Journal of Extracellular Vesicles最新文献

{"title":"Roadblocks of Urinary EV Biomarkers: Moving Toward the Clinic","authors":"Marvin Droste,&nbsp;Maija Puhka,&nbsp;Martin E. van Royen,&nbsp;Monica S. Y. Ng,&nbsp;Charles Blijdorp,&nbsp;Gloria Alvarez-Llamas,&nbsp;Francesc E. Borràs,&nbsp;Anja K. Büscher,&nbsp;Benedetta Bussolati,&nbsp;James W. Dear,&nbsp;Juan M. Falcón-Pérez,&nbsp;Bernd Giebel,&nbsp;Cristina Grange,&nbsp;Ewout J. Hoorn,&nbsp;Janne Leivo,&nbsp;Metka Lenassi,&nbsp;Alicia Llorente,&nbsp;Fabrice Lucien,&nbsp;Inge Mertens,&nbsp;Harald Mischak,&nbsp;Desmond Pink,&nbsp;Tobias Tertel,&nbsp;Swasti Tiwari,&nbsp;Dolores Di Vizio,&nbsp;Peter S. T. Yuen,&nbsp;Natasa Zarovni,&nbsp;Guido Jenster,&nbsp;Dylan Burger,&nbsp;Elena S. Martens-Uzunova,&nbsp;Uta Erdbrügger","doi":"10.1002/jev2.70120","DOIUrl":"10.1002/jev2.70120","url":null,"abstract":"<p>Despite remarkable interest in the biomarker potential of urinary extracellular vesicles (uEVs) and the identification of numerous promising candidates, their clinical translation still presents multiple challenges. The opportunities for successful translation are obvious, yet the main roadblocks on the way have hardly been systematically considered and more coordinated approaches are needed to overcome them. In the present review article, we have identified the most relevant roadblocks of clinical translation of urinary EV-based biomarkers and discuss possible solutions to overcome them. These roadblocks are categorized as fundamental and technical but also related to development of novel biomarker assays and clinical acceptance. In addition, hurdles within the regulatory approval process are discussed. It is clear that various roadblocks to clinical translation of urinary EV biomarkers exist; however, they are addressable by promoting rigor and reproducibility as well as collaboration between basic and clinical scientists, clinicians, industry and regulatory bodies. Moreover, knowledge of obstacles for assay development and regulatory requirements should already be considered when developing a new biomarker to maximize the chance of successful translation. This review presents not only a status quo, but also a roadmap for the further development of the field.</p>","PeriodicalId":15811,"journal":{"name":"Journal of Extracellular Vesicles","volume":"14 7","pages":""},"PeriodicalIF":14.5,"publicationDate":"2025-07-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/jev2.70120","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144647682","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":"ADAM Sheddase Activity Promotes the Detachment of Small Extracellular Vesicles From the Plasma Membrane","authors":"Chloé Bizingre,&nbsp;Zaira Arellano-Anaya,&nbsp;Flavien Picard,&nbsp;Mathéa Pietri,&nbsp;Anne Baudry,&nbsp;Florence Roussel,&nbsp;Clara Bianchi,&nbsp;Aurélie Alleaume-Butaux,&nbsp;Hector Ardila-Osorio,&nbsp;Maryse Romao,&nbsp;Grégory Lavieu,&nbsp;Graça Raposo,&nbsp;Benoit Schneider","doi":"10.1002/jev2.70114","DOIUrl":"10.1002/jev2.70114","url":null,"abstract":"<p>Small extracellular vesicles (SEVs) are involved in diverse functions in normal and pathological situations, including intercellular communication, immunity, metastasis and neurodegeneration. Cell release of SEVs is assumed to occur passively right after multivesicular bodies of the endocytic pathway fuse with the plasma membrane. We show here that the completion of SEV release depends on membrane-bound ADAM10 and ADAM17 sheddases that promote the detachment of SEVs from the cell surface by catalysing the cleavage of adhesion proteins of the SEV membrane. The intensity of ADAM10/17-mediated release of SEVs depends on a balanced control of 3-phosphoinositide–dependent kinase 1 (PDK1) and ERK1/2 signalling pathways converging on 90-kDa ribosomal S6 kinase-2 (RSK2), which, in turn, fine-tunes ADAM17 bioavailability and ADAM10/17 enzymatic activities at the plasma membrane, according to a mechanism that relies, at least in part, on variation of the rhomboid-like pseudoprotease iRhom2 cell surface level. By identifying a new proteolytic step involved in the basal release of SEVs, our work may help understand how the deregulation of ADAM10/17-mediated discharge of SEVs contributes to several pathological states.