Journal of extracellular biology最新文献

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Proteomics of Urinary Extracellular Vesicles Highlight the Involvement of Vitronectin and the Fibrinolytic and TNF Pathways as Mechanisms Underlying Renal Fibrosis in Kidney Transplant Patients 尿细胞外囊泡的蛋白质组学强调了玻璃体连接蛋白、纤溶蛋白和肿瘤坏死因子通路作为肾移植患者肾纤维化的机制
Journal of extracellular biology Pub Date : 2025-06-02 DOI: 10.1002/jex2.70056
Marta Clos-Sansalvador, Sergio G. Garcia, Paula Rodríguez-Martínez, Marta Sanroque-Muñoz, Miriam Font-Morón, Cristina Grange, Benedetta Bussolati, Marcella Franquesa, Javier Juega, Francesc E. Borràs
{"title":"Proteomics of Urinary Extracellular Vesicles Highlight the Involvement of Vitronectin and the Fibrinolytic and TNF Pathways as Mechanisms Underlying Renal Fibrosis in Kidney Transplant Patients","authors":"Marta Clos-Sansalvador,&nbsp;Sergio G. Garcia,&nbsp;Paula Rodríguez-Martínez,&nbsp;Marta Sanroque-Muñoz,&nbsp;Miriam Font-Morón,&nbsp;Cristina Grange,&nbsp;Benedetta Bussolati,&nbsp;Marcella Franquesa,&nbsp;Javier Juega,&nbsp;Francesc E. Borràs","doi":"10.1002/jex2.70056","DOIUrl":"https://doi.org/10.1002/jex2.70056","url":null,"abstract":"<p>Vitronectin (VTN) is a potential non-invasive biomarker for renal fibrosis, originally described in urinary extracellular vesicles (uEV) from kidney transplant patients (KTx). However, VTN's specific role in renal fibrosis is unclear, as it is involved in various physiological processes. This study aims to identify other uEV-associated proteins linked to renal fibrosis to clarify which pathways involve VTN. uEV were isolated from 33 KTx patients and five healthy controls. uEV proteins were analysed using proximity extension assay (PEA), and data were normalized and compared using Welch's two-sided <i>t</i>-test to identify differentially expressed proteins between fibrotic (<i>n</i> = 31) and non-fibrotic patients (<i>n</i> = 7). Urinary VTN levels and monocyte chemoattractant protein-1 (MCP-1) were measured by ELISA. PEA analysis identified 33 proteins overexpressed in the fibrotic group. These proteins clustered in STRING analysis, primarily associating with coagulation, fibrinolysis and TNF-inflammation involving macrophages. ELISA detection of MCP-1 further validated the results. High levels of VTN in the fibrotic group were accompanied by the upregulation of fibrinolytic pathway components (PAI-1, tPA and uPAR), which are well-known to interact with VTN. This study highlights TNF-induced inflammation involving macrophages and fibrinolysis as key mechanisms underlying renal fibrosis with direct implications of VTN, which support VTN's potential as a biomarker for this pathological process.</p>","PeriodicalId":73747,"journal":{"name":"Journal of extracellular biology","volume":"4 6","pages":""},"PeriodicalIF":0.0,"publicationDate":"2025-06-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/jex2.70056","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144191005","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}
引用次数: 0
Enolase and 16.5-kDa Tegument-Associated Protein in Fasciola hepatica Extracellular Vesicles: Clues to Their Role in Pathogenesis 肝片形吸虫细胞外泡的烯醇化酶和16.5 kda被膜相关蛋白:它们在发病机制中的作用线索
Journal of extracellular biology Pub Date : 2025-06-02 DOI: 10.1002/jex2.70055
Aránzazu González-Arce, Christian M. Sánchez-López, Liz F. Sánchez-Palencia, Antonio Marcilla, Dolores Bernal
{"title":"Enolase and 16.5-kDa Tegument-Associated Protein in Fasciola hepatica Extracellular Vesicles: Clues to Their Role in Pathogenesis","authors":"Aránzazu González-Arce,&nbsp;Christian M. Sánchez-López,&nbsp;Liz F. Sánchez-Palencia,&nbsp;Antonio Marcilla,&nbsp;Dolores Bernal","doi":"10.1002/jex2.70055","DOIUrl":"https://doi.org/10.1002/jex2.70055","url":null,"abstract":"<p>Fasciolosis, caused by <i>Fasciola hepatica</i>, is a parasitic zoonosis that induces liver fibrosis in infected hosts, including ruminants and humans. Extracellular vesicles secreted by <i>F. hepatica</i> (<i>Fh</i>EVs) play a crucial role in modulating host immune responses and promoting tissue