Extracellular vesicle最新文献

{"title":"Elevated TRPP2 promotes malignant behavior of head and neck cancer via enhancing exosome biogenesis","authors":"Suwen Bai ,&nbsp;Hexing Sun ,&nbsp;Xiling Xu ,&nbsp;Yuan Wei ,&nbsp;Yuxin Zhang ,&nbsp;Chenxi Zhang ,&nbsp;Qing Peng ,&nbsp;Junwei Zhu ,&nbsp;Juan Du","doi":"10.1016/j.vesic.2026.100105","DOIUrl":"10.1016/j.vesic.2026.100105","url":null,"abstract":"<div><h3>Background</h3><div>Exosomes promote cancer progression by mediating intercellular communication via oncogenic cargos. These vesicles originate as intraluminal vesicles (ILVs) within multivesicular bodies (MVBs), with their secretion dependent on MVB trafficking. The endosomal sorting complex required for transport (ESCRT) complex plays a key role in sorting proteins into exosomes, while the calcium channel Transient Receptor Potential Polycystin-2 (TRPP2) may regulate exosome biogenesis and amplify their oncogenic potential.</div></div><div><h3>Methods</h3><div>In this study, we analyzed TRPP2 level in HNC tissues using immunohistochemistry (IHC) and correlated them with clinical outcomes via UALCAN data. We characterized exosomes from medium, by nanoparticle tracking analysis (NTA), transmission electron microscopy, and exosome quantification. We measured intracellular calcium levels of HNC cells using Fluo-8 AM. Additionally, protein interactions were assessed via co-immunoprecipitation (co-IP) and immunofluorescence. Functional assays included <em>in vitro</em> proliferation, migration, cell cycle analysis, and in vivo xenograft models.</div></div><div><h3>Results</h3><div>The expression of TRPP2 was significantly higher in HNC tissues, correlating with advanced tumor stages and poor prognosis. Knockdown of TRPP2 in HNC cells diminished exosome secretion by impairing MVB-Rab11a interaction, a process dependent on TRPP2-mediated calcium influx. Exosomes derived from TRPP2 overexpression cells exhibited enriched oncogenic proteins, driven by TRPP2's regulation of ESCRT subunits VPS37B and CHMP6. These exosomes enhanced HNC cell proliferation, migration, and xenograft tumor growth.</div></div><div><h3>Conclusions</h3><div>Our study demonstrates that TRPP2 drives HNC malignancy via two pivotal mechanisms: calcium-dependent regulation of MVBs trafficking to enhance exosome secretion, and modulation of exosomal cargo through ESCRT components. These findings establish TRPP2 as a critical regulator of exosome biogenesis and underscore its promise as a therapeutic target for HNC. Targeted TRPP2 inhibition may disrupt tumor-stromal crosstalk, thereby improving therapeutic outcomes in HNC and other exosome-driven malignancies.</div></div>","PeriodicalId":73007,"journal":{"name":"Extracellular vesicle","volume":"7 ","pages":"Article 100105"},"PeriodicalIF":0.0,"publicationDate":"2026-01-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146077659","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Injected extracellular vesicles and other nanoparticles hitchhike on erythrocytes and platelets from circulation towards organ clearance","authors":"Svetlana Pavlova ,&nbsp;Doste R. Mamand ,&nbsp;Daniel W. Hagey ,&nbsp;Xiuming Liang ,&nbsp;H. Yesid Estupiñán ,&nbsp;Wenyi Zheng ,&nbsp;Guannan Zhou ,&nbsp;Risul Amin ,&nbsp;Antje M. Zickler ,&nbsp;Scott Bonner-Harris ,&nbsp;Zankruti Dave ,&nbsp;Miina Ojansivu ,&nbsp;Manuela O. Gustafsson ,&nbsp;Oliver G. Hayes ,&nbsp;Oscar P.B. Wiklander ,&nbsp;Manuchehr Abedi‐Valugerdi ,&nbsp;Samantha Roudi ,&nbsp;Antonin Marquant ,&nbsp;Joel Z. Nordin ,&nbsp;Molly M. Stevens ,&nbsp;Samir EL Andaloussi","doi":"10.1016/j.vesic.2025.100103","DOIUrl":"10.1016/j.vesic.2025.100103","url":null,"abstract":"<div><div>The application of extracellular vesicles (EVs) for targeted delivery of drugs, proteins, and RNA to specific cell types or organs is a promising approach in molecular therapy. However, to achieve efficient and targeted delivery, it is crucial to improve our understanding of the fate of injected EVs in terms of tissue and cellular biodistribution. Studies by us and others in mouse models have demonstrated that EVs