Journal of Extracellular Vesicles最新文献

{"title":"Extracellular RNA communication: A decade of NIH common fund support illuminates exRNA biology","authors":"Sara M. Amolegbe,&nbsp;Nicolas C. Johnston,&nbsp;Angela Ambrosi,&nbsp;Aniruddha Ganguly,&nbsp;T. Kevin Howcroft,&nbsp;Lillian S. Kuo,&nbsp;Patricia A. Labosky,&nbsp;Dobrila D. Rudnicki,&nbsp;John S. Satterlee,&nbsp;Danilo A. Tagle,&nbsp;Christine Happel","doi":"10.1002/jev2.70016","DOIUrl":"10.1002/jev2.70016","url":null,"abstract":"<p>The discovery that extracellular RNAs (exRNA) can act as endocrine signalling molecules established a novel paradigm in intercellular communication. ExRNAs can be transported, both locally and systemically in virtually all body fluids. In association with an array of carrier vehicles of varying complexity, exRNA can alter target cell phenotype. This highlights the important role secreted exRNAs have in regulating human health and disease. The NIH Common Fund exRNA Communication program was established in 2012 to accelerate and catalyze progress in the exRNA biology field. The program addressed both exRNA and exRNA carriers, and served to generate foundational knowledge for the field from basic exRNA biology to future potential clinical applications as biomarkers and therapeutics. To address scientific challenges, the exRNA Communication program developed novel tools and technologies to isolate exRNA carriers and analyze their cargo. Here, we discuss the outcomes of the NIH Common Fund exRNA Communication program, as well as the evolution of exRNA as a scientific field through the analysis of scientific publications and NIH funding. ExRNA and associated carriers have potential clinical use as biomarkers, diagnostics, and therapeutics. Recent translational applications include exRNA-related technologies repurposed as novel diagnostics in response to the COVID-19 pandemic, the clinical use of extracellular vesicle-based biomarker assays, and exRNA carriers as drug delivery platforms. This comprehensive landscape analysis illustrates how discoveries and innovations in exRNA biology are being translated both into the commercial market and the clinic. Analysis of program outcomes and NIH funding trends demonstrate the impact of this NIH Common Fund program.</p>","PeriodicalId":15811,"journal":{"name":"Journal of Extracellular Vesicles","volume":"14 1","pages":""},"PeriodicalIF":15.5,"publicationDate":"2025-01-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11735951/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143006705","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":"Helminth extracellular vesicles co-opt host monocytes to drive T cell anergy","authors":"Anne Borup,&nbsp;Mohammad Farouq Sharifpour,&nbsp;Litten S. Rossen,&nbsp;Bradley Whitehead,&nbsp;Anders T. Boysen,&nbsp;Rikke Olesen,&nbsp;Anja B. Bohn,&nbsp;Andrea Ridolfi,&nbsp;Marco Brucale,&nbsp;Francesco Valle,&nbsp;Lucia Paolini,&nbsp;Annalisa Radeghieri,&nbsp;Paolo Bergese,&nbsp;Kim Miles,&nbsp;Margaret Veitch,&nbsp;Tamara Thomas,&nbsp;Roland Ruscher,&nbsp;Phurpa Wangchuk,&nbsp;Paul Giacomin,&nbsp;Alex Loukas,&nbsp;Peter Nejsum","doi":"10.1002/jev2.70027","DOIUrl":"10.1002/jev2.70027","url":null,"abstract":"<p>Parasitic helminths secrete extracellular vesicles (EVs) into their host tissues to modulate immune responses, but the underlying mechanisms are poorly understood. We demonstrate that <i>Ascaris</i> EVs are efficiently internalised by monocytes in human peripheral blood mononuclear cells and increase the percentage of classical monocytes. Furthermore, EV treatment of monocytes induced a novel anti-inflammatory phenotype characterised by CD14⁺, CD16⁻, CC chemokine receptor 2 (CCR2⁻) and programmed death-ligand 1 (PD-L1)⁺ cells. In addition, <i>Ascaris</i> EVs induced T cell anergy in a monocyte-dependent mechanism. Targeting professional phagocytes to induce both direct and indirect pathways of immune modulation presents a highly novel and efficient mechanism of EV-mediated host-parasite communication. Intra-peritoneal administration of EVs induced protection against gut inflammation in the dextran sodium sulphate model of colitis in mice. <i>Ascaris</i> EVs were shown to affect circulating immune cells and protect against gut inflammation; this highlights their potential as a subject for further investigation in inflammatory conditions driven by dysregulated immune responses. However, their clinical translation would require further studies and careful consideration of ethical implications.