicalId":73747,"journal":{"name":"Journal of extracellular biology","volume":"4 3","pages":""},"PeriodicalIF":0.0,"publicationDate":"2025-03-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/jex2.70039","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143639150","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Extracellular Vesicles Derived From Regenerating Tissue Promote Stem Cell Proliferation in the Planarian Schmidtea mediterranea","authors":"Priscilla N. Avalos Najera,&nbsp;Lily L. Wong,&nbsp;David J. Forsthoefel","doi":"10.1002/jex2.70040","DOIUrl":"https://doi.org/10.1002/jex2.70040","url":null,"abstract":"<p>Extracellular vesicles (EVs) are secreted nanoparticles composed of a lipid bilayer that carry lipid, protein, and nucleic acid cargo between cells as a mode of intercellular communication. Although EVs can promote tissue repair in mammals, their roles in animals with greater regenerative capacity are not well understood. Planarian flatworms are capable of whole-body regeneration due to pluripotent somatic stem cells called neoblasts that proliferate in response to injury. Here, using transmission electron microscopy, nanoparticle tracking analysis, and protein content examination, we showed that EVs enriched from the tissues of the planarian <i>Schmidtea mediterranea</i> had similar morphology and size as other eukaryotic EVs, and that these EVs carried orthologs of the conserved EV biogenesis regulators ALIX and TSG101. PKH67-labeled EVs were taken up efficiently by planarian cells, including S/G2 neoblasts, G1 neoblasts/early progeny, and differentiated cells. When injected into living planarians, EVs from regenerating tissue fragments enhanced the upregulation of neoblast-enriched and proliferation-related transcripts. In addition, EV injection increased the number of <i>F-ara</i>-EdU-labelled cells by 49% as compared to buffer injection only. Our findings demonstrate that regenerating planarians produce EVs that promote stem cell proliferation, and suggest the planarian as an amenable in vivo model for the study of EV function during regeneration.</p>","PeriodicalId":73747,"journal":{"name":"Journal of extracellular biology","volume":"4 3","pages":""},"PeriodicalIF":0.0,"publicationDate":"2025-03-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/jex2.70040","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143581574","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Comparative and Functional Analysis of Exosomal microRNAs During Semelparous Reproduction in Ayu Fish (Plecoglossus altivelis)","authors":"Yida Pan,&nbsp;Lingxin Meng,&nbsp;Kazuma Yoshida,&nbsp;Liangjie Qiu,&nbsp;Taichi Ito,&nbsp;Ryo Yonezawa,&nbsp;Kazutoshi Yoshitake,&nbsp;Shunsuke Saito,&nbsp;Nahoko Bailey-Kobayashi,&nbsp;Tetsuhiko Yoshida,&nbsp;Shiheharu Kinoshita,&nbsp;Shuichi Asakawa","doi":"10.1002/jex2.70038","DOIUrl":"https://doi.org/10.1002/jex2.70038","url":null,"abstract":"<p>As a life history strategy, some semelparous organisms, such as the ayu fish (<i>Plecoglossus altivelis</i>), reproduce only once in their lifetime and then die. They invest heavily in their single reproductive event, producing many offspring. However, the regulatory mechanisms that trigger mortality after reproduction are not well understood. Exosomes serve as an essential pathway for intercellular communication, with internal microRNA (miRNA) playing a crucial role in regulating physiological activities within the organism. This study aimed to elucidate the function of exosomal miRNA during the semelparous reproduction period in <i>P. altivelis</i>. Exosomes were successfully extracted from <i>P. altivelis</i> plasma during reproduction, and abundant miRNA molecules were discovered through small RNA sequencing. The miRNA expression patterns in ayu fish during reproduction exhibited notable differences between females and males. Furthermore, it was observed that key cellular processes and signalling pathways associated with intercellular transmission and intracellular stress are regulated by miRNA expression, and these changes in regulation may be responsible for post-breeding mortality. This study enhances our understanding of the function of exosomal miRNA during semelparous reproduction in ayu fish and provides further insight into the intrinsic mechanisms of ageing and mortality in organisms.</p>","PeriodicalId":73747,"journal":{"name":"Journal of extracellular biology","volume":"4 3","pages":""},"PeriodicalIF":0.0,"publicationDate":"2025-03-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/jex2.70038","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143581573","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Extracellular Vesicles From Follicular Fluid in Infertile Women: Size, Morphology and miRNA Content Analysis","authors":"Solène Ducarre,&nbsp;Regina Maria Chiechio,&nbsp;Gregory Moulin,&nbsp;Ester Butera,&nbsp;Aurélien Dupont,&nbsp;Christophe Penno,&nbsp;Abdelhak El Amrani,&nbsp;Claire Heichette,&nbsp;Pascale Even-Hernandez,&nbsp;Valérie Marchi,&nbsp;Célia Ravel","doi":"10.1002/jex2.70041","DOIUrl":"https://doi.org/10.1002/jex2.70041","url":null,"abstract":"<p>The declining birth rates and fertility challenges in Europe have intensified global concerns over rising