are rapidly cleared from the blood circulation following distribution to different organs, with large fractions ending up in liver, spleen and lungs within minutes after injection. In this study, we investigated the key steps and kinetics of this clearance process and particularly focused on understanding how intra-venously injected EVs would interact with circulating blood cells shortly after injection. Following the injection of fluorescently tagged EVs into mice, we utilized high sensitivity imaging flow cytometry to study the binding or association of EVs to different blood cell types, including platelets (PLTs) and red blood cells (RBCs), both <em>in vitro</em> and <em>in vivo</em>. Notably, PLTs and RBCs are regularly excluded in EV biodistribution and targeting studies even though they account for more than 99 % of all blood cells. We demonstrate that a significant proportion of injected EVs binds to PLTs and RBCs within minutes, and we further show that EVs ‘hitchhike’ on both PLTs and RBCs to the liver and spleen, where they are collectively cleared by macrophages. Detailed assessment of this process in a newly established <em>ex vivo</em> PLT/RBC binding assay revealed that this process is independent of EV purity, EV source and species, and highlights that it also applies to other nanoparticles such as lipid nanoparticles (LNPs) and liposomes. Interestingly, EVs engineered to bind albumin show clearly reduced PLT/RBC association. In summary, we report a highly relevant yet largely overlooked mechanism that significantly contributes to clearance of EVs and impacts their biodistribution and pharmacokinetics.</div></div>","PeriodicalId":73007,"journal":{"name":"Extracellular vesicle","volume":"7 ","pages":"Article 100103"},"PeriodicalIF":0.0,"publicationDate":"2025-12-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145841264","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Discriminating extracellular vesicles by their membranes","authors":"Carlo Guardiani ,&nbsp;Matteo Riboli ,&nbsp;Flavio Costa ,&nbsp;Massimiliano Bruno ,&nbsp;Giulia Rossi ,&nbsp;Alberto Giacomello","doi":"10.1016/j.vesic.2025.100101","DOIUrl":"10.1016/j.vesic.2025.100101","url":null,"abstract":"<div><div>Extracellular Vesicles (EVs) are increasingly employed for targeted drug delivery and advanced diagnostic assays. However, the diagnostic application of EVs so far only relied on biochemical markers as in the case of prostate cancer. Pioneering work by Whitehead and by LeClaire showed that the diagnostic power of EVs can be significantly expanded by using the mechanical properties of EV membranes for mechanophenotyping. Inspired by this idea, we computationally characterised the mechanical properties of EVs released by the prostate cancer cell line PC-3 and their healthy counterparts, the prostasomes. Our work confirms that healthy and cancer EVs exhibit different mechanical properties. However, while Whitehead and LeClaire observed a softening of the membrane in cancer EVs, our analysis reveals that the membrane of PC-3 EVs is stiffer than that of healthy prostasomes. This suggests that the diagnostic interpretation of mechanical data will have to be done on a case-by-case basis. As experimental evidence suggests that EV membranes are asymmetric, we also explored the role of numerical and compositional asymmetry starting from the PC-3 composition. Our work shows that the tilt and splay moduli of the asymmetric PC-3 membrane are the average of the moduli of the corresponding symmetric membranes. This result confirms the predictions of the elastic theory of membranes, which have been recently challenged by experimental data. Finally, our work reveals coupling effects such that the moduli of a monolayer depend not only on its composition but also on the features of the companion leaflet.