</p>","PeriodicalId":15811,"journal":{"name":"Journal of Extracellular Vesicles","volume":"14 1","pages":""},"PeriodicalIF":15.5,"publicationDate":"2025-01-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11735955/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143006709","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":"Presenilins as hub proteins controlling the endocytic and autophagic pathways and small extracellular vesicle secretion","authors":"Inger Lauritzen,&nbsp;Anaïs Bini,&nbsp;Anaïs Bécot,&nbsp;Anne-Sophie Gay,&nbsp;Céline Badot,&nbsp;Sophie Pagnotta,&nbsp;Mounia Chami,&nbsp;Frédéric Checler","doi":"10.1002/jev2.70019","DOIUrl":"10.1002/jev2.70019","url":null,"abstract":"<p>Emerging evidence indicates that autophagy is tightly connected to the endocytic pathway. Here, we questioned the role of presenilins (PSENs 1 and 2), previously shown to be involved in autophagy regulation, in the secretion of small endocytic-originating extracellular vesicles known as exosomes. Indeed, while wild-type cells responded to stimuli promoting both multivesicular endosome (MVE) formation and secretion of small extracellular vesicles (sEVs) enriched in canonical exosomal proteins, PSEN-deficient cells were almost unaffected to these stimuli. Moreover, in PSEN-deficient cells, the re-expression of either PSEN1 or the functional active PSEN1delta9 mutant led to a rescue of most sEV secretion, while the deletion of PSEN1 alone almost fully phenocopied total PSEN invalidation. We found that the lack of sEV secretion in PSEN-deficient cells was also due to overactivated autophagy promoting MVEs to degradation rather than to plasma membrane fusion. Hence, in these cells, the autophagic blocker bafilomycin A1 (BafA1) not only increased the intracellular levels of the MVE protein CD63, but also turned on sEV secretion by stimulating autophagy-dependent unconventional secretion. In that case, sEVs arised from amphisomes and were enriched in both canonical exosomal proteins and lysosomal-autophagy-associated cargo. Altogether, we here demonstrate that PSENs, and particularly PSEN1, act as hub proteins controlling the balance between endosomal/autophagic degradation and secretion. More generally, our findings strengthen the view of a strong interconnection between the endocytic and autophagic pathways and their complementary roles in sEV secretion.</p>","PeriodicalId":15811,"journal":{"name":"Journal of Extracellular Vesicles","volume":"14 1","pages":""},"PeriodicalIF":15.5,"publicationDate":"2025-01-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11735957/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143006333","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":"Distinct molecular properties and functions of small EV subpopulations isolated from human umbilical cord MSCs using tangential flow filtration combined with size exclusion chromatography","authors":"Wei Liu,&nbsp;Xinyu Wang,&nbsp;Yating Chen,&nbsp;Jiapei Yuan,&nbsp;Huiyu Zhang,&nbsp;Xin Jin,&nbsp;Yuying Jiang,&nbsp;Junjing Cao,&nbsp;Zibin Wang,&nbsp;Shuo Yang,&nbsp;Bingwei Wang,&nbsp;Tinghe Wu,&nbsp;Jing Li","doi":"10.1002/jev2.70029","DOIUrl":"10.1002/jev2.70029","url":null,"abstract":"<p>As functional derivatives of mesenchymal stem cells (MSCs), small extracellular vesicles (sEVs) have garnered significant attention and application in regenerative medicine. However, the technical limitations for large-scale isolation of sEVs and their heterogeneous nature have added complexity to their applications. It remains unclear if the heterogeneous sEVs represent different aspects of MSCs functions. Here, we provide a method for the large-scale production of sEVs subpopulations derived from human umbilical cord mesenchymal stem cells (HucMSCs), utilizing tangential flow filtration combined with size exclusion chromatography. The resulting subpopulations, S1-sEVs and S2-sEVs, exhibited stable variations in size, membrane-marked proteins, and carrying cargos, thereby displaying distinct functions both in vitro and in animal disease models. S1-sEVs, that highly expressed CD9, HRS and GPC1, demonstrated a greater immunomodulatory impact, while S2-sEVs with enriched expression of CD63 and FLOT1/2 possessed enhanced capacities in promoting cell proliferation and angiogenesis. These discrepancies are attributed to the specific proteins and miRNAs they contain. Further investigation revealed that the two distinct sEVs subpopulations corresponded to different biological processes: the ESCRT pathway (S1-sEVs) and the ESCRT-independent pathway represented by lipid rafts (S2-sEVs). Therefore, we propose the potential for large-scale isolation and purification of sEVs subpopulations from HucMSCs with distinct functions. This approach may provide advantages for targeted therapeutic interventions in various MSC indications.