infertility, particularly among women. This study decisively investigates follicular fluid-related extracellular vesicles (FF-EVs) from infertile patients with polycystic ovary syndrome (PCOS) or diminished ovarian reserve (DOR) undergoing in vitro fertilization (IVF), comparing them to a healthy control group. We have identified significant variations in protein content and polydispersity in crude follicular fluid using UV-Vis absorption and dynamic light scattering (DLS) techniques. Furthermore, the morphology of the extracellular vesicles (EVs) and the patterns of non-coding RNA content, including miRNAs, reveal distinct differences in infertile patients. These findings offer critical insights into the molecular signatures associated with these conditions. This study plays a vital role in advancing reproductive healthcare by pinpointing potential targets that can enhance diagnosis and deepen our understanding of ovarian disorders.</p>","PeriodicalId":73747,"journal":{"name":"Journal of extracellular biology","volume":"4 3","pages":""},"PeriodicalIF":0.0,"publicationDate":"2025-03-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/jex2.70041","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143554386","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Establishment of an experimental system to analyse extracellular vesicles during apoplastic fungal pathogenesis","authors":"Nathaniel Hearfield,&nbsp;Dominik Brotherton,&nbsp;Zedi Gao,&nbsp;Jameel Inal,&nbsp;Henrik U. Stotz","doi":"10.1002/jex2.70029","DOIUrl":"https://doi.org/10.1002/jex2.70029","url":null,"abstract":"<p>Phoma stem canker disease of oilseed rape (<i>Brassica napus</i>) is caused by the extracellular fungal pathogen <i>Leptosphaeria maculans</i>. Although this pathogen resides exclusively in apoplastic spaces surrounding plant cells, the significance of extracellular vesicles (EVs) has not been assessed. Here, we show a method to collect apoplastic fluids (AFs) from infected leaves or cotyledons for collection of EVs during the process of host colonisation. The 15,000 × <i>g</i> supernatants of AFs were shown to contain ribulose-bisphosphate carboxylase (RuBisCO) at 7 days post-inoculation with <i>L. maculans</i>, a protein that was absent from unchallenged cotyledons. RuBisCO release coincided with the switch from biotrophy to necrotrophy, suggesting the involvement of host cell death. However, RuBisCO release did not differ between compatible and incompatible interactions, suggesting necrotrophic host cell death might not be the only process involved. EVs were also collected from axenic fungal cultures and characterised for their particle size distribution using nanoparticle tracking analysis and transmission electron microscopy. The protein composition of EV-enriched fractions was analysed using SDS-PAGE and proteomics. Enrichment analysis of gene ontology terms provided evidence for involvement of glucan and chitin metabolism as well as catalase and peptidase activities. Most of the proteins identified have previously been found in EV studies and/or EV databases, and for most of the proteins evidence was found for an involvement in pathogenicity/virulence.</p>","PeriodicalId":73747,"journal":{"name":"Journal of extracellular biology","volume":"4 2","pages":""},"PeriodicalIF":0.0,"publicationDate":"2025-02-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/jex2.70029","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143424174","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Comparison of Methods for Isolation and Characterization of Total and Astrocyte-Enriched Extracellular Vesicles From Human Serum and Plasma","authors":"Leandra K. Figueroa-Hall,&nbsp;Kaiping Burrows,&nbsp;Ahlam M. Alarbi,&nbsp;Bethany N. Hannafon,&nbsp;Cole Hladik,&nbsp;Chibing Tan,&nbsp;Rajagopal Ramesh,&nbsp;Jennifer L. Stewart,&nbsp;Victoria B. Risbrough,&nbsp;Martin P. Paulus,&nbsp;T. Kent Teague","doi":"10.1002/jex2.70035","DOIUrl":"https://doi.org/10.1002/jex2.70035","url":null,"abstract":"<p>Extracellular vesicles (EV) which play critical roles in intercellular communication, have garnered interest as biomarkers with researchers studying brain-related disease processes due to their ability to be isolated from various biofluids. Astrocytes, a type of glial cell, play a critical role in neuronal regulation and function. As such, EV enriched from astrocytes can be used to interrogate cargo and identify mechanisms by which astrocytes communicate with other cells of the central nervous system or shed light on pathophysiological conditions. This manuscript compared five EV isolation methods (differential ultracentrifugation [dUC], precipitation, precipitation + purification, silicon carbon resin and size exclusion chromatography [SEC]) using small volumes of human plasma and serum with a focus on immunocapture of astrocyte-enriched EV (AEEV), with the excitatory amino acid transporter 1, or GLAST. Methods were evaluated on yield, purity, recovery and downstream application to include immunoassays for tetraspanin, immune and astrocyte markers. Results revealed that whilst precipitation-based methods such as ExoQuick yielded higher EV concentrations, size exclusion (SmartSEC, qEV) provided greater purity, emphasizing a trade-off between yield and purity. This study provides a comprehensive resource for researchers in selecting EV isolation methods tailored to small biobanked clinical samples, with the goal of advancing biomarker discovery in Neuroscience.