re-modelling. This work explores the effects of two proteins found in <i>Fh</i>EVs, enolase (<i>Fh</i>enolase), enriched in the vesicular lumen, as well as the 16.5-kDa tegument-associated protein (<i>Fh</i>16.5TP), highly abundant in the EV membrane, on hepatic and liver-associated immune cells. Recombinant proteins (r-<i>Fh</i>enolase and r-<i>Fh</i>16.5TP) were produced to evaluate their impact on cell viability, inflammatory responses, proteomic profiles and EV secretion in THP1-XBlue CD14 macrophages, HepG2 hepatocytes and LX-2 hepatic stellate cells (HSCs). Interestingly, r-<i>Fh</i>enolase, but not r-<i>Fh</i>16.5TP, showed anti-inflammatory properties in lipopolysaccharide (LPS)–activated macrophages, by reducing NF-κB activation and inducing significant changes in the protein cargo of macrophage-derived EVs, which contained lower levels of the pro-inflammatory cytokines IL-1β, TNF-α and IL-6. Proteomic analysis of cells treated with r-<i>Fh</i>enolase revealed distinct alterations in proteins related to fibrotic and inflammatory pathways, including a reduction in extracellular matrix (ECM) proteins and suggesting a potential role in mitigating liver fibrosis. Furthermore, r-<i>Fh</i>enolase reduced EV production and fibrotic markers in hepatic cells, but not in macrophages. In contrast, r-<i>Fh</i>16.5TP increased pro-fibrotic proteins in both, cells and EVs, and increased EV production specifically in LX-2 cells, indicating its possible contribution to fibrosis progression in fasciolosis. These findings represent a first approach to analyse EV-associated proteins and study their potential role in the molecular mechanisms of <i>F. hepatica</i>–host interactions.</p>","PeriodicalId":73747,"journal":{"name":"Journal of extracellular biology","volume":"4 6","pages":""},"PeriodicalIF":0.0,"publicationDate":"2025-06-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/jex2.70055","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144191152","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}
引用次数: 0
Enhanced Anti-Tumour Efficacy of iRGD-Modified Cell-Bound Membrane Vesicles (iRGD-CBMVs) as a Novel Drug Carrier irgd修饰的细胞结合膜囊泡(iRGD-CBMVs)作为一种新型药物载体的抗肿瘤效果增强
Journal of extracellular biology Pub Date : 2025-06-02 DOI: 10.1002/jex2.70052
Haonan Zhao, Zhendong Huang, Qinghua Sheng, Wenxiang Shao, Min Zeng, Kun Wang, Yang Zhang, Ying Qin, Zhihao Xiong, Lizhen Chen, Huaying Wang, Tong Rong, Zhitao Qiu, Hongda Zhuang, Zhiwen Wu, Yuan Zhang, Wendiao Zhang, Yong Chen
{"title":"Enhanced Anti-Tumour Efficacy of iRGD-Modified Cell-Bound Membrane Vesicles (iRGD-CBMVs) as a Novel Drug Carrier","authors":"Haonan Zhao,&nbsp;Zhendong Huang,&nbsp;Qinghua Sheng,&nbsp;Wenxiang Shao,&nbsp;Min Zeng,&nbsp;Kun Wang,&nbsp;Yang Zhang,&nbsp;Ying Qin,&nbsp;Zhihao Xiong,&nbsp;Lizhen Chen,&nbsp;Huaying Wang,&nbsp;Tong Rong,&nbsp;Zhitao Qiu,&nbsp;Hongda Zhuang,&nbsp;Zhiwen Wu,&nbsp;Yuan Zhang,&nbsp;Wendiao Zhang,&nbsp;Yong Chen","doi":"10.1002/jex2.70052","DOIUrl":"https://doi.org/10.1002/jex2.70052","url":null,"abstract":"<p>Cancer continues to be the foremost cause of mortality in humans. Persistent challenges in cancer treatment include inadequate drug targeting, severe toxicological side effects and uncontrolled drug distribution. The bioinspired membrane vesicle drug delivery systems have been emerging as promising therapeutic strategies. This study characterises unique cell-bound membrane vesicles (CBMVs), which are impervious to standard cleaning agents and effectively loaded with doxorubicin (DOX). For the first time, we used iRGD peptide to modify the CBMVs to enhance the CBMVs' targeting capabilities for cancer cells. Laser confocal microscopy and <sup>1</sup>H Nuclear Magnetic Resonance Spectra (<sup>1</sup>H NMR) have confirmed the CBMVs' iRGD modification and effective encapsulation with DOX (iRGD-CBMVs-DOX). Then, we used the iRGD-CBMVs-DOX to treat tumour cell lines and tumour-bearing mouse models. Our research identified that iRGD-CBMVs-DOX proves effective in inhibiting cell growth and migration for tumour cell lines, significant anti-tumour ability, reduced organ toxicity and continuous drug administration were revealed in tumour-bearing mouse models. Additionally, the iRGD-CBMVs-DOX demonstrated sustained drug release, indicating their potential for prolonged circulation. These findings are pivotal in enhancing cancer treatment through novel nanomedicine strategies, and highlight the potential of iRGD-modified vesicles (e.g., iRGD-CBMVs) as efficient drug carriers, contributing to targeted and biocompatible drug delivery advancements for cancer treatment.