</p>","PeriodicalId":15811,"journal":{"name":"Journal of Extracellular Vesicles","volume":"14 7","pages":""},"PeriodicalIF":14.5,"publicationDate":"2025-07-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/jev2.70114","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144647578","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":"124I-labelled BMSC-Derived Extracellular Vesicles Deliver CRISPR/Cas9 Ribonucleoproteins With a GFP-Reporter System to Inhibit Osteosarcoma Proliferation and Metastasis","authors":"Yujie Pan,&nbsp;Xianteng Yang,&nbsp;Zhirui Zeng,&nbsp;Futao Liu,&nbsp;Jin Luo,&nbsp;Mao Shen,&nbsp;Wei Zhou,&nbsp;Jianyang Li,&nbsp;Guangfu Jiang,&nbsp;Li Sun,&nbsp;Haifeng Huang,&nbsp;Runsang Pan","doi":"10.1002/jev2.70130","DOIUrl":"10.1002/jev2.70130","url":null,"abstract":"<p>Metastasis constitutes the principal factor leading to the unfavourable prognosis of osteosarcoma patients. Hypoxia, as the inherent microenvironment of osteosarcoma, can upregulate HIF-1α via multiple pathways, thereby facilitating osteosarcoma proliferation and metastasis. Our previous research indicated that the inwardly rectifying potassium channel subfamily J member 2 (KCNJ2) inhibits the degradation of HIF-1α in osteosarcoma. Concurrently, HIF-1α upregulates the expression of KCNJ2 through a positive feedback regulatory mechanism. This positive regulatory mechanism significantly promotes the proliferation and metastasis of osteosarcoma. Therefore, the development of a KCNJ2-targeted therapeutic strategy capable of disrupting this reciprocal regulatory loop represents a crucial intervention for impeding osteosarcoma progression. The CRISPR/Cas9 targeted gene editing technology has garnered extensive attention in the field of tumour treatment due to its high efficiency and low off-target rate. Nevertheless, the relative lag of the delivery systems has restricted its application. The extracellular vesicles (EVs) secreted by bone marrow mesenchymal stem cells (BMSCs) have a natural targeting specificity for osteosarcoma and possess superior biocompatibility, making them ideal carriers for in vivo delivery. However, it is essential to confirm whether the CRISPR/Cas9 system mediated by EVs can accurately function intracellularly. Hence, we developed a fluorescence-based Cas9 editing efficiency reporter system. When CRISPR/Cas9 system induces double-strand breaks at specific target sites and results in frameshift mutations, osteosarcoma cells will stably express GFP. This system enables the transformation of gene editing events into quantifiable fluorescence signals. Furthermore, we engineered radiolabelled EVs derived from BMSCs to deliver the CRISPR/Cas9 system targeting KCNJ2. Using this reporter system, we confirmed their efficient gene-editing capabilities in vitro. Additionally, leveraging their radiolabelling properties, we validated their targeted distribution in vivo. Subsequent investigations revealed that our constructed ¹²⁴I@EVs-Cas9 effectively suppresses the proliferation and metastasis of osteosarcoma by targeting the inhibition of KCNJ2 expression and promoting HIF-1α ubiquitin-dependent degradation (as depicted in Graphical Abstract).