</div></div>","PeriodicalId":73007,"journal":{"name":"Extracellular vesicle","volume":"7 ","pages":"Article 100101"},"PeriodicalIF":0.0,"publicationDate":"2025-12-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145841262","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Mitochondria-containing large extracellular vesicles target mouse motor neurons upon intramuscular injection","authors":"Paromita Paul Pinky ,&nbsp;Zhong-Min Wang ,&nbsp;Purva Khare ,&nbsp;Jhanvi R. Jhaveri ,&nbsp;Abigail Sullivan ,&nbsp;Vivek Basudkar ,&nbsp;Krithika S. Rao ,&nbsp;Audrey Lawrence ,&nbsp;Adithri Pingali ,&nbsp;Kandarp M. Dave ,&nbsp;Donna B. Stolz ,&nbsp;Ming Sun ,&nbsp;Si-yang Zheng ,&nbsp;Sruti S. Shiva ,&nbsp;Carol Milligan ,&nbsp;Osvaldo Delbono ,&nbsp;Devika S. Manickam","doi":"10.1016/j.vesic.2025.100100","DOIUrl":"10.1016/j.vesic.2025.100100","url":null,"abstract":"<div><div>Amyotrophic Lateral Sclerosis (<strong>ALS</strong>) is a neurological disorder that causes progressive degeneration of motor neurons. Mitochondrial dysfunction accelerates neurodegeneration, aggravating the severity of ALS. We hypothesized that increasing the mitochondrial function of motor neurons may promote neuronal survival. Therefore, we investigated the potential of neuron-derived mitochondria containing extracellular vehicles (<strong>EVs</strong>) as a novel therapeutic approach for ALS using differentiated NSC-34 cells as a surrogate for neurons. Neuron derived-large EVs (<strong>lEVs</strong>) but not small EVs (<strong>sEVs</strong>) contained mitochondria. However, we observed increased cell viability and oxygen consumption rates in heat-stressed neurons treated with both sEVs and lEVs suggesting improved mitochondrial function in the recipient neurons. The increased oxygen consumption rates in sEV-treated heat-stressed neurons was accompanied by a <em>greater proton leak</em> compared to lEV treatment. The greater proton leak observed with sEVs likely suggests a lower efficiency of oxidative phosphorylation compared to that achieved by cells treated with mitochondria-containing lEVs. These findings suggest that mitochondrial components present in sEVs, such as proteins and mitochondrial DNA, may too contribute to improving cellular respiration. Furthermore, we demonstrated that lEV mitochondria are transported into the lumbar spinal cord motor neurons following intramuscular injection in C57BL/6 mice in an EV dose-dependent manner. Collectively, for the first time, we have demonstrated the therapeutic effects of neuronal EVs in recipient heat-stressed neurons and the delivery of lEV mitochondria to spinal cord motor neurons <em>in vivo</em> without any EV surface modifications for neuronal targeting. Further studies will determine the therapeutic efficacy of mitochondria-containing EVs in the SOD1^G93A transgenic mouse model of ALS.</div></div>","PeriodicalId":73007,"journal":{"name":"Extracellular vesicle","volume":"7 ","pages":"Article 100100"},"PeriodicalIF":0.0,"publicationDate":"2025-12-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145841263","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Harnessing extracellular vesicles for stabilized and functional IL-10 delivery in macrophage immunomodulation","authors":"Najla A. Saleh ,&nbsp;Matthew A. Gagea ,&nbsp;Xheneta Vitija ,&nbsp;Sadhana Kilangodi ,&nbsp;Ahmed A. Zarea ,&nbsp;Tomas Janovic ,&nbsp;Jens C. Schmidt ,&nbsp;Cheri X. Deng ,&nbsp;Masamitsu Kanada","doi":"10.1016/j.vesic.2025.100102","DOIUrl":"10.1016/j.vesic.2025.100102","url":null,"abstract":"<div><div>Extracellular vesicles (EVs) are gaining recognition as promising therapeutic carriers for immune modulation. We investigated the potential of EVs derived from HEK293FT cells to stabilize and deliver interleukin-10 (IL-10), a key anti-inflammatory cytokine. Using minicircle (MC) DNA vectors, we achieved IL-10 overexpression and efficient incorporation into filter-isolated small EVs (F-sEVs), resulting in superior stability compared to free recombinant IL-10. Detailed biophysical and functional analyses revealed that IL-10⁺ F-sEVs contain both monomeric and oligomeric IL-10 on their external surface and encapsulated within vesicles. Size-based fractionation of IL-10⁺ large EVs (lEVs), small EVs (UC-sEVs), and non-vesicular extracellular particles (NVEPs) revealed IL-10 presence across all fractions, predominantly