</p>","PeriodicalId":73747,"journal":{"name":"Journal of extracellular biology","volume":"4 6","pages":""},"PeriodicalIF":0.0,"publicationDate":"2025-06-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/jex2.70052","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144191154","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}
引用次数: 0
Response to ‘Purification of Mesenchymal Stromal Cell-Derived Small Extracellular Vesicles Using Ultrafiltration’ 对“超滤纯化间充质基质细胞来源的细胞外小泡”的响应
Journal of extracellular biology Pub Date : 2025-06-02 DOI: 10.1002/jex2.70057
Anders Toftegaard Boysen PhD
{"title":"Response to ‘Purification of Mesenchymal Stromal Cell-Derived Small Extracellular Vesicles Using Ultrafiltration’","authors":"Anders Toftegaard Boysen PhD","doi":"10.1002/jex2.70057","DOIUrl":"https://doi.org/10.1002/jex2.70057","url":null,"abstract":"&lt;p&gt;Dear Editor,&lt;/p&gt;&lt;p&gt;I read the recent article ‘&lt;i&gt;Purification of mesenchymal stromal cell-derived small extracellular vesicles using ultrafiltration&lt;/i&gt;’ by Lei et al. (&lt;span&gt;2025&lt;/span&gt;) with great interest, as they have thoroughly characterised an ultrafiltration method also used by me (Boysen et al. &lt;span&gt;2024&lt;/span&gt;). I would like to weigh in with some of my own experiences and findings on their conclusions. The extracellular vesicle (EV) characterisation techniques (TEM and NTA) used by the authors have size and concentration determination limitations. While TEM does a great job of identifying EVs in their cup-shaped appearance, it is not a great tool in size and concentration determination, as the amount of EVs present is few and they have been dried, thereby having lost their true size and morphology. Similarly, NTA has drawbacks as EVs have a low refractive index, the limit of detection is regarded to be around 50 nm (Dragovic et al. &lt;span&gt;2011&lt;/span&gt;), leading to a potentially skewed size profile and reduced particle concentration. The study of Lei et al. is therefore at risk of not showing a potential loss of sub-50 nm EVs using ultrafiltration and their specified molecular weight cut-off (MWCO) values. While they argue that the pore size of the membranes used is 6.13, 8.84 and 10.48 nm for 100, 300 and 500 kDa MWCO, respectively, this is only true from a theoretical perspective based on the hydrodynamic radius of molecules. The pore size of commercially available filters is &lt;span&gt;not&lt;/span&gt; based on theoretical values but trial and error and the &lt;span&gt;average&lt;/span&gt; pore size of the selected filters by Lei et al. are 10, 35 and 55 nm for 100, 300 and 500 kDa MWCO respectively as seen in Figure 1 (determined using cryo-EM, personal correspondence with Merck Millipore and as reported by Pall corporation (Pall Corporation &lt;span&gt;2022&lt;/span&gt;)). We have translated a copious amount of the knowledge from virology to EV research, as EVs and viruses have many biophysical similarities, and we could also learn from virology in this case. MS2, a icosahedral ∼27 nm bacteriophage, is not retained on a 300 kDa MWCO filter, and can be found in the permeate (Wick and McCubbin &lt;span&gt;1999&lt;/span&gt;). While fouling, particle deposition, and potential loss of EVs can be reduced by adding shear force to the membrane, this force will not be homogeneous, and loss will occur (Hwang et al. &lt;span&gt;2016&lt;/span&gt;; Zhao et al. &lt;span&gt;2023&lt;/span&gt;). Cryo-EM, in contrast to TEM and NTA, can show sub-50 nm particles in their native state (Yuana et al. &lt;span&gt;2013&lt;/span&gt;). As a true size distribution of an EV sample, I have made individual measurements of 130 EVs derived from human embryonic kidney cells (HEK293T) cultured in chemically defined media (HEK VIP NX, Satorius) and enriched using size exclusion chromatography (35 nm, Izon) on a 100 kDa MWCO spin filter (Thermo Fisher) as shown below.