</p>","PeriodicalId":15811,"journal":{"name":"Journal of Extracellular Vesicles","volume":"14 7","pages":""},"PeriodicalIF":14.5,"publicationDate":"2025-07-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/jev2.70130","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144649521","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":"Hypoxic Neural Stem Cells Enhance Spinal Cord Repair Through HIF-1a/RAB17-Driven Extracellular Vesicle Release","authors":"Tian Qin,&nbsp;Yiming Qin,&nbsp;Haicheng Wen,&nbsp;Tianding Wu,&nbsp;Chunyue Duan,&nbsp;Yong Cao,&nbsp;Yi Sun,&nbsp;Hongkang Zhou,&nbsp;Hongbin Lu,&nbsp;Liyuan Jiang,&nbsp;Jianzhong Hu,&nbsp;Chengjun Li","doi":"10.1002/jev2.70126","DOIUrl":"10.1002/jev2.70126","url":null,"abstract":"<p>Spinal cord injury (SCI) is a severe and complex condition that can lead to significant physical impairments and affect the life quality of patients. Neural stem cells (NSCs) transplantation holds as a promising therapeutic approach for SCI. However, the challenging post-SCI microenvironment limits NSCs effectiveness. Our current research has found that transplanted NSCs, though with lower survival and differentiation, still aided in injury repair. Hypoxia was identified as a stressor inducing the release of extracellular vesicles (EVs) from NSCs through HIF-1α/RAB17 enhancing SCI repair. By extracting and modifying these EVs derived from hypoxia treated NSCs with CAQK/Angiopep2 peptides, we were able to accurately deliver them to the injury site, enhancing recovery without relying on cell survival or differentiation. This study delved into the reparative role and underlying mechanisms of transplanted NSCs in SCI, focusing on their non-cellular contributions and developed an innovative, targeted strategy for the transplantation of EVs derived from NSCs, offering a cell-free, precision therapeutic intervention for the treatment of SCI.</p>","PeriodicalId":15811,"journal":{"name":"Journal of Extracellular Vesicles","volume":"14 7","pages":""},"PeriodicalIF":14.5,"publicationDate":"2025-07-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/jev2.70126","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144624689","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":"Improved Accessibility of Extracellular Vesicle Surface Molecules Upon Partial Removal of the Protein Corona by High Ionic Strength","authors":"András I. Försönits,&nbsp;Eszter Á. Tóth,&nbsp;Sára Jezsoviczky,&nbsp;Tünde Bárkai,&nbsp;Delaram Khamari,&nbsp;Alicia Galinsoga,&nbsp;Panna Királyhidi,&nbsp;Ágnes Kittel,&nbsp;Júlia Fazakas,&nbsp;Dorina Lenzinger,&nbsp;Hargita Hegyesi,&nbsp;Xabier Osteikoetxea,&nbsp;Tamás Visnovitz,&nbsp;Krisztina Pálóczi,&nbsp;Szilvia Bősze,&nbsp;Edit I. Buzás","doi":"10.1002/jev2.70124","DOIUrl":"10.1002/jev2.70124","url":null,"abstract":"<p>Recent studies have confirmed that a biomolecular corona forms around extracellular vesicles (EVs) in biofluids. However, there is limited data on how this adsorbed corona affects the accessibility of EV surface molecules. Here, we investigated various potential corona-stripping conditions for their ability to affect the immune detection of EVs. First, we artificially formed an EV corona around nascent HEK293T-PalmGFP cell-derived large EVs (lEVs) by incubating them with Cy5-labelled human plasma proteins. The co-localisation rate of plasma proteins and lEVs decreased significantly upon high-salt washing with NaCl, LiCl and KCl solutions, suggesting a considerable removal of the corona components. Additional evidence for corona modification was a significantly increased fluorescent annexin V binding to plasma lEVs and annexin V affinity capture of both THP1- and blood plasma-derived lEVs upon high-salt washing. A similar effect of high ionic strength was observed when THP1 lEVs were separated from a serum-containing medium, which allowed for corona formation, but not when EVs were produced under serum-free conditions. Using a MACSPlex kit and high-salt washing for small EVs from plasma and THP1 conditioned medium, we also demonstrated significantly improved immunodetection of 15 and 9 out of 37 surface markers, respectively. In this Technical Note, we present evidence that modifying the protein corona around EVs can significantly affect the immune detection of specific EV markers.