in monomeric form. Anion exchange chromatography successfully enriched IL-10⁺ exosomes that efficiently associated with both IL-10 monomers and oligomers. IL-10⁺ F-sEVs suppressed inflammatory cytokine expression in pro-inflammatory macrophages (two-to 14-fold more effectively than naïve F-sEVs) without inducing anti-inflammatory repolarization. However, detailed analysis of IL-10-loaded EV subpopulations revealed that anti-inflammatory activity was distributed across multiple fractions. Moreover, naïve F-sEVs derived from non-transfected cells also exhibited anti-inflammatory effects, suggesting that endogenous EV cargo contributes to their immunomodulatory activity and complicates attribution of effects specifically to IL-10. These findings highlight the therapeutic potential of EVs while emphasizing the need to disentangle contributions of engineered cytokines from endogenous vesicular components.</div></div>","PeriodicalId":73007,"journal":{"name":"Extracellular vesicle","volume":"7 ","pages":"Article 100102"},"PeriodicalIF":0.0,"publicationDate":"2025-12-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145718983","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Systematic characterization of mammalian extracellular vesicles using nano-flow cytometry","authors":"Benjamin T. Vyzourek ,&nbsp;Dirk Anderson ,&nbsp;Luke Skrabal ,&nbsp;Christine E. Humphrey ,&nbsp;Eduardo Romero ,&nbsp;Brittany Schweiger ,&nbsp;Forrest Kievit ,&nbsp;Jeremy R. Miles ,&nbsp;Angela K. Pannier","doi":"10.1016/j.vesic.2025.100098","DOIUrl":"10.1016/j.vesic.2025.100098","url":null,"abstract":"<div><div>Extracellular vesicles (EVs) are nanoscale, membrane-enclosed particles that transport bioactive cargo between cells and are increasingly studied for their potential in diagnostic and therapeutic applications. Advancing EV-based technologies for these applications depend on the ability to consistently isolate and characterize vesicle populations with defined biophysical and molecular properties. Efforts to obtain pure EV populations from cell culture systems are limited by inherent EV heterogeneity, exogenous particle contamination introduced by media supplements, and the co-isolation of non-vesicular contaminants. These challenges are further compounded by the limitations of conventional EV characterization platforms, which often lack the resolution to distinguish EVs from similarly sized non-vesicular particles or to capture molecular heterogeneity at the single-vesicle scale. Together, these limitations highlight the need for analytical approaches capable of resolving EV heterogeneity and enabling comparisons across EV production conditions and isolation strategies. In this study, we used nano-flow cytometry (nFCM) for high-resolution analysis of individual EVs, enabling simultaneous measurement of particle size, concentration, and tetraspanin expression. This approach revealed substantial amounts of exogenous particle contamination in media supplements commonly used to culture EV-producing cells, and quantified differences in EV purity and yield between methods used to isolate EVs from the media of the producing cells. Additionally, analysis of EVs derived from HEK293T, U-87 MG, and hMSC mammalian cell cultures revealed cell type-specific differences in EV production and expression of tetraspanin markers CD9, CD63, and CD81. Collectively, these results demonstrate that careful selection of media compositions and isolation strategies, combined with nFCM analytical techniques can resolve biological differences in EV populations.