</p>","PeriodicalId":73747,"journal":{"name":"Journal of extracellular biology","volume":"4 2","pages":""},"PeriodicalIF":0.0,"publicationDate":"2025-02-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/jex2.70035","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143404366","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Molecular insights into the unique properties of the blood-circulating proteasome","authors":"Yegor Leushkin,&nbsp;David Morgenstern,&nbsp;Shifra Ben-Dor,&nbsp;Rebecca Haffner-Krausz,&nbsp;Katharina Zittlau,&nbsp;Gili Ben-Nissan,&nbsp;Michal Sharon","doi":"10.1002/jex2.70034","DOIUrl":"10.1002/jex2.70034","url":null,"abstract":"<p>Proteasomes are essential for protein degradation and maintaining cellular balance, yet their roles in extracellular fluids are not well understood. Our study investigates the freely circulating proteasome in blood, to uncover its unique molecular characteristics, compared to its intracellular counterparts. Using a transgenic mouse model, mass spectrometry, and biochemical tools, we show that the predominant proteasome in serum is the free uncapped 20S particle, which seems to assemble intracellularly before entering the bloodstream. This serum proteasome is composed of constitutive and immuno subunits and exhibits all three catalytic activities. Moreover, the complex displays distinct post-translational modifications, indicating specialization for extracellular roles, as demonstrated by its enhanced caspase-like activity. We also found that physiological stress significantly upregulates serum 20S proteasome levels, paralleling human data. This research highlights the specialized characteristics of circulating proteasomes, offering new insights into protein turnover in the blood with significant implications for understanding proteostasis beyond the intracellular environment.</p>","PeriodicalId":73747,"journal":{"name":"Journal of extracellular biology","volume":"4 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2025-01-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11770374/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143054383","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Mesenchymal stem cell-derived exosomes mitigate amyloid β-induced retinal toxicity: Insights from rat model and cellular studies","authors":"Amanda Qarawani,&nbsp;Efrat Naaman,&nbsp;Rony Ben-Zvi Elimelech,&nbsp;Michal Harel,&nbsp;Shahaf Sigal-Dror,&nbsp;Tali Ben-Zur,&nbsp;Tamar Ziv,&nbsp;Daniel Offen,&nbsp;Shiri Zayit-Soudry","doi":"10.1002/jex2.70024","DOIUrl":"10.1002/jex2.70024","url":null,"abstract":"<p>Amyloid β (Aβ) has emerged as a pathophysiological driver in age-related macular degeneration (AMD), emphasizing its significance in the aetiology of this prevalent sight-threatening condition. The multifaceted nature of AMD pathophysiology, presumably involving diverse retinal cascades, corresponds with the complexity of Aβ-induced retinopathy. Therefore, targeting a broad array of pathogenic processes holds promise for therapeutic intervention in AMD-associated retinal pathology. This study investigates the potential of exosomes derived from adipose tissue mesenchymal stem cells (AT-MSC-Exosomes) in alleviating Aβ-induced retinotoxicity. Through intravitreal injections in wild-type rats and RPE-like cell culture experiments, we examined the protective effects of AT-MSC-Exosomes against Aβ42 retinotoxicity. Our findings reveal that pre-treatment with AT-MSC-Exosomes enabled nearly-intact retinal function in vivo and maintained retinal cell viability in vitro, evidenced by longitudinal electroretinography (ERG) and XTT proliferation assays, respectively. Fluorescent labelling demonstrated increased migration of AT-MSC-Exosomes towards retinal cells under conditions of amyloid-related toxicity. Proteomic analysis indicated a decrease in the retinal levels of heat-shock proteins activated by pathogenic Aβ fibrils following AT-MSC-Exosome treatment. Similarly, immunostaining highlighted the modulation of α-crystallin expression in retinal astrocytes by AT-MSC-Exosomes. These results suggest the potential therapeutic relevance of AT-MSC-Exosomes in Aβ-related retinal pathology, offering a promising avenue for future AMD treatment strategies.