</p>","PeriodicalId":15811,"journal":{"name":"Journal of Extracellular Vesicles","volume":"14 7","pages":""},"PeriodicalIF":14.5,"publicationDate":"2025-07-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/jev2.70124","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144624691","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":"STING Agonist Drug Delivery by Bacterial Extracellular Vesicles Induces Synergistic Immuno-Oncology Responses and Efficient Inhibition of Tumour Growth","authors":"Jan Lötvall,&nbsp;Negar Ordouzadeh,&nbsp;Rossella Crescitelli,&nbsp;Meghshree Deshmukh,&nbsp;Tao Jin,&nbsp;Kyong-Su Park","doi":"10.1002/jev2.70117","DOIUrl":"10.1002/jev2.70117","url":null,"abstract":"<p>Bacterial extracellular vesicles are spherical, nanosized structures with lipid bilayer membranes and can suppress tumour growth in cancer models. However, the efficacy of some of these models is limited. One potential way to enhance their effects is by loading the bacterial vesicles with immunostimulatory molecules. We have here utilised synthetic bacterial vesicles (SyBV), previously shown to have anti-tumour effects but with reduced side effects. We hypothesized that loading SyBV with a STimulator of InterferoN Genes (STING) agonist can enhance anti-tumour effects. SyBV were generated from <i>Escherichia coli</i> membranes through cell breakdown induced by lysozyme and ionic stress. The produced nanovesicles encapsulated the STING agonist (SyBV^STING). SyBV^STING synergistically activated dendritic cells, leading to enhanced production of Interferon-β. Furthermore, in vivo experiments showed that immunisation with SyBV^STING synergistically suppresses melanoma and colon cancer growth by increasing the tumour infiltration of T cells. Intratumoural or subcutaneous injection of the SyBV resulted in retention in the tumour tissue over 24 h, but with some distribution to local lymph nodes. A toxicology experiment resulted in no histopathological concerns with SyBV^STING. These findings show that SyBV loaded with a STING agonist synergistically enhance anti-tumour immunity and may be a promising clinical immuno-oncology tool.</p>","PeriodicalId":15811,"journal":{"name":"Journal of Extracellular Vesicles","volume":"14 7","pages":""},"PeriodicalIF":14.5,"publicationDate":"2025-07-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/jev2.70117","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144573145","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":"Small and Large Extracellular Vesicles From Human Preovulatory Follicular Fluid Display Distinct ncRNA Cargo Profiles and Differential Effects on KGN Granulosa Cells","authors":"Inge Varik,&nbsp;Katariina Johanna Saretok,&nbsp;Kristine Rosenberg,&nbsp;Ileana Quintero,&nbsp;Maija Puhka,&nbsp;Nataliia Volkova,&nbsp;Aleksander Trošin,&nbsp;Paolo Guazzi,&nbsp;Agne Velthut-Meikas","doi":"10.1002/jev2.70119","DOIUrl":"10.1002/jev2.70119","url":null,"abstract":"<p>Follicular fluid extracellular vesicles (FF EVs) facilitate communication between oocytes and somatic cells within the ovarian follicle, playing a pivotal role in follicular development. This study highlights the molecular and functional distinctions between small (SEV) and large (LEV) FF EV subpopulations, revealing their specialised regulatory roles in granulosa cell (GC) biology and their consequential impact on ovarian function. Single-EV profiling uncovered distinct tetraspanin distributions, with LEVs containing a lower proportion of CD9/CD63/CD81-positive particles compared to SEVs. Fluorescent labelling confirmed uptake of both SEVs and LEVs by GCs, supporting their capacity to impact cellular behaviour. Functionally, LEVs increased testosterone production by GCs, whilst SEVs had no effect on steroid hormone secretion, suggesting a specific role for LEVs in androgen biosynthesis. Transcriptomic analysis revealed extensive SEV-induced changes in GC gene expression, affecting