</p>","PeriodicalId":15811,"journal":{"name":"Journal of Extracellular Vesicles","volume":"14 1","pages":""},"PeriodicalIF":15.5,"publicationDate":"2025-01-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11714183/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142949837","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 promote the infection and pathogenicity of Japanese encephalitis virus","authors":"Junyao Xiong,&nbsp;Ling'en Yang,&nbsp;Xiaowei Nan,&nbsp;Shuo Zhu,&nbsp;Mengxue Yan,&nbsp;Shengxian Xiang,&nbsp;Luping Zhang,&nbsp;Qi Li,&nbsp;Chengjie Yang,&nbsp;Xugang Wang,&nbsp;Ning Wei,&nbsp;Huanchun Chen,&nbsp;Youhui Si,&nbsp;Shengbo Cao,&nbsp;Jing Ye","doi":"10.1002/jev2.70033","DOIUrl":"10.1002/jev2.70033","url":null,"abstract":"<p>Japanese encephalitis virus (JEV) is a neurotropic zoonotic pathogen that poses a serious threat to public health. Currently, there is no specific therapeutic agent available for JEV infection, primarily due to the complexity of its infection mechanism and pathogenesis. Extracellular vesicles (EVs) have been known to play an important role in viral infection, but their specific functions in JEV infection remain unknown. Here, ultracentrifugation in combination with density gradient centrifugation was conducted to purify EVs from JEV-infected cells. The purified EVs were found to be infectious, with virions observed inside the EVs. Furthermore, our study showed the formation process of virion-containing EVs both in vitro and in vivo, which involved the fusion of multivesicular bodies with the cell membrane, leading to the release of virion-containing intraluminal vesicles into the extracellular space. Further studies revealed that EVs played a crucial role in JEV propagation by facilitating viral entry and assembly-release. Furthermore, EVs assisted JEV in evading the neutralizing antibodies and promoted viral capability to cross the blood-brain and placental barriers. Moreover, in vivo experiments demonstrated that EVs were beneficial for JEV infection and pathogenicity. Taken together, our findings highlight the significant contribution of EVs in JEV infection and provide valuable insights into JEV pathogenesis.</p>","PeriodicalId":15811,"journal":{"name":"Journal of Extracellular Vesicles","volume":"14 1","pages":""},"PeriodicalIF":15.5,"publicationDate":"2025-01-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11714208/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142949840","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 simple polydopamine-based platform for engineering extracellular vesicles with brain-targeting peptide and imaging probes to improve stroke outcome","authors":"Xiaojing Shi,&nbsp;Lu Zhang,&nbsp;Shengju Wu,&nbsp;Chunfu Zhang,&nbsp;Muyassar Mamtilahun,&nbsp;Yongfang Li,&nbsp;Zhijun Zhang,&nbsp;Changjing Zuo,&nbsp;Fengzhen Cui,&nbsp;Wanlu Li,&nbsp;Guo-Yuan Yang,&nbsp;Yaohui Tang","doi":"10.1002/jev2.70031","DOIUrl":"10.1002/jev2.70031","url":null,"abstract":"<p>Extracellular vesicles (EVs) have shown great potential for treating various diseases. Translating EVs-based therapy from bench to bedside remains challenging due to inefficient delivery of EVs to the injured area and lack of techniques to visualize the entire targeting process. Here we developed a dopamine surface functionalization platform that facilitates easy and simultaneous conjugation of targeting peptide and multi-mode imaging probes to the surface of EVs. Utilizing this platform we concurrently modified M2 microglia-derived EVs (M2-EVs) with neuronal targeting peptide rabies virus glycoprotein peptide 29 (RVG29) and multi-modal imaging tracers, resulting in the targeted delivery of M2-EVs to stroke mice brain and enabled the dynamic visualization of the targeting process from whole-body to cellular levels. We determined that intra-arterial injection achieved the highest efficiency of targeted delivery of engineered EVs to the stroke mice brain, improved therapeutic efficacy by reducing neuronal apoptosis. Mechanistically, EVs miRNA array revealed that a number of anti-apoptosis related miRNAs were significantly up-regulated, including miR-221-3p and miR-423-3p, both exerted anti-apoptotic effects through p38/ERK signalling pathways in stroke. Overall, this platform provides a facile and powerful tool for multifunctional engineering of EVs for multiscale therapeutic evaluation and enhancement of EV-based therapy, with valuable prospects for clinical translation.