</div></div>","PeriodicalId":73007,"journal":{"name":"Extracellular vesicle","volume":"6 ","pages":"Article 100098"},"PeriodicalIF":0.0,"publicationDate":"2025-11-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145519278","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Proteomic profiling of tissue extracellular vesicles (EVs) identifies tissue EV markers and estimates the abundance of tissue EVs in the circulation","authors":"Sho Watanabe ,&nbsp;Takumi Makino ,&nbsp;Kahori Hiro ,&nbsp;Takashi Sasaki ,&nbsp;Yu Takahashi ,&nbsp;Kei Futagawa ,&nbsp;Michio Suzuki ,&nbsp;Ryuichiro Sato ,&nbsp;Yoshio Yamauchi","doi":"10.1016/j.vesic.2025.100097","DOIUrl":"10.1016/j.vesic.2025.100097","url":null,"abstract":"<div><div>Extracellular vesicles (EVs) mediate cell-to-cell communication in endocrine, paracrine, and autocrine fashions, but their roles and transport in the body remain incompletely understood. It is suggested that EVs circulate in the body through the bloodstream, where EVs derived from various tissues are present. However, how much each tissue contributes to circulating EVs is largely unknown. Therefore, identifying tissue-specific EV markers is of importance for elucidating <em>in vivo</em> dynamics of tissue EVs. A key issue that hampers studying EVs <em>in vivo</em> is the lack of methodologies for collecting them from tissues. Here, we developed methods to isolate EVs from four different tissues, skeletal muscle, heart, liver, and adipose tissue, and performed proteomic analysis on their EVs. Unbiased and quantitative proteomic analysis revealed protein signatures of EVs from the four tissues and identified marker proteins specific to or highly enriched in EVs of each tissue. Furthermore, through comprehensive comparisons of the proteome from tissue and plasma EVs, we estimated the relative contribution of tissue EVs to circulating EVs. Our data suggest that adipose tissue-derived EVs are highly enriched, while skeletal muscle-derived EVs are only a minor population in circulating EVs. Collectively, our tissue EV proteome identified potential tissue EV markers and suggested that the abundance of tissue EVs in circulation highly depends on the original tissues.</div></div>","PeriodicalId":73007,"journal":{"name":"Extracellular vesicle","volume":"6 ","pages":"Article 100097"},"PeriodicalIF":0.0,"publicationDate":"2025-11-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145465463","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Therapeutic application of mesenchymal stem cell-derived extracellular vesicles in radiation injury: research status, challenges, and prospects","authors":"Runxuan Wang,&nbsp;Tong An,&nbsp;Daqin Li,&nbsp;Huijuan Song,&nbsp;Ningning He,&nbsp;Qiang Liu","doi":"10.1016/j.vesic.2025.100096","DOIUrl":"10.1016/j.vesic.2025.100096","url":null,"abstract":"<div><div>Radiation injury treatment represents a longstanding scientific challenge in radiotherapy research and an urgent medical problem. As the number of cancer patients in China increases, so does the number of radiotherapy recipients and the frequency of treatments. Concurrently, nuclear accidents continue to occur. Under these circumstances, public attention is increasingly focused on various types of radiation-induced bodily damage. However, the multifaceted damaging effects of radiation impede the activation of organ regeneration responses. With the deepening research on mesenchymal stem cells (MSCs) and their extracellular vesicles (EVs), accumulating evidence suggests that Mesenchymal Stem Cell-Derived Extracellular Vesicles (MSC-EVs) are ideal candidates for cell-free therapy of radiation injury. In recent years, MSC-EVs have become a hot research topic in the biomedical field, and more and more studies have proven that they are excellent nanocarriers. Although significant gaps remain in understanding the therapeutic mechanisms of MSC-EVs for radiation injury repair, current research achievements sufficiently demonstrate their application value in treating radiation damage. Therefore, this paper summarizes and discusses recent advancements to provide new insights into cell-free EV therapy for radiation injury.