</p>","PeriodicalId":73747,"journal":{"name":"Journal of extracellular biology","volume":"4 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2025-01-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11752158/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143025907","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Purification of mesenchymal stromal cell-derived small extracellular vesicles using ultrafiltration","authors":"Rui Lei,&nbsp;Shuai Ren,&nbsp;Hua Ye,&nbsp;Zhanfeng Cui","doi":"10.1002/jex2.70030","DOIUrl":"10.1002/jex2.70030","url":null,"abstract":"<p>Mesenchymal stromal cell-derived small extracellular vesicles (MSC-sEVs) are pivotal for the curative effects of mesenchymal stromal cells, but their translation into clinical products is hindered by the technical challenges of scaled production and purification. Ultrafiltration, a pressure-driven membrane separation method, is well known as an efficient, scalable, and cost-effective approach for bioseparation. However, there has been little study so far that comprehensively evaluates the potential application of ultrafiltration for scaled sEV isolation and purification. In this study, the feasibility and effectiveness of ultrafiltration for MSC-sEV isolation and purification are studied, and the effects of key process design and operational parameters, including the membrane pore size, transmembrane pressure (TMP), stirring speed (shear rate), feed concentration, are quantified using a stirred cell setup. Results revealed that 500 kDa molecular weight cut-off (MWCO) polyethersulfone membrane demonstrated superior suitability for MSC-sEV separation, yielding higher purity and productivity compared to 100 and 300 kDa MWCO membranes of the same material. The MSC-sEV productivity and purity could also be improved by applying a moderate stirring speed and lower operational pressure, respectively. Isovolumetric diafiltration was incorporated to enhance the purity of MSC-sEVs, successfully removing about 99% of protein contaminants by six diafiltration volumes (DVs). Subsequently, a fed-batch ultra-diafiltration (UF/DF) process with optimised filtration parameters was developed and compared with the currently most used ultracentrifugation (UC) method, showing exceptional effectiveness and performance in the isolation of MSC-sEVs: it increased the recovery of MSC-sEV from 20.59% to 60.88% (about three folds increase) and nearly doubled the purity, while also reducing processing time from over 4 h to 3.5 h, with a potential further reduction to less than 2.5 h through automation. The study concludes that ultrafiltration could be a promising method for both lab-scale preparation and industrial-scale manufacture of MSC-sEVs, offering advantages of high recovery, scalability, fast, and cost-effectiveness.</p>","PeriodicalId":73747,"journal":{"name":"Journal of extracellular biology","volume":"4 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2025-01-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11739894/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143017722","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Brain penetration of peripheral extracellular vesicles from Alzheimer's patients and induction of microglia activation","authors":"Hermine Counil,&nbsp;Rummenigge Oliveira Silva,&nbsp;Jean-Michel Rabanel,&nbsp;Charlotte Zaouter,&nbsp;Mohamed Haddad,&nbsp;Mohamed Raâfet Ben Khedher,&nbsp;Davide Brambilla,&nbsp;Tamas Fülöp,&nbsp;Shunmoogum A. Patten,&nbsp;Charles Ramassamy","doi":"10.1002/jex2.70027","DOIUrl":"10.1002/jex2.70027","url":null,"abstract":"<p>Alzheimer's disease (AD) is an age-related neurodegenerative pathology. Brain-derived extracellular vesicles (EVs) have been demonstrated to be implicated in AD pathogenesis by facilitating the propagation of Tau, amyloid-β and inflammatory cytokines. However, the impact of peripheral EVs (pEVs) in AD pathogenesis remains poorly investigated. The objective of our study was to compare the passage of pEVs from adults, cognitively healthy elderly, and AD patients through the blood-brain barrier (BBB), to evaluate their uptake in the brain and to assess their impact on the microglia activity using in vitro and in vivo models. To this end, pEVs were enriched, characterized, and fluorescently labelled. The passage of pEVs through the endothelial bEnd.3 cells was studied in a Transwell device with either neuronal or microglia cells seeded at the bottom of the well. Following the internalization of pEVs from AD patients, microglia adopted an amoeboid morphology and released a heightened level of pro-inflammatory cytokine IL-6. To further assess their in vivo transport across the BBB, pEVs were injected into the blood circulation of 2-days post-fertilization Tg(<i>flk1:EGFP</i>) zebrafish. The biodistribution of pEVs was monitored at 1 and 24 h post-injection using confocal microscopy. We demonstrated that pEVs traverse the BBB by transcytosis and subsequently diffuse progressively into the brain. pEVs were then internalized by neuronal and radial glial cells as seen in <i>Tg(huc:EGFP)</i> and <i>Tg(gfap:EGFP)</i> zebrafish, respectively. Additional experiments were performed with the intrahippocampal injection of pEVs in the mouse, indicating their spreading throughout the brain and their uptake by neuronal and glial cells. These findings contribute to novel insights into the fate of pEVs following their passage through the BBB in vitro and in vivo, and demonstrate for the first time that pEVs from AD patients affect microglia activity. This suggests a potential mechanism through which peripheral tissue cues may contribute to AD pathogenesis.</p>","PeriodicalId":73747,"journal":{"name":"Journal of extracellular biology","volume":"4 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2025-01-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11740088/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143017715","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}