\u0000\u0000 &lt;/p&gt;&lt;p&gt;Cryo-EM tends to exclude some of the larger EVs (Welsh et al.","PeriodicalId":73747,"journal":{"name":"Journal of extracellular biology","volume":"4 6","pages":""},"PeriodicalIF":0.0,"publicationDate":"2025-06-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/jex2.70057","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144191153","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}
引用次数: 0
Correction to “Exploring the Potential of in Vitro Extracellular Vesicle Generation in Reproductive Biology” 对“探索体外细胞外囊泡生成在生殖生物学中的潜力”的更正
Journal of extracellular biology Pub Date : 2025-05-29 DOI: 10.1002/jex2.70061
{"title":"Correction to “Exploring the Potential of in Vitro Extracellular Vesicle Generation in Reproductive Biology”","authors":"","doi":"10.1002/jex2.70061","DOIUrl":"https://doi.org/10.1002/jex2.70061","url":null,"abstract":"<p>R. Franko, and M. de Almeida Monteiro Melo Ferraz, “Exploring the Potential of In vitro Extracellular Vesicle Generation in Reproductive Biology,” <i>Journal of Extracellular Biology</i> 3 (2024): e70007. https://doi.org/10.1002/jex2.70007</p><p>In the originally published article, the in-text citation for the reference ‘Montecalvo et al., 2008’ was given incorrectly as ‘Montecalvo et al., 1950’. This has been corrected in the online version of the article.</p><p>We apologise for this error.</p>","PeriodicalId":73747,"journal":{"name":"Journal of extracellular biology","volume":"4 6","pages":""},"PeriodicalIF":0.0,"publicationDate":"2025-05-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/jex2.70061","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144171944","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}
引用次数: 0
Correction to “Prostate-Derived Circulating microRNAs Add Prognostic Value to Prostate Cancer Risk Calculators” 对“前列腺衍生循环microrna增加前列腺癌风险计算器的预后价值”的更正
Journal of extracellular biology Pub Date : 2025-05-29 DOI: 10.1002/jex2.70060
{"title":"Correction to “Prostate-Derived Circulating microRNAs Add Prognostic Value to Prostate Cancer Risk Calculators”","authors":"","doi":"10.1002/jex2.70060","DOIUrl":"https://doi.org/10.1002/jex2.70060","url":null,"abstract":"<p>M. L. Zenner, B. Kirkpatrick, T. R. Leonardo, et al., “Prostate-Derived Circulating microRNAs Add Prognostic Value to Prostate Cancer Risk Calculators,” <i>Journal of Extracellular Biology</i> 2 (2023): e122, https://doi.org/10.1002/jex2.122.</p><p>In the originally published article, the in-text citation for the reference ‘Cancer Statistics, 2023 (2023)’ was given incorrectly as ‘Cancer Facts &amp; Figures 2023, 1930’. This has been corrected in the online version of the article.</p><p>We apologise for this error.</p>","PeriodicalId":73747,"journal":{"name":"Journal of extracellular biology","volume":"4 6","pages":""},"PeriodicalIF":0.0,"publicationDate":"2025-05-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/jex2.70060","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144171945","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}
引用次数: 0
Scalable, High-Throughput Isolation of Extracellular Vesicles Using Electrokinetic-Assisted Mesh Filtration: ExoFilter 可扩展的,高通量分离细胞外囊泡使用电动辅助网状过滤:exfilter
Journal of extracellular biology Pub Date : 2025-05-28 DOI: 10.1002/jex2.70054
KangMin Lee, Minju Bae, YongWoo Kim, SoYoung Jeon, Sujin Kang, Wonjong Rhee, Sehyun Shin