pathways involved in transcription, TGF-β signalling, extracellular matrix (ECM) remodelling and cell cycle regulation. In contrast, LEVs elicited minimal transcriptional changes, primarily modulating genes associated with immune regulation and oxidative stress defence. Small RNA sequencing further revealed distinct non-coding RNA (ncRNA) profiles, with SEVs enriched in miRNAs targeting pathways critical for GC differentiation, whilst LEVs carried higher levels of piRNAs implicated in maintaining genomic stability. These findings advance our understanding of FF EV-mediated intercellular communication and underscore the importance of investigating EV subpopulations independently.</p>","PeriodicalId":15811,"journal":{"name":"Journal of Extracellular Vesicles","volume":"14 7","pages":""},"PeriodicalIF":14.5,"publicationDate":"2025-07-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/jev2.70119","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144573149","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":"Umbilical Cord-Mesenchymal Stromal Cell-Derived Extracellular Vesicles Target the Liver to Improve Neurovascular Health in Type 2 Diabetes With Non-Alcoholic Fatty Liver Disease","authors":"Minghao Du,&nbsp;Hao Yang,&nbsp;Jinyun Niu,&nbsp;Jing Huang,&nbsp;Lihong Wang,&nbsp;Junxiu Xi,&nbsp;Panpan Meng,&nbsp;Zhiyong Liu,&nbsp;Guaiguai Ma,&nbsp;Jiani Li,&nbsp;Xiaoyan Liu,&nbsp;Liang Guo,&nbsp;Mingjun Hu,&nbsp;Zhufang Tian,&nbsp;Bin Liu,&nbsp;Weiping Liu,&nbsp;Ashok K. Shetty,&nbsp;Shengxi Wu,&nbsp;Andrius Baskys,&nbsp;Qianfa Long","doi":"10.1002/jev2.70125","DOIUrl":"10.1002/jev2.70125","url":null,"abstract":"<p>Type 2 diabetes mellitus (T2DM) combined with non-alcoholic fatty liver disease (NAFLD) exacerbates metabolic dysregulation and neurovascular complications, presenting significant therapeutic challenges. We demonstrate, using SPECT/CT imaging, that extracellular vesicles (EVs) from mesenchymal stromal cells (MSCs) predominantly accumulate in the liver, where they deliver miR-31-5p to suppress platelet-derived growth factor B (PDGFB) produced by hepatic macrophages. This intervention impedes NAFLD progression and establishes a mechanistic link between liver repair and neurovascular improvement. Specifically, single-nucleus RNA sequencing reveals that PDGFB suppression enhances hippocampal pericyte recovery via the PDGFB-PDGFRβ axis and orchestrates the activation of growth differentiation factor 11 (GDF11), thus promoting neuroplasticity. Furthermore, AAV injections indicate that hepatic PDGFB modulation recalibrates transthyretin (TTR) dynamics, thereby restoring its neuroprotective functions and preventing its pathological deposition in the brain. These findings position MSC-EVs as a transformative therapeutic platform that leverages the liver-brain axis to address the intertwined metabolic and neurovascular complications of T2DM, offering a promising avenue for clinical translation.</p>","PeriodicalId":15811,"journal":{"name":"Journal of Extracellular Vesicles","volume":"14 7","pages":""},"PeriodicalIF":14.5,"publicationDate":"2025-07-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/jev2.70125","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144573150","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 Administered via Intrathecal Injection Mediate Safe Delivery of Nucleic Acids to the Central Nervous System for Gene Therapy","authors":"Brenda Wan Shing Lam,&nbsp;Melissa Tan,&nbsp;Chang Gao,&nbsp;Thach Tuan Pham,&nbsp;Lan Thi Ngoc Tran,&nbsp;Lan Ngoc Nguyen,&nbsp;Harwin Sidik,&nbsp;Matthias B. H. Lim,&nbsp;Anh Hong Le,&nbsp;Tram T. T. Nguyen,&nbsp;Nicole Lim,&nbsp;Lap Nguyen,&nbsp;Thuy Huong Nguyen,&nbsp;Yuin-Han Loh,&nbsp;Muhammad Waqas Usman,&nbsp;Minh T. N. Le","doi":"10.1002/jev2.70116","DOIUrl":"10.1002/jev2.70116","url":null,"abstract":"<p>Gene therapy holds great potential for treating neurological disorders, but its implementation is