</div></div>","PeriodicalId":73007,"journal":{"name":"Extracellular vesicle","volume":"6 ","pages":"Article 100096"},"PeriodicalIF":0.0,"publicationDate":"2025-10-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145320070","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Encapsulating extracellular vesicles with a minimal RISC complex as novel gene silencing tool","authors":"Tao Qiu,&nbsp;Yu Yan,&nbsp;Rui Hu,&nbsp;Yuan Yi,&nbsp;Guowu Liu,&nbsp;Wenqiang Lu,&nbsp;Xin Zhou,&nbsp;Ke Xu","doi":"10.1016/j.vesic.2025.100094","DOIUrl":"10.1016/j.vesic.2025.100094","url":null,"abstract":"<div><div>Gene silencing modalities including small interfering RNAs (siRNAs) and antisense oligonucleotides (ASOs) have prospered in both fundamental research and clinical translations in recent years, with delivery platform being one of the key elements for success. Extracellular vesicles (EVs) as natural carriers for cell-cell communication have been engineered in a variety of ways as delivery platform for gene silencing, yet facing limited efficiency and reproducibility. In this study, we developed a new strategy for engineering EVs as gene silencing tool. A minimal RNA-induced silencing complex (RISC), composing of modified Argonaute 2 (AGO2) protein and specially designed guide strand RNAs, were encapsulated into EVs and elicited prominent EGFP silencing in proof-of-concept study. This modular EVs platform, which we named as minRISC-EVs, efficiently silenced iNOS expression in M1 macrophages as well as STAT6/A20 expression in M2 macrophages, enabling macrophages polarization towards desired directions. The macrophage modulating ability was further validated <em>in vivo</em>, as minRISC-EVs against iNOS alleviated mice lung inflammation in lipopolysaccharide (LPS)-induced acute lung injury model, and minRISC-EVs against STAT6/A20 inhibited B16F10 tumor progression in the tumor xenograft model. In summary, minRISC-EVs can be utilized as novel gene silencing tool, and hold great promise for clinical translation in the future.</div></div>","PeriodicalId":73007,"journal":{"name":"Extracellular vesicle","volume":"6 ","pages":"Article 100094"},"PeriodicalIF":0.0,"publicationDate":"2025-09-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145094817","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Generation and characterization of platelet-derived extracellular vesicle analogues from lyophilized platelets","authors":"Xin Xin,&nbsp;Ingrid Dijkgraaf,&nbsp;Tilman M. Hackeng,&nbsp;Rory R. Koenen","doi":"10.1016/j.vesic.2025.100095","DOIUrl":"10.1016/j.vesic.2025.100095","url":null,"abstract":"<div><div>Platelet-derived extracellular vesicles (P-EV) are thought to facilitate the transfer of information from platelets to target cells, playing a role in both physiologic and pathophysiologic processes, particularly in regulating immune responses and healing processes. In addition, P-EV show promise as drug carriers and biomarkers for disease. However, the procedures for isolation, purification and fluorescent labeling of P-EV remain unstandardized. Moreover, the requirement to use freshly obtained platelets for generating EV presents a logistical challenge for their study. In this study, we isolated, characterized, and compared P-EV analogues by sonication of freshly obtained and lyophilized platelets, investigated fluorescent labeling methods, and monitored cellular uptake. We found that P-EV analogues derived from fresh or lyophilized platelets showed similar characteristics regarding size, surface proteins and content. Among the fluorescent labeling methods, CFSE and DiO-C6 were most effective in labeling P-EV analogues from both fresh and lyophilized platelets. All labeling methods led to an increase in P-EV analogue's size, with CFSE and DiO-C6 resulting in the smallest increase. The addition of P-EV analogues to cultured immortal endothelial cells revealed that P-EV analogues were effectively internalized and directed to the lysosomal compartment. The results indicate that P-EV analogues from lyophilized platelets have similar functional properties as those from freshly isolated platelets and these are retained after labeling with CFSE. Thus, lyophilized platelets can serve as a source of P-EV analogues for functional studies.</div></div>","PeriodicalId":73007,"journal":{"name":"Extracellular vesicle","volume":"6 ","pages":"Article 100095"},"PeriodicalIF":0.0,"publicationDate":"2025-09-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145048800","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}