{"title":"Scalable, High-Throughput Isolation of Extracellular Vesicles Using Electrokinetic-Assisted Mesh Filtration: ExoFilter","authors":"KangMin Lee,&nbsp;Minju Bae,&nbsp;YongWoo Kim,&nbsp;SoYoung Jeon,&nbsp;Sujin Kang,&nbsp;Wonjong Rhee,&nbsp;Sehyun Shin","doi":"10.1002/jex2.70054","DOIUrl":"https://doi.org/10.1002/jex2.70054","url":null,"abstract":"<p>As extracellular vesicles (EVs) are increasingly recognized for their superior functions for therapeutics, the need for large-scale EV isolation technology is becoming more critical for clinical and industrial applications. Most existing EV isolation methods are optimized for small-scale laboratory samples, limiting their efficiency and scalability for large-scale production. Here, an electrokinetic-assisted filtration system (ExoFilter), which introduces charge interaction into physical mesh flow filtration, is proposed as a new candidate to address the challenges of scalable EV isolation. The hybrid filtration system demonstrates outstanding high-throughput EV isolation performance (a flux of ∼750 mL/min) using only a coarse physical filter by electrokinetically arresting EVs flowing through the filter lattice. Furthermore, the recovery efficiency of ExoFilter, analysed based on the ELISA results, was found to be approximately 98%, demonstrating the filter's exceptional efficiency in EV isolation. Additionally, ExoFilter enables the rapid isolation of EVs from small samples as little as 200 µL, facilitating quick and easy blood-based EV research. Furthermore, low-molecular-weight albumin from plasma samples was effectively removed. The high-throughput and high-efficiency characteristics of ExoFilter make it well-suited for scalable EV production, offering greater convenience for various clinical applications.</p>","PeriodicalId":73747,"journal":{"name":"Journal of extracellular biology","volume":"4 6","pages":""},"PeriodicalIF":0.0,"publicationDate":"2025-05-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/jex2.70054","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144171470","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}
引用次数: 0
Conference & School on Extracellular vesicles and Nanoparticles 2024 (CSEVP-2024) 2024年细胞外囊泡和纳米颗粒会议与学校(CSEVP-2024)
Journal of extracellular biology Pub Date : 2025-05-22 DOI: 10.1002/jex2.70046
{"title":"Conference & School on Extracellular vesicles and Nanoparticles 2024 (CSEVP-2024)","authors":"","doi":"10.1002/jex2.70046","DOIUrl":"https://doi.org/10.1002/jex2.70046","url":null,"abstract":"","PeriodicalId":73747,"journal":{"name":"Journal of extracellular biology","volume":"4 S1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2025-05-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/jex2.70046","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144118142","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}
引用次数: 0
Control of Physical and Biochemical Parameters Influencing Exogeneous Cargo Protein Association to Extracellular Vesicles Using Lipid Anchors Enables High Loading and Effective Intracellular Delivery 利用脂质锚定控制影响外源货物蛋白与细胞外囊泡关联的物理和生化参数,实现高负载和有效的细胞内递送
Journal of extracellular biology Pub Date : 2025-05-14 DOI: 10.1002/jex2.70048
Antonin Marquant, Jade Berthelot, Claudia Bich, Zeineb Ibn Elfekih, Laurianne Simon, Baptiste Robin, Joël Chopineau, David Tianpei Wang, Samuel Jay Emerson, Aijun Wang, Clément Benedetti, Simon Langlois, Laurence Guglielmi, Pierre Martineau, Anne Aubert-Pouëssel, Marie Morille
{"title":"Control of Physical and Biochemical Parameters Influencing Exogeneous Cargo Protein Association to Extracellular Vesicles Using Lipid Anchors Enables High Loading and Effective Intracellular Delivery","authors":"Antonin Marquant,&nbsp;Jade Berthelot,&nbsp;Claudia Bich,&nbsp;Zeineb Ibn Elfekih,&nbsp;Laurianne Simon,&nbsp;Baptiste Robin,&nbsp;Joël Chopineau,&nbsp;David Tianpei Wang,&nbsp;Samuel Jay Emerson,&nbsp;Aijun Wang,&nbsp;Clément Benedetti,&nbsp;Simon Langlois,&nbsp;Laurence Guglielmi,&nbsp;Pierre Martineau,&nbsp;Anne Aubert-Pouëssel,&nbsp;Marie Morille","doi":"10.1002/jex2.70048","DOIUrl":"https://doi.org/10.1002/jex2.70048","url":null,"abstract":"<p>Despite biomolecule delivery is a natural function of extracellular vesicles (EVs), low loading of exogenous macromolecules such as proteins into EVs limits their interest as convincing protein delivery systems for health applications. In this