</p>","PeriodicalId":15811,"journal":{"name":"Journal of Extracellular Vesicles","volume":"14 1","pages":""},"PeriodicalIF":15.5,"publicationDate":"2025-01-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11714163/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143056036","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":"DetectEV: A functional enzymatic assay to assess integrity and bioactivity of extracellular vesicles","authors":"Giorgia Adamo,&nbsp;Sabrina Picciotto,&nbsp;Paola Gargano,&nbsp;Angela Paterna,&nbsp;Samuele Raccosta,&nbsp;Estella Rao,&nbsp;Daniele Paolo Romancino,&nbsp;Giulio Ghersi,&nbsp;Mauro Manno,&nbsp;Monica Salamone,&nbsp;Antonella Bongiovanni","doi":"10.1002/jev2.70030","DOIUrl":"10.1002/jev2.70030","url":null,"abstract":"<p>The application of extracellular vesicles (EVs) as therapeutics or nanocarriers in cell-free therapies necessitates meticulous evaluations of different features, including their identity, bioactivity, batch-to-batch reproducibility, and stability. Given the inherent heterogeneity in EV preparations, this assessment demands sensitive functional assays to provide key quality control metrics, complementing established methods to ensure that EV preparations meet the required functionality and quality standards. Here, we introduce the detectEV assay, an enzymatic-based approach for assessing EV luminal cargo bioactivity and membrane integrity. This method is fast, cost-effective, and quantifiable through enzymatic units. Utilizing microalgae-derived EVs, known as nanoalgosomes, as model systems, we optimised the assay parameters and validated its sensitivity and specificity in quantifying the enzymatic activity of esterases within the EV lumen while also evaluating EV membrane integrity. Compared to conventional methods that assess physicochemical features of EVs, our single-step analysis efficiently detects batch-to-batch variations by evaluating changes in luminal cargo bioactivity and integrity across various EV samples, including differences under distinct storage conditions and following diverse isolation and exogenous loading methods, all using small sample sizes. The detectEV assay's application to various human-derived EV types demonstrated its versatility and potential universality. Additionally, the assay effectively predicted EV functionality, such as the antioxidant activity of different nanoalgosome batches. Our findings underscore the detectEV assay's utility in comprehensive characterization of EV functionality and integrity, enhancing batch-to-batch reproducibility and facilitating their therapeutic applications.</p>","PeriodicalId":15811,"journal":{"name":"Journal of Extracellular Vesicles","volume":"14 1","pages":""},"PeriodicalIF":15.5,"publicationDate":"2025-01-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11705427/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142949836","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":"TGF-β regulates the release of breast cancer cell-derived extracellular vesicles and the sorting of their protein cargo by downregulating RAB27B expression","authors":"Chao Li,&nbsp;Agustin Enciso-Martinez,&nbsp;Roman I. Koning,&nbsp;Mona Shahsavari,&nbsp;Peter ten Dijke","doi":"10.1002/jev2.70026","DOIUrl":"10.1002/jev2.70026","url":null,"abstract":"<p>Extracellular vesicles (EVs) are important mediators of intercellular communication in the tumour microenvironment. The cytokine transforming growth factor-β (TGF-β) facilitates cancer progression via EVs secreted by cancer cells, which act on recipient cells in the tumour microenvironment. However, the mechanisms of how TGF-β affects cancer cell EV release and composition are incompletely understood. Here, we systematically investigate the effects of TGF-β on the release and protein composition of EVs from breast cancer cells. TGF-β suppresses the transcription of <i>RAB27B</i> mediated by SMAD3 and thereby hampers EV release. Using click chemistry and quantitative proteomics, we found that TGF-β increases the quantity of protein cargo and changes the composition of EVs by downregulating RAB27B expression. The recomposed EVs, induced by TGF-β or RAB27B depletion, inhibit CD8⁺ T cell-mediated breast cancer killing. Our findings reveal the critical roles of TGF-β and RAB27B in cancer development by regulating EV release and composition and thus provide potential targets to improve cancer immunotherapy.