limited by the challenge of developing a safe and effective delivery method to the central nervous system (CNS). Red blood cell-derived extracellular vesicles (RBCEVs) have the potential to address these challenges due to their non-immunogenicity, non-cytotoxicity, ability to be redosed, and suitability for nucleic acid loading. In this study, we demonstrate the efficacy and safety of RBCEV-mediated nucleic acid delivery to the CNS. We found that RBCEVs administered through intrathecal injection are widely distributed across the CNS and efficiently taken up by neuronal cells. Delivery of RBCEVs loaded with GFP-encoding plasmids results in GFP expression in neurons. Our data also highlight the potential of RBCEVs to deliver plasmids encoding secretory proteins, resulting in protein secretion within the cerebrospinal fluid. Furthermore, experiments conducted in both mouse and non-human primate models indicate that intrathecal injection of plasmid-loaded RBCEVs do not lead to any systemic or local acute toxicity. In summary, our findings illustrate the potential of the RBCEV-based platform as a viable and safe approach for nucleic acid delivery to the CNS, facilitating further development of gene therapy for neurological disorders.</p>","PeriodicalId":15811,"journal":{"name":"Journal of Extracellular Vesicles","volume":"14 7","pages":""},"PeriodicalIF":14.5,"publicationDate":"2025-07-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/jev2.70116","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144573151","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":"Defining the Parameters for Sorting of RNA Cargo Into Extracellular Vesicles","authors":"Ahmed Abdelgawad,&nbsp;Yiyao Huang,&nbsp;Olesia Gololobova,&nbsp;Yanbao Yu,&nbsp;Kenneth W. Witwer,&nbsp;Vijay Parashar,&nbsp;Mona Batish","doi":"10.1002/jev2.70113","DOIUrl":"10.1002/jev2.70113","url":null,"abstract":"<p>Extracellular vesicles (EVs) are small particles that are released by cells and mediate cell–cell communication by transferring bioactive molecules such as RNA. RNA cargo of EVs, including coding and non-coding RNAs, can change the behaviour of recipient cells, affecting processes including gene expression, proliferation, and Fapoptosis. CircRNAs are stable and resistant to degradation and have been shown to be enriched in EVs. They play key roles in gene regulation and are also emerging as promising biomarkers for disease diagnosis due to their stability and disease-specific expression. Although microRNAs (miRNAs) are the most well studied RNA cargo of EVs, very little is known about the mechanisms of enrichment of circular RNAs (circRNAs) as well as long linear RNAs. Here, we take a comprehensive genome-wide approach to investigate the role of structuredness and shape along with GC%, size, exon count and coding potential, in the sorting and enrichment of circular and long linear RNAs into EVs. We developed a model using these parameters to predict the likelihood of EV packaging of RNA and it was validated by using single molecule RNA imaging of EV bound RNAs. Furthermore, we found that structuredness could explain the relative enrichment of circRNAs over their linear counterparts. These results were validated on existing public databases of circular and linear RNAs in EVs. By identifying and analysing these factors, we aim to better understand the complex mechanisms behind EV-mediated RNA transfer and its impact on cell communication in both health and disease. This mechanistic understanding of RNA enrichment in EVs is crucial for engineering EVs with selective RNA cargo.</p>","PeriodicalId":15811,"journal":{"name":"Journal of Extracellular Vesicles","volume":"14 7","pages":""},"PeriodicalIF":14.5,"publicationDate":"2025-07-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/jev2.70113","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144573147","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}