context, lipid-anchorage of exogenous cargo into EV membrane recently emerged as a promising option to enable their vectorisation into cells. Nevertheless, this option was not explored for protein intracellular delivery, and further characterisation of critical parameters governing the association of a lipid-anchored cargo protein to EVs is still needed to confirm the relevance of this anchorage strategy. Therefore, we sought to identify these parameters in a precise and quantitative manner, using bulk and single nanoparticle analysis methods to identify protein loading capacity and subsequent intracellular delivery. We identified incubation temperature, cargo concentration, lipid anchor (LA) structure (lipid nature, linker) and EV origin as critical factors influencing maximal EV loading capacity. Precise control of these parameters enabled to load cargo protein close to EV saturation without hindering cellular delivery. The structural properties of LA influenced not only cargo protein/EV association but also intracellular delivery into different carcinoma cell lines. By thoroughly characterising Lipid-PEG-protein anchorage, this study evidences the interest of this tunable and controllable approach for efficient EV protein delivery.</p>","PeriodicalId":73747,"journal":{"name":"Journal of extracellular biology","volume":"4 5","pages":""},"PeriodicalIF":0.0,"publicationDate":"2025-05-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/jex2.70048","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143950007","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}
引用次数: 0
Revisiting the Nanoflow Cytometric Quantification of Extracellular Vesicles Under the Framework of ICH Q14 Guidelines 在ICH Q14指南框架下重新审视细胞外囊泡的纳米流式细胞术定量
Journal of extracellular biology Pub Date : 2025-05-08 DOI: 10.1002/jex2.70050
Ganghui Li, Qizhe Cai, Yanan Dong, Xiang Li, Xi Qin, Miaomiao Xue, Haifeng Song, Yi Wang
{"title":"Revisiting the Nanoflow Cytometric Quantification of Extracellular Vesicles Under the Framework of ICH Q14 Guidelines","authors":"Ganghui Li,&nbsp;Qizhe Cai,&nbsp;Yanan Dong,&nbsp;Xiang Li,&nbsp;Xi Qin,&nbsp;Miaomiao Xue,&nbsp;Haifeng Song,&nbsp;Yi Wang","doi":"10.1002/jex2.70050","DOIUrl":"https://doi.org/10.1002/jex2.70050","url":null,"abstract":"<p>Nanoflow cytometry (nanoFCM) is an increasingly important analytical procedure in every aspect of extracellular vesicle (EV) research, particularly in the development of EV-based therapeutics. The main objective of this study was to evaluate and optimise the key determinant factors of nanoFCM in the quantification analysis of EVs to ensure its consistency and reliability in the development of EV therapeutic drugs, thereby serving as a potential quality control measure. Our investigation followed the International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human Use (ICH) Q14 guideline. We revisited the day-to-day practice of nanoFCM measurement for HEK293 cell-derived and milk-derived EVs (mEVs), focusing on optimising particle quantification and identifying risk factors. Initial evaluation of the procedure revealed a considerable lack of consistency and reliability, which was then subjected to extensive optimisation. The key outcomes of this study include: (1) an optimised analytic procedure incorporating Tween-20, which significantly enhanced the precision and accuracy of the nanoFCM measurement and expanded the reportable range; (2) an analytical target profile (ATP) which provides a preliminary standard for future validation of nanoFCM procedures. Overall, this study serves as a foundation for future efforts towards the standardisation of analytical procedures for EV therapeutics.</p>","PeriodicalId":73747,"journal":{"name":"Journal of extracellular biology","volume":"4 5","pages":""},"PeriodicalIF":0.0,"publicationDate":"2025-05-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/jex2.70050","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143919569","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}
引用次数: 0
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