</p>","PeriodicalId":15811,"journal":{"name":"Journal of Extracellular Vesicles","volume":"13 12","pages":""},"PeriodicalIF":15.5,"publicationDate":"2024-12-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11669950/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142895070","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":"Fused extracellular vesicles from M2 macrophages and human umbilical cord mesenchymal stem cells for the targeted regulation of macrophage pyroptosis in periprosthetic osteolysis","authors":"Qimeng Liu,&nbsp;Tianliang Ma,&nbsp;Zheyu Zhang,&nbsp;Jiangyu Nan,&nbsp;Guanzhi Liu,&nbsp;Yute Yang,&nbsp;Yihe Hu,&nbsp;Jie Xie","doi":"10.1002/jev2.70028","DOIUrl":"10.1002/jev2.70028","url":null,"abstract":"<p>The development of strategies for the prevention and treatment of aseptic loosening of prostheses stands as a critical area of global research interest. The pyroptosis of local macrophages triggered by wear particles plays a pivotal role in the onset of periprosthetic osteolysis and subsequent loosening. Extracellular vesicles, carrying the surface components and regulatory molecules of their parent cells, embody the cellular characteristics and biological functions of these progenitors. In a pioneering approach to precisely inhibit the pyroptosis of local macrophages induced by wear particles, we have engineered fused extracellular vesicles (fEV) from M₂ macrophages and human umbilical cord mesenchymal stem cells. These fEV boast the distinctive capability for targeted transport and immune evasion, collectively enhancing the anti-pyroptosis effect of the therapeutic extracellular vesicles. Our research demonstrates the targeted, significant preventive and therapeutic potential of fEVs against periprosthetic osteolysis prompted by wear particles, highlighting its crucial clinical significance and application prospects. These findings suggest that extracellular vesicle fusion technology heralds a novel paradigm in the design and development of targeted extracellular vesicle-based drug delivery systems.</p>","PeriodicalId":15811,"journal":{"name":"Journal of Extracellular Vesicles","volume":"13 12","pages":""},"PeriodicalIF":15.5,"publicationDate":"2024-12-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11664327/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142877040","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":"Challenges of MS-based small extracellular vesicles proteomics","authors":"Daniel Fochtman,&nbsp;Lukasz Marczak,&nbsp;Monika Pietrowska,&nbsp;Anna Wojakowska","doi":"10.1002/jev2.70020","DOIUrl":"10.1002/jev2.70020","url":null,"abstract":"<p>Proteomic profiling of small extracellular vesicles (sEV) is a powerful tool for discovering biomarkers of various diseases. This process most often assisted by mass spectrometry (MS) usually lacks standardization and recognition of challenges which may lead to unreliable results. General recommendations for sEV MS analyses have been briefly given in the MISEV2023 guidelines. The present work goes into detail for every step of sEV protein profiling with an overview of factors influencing such analyses. This includes reporting and defining the sEV source and vesicle isolation, protein solubilization and digestion, ‘offline’ and ‘online’ sample complexity reduction, the analysis type itself, and subsequent data analysis. Every stage in this process affects the others, which could result in different outcomes. Although characterization and comparisons of different sEV isolation methods are known and accessible and MS-based profiling details are provided for cell or tissue samples, no consensus work has been ever published to describe the whole process of sEV proteomic analysis. Reliable results can be obtained from sEV profiling provided that the analysis is well planned, prepared for, and backed by pilot studies or appropriate research.</p>","PeriodicalId":15811,"journal":{"name":"Journal of Extracellular Vesicles","volume":"13 12","pages":""},"PeriodicalIF":15.5,"publicationDate":"2024-12-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/jev2.70020","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142846789","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}